MAIN LIBRARY AGRIC. DEPT. 
 
BACTERIOLOGY 
 
 AND 
 
 INFECTIVE DISEASES 
 
A TEXT-BOOK 
 
 OP 
 
 BACTERIOLOGY 
 
 INCLUDING THE 
 
 ETIOLOGY AND PREVENTION 
 
 OF 
 
 INFECTIVE DISEASES 
 
 AND A SHORT ACCOUNT OF 
 
 YEASTS AND MOULDS, H^EMATOZOA, AND 
 PSOROSPERMS 
 
 BY 
 
 EDGAR M. CROOKSHANK, M.B. 
 
 PROFESSOR OF COMPARATIVE PATHOLOGY AND BACTERIOLOGY, AND FELLOW OF KING 
 
 FOURTH EDITION 
 RECONSTRUCTED, REVISED AND GREATLY ENLARGED 
 
 LONDON 
 H. K. LEWIS, 136, GOWER STREET, W.C. 
 
 1896 
 
\ ^ r i b 
 
 B10LOSY 
 
 LIBRARY 
 
 G 
 
 LONDON : 
 U. K. LEWIS. 136, GOWER STREET. W.C. 
 
SIR JOSEPH LISTER, BART., M.B., P.R.S., 
 
 WHO HAS CREATED A NEW EPOCH IN 
 
 MEDICINE AND SURGERY. 
 
 BY APPLYING A KNOWLEDGE OF MICRO-ORGANISMS 
 TO THE TREATMENT OF DISEASE, 
 
 This eHork is, toith permission, ^pebicatcb 
 BY THE AUTHOR 
 AS A 
 
 TOKEN OF ADMIRATION AM. RESPECT 
 
PREFACE 
 
 TO THE 
 
 FOURTH EDITION. 
 
 THIS book, though nominally a fourth edition, is practically 
 speaking a new work. The progress of Bacteriology has been 
 very rapid, and many new investigations have been made in 
 connection with the etiology, prevention and treatment of 
 communicable diseases. It has been necessary to reconstruct, 
 enlarge and thoroughly revise the text of the third edition, 
 and I have added twenty-six chapters. 
 
 The most important researches conducted in bacteriological 
 laboratories are those relating to the contagia. In many 
 diseases of man and animals it has not been possible to 
 identify the contagium with a bacterium, or indeed with any 
 micro-organism ; but when the virus is chemically examined, 
 or investigated with a view to protective inoculation, or utilised 
 for experiments in serum-therapeutics, such researches are 
 within the province of the bacteriologist. 
 
 The recognition of the fact that in so many diseases the 
 nature of the contagium has not yet been determined will 
 have the effect of encouraging continued activity in this 
 important Hi-Id of scientific investigation. 
 
 I hope that this work will continue to be of use as a text- 
 book for the bacteriological laboratory, and that the chapters 
 on the etiology and prevention of the communicable diseases 
 
viii PREFACE TO THE FOURTH EDITION. 
 
 of man and animals will be not only of scientific interest, 
 but of practical value to Medical Officers of Health and 
 Veterinary Inspectors. 
 
 I have divided the book into three parts. Part I. is 
 mainlv technical, and includes the most recent methods 
 employed in studying bacteria and investigating the etiology 
 of disease. Part II. deals with infective diseases and the 
 bacteria associated with them. Any clinical or pathological 
 evidence which may help to throw light on the nature and 
 origin of the contagia is taken into account. The most 
 effectual measures for stamping out these diseases are 
 referred to, as they are intimately connected with a 
 knowledge of the life-history of micro-organisms. Part III. 
 contains descriptions of about five hundred bacteria. Many 
 are of no practical importance and of very little scientific 
 interest, but a text-book for the laboratory cannot be con- 
 sidered complete unless an account is given of all bacteria 
 which have been more or less completely investigated. I 
 have endeavoured to refer to the original descriptions and 
 to verify them by comparison with actual cultivations, but 
 in a very great number of instances this has been quite 
 impossible, and I desire to acknowledge the assistance I have 
 received from the works of several authors, especially those 
 of Fltigge, Frankel, Eisenberg, Baumgarten, Frankland, 
 Sternbenr, Lehmanu and Neumann. 
 
 I have rearranged the bibliography according to the 
 chapters, and the names of authors are given in alphabetical 
 order. With the aid of the current numbers of the Annales 
 </, rinxtitut Pasteur, the Zeitschrift far Hygiene, the 
 Centralblatt fur Bakterioloyie und r<u-(tsitenkunde, and the 
 Journal of Comparative Pathology and Bacteriology, it is 
 possible to become acquainted witli the most recent litera- 
 ture of the subject. 
 
 Many of the coloured plates illustrating the last edition 
 have not been reproduced. Those substituted for them have 
 been drawn from my own preparations, and most of them 
 
PREFACE TO THE FOURTH EDITION. IX 
 
 have already appeared in my Reports to the Board of 
 Agriculture and in other publications. 
 
 One hundred and thirty-three woodcuts and photographs 
 have been added in the text, and I have reverted to the plan 
 which I adopted in the second edition, of having many of 
 them printed in colours. 
 
 I take this opportunity of thanking Professor Frankel for 
 kindly permitting me to reproduce some of the photographs 
 in his excellent Atlas. 
 
 1 am particularly indebted to Professor Hamilton for 
 the use of c//W//.v of figures in his classical treatise on 
 Pathology, and to the New Syclenham Society for several 
 from the English translation of Professor Fliigge's well- 
 known work on micro-organisms. 
 
 To my Demonstrator, Dr. George Xewman, D.P.H., I am 
 indebted for much assistance in correcting the proof-sheets, 
 and for the preparation of an index. 
 
 EDGAR M. CROOKSHAXK. 
 
 SAINT HILL, EAST (THIN-STEAD. Srssj-jx. 
 August 1st, 1896. 
 
 P.S. Since this work was finally passed for press the con- 
 clusions of the Royal Vaccination Commissioners have been 
 published. I have at the last moment added extracts in the 
 form of a supplementary Appendix. E. M. C. 
 
CONTENTS. 
 
 PAKT I. 
 
 THEORETICAL AND TECHNICAL. 
 
 PAGE 
 
 CHAPTER I. 
 HISTORICAL INTRODUCTION 1 
 
 CHAPTER II. 
 
 MORPHOLOGY AND PHYSIOLOGY OF BACTERIA . . .11 
 
 CHAPTER III. 
 EFFECT OF ANTISEPTICS AND DISINFECTANTS ON BACTERIA . 30 
 
 CHAPTER IV. 
 
 CHEMICAL PRODUCTS OF BACTERIA . . . . .39 
 
 CHAPTER V. 
 IMMUNITY .......... 49 
 
 CHAPTER VI. 
 
 ANTITOXINS AND SERUM THERAPY ..... 56 
 
 CHAPTER VII. 
 
 THE BACTERIOLOGICAL MICROSCOPE 65. 
 
 CHAPTER VIII. 
 
 MICROSCOPICAL EXAMINATION OF BACTERIA . . 83 
 
 CHAPTER IX 
 
 PREPARATION OF NUTRIENT MEDIA AM> METHODS OF CULTIVA- 
 TION . 99 
 
Xii CONTENTS. 
 
 PAGE 
 
 CHAPTER X. 
 
 EXPERIMENTS UPON THE LIVING ANIMAL . .134 
 
 CHAPTER XI. 
 
 EXAMINATION OF AIR, SOIL, AND WATER . . .140 
 
 CHAPTER XII. 
 
 PHOTOGRAPHY OF BACTERIA . . . . .150 
 
 PAKT II. 
 
 ETIOLOGY AND PREVENTION OF INFECTIVE 
 DISEASES. 
 
 CHAPTER XIII. 
 SUPPURATION, PYAEMIA, SEPTICAEMIA, ERYSIPELAS . .173 
 
 CHAPTER XIV. 
 ANTHRAX .191 
 
 CHAPTER XV. 
 
 QUARTER-EVIL. MALIGNANT CEDEMA. RAG-PICKERS' SEPTI- 
 CAEMIA. SEPTICAEMIA OF GUINEA-PIGS. SEPTICAEMIA OF 
 MICE. . . .217 
 
 CHAPTER XVI. 
 
 SEFIICjEMIA OF BUFFALOES.- SEPTIC PLEURO-PNEUMONIA OF 
 CALVES. SWINE FEVER. SEPTICAEMIA OF DEER. SEPTI- 
 CAEMIA OF RABBITS. FOWL CHOLERA. FOWL ENTERITIS. 
 DUCK CHOLERA. GROUSE DISEASE . . . 226 
 
 CHAPTER XVII. 
 
 I'M. r.MONIA. INFECTIOUS PLEURO-PNEUMONIA OF CATTLE. 
 
 INFLUEN/A . . . .233 
 
 CHAPTER XVIII. 
 
 ORIENTAL PLAGUE. RELAPSING FEVER. TYPHUS FEVER. 
 
 YELLOW FKVKU . .250 
 
CONTENTS. XI 11 
 
 PAC.K 
 
 CHAPTER XIX. 
 
 SCARLKT FEVER. MEASLES 261 
 
 CHAPTER XX. 
 
 SMALL-POX. CATTLE PLAGUE 284 
 
 CHAPTER XXI. 
 SHEEP-POX. FOOT-AND-MOUTH DISEASE . . . 297 
 
 CHAPTER XXII. 
 HORSE-POX. COW-POX 303 
 
 CHAPTER XXIII. 
 DIPHTHERIA 330 
 
 CHAPTER XXIV. 
 
 TYPHOID FEVER ......... 340 
 
 CHAPTER XXV. 
 
 SWINE FEVER . . . . . . . . .347 
 
 CHAPTER XXVI. 
 
 >WINE MEASLES. DISTEMPER IN DOGS. EPIDEMIC DISEASE OF 
 
 FERRETS. EPIDEMIC DISEASE OF MICE . . . .355 
 
 CHAPTER XXVII. 
 
 ASIATIC CHOLERA. CHOLERA NOSTRAS. CHOLERAIC DIARRHCEA 
 FROM MEAT POISONING. DYSENTERY. CHOLERAIC DIAR- 
 RHCEA IN FOWLS . 360 
 
 CHAPTER XXVIII. 
 
 TUBERCULOSIS 375 
 
 CHAPTER XXIX. 
 LEPROSY. SYPHILIS. RHINOSCLEROMA. TRACHOMA . . 406 
 
 CHAPTER XXX. 
 
 ACTINOMYCOSIS. MADURA DISEASE . . . .413 
 
 CHAPTER XXXI. 
 
 (.LANDERS . -451 
 
xiv CONTENTS. 
 
 I'AGE 
 
 CHAPTER XXXII. 
 
 TETANUS. RABIES. LOUPING-ILL . 457 
 
 CHAPTER XXXIII. 
 FOOT-ROT 464 
 
 CHAPTER XXXIV. 
 
 FOUL-BROOD. INFECTIOUS DISEASE OF BEES IN ITALY. 
 P^BRINE. FLACHERIE. INFECTIOUS DISEASE OF CATER- 
 PILLARS .469 
 
 PAET III. 
 
 SYSTEMATIC AND DESCRIPTIVE 
 
 CHAPTER XXXV. 
 
 CLASSIFICATION AND DESCRIPTION OF SPECIES . . .475 
 
 APPENDICES. 
 
 APPENDIX I. 
 
 YEASTS AND MOULDS . 577 
 
 APPENDIX II. 
 
 ILEMATOZOA IN MAN, BIRDS, AND TURTLES. ILEMATOZOA IN 
 
 EQUINES, CAMELS AND FISH. ILEMATOZOA IN FROGS . 589 
 
 APPENDIX III. 
 
 PSOROSPERMS OR COCCIDIA. AMCEBA COLI .... 609 
 
 APPENDIX IV. 
 
 APPARATUS, MATERIAL AND REAGENTS EMPLOYED IN A BAC- 
 TERIOLOGICAL LABORATORY . . . . . .612 
 
 APPENDIX V. 
 I'-IULIOGRAPHY . . 639 
 
 SUPPLEMENTARY APPENDIX. 
 
 EXTRACTS FROM THE FINAL REPORT OF THE ROYAL VACCINA- 
 TION COMMISSION . 667 
 
LIST OF ILLUSTRATIONS. 
 
 WOOD ENGRAVINGS AND PHOTOGRAPHS. 
 
 FIG. PAGE 
 
 1. Ascococcus Billrothii, x 65 (Cobn) . . . . .14 
 
 _'. Spirochseta from Sewage Water, x 1200 (E.M.C.) . . 15 
 
 a. Flagella (Koch, Brefeld, Warming, Zopf) . .16 
 
 4. Bacillus Megatherium (De Bary) . . . . .17 
 
 .">. riostridium Butyricum, x 1020 (Prazmowski) . . . .18 
 
 6. Leuconostoc Meseuteroides ; Cocci-chains with Arthrospores (Van 
 
 Tieghem and Cienkowski) . . . . . .19 
 
 7. >\ (ore-bearing Threads of Bacillus Anthracis, double-stained with 
 
 Fucbsine and Methylene Blue, x 1200 (E.M.C.) . . 20 
 
 >. I'.acilli of Tubercle in Sputum, x 2500 (E.M.C.) . . 21 
 
 '.'. Comma Bacilli in Sewage Water, stained with Gentian Violet, x 1200 
 
 (E.M.C.) 22 
 
 10. Vibrios in Water contaminated with Sewage, x 1200 (E.M.C.) . 22 
 
 11. Refraction of Light (Carpenter) . . . . . .66 
 
 12. Spherical Aberration (Carpenter) . . . .67 
 
 13. Combination of Lenses in Abbe's Homogeneous Immersion (Car- 
 
 penter) ......... 67 
 
 14. Chromatic Aberration (Carpenter) . . . . .68 
 1.",. Objective with Collar Correction (Zeiss) . . .69 
 !'.. Microscope English Model (Swift) . . 71 
 17. Removable Mechanical Stage (Swift) . ... 72 
 1*. Microscope Continental Model (Zeiss) . . . . 73 
 1!. Iris Diaphragm (Zeiss) . . .71 
 L'II. A t.be's Condenser (Zeiss) . . . . . . .75 
 
 21. Microscope Lamp (Baker) ...... 6 
 
 22. I. nine Microscope Lamp (Swift) . , . . . .77 
 
 rangement of Powell and Lealand's Microscope in working directly 
 on the Edge of the Flame, with Stand for Micrometer Eye-piece to 
 secure Steadiness and Accuracy of Measurement (Carpenter after 
 Nelson) ......... 79 
 
 24. Ramsden Micrometer Eye-piece (Swift) . . . 80 
 
 2.">. Micrometer Eye-piece (Zeiss) . . . SI 
 
 2;. Inoculating Needles (E.M.C.) . . 84 
 
 27. Freezing Microtome (Swift) . . . . . .04 
 
 2 s . Microtome (Jung) . . . . . . .95 
 
 29. Wire-cage for Teat-tubes (Mnencke) . ... 100 
 
 ::<J. Hot-air Steriliser (E.M.C.) . . 101 
 
Xvi LIST OF ILLUSTRATIONS. 
 
 FK;. 
 
 A(,K 
 
 31. Hot-water Filtering Apparatus (Muencke) . .102 
 
 32. Method of making a Folded Filter (E.M.C.) . 103 
 
 33. Steam Steriliser (Baird and Tatlock) . ... 103 
 
 34. Incubator (Muencke) . ... 104 
 
 35. Method of Inoculating a Test-tube containing Sterile Nutrient Jelly 
 
 (E.M.C.) 105 
 
 36. Levelling Apparatus (E.M.C) . ... 107 
 
 37. Iron box for Glass Plates (Muencke) . . . . .108 
 
 38. Method of Inoculating Test-tubes in the Preparation of Plate-cultiva- 
 
 tions (E.M.C.) .... .108 
 
 39. Damp-chamber containing Plate-cultivations (E.M.C) . .110 
 
 40. Pasteur's Large Incubator (Becker) . . . . .111 
 
 41. Petri's Dish (Becker) . . . . . . .112 
 
 42. Glass Benches and Slides (Becker) . . . . .112 
 
 43. Koch's Serum Steriliser (Muencke) . . . . .114 
 
 44. Hueppe's Serum Inspissator (Baird and Tatlock) . . .115 
 
 45. Box for Sterilising Instruments (Becker) .... 116 
 
 46. Damp Chamber for Potato-cultivations (E.M.C.) . . .117 
 
 47. Apparatus for Sterilisation by Steam under pressure (Baird and 
 
 Tatlock) ........ 119 
 
 48. Drop Cultivation (Fliigge) . . . . . .121 
 
 49. Simple Method of forming a Moist Cell (Schafer) . . .122 
 
 50. Warm Stage (Schafer) . . . . . . .123 
 
 51. Warm Stage shown in Operation (Schafer) .... 123 
 
 52. Warming Apparatus in Operation (Israel) . . . -.124 
 
 53. Section of Warming Apparatus and Drop-culture Slide (Israel) 125 
 
 54. Israel's Warming Apparatus . . . . . .125 
 
 55. Gas Chamber in use with Apparatus ior generating Carbonic Acid 
 
 (Schafer) . . ... 126 
 
 56. Gas Chamber (Schafer) . . ... 126 
 ">7. Moist Cell adapted for Transmission of Electricity (Schafer) . .127 
 58. Apparatus arranged for Transmitting Electricity (Schafer) . . 127 
 r><). Slide with Gold-leaf Electrodes (Schafer) . . . . " 128 
 tin. List f-r's Flask (Becker) . .128 
 r,l. Sternberg's Bulb (Becker) . . 128 
 (\-2. Aitken's Tube (Becker) . . . 129 
 I',:;. Miquel's Bulb (Becker) . 129 
 
 64. Pasteur's Flask (Baird and Tatlock) . . . 130 
 
 65. Pasii-ur's Double Tube (Baird and Tatlock) . . 130 
 
 66. F ranker's Anaerobic Tube-culture (Frankland) . . .131 
 
 67. Anaerobic Culture Tube (Liborius) . . . 132 
 
 68. Apparatus for Anaerobic Cultures (Roscoe and Lunt) . . 133 
 <;'.. Korir.< Swinge (Baird and Tatlock) . . . 135 
 7(>. Syringe with Asbestos Plug (Baird and Tatlock) . 135 
 
 71. Hesse's Apparatus (Muencke) . . . . .142 
 
 72. Sedgwick and Tucker's Tube (Baird and Tatlock) . . 143 
 
 73. Pouchet's Aeroscope (Hamilton) . . . . .143 
 
 74. Apparatus for Estimating the number -!' Colonies in a Plate-cultiva- 
 
 tion (Muencke) ....... 14(5 
 
 7.". K>ni;i)-cli's Roll-culture (Frankland) . . . 147 
 
 76. Apparatus for Counting Colonies in a Roll-culture (Becker) . . 148 
 
 77. Horizontal Micro-photographic Apparatus (Swift) . . 150 
 
LIST OF ILLUSTRATIONS. XV11 
 
 EAGE 
 
 78. Reversible Micro-photographic Apparatus (E.M.C.) . . . 157 
 
 79. Reversible Micro-photographic Apparatus arranged in the Vertical 
 
 Position (E.M.C.) . . .158 
 
 80. Large Micro-photographic Apparatus (Swift) .... 160 
 *1. Photograph of an Impression Preparation (E.M.C.) . . . 162 
 Bft Photograph of a Cultivation of Bacillus anthracls (E.M.C.) . . 168 
 
 ^uppuration of Subcutaneous Tissue (Cornil and Ranvier) . . 174 
 -4. I'us with Staphylococci, x 800 (Fliigge) . . . .177 
 So. Subcutaneous Tissue of a Rabbit forty-eight hours after an Injection 
 
 of Staphylococci, x 950 (Baumgarten) .... 177 
 9ft Ulcerative Endocarditis : Section of Cardiac Muscle, x 700 (Koch) 183 
 X 7. Pure-cultures of Streptococcus Pyogenes (E.M.C.) . . . 184 
 B8 ^-ct ion of Skin in Erysipelas .(Cornil and Ranvier) . . . 185 
 -reptococcus Pyogenes Hominis ; Pure-cultures on Nutrient Gela- 
 tine (E.M.C.) 1*7 
 
 - 1 reptococcus Pyogenes Bovis ; Pure-cultures on Nutrient Gelatine. 
 
 (E.M.C.) .... . 188 
 
 11. Gonococcns, x 800 (Bumm) ... . . 190 
 
 92. Bacillus Anthracis, x 1200. Blood Corpuscles and Bacilli unstained ; 
 
 . from an Inoculated Mouse (Frankel and Pfeiffer) . . . 192 
 
 93. Pure-cultivation of Bacillus anthracis in Nutrient Gelatine (E.M.C.) 193 
 
 94. Colonies of Bacillus ant /tracts, x 8<> (Flttgge) . . . 194 
 
 95. Impression-preparation of a Colon}', x 70 (E.M.C.) . . . 194 
 
 .;. Margin of a Colony, x 250 (E.M.C.) 195 
 
 97. Filaments with Oval and Irregular Elements, x 800 (E.M.C.) . 195 
 
 -pores of BacUlu* ant/iracis stained with Gentian Violet, x 1500 
 
 (E.M.C.) 197 
 
 '.''.i. Anthrax in Swine (E.M.C.) . . . . . .203 
 
 100. Anthrax in Swine (E.M.C.) ...... 205 
 
 101. Bacilli of Quarter-evil, x 1000 (Frankel and Pfeiffer) . . 218 
 1<2. Pure-culture of Bacilli of Quarter-evil in Grape-sugar Gelatine 
 
 (Frankel and Pfeiffer) . . . . . . .218 
 
 K>3. Bacilli of Malignant (Edema, x 9CO (Baumgarten) . . . 221 
 
 lo4. Pure-culture of Bacillus of Malignant (Edema in Grape-Sugar 
 
 Gelatine (Frankel and Pfeiffer) . . . . .222 
 
 In5. Bacilli of Malignant (Edema, x 1000 (Frankel and Pfeiffer) - . 223 
 
 lor,. Pure-cultivation of the Bacillus of Septicaemia of Mice in Nutrient 
 
 Gelatine (E.M.C.) .... . 225 
 
 lo7. Bacterium of Rabbit Septicaemia ; Blood of Sparrow, x 700 (Koch) 228 
 lo-. Bacterium of Fowl-cholera, x 1200 (E.M.C.) . . . . 22S 
 
 IM'.I. Bacterium of Fowl-cholera, x 2500 (E.M.C.) . . . .228 
 
 110. Bacterium of Fowl-cholera; Section from Liver of Fowl, x 700 
 
 (Fliigge) .... . 229 
 
 111. Bacillus of Haemorrhagic Septicaemia, x 950 (Baumgarten) . . 231 
 
 112. Bacillus of Haemorrhagic Septicaemia: Pure-culture in Gelatine 
 
 (Baumgarten) . . . . . . . .231 
 
 113. Bacterium Pneumoniae Crouposae from Pleural Cavity of a Mouse, 
 
 x l.VK)(Zopf) .... .234 
 
 114. Friedliinders Pneumococcus ; Pure-culture in Nutrient Gelatine 
 
 (Baumgarten) .... ... 234 
 
 115. < ; q,suk- < <><-<-i from Pneumonia, x 1500 (Baumgarten) . 235 
 11G. Microcoecus of Sputum Septicaemia, -x 10CO (Frankel and Pfeiffer) . 23*5 
 
 b 
 
xviii LIST OF ILLUSTRATIONS. 
 
 FIG. PAGE 
 
 117. Colonies of Steinberg's Micrococcus, x 100 (Frankel and PfeifEer) . 237 
 
 118. Acute Catarrhal Pneumonia, x 480 (Hamilton) . . . 239 
 
 119. Infectious Pleuro-pneumonia of Cattle (Hamilton) . . . 240 
 
 120. Infectious Pleuro-pneumonia of Cattle (Hamilton) . . . 241 
 
 121. Bacillus of Influenza, x 1000 (Itzerott and Niemann) . . 248 
 
 122. Bacillus of Influenza, x 1200 (E.M.C) .... 249 
 
 123. Bacilli of Plague and Phagocytes, x 800 (Aoyama) . . . 253 
 
 124. Spirillum Obermeieri in Blood of Monkey inoculated with Spirilla 
 
 after Removal of the Spleen (Soudakewitch) . . . 258 
 
 125. Pure-cultivations of Streptococcus Pyogenes (E.M.C.) . . 263 
 
 126. Free Surface of Diphtheritic Larynx, x 350 (Hamilton) . . 331 
 
 127. Bacillus of Diphtheria ; from a Cultivation on Blood Serum, x 1000 
 
 (Frankel and Pfeiffer) ....... 332 
 
 128. Pure-cultures of Bacillus Diphtheria? on Glycerine- Gelatine (E.M.C.) 333 
 
 129. Typhoid Fever. Ileum of Adult, showing Sloughy and Infiltrated 
 
 Patches (Hamilton) . . . . . . .341 
 
 130. Typhoid Bacilli from a Colony on Nutrient Gelatine, x 1000 
 
 (Frankel and Pfeiffer) . ... . . . .342 
 
 131. Typhoid Bacilli, x 950 (Baumgarten) . . . . .342 
 
 132. Flagella of Typhoid Bacilli, x 1000 (Frankel and Pfeiffer) . . 343 
 
 133. Colonies of the Typhoid Bacillus (Frankel and Pfeiffer) . . 344 
 
 134. Pure-culture of Typhoid Bacilli inoculated in the Depth of Nutrient 
 
 Gelatine (Baumgarten) ....... 344 
 
 135. Typhoid Bacilli in a Section of Spleen, x 800 (Flugge) . . 345 
 
 136. Typhoid Bacilli in a Section of Intestine invading the Submucous 
 
 and Muscular Layers, x 950 (Baumgarten) .... 346 
 
 137. Ulceration of the Intestine in a Typical Case of Swine-fever (E.M.C.) 348 
 
 138. Klein's Bacillus of Swine-fever (No. .1) .... 349 
 
 139. From a Preparation of Bronchial Mucus of Klein's Swine-fever 
 
 Bacillus (No. 2) 349 
 
 140. Bacilli from an Artificial Culture with Spores, Bacillus No. 2 (Klein) 349 
 
 141. Blood of Fresh Spleen of a Mouse after Inoculation with Swine-fever 
 
 Bacillus No. 2 (Klein) 350 
 
 142. Bacilli of Swine Erysipelas (Baumgarten) .... 356 
 
 143. Blood of Pigeon inoculated with Bacilli of Swine Erysipelas, x 600 
 
 (fcchiitz) 356 
 
 144. Pure-culture in Nutrient Gelatine of Bacilli from Swine Erysipelas 
 
 (Baumgarten) ... 357 
 
 145. Cover-glass Preparation of a Drop of Meat Infusion containing a 
 
 Pure-cultivatipn of Comma-bacilli (Koch) . . . .361 
 
 146. Arthrospores of Comma-bacilli (Hueppe) .... 361 
 
 147. Flagella of Comma-bacilli ; stained by Loffler's Method (Franfcel 
 
 and Pfeiffer) 3 62 
 
 148. Involution Forms of Comma-bacilli, x 700 (Van Ermengem) . 362 
 
 149. Colonies of Comma- bacilli on Nutrient Gelatine ; natural size (Koch) 362 
 
 150. Colonies of Koch's Comma-bacilli, x 60 (E.M.C.) . . . 363 
 
 151. Cover-glass Preparation from the Contents of a Cholera Intestine, 
 
 x 600 (Koch) ........ ^63 
 
 152. Cover-glass Preparation of Cholera Dejecta on Damp Linen, x 600 
 
 (Koch) . . .... 363 
 
 -Action of the Mucous Membrane of a Cholera Intestine, x 600 
 
 ... ,364 
 
LIST OF ILLUSTRATIONS. 
 
 I'll. 1 'ure-cultivations; in Nutrient Gelatine of Koch's and of Tinkler's 
 
 Comma-bacilli (E.M.C.) ...... 365 
 
 15.5. Comma-shaped Organisms with other Bacteria in Sewage-con- 
 
 taminated Water, x . 1200 . . . . . 366 
 
 156. Comma-bacilli of the Mouth, x 700 (Van Ermengem) . . 367 
 
 157. Tinkler's Comma-bacilli, from Cholera nostras, x 700 (Fliigge) . 367 
 l.>. Deneke's Comma-bacilli, from Cheese, x 700 (Fliigge) . . 367 
 159. Pure-cultivation of the Spirillum Finkler- Prior, in Nutrient Gelatine 
 
 (E.M.C.) ........ 370 
 
 1 50. Tropical Dysentery ; Mucous Membrane of Large Intestine (Hamilton) 372 
 
 161. Tubercle of the Lung in a very Early Stage, x 400 (Hamilton) . 376 
 
 162. Primary Tubercle of Lung two to three weeks old, x 50 (Hamilton) 377 
 
 163. Large Oval Giant Cell from Tubercle of Lung, x 300 (Hamilton) . 377 
 
 164. Bacillus Tuberculosis, from Tubercular Sputum, x 2500 (E.M.C.) . 379 
 
 165. Pure-cultivation of the Tubercle Bacillus on Glycerine Agar-agar 
 
 (E.M.C.) . ..... 380 
 
 166. Pure-cultivation in Glycerine Agar-agar after ten months' growth 
 
 (E.M.C.) ..." ....... 381 
 
 167. Pure-cultivations of Tubercle Bacillus in Glycerine Agar-agar ; a sub- 
 
 culture from a Pure-culture in Glycerine Milk (E.M.C.) . . 381 
 
 16H. Section through a Lupus Nodule of the Nose (Hamilton) . . 387 
 
 169. Tubercular Ulceration of Mucosa of Ileum (Hamilton) . . 388 
 
 170. Section of Lupus of the Skin ; Giant Cell containing Tubercle 
 
 Bacillus (Fliigge) . ... . . . .389 
 
 171. Tuberculosis of Pleura; "Grape-disease" (E.M.C.) . . . 390 
 
 172. Tubercular Ulceration of the Intestine of a Cow (E.M.C.) . . 393 
 
 173. Tubercular Ulceration of the Intestine of a Rabbit (E.M.C.) . . 395 
 
 174. Tubercular Lungs of Rabbit (E.M.C.) ..... 396 
 
 175. Cover-glass Preparation of Pus from a Chancre, x 1050 (Lustgarten) 410 
 lit). Wandering Cell containing Bacilli (Lustgarten) . . . 410 
 177. Section of Liver from a Case of Actinomycosis in Man (E.M.C.) . 417 
 17*. Actinomycotic Tumour in the Throat of a Steer (E.M.C) . . 424 
 
 179. Actinomycotic Tumour of the Cheek (E.M.C.) . . . . 424 
 
 180. Steer with Emaciation the Result of Actinomycosis (E.M.C.) .. 425 
 
 181. Actinomycotic Growths from the Pleura resembling * Grape- 
 
 Disease" (E.M.C.) ....... 425 
 
 1 >2. Actinomycosis of the Skin (E.M.C.) . . . . .430 
 
 !*.">. Part of Human Foot with Madura Disease (E.M.C.) . . . 448 
 1*4. Bacilli of Glanders, x 700 (Flugge) . . . . .452 
 
 \^~>. Section of a Branch of the Pulmonary Artery showing Glanders 
 
 Bacilli penetrating the Wall (Hamilton) .... 453 
 1 *6. Pure-culture of the Tetanus Bacillus in Grape-sugar Gelatine (Frankel 
 
 and Pfeiffer) . . . . . . . .458 
 
 1 S 7. Foot of Sheep showing Disease of Horn (Brown) . . . 465 
 "ction through the Foot showing a Crack extending through the 
 
 Wall ........ .465 
 
 l^'.i. s, c-ret inir Membrane covered with Fungoid Growths (Brown) . 466 
 
 1W. Advanced Form of Disease of Skin between the Claws (Brown) . 466 
 
 191. Distortion of Hoof in an Advanced Form of Foot-rot (Brown) . 467 
 I'.'L'. Diseased Comb (Cowan) . . . . . .469 
 
 I!*: 1 ,. Spores of Bacillus Alvei (E.M.C.) . . . . .470 
 
 191. Pure-culture in Nutrient Gelatine (Cheshire and Cheyne) . . 470 
 
XX LIST OF ILLUSTRATIONS. 
 
 FH;. PAGE 
 
 195. Cultivation on the Surface of Gelatine (Cheshire and Cheyne) . 471 
 
 1%. Cladothrix Dichotoma (Zopf) ... . . . . 479 
 
 197. Friedlander's Pneumococcus, x 1500 (Zopf) . . . .483 
 
 19S. Ascococcus Billrothii (Cohn) . . . . . 498 
 
 199. Clostridium Butyricum (Prazmowski) ..... 503 
 
 200. Bacillus Cyanogenus, x 650 (Neelsen) .... 507 
 
 201. Pure -cultivation of Bacillus figurans on the Surface of Nutrient 
 
 Agar-agar (E.M.C.) ....... 509 
 
 202. Photograph of Part .of an Impression Preparation of. Bacillus 
 
 figurans on Nutrient Gelatine, x 50 (E.M.C.) . . . 510 
 
 203. Part of the same Specimen, x 200 . . . .510 
 
 204. Bacillus Indicus : Colonies in Agar, x 60 . . . 518 
 
 205. Bacillus Neapolitans, x 700 (Emmerich) .... .522 
 
 206. Bacillus Megatherium (De Bary) ..... 524 
 
 207. Pure-culture of Bacillus Megatherium in Gelatine (E.M.C.) . . 524 
 
 208. Bacillus Putrificus Coli, x 1000 (Bienstock) ... . .529 
 
 209. Bacillus Pyogenes Fcetidus, x 790 (Passet) .... 530 
 
 210. Bacillus Saprogenes, No. 1 (R,osenbach) . . . . . 531 
 
 211. Bacillus Subtilis with Spores (Baumgarten) .... 535 
 
 212. Pure-culture of Bacillus Subtilis in Nutrient Gelatine (Baumgarten) . 535 
 
 213. Pure-culture of Bacillus Subtilis on the Surface of Nutrient Agar 
 
 (E.M.C) 53(5 
 
 214. Bacterium Zopfii (Kurth) . . . . . . 542 
 
 215. Beggiatoa Alba (Zopf). . ..... . . .543 
 
 216. Phase-forms of . Beggiatoa Persicina (Warming) . . . 544 
 
 217. Cladothrix Dichotoma (Zopf) ...... 545 
 
 21 s. Crenothrix Kiihniana (Zopf) ...... 546 
 
 219. Leuconostoc Mesenteroides (Van Tieghem and Cienkowski) . . 550 
 
 220. Micrococcus in Pyaemia in Rabbits (Koch) .... 556 
 
 221. Proteus Mirabilis ; Swarming Islands on the Surface of Gelatine, 
 
 x 285 (Hauser) ....... 561 
 
 222. Proteus Mirabilis ; Involution Forms, x 524 (Hauser) . 561 
 
 223. Proteus Vulgaris, x 285 (Hauser) ... 5(52 
 
 224. Sarrina. x 60O (Fliigge) ... '. 5(53 
 
 225. Spirochjeta Plicatile (E.M.C.) . . [ 5<j6 
 
 226. Comma : bacilli. in Water contaminated with. Sewage . . . 567 
 
 227. Comma : bacilli of the Mouth, x 700 (Van Ermengem) . . 568 
 
 228. peneke's Comma-bacilli, from Cheese, x 700 (Fliigge) . . 5(58 
 
 229. Streptococcus in Progressive Tissue Necrosis in Mice (Koch) . 571 
 
 230. Vibrio Rugula, x 1020 (Prazmowski) ... . 574 
 
 231. Black Torula ; Pure-cultivation on Potato (E.M.C.) . . 581 
 Head and Neck of Calf with Advanced Ringworm (Brown) . . 585 
 Non-pigmented Amoeboid Forms (Marchiafava and Celli) . . 590 
 
 I. I'iirmented Anusboid Forms (Golgi) . . 590 
 
 286. Semi-lunar Bodies of Laveran (Golgi) . 590 
 
 236. Kosi-tte Forms with Segmentation (Golgi) . .V.ll 
 
 2:;7. Flagcllat.-d Forms (Vandyke Carter) . . . . 593 
 
 ' Su i ra " I'arasites, occurring Singly and Fused, x 1200 . , 59* 
 
 '. I'Juasites in the Blood of Rats (Lewis) . . ' 599 
 
 2 HI. A Monad in Hat's Blood, x :-{(XK) (E.M.C.) . [ tfOl 
 
 l 1 1 . Monads in Rat's Blood, x 1200 (E. M. ( '. ) . ' <H)2 
 
 24& M""a.l> in Kats Bloodstained with Methyl Violet, x 1200 (E.M.C.) 603 
 
LIST OF ILLUSTRATIONS. XXI 
 
 Hf.. I'AGK 
 
 21::. Organisms in the Blood of Mud-fish (Mitrophanow) . . . 604 
 
 2J \. Organisms in the Blood of the Carp (Mitrophanow) . . . 605 
 
 24.'.. Amoeba cola in Intestinal Mucus (Lb'sch) . . 611 
 J1H. Warm Stage (Schafer) . . x . . . .613 
 
 217. Warm Stage (Strieker) ....... 614 
 
 2K Combined Gas Chamber and Warm Stage (Strieker) . . . 614 
 
 2i!. Vertical Micro-photographic Apparatus (Leitz) . . 622 
 
 _'.")(. Koch's Steam Steriliser (Muencke) . . . 623 
 
 2r>l. Hot-air Steriliser (Muencke) .... . 623 
 
 2.~>2. Section of Hot-air Steriliser (Muencke) .... 623 
 
 J.VJ. Hot-water Filtering Apparatus with Ring Burner (Rohrbeck) . 624 
 
 2.~>4. Wire -cage for Test-tubes (Muencke) , ... 626 
 
 2.-.V I'latinum-needles (E.M.C.) . . . - 626 
 
 2:,t; Damp Chamber for Plate-cultivations (E.M.C.) . 626 
 
 2.~i7. Apparatus for Plate-cultivations (E.M.C.) .... 627 
 
 i'."):'). j '.ox for Glass-plates (Muencke) . . . . 627 
 
 2.~>t. Glass Benches for Glass-plates (Becker) . 627 
 
 i>'.0. Israel's Case (Becker) . . 628 
 
 261. Damp Chamber for Potato Cultivations (E.M.C.) . . . 628 
 2f,2. Koch's Serum Steriliser (Muencke) . .629 
 
 2t>3. Serum Inspissator (Muencke) .... . 629 
 
 2i)4. D'Arsonval's Incubator (Muencke) , 631 
 
 2f;r>. Scblosing's Membrane Regulator (Muencke) . . 632 
 
 2f>i'.. Gas Burner protected with Mica Cylinder (Muencke) . 633 
 
 2>7. Koch's Safety Burner (Muencke) . 633 
 
 2>'. x . Babes' Incubator (Muencke) ... . 634 
 
 2f,!). Moitessier's Gas-pressure Regulator (Muencke) 634 
 
 270. Reichert's Thermo-regulator (Muencke) .. . 635 
 
 271. Meyer's Thermo-regulator (Muencke) ... 636 
 
 272. Siphon Bottle with Flexible Tube, Glass Nozzle, and a Mohr's Pinch - 
 
 cock (E.M.C.) .... -637 
 
 27:'.. Desiccator (E.M.C.) .....- 638 
 
DESCRIPTION OP PLATES. 
 
 DESCRIPTION OF PLATE I. 
 
 Bacteria, Schizomycetes, or Fission Fungi. 
 
 Following p. 14. 
 
 1. Cocci singly and varying in size. 2. Cocci in chains or rosaries (strepto- 
 coccus). 3. Cocci in a mass (staphylococcus). 4 and 5. Cocci in pairs 
 (diplococcus). 6. Cocci in groups of four (merismopedia). 7. Cocci in packets 
 (sarcina). 8. Bacterium termo. 9. Bacterium termo x 4000 (Dallinger and 
 Drysdale). 10. Bacterium septictemite hcemorrJiagicce. 11. Bacterium pneu- 
 monia crouposce. 12. Bacillus subtilis. 13. Bacillus murisepticus. 14. 
 Bacillus diphtherien. 15. Bacillus typhosus (Eberth). 16. Spirillum undula 
 (Cohn). 17. Spirillum volutans (Cohn). 18. Spirillum cholerce Asiatics. 
 19. Spirillum Obermeieri (Koch). 20. Spirocliceta plicatilis (Flugge). 21. 
 Vibrio rugula (Prazmowski). 22. Cladothrix Forsteri (Cohn). 23. Cladothrix 
 dichotoma (Cohn). 24. Nonas Okenii (Cohn). 25. Nonas Warmingii (Cohn). 
 26. Rhabdomonas rosea (Cohn). 27. Spore-formation {Bacillus alvei). 28. 
 Spore-formation (Bacillus anthracis). 29. Spore-formation in bacilli cultivated 
 from a rotten melon (Frankel and Pfeiffer). 30. Spore-formation in bacilli 
 cultivated from earth (Frankel and Pfeiffer). 31. Involution-form of Crenotkrix 
 (Zopf). 32. Involution-forms of Vibrio serpens (Warming). 33. Involution- 
 forms of Vibrio rugula (Warming). 34. Involution-forms of Clostridium 
 polymyxa (after Prazmowski). 35. Involution-forms of Spirillum cholerce 
 Asiatics. 36. Involution-forms of Bacterium aceti (Zopf and Hansen). 
 37. Spirulina-form of Beggiatoa alba (Zopf). 38. Various thread-forms of 
 Bacterium merismopedioides (Zopf). 39. False-branching of Cladatkrifo(Zopi). 
 
 DESCRIPTION OF PLATE II. 
 
 Pure-cultivations of Bacteria. 
 
 Following p. 100. 
 
 FIG. \.-ln the depth of Nutrient Gelatine. A pure-cultivation of Koch's 
 comma-bacillus (Spirillum cholerae Asiatics) showing in the track of 
 the needle a funnel-shaped area of liquefaction enclosing an air-bubble, 
 and a white thread. Similar appearances are produced in cultivations of 
 the comma-bacillus of Metchnikoff. 
 
 FIG. 2. On the surface of Nutrient Gelatine. A pure-cultivation of Bacillus 
 typhosus on the surface of obliquely solidified nutrient gelatine. 
 
 xxii 
 
DESCRIFflON OF PLATES. xxiii 
 
 Kn;. 'A. On the surface of Nutrient Agar-agar. Pure-cultivation of Bacillus 
 indicus on the surface of obliquely solidified nutrient agar-agar. The 
 growth has the colour of red sealing-wax, and a peculiar crinkled 
 appearance. After some days it loses its bright colour and becomes 
 purplish, like an old cultivation of Micrococcus prodigiosus. 
 
 FIG. 4. On the surface of Nutrient Agar-agar. A pure-cultivation obtained 
 from an abscess (Staphylococcus pyogenes aureus). 
 
 FIG. 5. On the surface of Nutrient Agar-agar. A pure-cultivation obtained 
 from green pus (Bacillus pyocyaneus). The growth forms a whitish, 
 transparent layer, composed of slender bacilli, and the green pigment 
 is diffused throughout the nutrient jelly. The growth appears green by 
 transmitted light, owing to the colour of the jelly behind it. 
 
 FIG. 0. On the surface of Potato. A pure-cultivation of the bacillus of 
 glanders on the surface of sterilised potato. 
 
 DESCRIPTION OF PLATE III. 
 
 Plate-cultivation. 
 
 Following p. 108. 
 
 This represents the appearance of a plate-cultivation of the comma-bacillus 
 of Cholera nostras, when it is examined over a slab of blackened plate-glass. 
 The drawing was made from a typical result of thinning out the colonies by 
 the process of plate -cultivation. At this stage they were completely isolated 
 one from the other ; but later they became confluent, and produced complete 
 liquefaction of the gelatine. 
 
 DESCRIPTION OF PLATE IV. 
 
 Streptococcus Pyogenes. 
 
 Following p. 178. 
 
 FIG. 1. From a cover-glass preparation of pus from a pyaemic abscess. 
 Stained with gentian-violet by the method of Gram, and contrast-stained 
 with eosin. x 1200. Powell and Lealand's apochromatic T \ Horn. imm. 
 E. P. 10. 
 
 FIG. 2. From cover-glass preparations of artificial cultivations of the strepto- 
 coccus in broth and in milk at different stages of growth, x 1200. Powell 
 and Lealand's apocbromatic ^ Horn. imm. E. P. 10. 
 
 In these preparations there is a great diversity in size and form of the 
 chains and their component elements. In the drawing examples are 
 figured of the following: 
 
 (a) Branched chains. 
 
 (b) Simple chains composed of elements much smaller than the 
 average size. 
 
 (c) Chains with spherical and spindle-shaped elements at irregular 
 intervals. These are conspicuous by their size, and are sometimes 
 terminal. 
 
 (d e) Chains in which the elements are more or less uniform in size. 
 (/) Complex chains with elements dividing both longitudinally and 
 
 transversely, and varying considerably in size in different lengths 
 
 of the same chain. 
 
XXIV DESCRIPTION OF PLATES. 
 
 DESCRIPTION OF PLATE V. 
 Bacillus Anthracis. 
 
 Following p. 102. 
 
 FIG. 1. From a cover-glass preparation of blood from the spleen of a guinea- 
 pig inoculated with blood from a sow. x 1200. Powell and Lealand's 
 apochromatic T \ Horn. imm. E. P. 10. 
 
 FIG. 2. From a section of a kidney of a mouse. Under a low power the 
 preparation has exactly the appearance of an injected specimen. Under 
 higher amplification the bacilli are seen to have threaded their way along 
 the capillaries between the tubules, and to have collected in masses in 
 the glomeruli. Stained with Gram's method (gentian-violet), and eosin. 
 x 500. 
 
 FIG. 3. Bacillus antkracis and Micrococcus tetragemis. From a section from 
 the lungs of a mouse which had been inoculated with anthrax three days 
 after inoculation with Micrococcus tetragenus. A double or mixed infection 
 resulted. Anthrax-bacilli occurred in vast numbers, completely filling the 
 small vessels and capillaries, and in addition there were great numbers 
 of tetrads. Stained by Gram's method (gentian-violet), and with eosin. 
 x 500. 
 
 DESCRIPTION OF PLATE VI. 
 
 Bacillus Murisepticus. 
 
 Following p. 224. 
 
 FIG. 1. From a section Of a kidney of a mouse which had died after inocula 
 tion with a pure-cultivation of the bacillus. With moderate amplification, 
 the white blood-corpuscles have a granular appearance, and irregular 
 granular masses are scattered between the kidney tubules. Stained by 
 Gram's method with eosin. x 200. 
 
 FIG. 2. Part of the same preparation with high amplification. The granular 
 appearances are found to be due to the presence of great numbers of 
 extremely minute bacilli, x 1500. 
 
 DESCRIPTION OF PLATE VII. 
 Casual Cow-pox. 
 
 Following p. 27*. 
 
 FIG. 1. Case of W. P , a milker, infected from the teats of a cow with 
 
 natural cow-pox. There was a large depressed vesicle with a small 
 central crust and a tumid margin, the whole being surrounded by a 
 well-marked areola and considerable surrounding induration. 
 
 !'!<;. 2. The same case a week later, showing a reddish-brown crust on a 
 reddened elevated and indurated base. 
 
DESGBIPTIOID OF PLATES. xxv 
 
 IKS< KUTION OF PLATE VII L 
 
 Bacillus diphtherias and Bacillus typhosus. 
 
 following p. 382. 
 
 FIG. 1. ( 'ovt-r-glass preparation from a pure-cultivation of Bacillus diph- 
 therias on blood serum ; obtained from the throat in a typical case of 
 diphtheria. Stained with gentian-violet, x 1200. 
 
 FIG. 2. Cover-glass preparation from a pure-cultivation of Bacillus typhosus 
 on nutrient-agar ; from the spleen in a case of typhoid fever. Stained 
 with gentian-violet, 
 
 DESCRIPTION OF PLATES IX. AND X. 
 
 Swine Fever. 
 
 Following p. 348. 
 
 PLATE IX. Part of intestine from a typical case of swine fever, showing 
 
 scattered ulcers and ulceration of the ileo-csecal valve. 
 PLATE X. From the same case of swine fever. The lungs were extensively 
 
 inflamed and partly consolidated, and the lymphatic glands were enlarged 
 
 and of a deep red or reddish -purple colour. 
 
 DESCRIPTION OF PLATE XL 
 Bacillus tuberculosis. 
 
 Following p. 378. 
 
 The figures in this plate represent the bacilli of tuberculosis in 
 different animals, examined under the same conditions of amplifica- 
 tion and illumination. X 1200. Lamp-light illumination. 
 
 FIG. 1. Bacilli in pus from the wall of a human tubercular cavity. In 
 this specimen the bacilli are shorter than those in tubercular sputum, 
 and are very markedly beaded. 
 
 FIG. 2. Bacilli in pus from a tubercular cavity from another case in man. 
 They are present in the preparation in enormous numbers. The proto- 
 plasm occupies almost the whole of the sheath, and the bacilli are 
 strikingly thin and long. 
 
 Fi<;. ".. J'acilli in sputum from an advanced case of phthisis, showing 
 the ordinary appearance of bacilli in sputum ; some beaded, others 
 stained in their entirety ; occurring both singly and in pairs, and 
 in groups resembling Chinese letters. 
 
 FKI. 4. Bacilli in a section from the lung in a case of tuberculosis in man. 
 The bacilli in human tuberculosis are found in, and between, the tissue 
 cells ; and sometimes, as in equine and bovine tuberculosis, in the 
 interior of giant cells, but not so commonly. 
 
 FH:. . From a cover-glass preparation of the deposit in a sample of milk 
 from a tubercular cow. The bacilli were longer than the average 
 length of bacilli in bovine tissue sections, and many were markedly 
 beaded. 
 
DESCRIPTION OF PLATES. 
 
 FIG. 6. From a section of the brain in a case of tubercular meningitis in a 
 
 calf, showing a giant cell containing bacilli with the characters usually 
 
 found in sections of bovine tuberculosis. 
 FIG. 7. From a section of the liver of a pig with tubercle bacilli at the 
 
 margin of a caseous nodule. 
 FIG. 8. From a cover-glass preparation of a crushed caseous mesenteric 
 
 gland from a rabbit infected by ingestion of milk from a cow with 
 
 tuberculosis of the udder. 
 FIG. 9. From a section of lung in a case of equine tuberculosis, showing a 
 
 giant cell crowded with tubercle Bacilli. 
 FIG. 10. From a section of lung from a case of tuberculosis in the cat, with 
 
 very numerous tubercle bacilli. 
 FIG. 11. From a cover-glass preparation of a crushed caseous nodule from 
 
 the liver of a fowl, with masses of bacilli. These are for the most part 
 
 short, straight rods ; but other forms, varying from long rods to mere 
 
 granules, are also found. 
 
 FIG. 12. From sections of the liver and of the lung in a case of tubercu- 
 losis of a Khea. Isolated bacilli are found, as well as bacilli packed in 
 
 large cells, colonies of sinuous bacilli, and very long forms with terminal 
 
 spore-like bodies and free oval grains. 
 
 The preparations from which these figures were drawn were all 
 stained by the Ziehl-Neelsen method, with the exception of the first, 
 which was stained by Ehrlich's method. 
 
 DESCRIPTION OF PLATE XII. 
 
 Tubercular Mammitis. 
 
 Following p. 394. 
 
 FIG. 1. From a section of the udder of a milch cow. The tubercular deposit 
 is seen to invade the lobules of the gland. Lobules comparatively healthy 
 are marked off, more or less sharply, from the diseased ones in which the 
 new growth in its progress compresses and obliterates the alveoli. Stained 
 by the Ziehl-Neelsen method and with metl^lene-blue. x 50. 
 
 FIG. 2. Part of the same preparation. On the right of the section part of a 
 healthy lobule is seen. On the left a lobule is invaded by tubercular new 
 growth composed of round cells, epithelioid cells and typical giant cells. 
 Tubercle bacilli can be seen both singly and collected in groups. They 
 are found in and between the cells, and in the interior of giant cells. 
 Bacilli may be seen between the cells lining an alveolus and projecting 
 into its lumen, x 800. 
 
 DESCRIPTION OF PLATE XIII. 
 
 Tuberculosis in Swine. 
 
 Following p. 400. 
 
 Section of liver of a pig with scattered tubercular nodules. Microscopical 
 sections of the liver showed tubercle bacilli in very small numbers. 
 
DESCRIPTION OF PLATES. XXvii 
 
 INSCRIPTION OF PLATE XIV. 
 
 Bacillus Leprae. 
 
 Following p. 408. 
 
 FIG. 1. From a section of the skin of a leper. The section is, almost in 
 its entirety, stained red, and, with moderate amplification, has a finely 
 granular appearance. Stained by the Ziehl-Neelsen method (carbolised 
 fuchsine and methylene-blue). x 200. 
 
 FIG. 2. Part of the same preparation with high amplification, showing that 
 the appearances described above are due entirely to an invasion of the 
 tissue by the bacilli of leprosy, x 1500. 
 
 DESCRIPTION OF PLATES XV. AND XVI. 
 
 Actinomyces. 
 
 Following p. 432. 
 
 PLATE XV. 
 
 FIG. 1. From a preparation of the grains from an actinomycotic abscess in 
 a boy; examined in glycerine. The drawing has been made of a com- 
 plete rosette examined by focussing successively the central and peripheral 
 portions. Towards the centre the extremities of the clubs are alone 
 visible ; they vary in size, and if pressed upon by the cover-glass give the 
 appearance of an irregular mosaic. Towards the periphery the clubs are 
 seen in profile, and their characteristic form recognised. At one part 
 there are several elongated elements, composed of separate links, x 1200. 
 FIG. 2. Different forms of clubs from preparations in which the rosettes have 
 been flattened out by gentle pressure on the cover-glass, x 2500. 
 
 (a) Single club. (*) Bifid club, (c) Club giving rise to four 
 secondary clubs, (d) Four clubs connected together, recalling 
 the form of a bunch of bananas. (<?) Mature club with a lateral 
 bud. (/) Apparently a further development of the condition 
 represented at (e). (^) Club with a lateral bud and transverse 
 segmentation. (#) Single club with double tranverse segmenta- 
 tion, (i) Club with oblique segmentation, (j) Collection of 
 four clubs, one with lateral gemmation, another with oblique 
 segmentation. (Jt) Club with lateral buds on both sides, and 
 cut off square at the extremity. () Club with a daughter club 
 which bears at its extremity two still smaller clubs, (m) Club 
 divided by transverse segmentation into four distinct elements. 
 () Elongated club composed of several distinct elements. (0) and 
 (p) Clubs with terminal gemmation. (</) Palmate group of clubs. 
 (r) Trilobed club. (*) Club with apparently a central channel. 
 () Filament bearing terminally a highly refractive oval body. 
 
 PLATE XVI. 
 FIG. 1. From a section of a portion of the growth removed from a boy 
 
 during life. The tissue was hardened in alcohol, and cut in celloidin. 
 
 The section was stained by Gram's method and with orange-rubin. x 50. 
 Fi<;. '2. From the *ame section. A mass of extremely fine filaments occupies 
 
 the central part of the rosette. Many of the filaments have a terminal 
 
 enlargement. The marginal part shows a palisade of clubs stained by the 
 
 orange-rubin. x 500. 
 
XXviii DESCRIPTION OF PLATES. 
 
 FIGS. 3 and 4. From cover-glass preparations of the fungus teased out of the 
 new growths produced by inoculation of a calf with pus from a boy 
 suffering from pulmonary actinomycosis. Stained by Gram's method and 
 orange-rubin. The threads' are stained blue and the clubs crimson () 
 In the younger clubs the thread can be traced into the interior of the 
 club (&). In some of the older clubs the central portion takes a yellowish 
 stain, and in others the protoplasm is not continued as a thread, but is 
 collected into a spherical or ovoid or pear-shaped mass. In others, again, 
 irregular grains stained blue are scattered throughout the central portion 
 (Fig. 4). x 120(1 
 
 FIG. 5. From a pure-culture on glycerine-agar. (a) branching filaments, 
 (b) a mass of entangled filaments. Gram's method, x 1200. 
 
 FIG. 6. From a similar but older cultivation, (a) a filament with spores, 
 (i) chains of spores simulating streptococci. Gram's method, x 1200. 
 
 DESCRIPTION OF ' PLATES XVII. AND XVIII. 
 
 Actinomycosis Bovis. 
 
 Following p. 434. 
 
 PLATE XVII. 
 
 Section of an actinomycotic tongue stained by the method of 
 Gram and with eosin. 
 
 FIG. 1. This illustrates the appearance which is usually seen under a low 
 power, when a section is stained by Gram's method and with eosin. The 
 central portion of a mass of the fungus is either unstained or tinged with 
 eosin, while the marginal portion is stained blue. The reverse is seen, as a 
 rule, in sections from man ; although under a low power the general appear- 
 ance of sections from these two sources is somewhat similar, x 50. 
 
 FIG. 2. a, b, c, d, represent the earliest recognisable forms of the ray fungus 
 in the interior of leucocytes. In e the club-forms can be recognised. In 
 /and g there are small stellate groups of clubs, x 500. 
 
 FIG. 3. A part of the section represented in Fig. 1, under a high power. The 
 marginal line of blue observed under a low power is now recognised as the 
 result of the stain being limited to the peripherally arranged clubs. At 
 (</) part of a rosette has undergone calcification ; the clubs are granular. 
 and have not retained the stain. At (&) and close to it there are the 
 remains of rosettes in which the process of calcification is almost complete. 
 x 500. 
 
 PLATE XVIII. 
 
 The figures in this plate are taken from sections of a case of 
 so-called " osteosarcoma," in which the growth of the fungus was 
 remarkably luxuriant. The specimens were stained by Plants' 
 method. 
 
 Flu. 1. Different forms of clubs in different specimens : x 1200. 
 (a) Very small club-shaped elements. 
 (&) A club with transverse segmentation. 
 (?) A club with lateral daughter clubs. 
 
DESCRIPTION OF PLATES. Xxix 
 
 (tl and e) Clubs with terminal offshoots resembling teleutospores. 
 (./) A club with developing daughter clubs on the left, and on the 
 
 right a mature secondary club. 
 (//) A segmental club with lateral offshoots. 
 (A) Two clubs undergoing calcification. 
 
 FK;. 2. A very remarkable stellate growth comprised of nine wedge-shaped 
 collections of clubs radiating from a mass of finely granular material, 
 x 500. 
 
 FIG. 3. A rosette undergoing central calcification, and consisting in part of 
 extremely elongated clubs resembling paraphyses. Calcareous matter is 
 also being deposited in the club-shaped structures, x 500. 
 FIG. 4. Part of a rosette with continuation of the club-shaped bodies 
 into transversely segmented branching cells apparently representing short 
 hyphae. x 500. 
 
 FIG. 5. A rosette from another section in which similar appearances are 
 observed as in Fig. 4. x 500. 
 
 DESCRIPTION OF PLATE XIX. 
 
 Pure-cultivations of Actinomyces. 
 
 Following p. 438. 
 
 These tubes were selected from a great number of cultivations 
 in which there were different appearances. In some instances the 
 growths had a faint tinge of pink. 
 
 FIG. 1. Pure-cultivation on the surface of potato, showing a luxuriant 
 sulphur-yellow growth entirely composed of entangled masses of fila- 
 ments. After three months' growth. 
 
 FIG. 2. Pure-culture from the same series, on glycerine-agar. In this case 
 the culture remained perfectly white. The jelly was coloured reddish- 
 brown. After fifteen months' growth. 
 
 FIG. 3. Pure-culture on glycerine-agar in which the growth was dark- 
 brown, in parts black, and the jelly stained dark-brown. After nearly 
 two years' growth. 
 
 DESCRIPTION OF PLATES XX. AND XXI. 
 
 Actinomycosis Bovis. 
 
 Following p. 440. 
 
 PLATE XX. 
 
 Fn;. 1. From a section of an actinomycofeic tongue stained by the triple 
 method (Ziehl-Neelsen, logwood and orange-rubin). In this section the 
 separate centres of growth are clearly shown. Each neoplasm consists of 
 a fungus system, im which the masses of the fungus, situated more or less 
 centrally, are surrounded with round cells, epithelioid cells, sometimes 
 giant cells, and lastly fibrous tissue forming a more or less distinct 
 capsule. In parts the fungi have fallen out of the section. x 50. 
 
 FIG. 2. From a section of a "tubercular" nodule from the lungs of a 
 Norfolk heifer with pulmonary actinomycosis. The nodule is a multiple 
 growth surrounding a bronchus, and is enclosed by a capsule, in the 
 
XXX DESCRIFHON OF PLATES. 
 
 vicinity of which the pulmonary alveoli are compressed. It is composed 
 of a number of separate neoplasms, and each of the latter is composed of 
 secondary centres of growth resembling the giant-cell systems of bacillary 
 tuberculosis. The new growth is composed of ray-fungi, large multi- 
 nucleated cells, sometimes distinct giant cells, round cells, epithelioid cells, 
 and, surrounding them, fibrous tissue. On examination of the same 
 specimen with a higher power the typical rosettes of clubs are sometimes 
 surrounded by multinucleated cells, and sometimes small rosettes are 
 found like tubercle bacilli, in the interior of giant cells. From a pre- 
 paration stained by Ziehl-Neelsen, logwood, and orange-rubin. x 50. 
 
 PLATE XXI. 
 
 FlG. !.(#) A leucocyte containing the fungus in its earliest recognisable 
 form. (&) A large multinucleated cell containing the fungus in an early 
 stage with the club-form already visible, (c) A leucocyte containing a 
 small stellate fungus, (d) A large cell containing clubs arranged in a 
 small rosette. (<?) A multinucleated cell with clubs arranged in a palmate 
 form. All the above are drawn from sections of actinomycotic tongues 
 stained by the triple method, x 500. 
 
 FIG. 2. A giant cell with large vesicular nuclei at the periphery, and in the 
 centre a fully formed rosette of actinomyces with a smaller growth within 
 a " daughter " cell. From a section of the tongue of an ox stained by 
 the triple method, x 500. 
 
 FIG. 3. A very large circular giant cell, with its ring of nuclei at the 
 periphery, enclosing several isolated tufts of actinomyces. From a section 
 of a nodule in the lung. Stained by the triple method, x 500. 
 
 FIG. 4. Three rosettes of actinomyces surrounded by a row of large, some- 
 what angular multinucleated cells. From a section of the tongue of an 
 ox stained by the triple method, x 430. 
 
 DESCRIPTION OF PLATE XXII. 
 
 Bacillus tetani. 
 
 Following p. 458. 
 
 FIG. 1. From a cover-glass preparation of a pure-cultivation of the tetanus 
 bacillus in broth; stained with Neelsen's carbolised fuchsine. x 1200. 
 Lamplight illumination. 
 
 FIG. 2. From a cover-glass preparation from the same source ; stained with 
 Neelsen's solution and methylene blue, x 1200. Lamplight illumination. 
 
PART I. 
 
 THEORETICAL AND TECHNICAL. 
 
BACTERIOLOGY 
 
 AND 
 
 INFECTIVE DISEASES. 
 
 CHAPTER I. 
 
 HISTORICAL INTRODUCTION. 
 
 THE researches of Pasteur into the rdle played by bacteria in the 
 processes of fermentation and putrefaction, and the investigations of 
 the practical inind of Lister, with the resulting evolution of antiseptic 
 surgery, demonstrated the necessity for a more intimate acquaint- 
 ance with the life- history of these micro-organisms. Further re- 
 searches in diseases such as anthrax, the silkworm malady, pyaemia, 
 septicaemia, and fowl-cholera, invested the science of Bacteriology 
 with universal interest and vast importance; while the investiga- 
 tions which established an intimate connection between bacteria 
 and other infective diseases, and more especially the discovery by 
 Koch of bacteria in tuberculosis and in Asiatic cholera, claimed the 
 attention of the whole thinking world. 
 
 Those bacteria which are connected with disease, and more 
 especially those which have been proved to be the causa causans^ are 
 of predominant interest and importance. 
 
 The first attempt to demonstrate the existence of a contagiwtn 
 rivum dates back almost to the discovery of the microscope. 
 Athanasius Kircher, nearly two and a half centuries ago, expressed 
 his belief that there were definite micro- organisms to which diseases 
 were attributable. The microscope had revealed that all decom- 
 posing substances swarmed with countless micro-organisms which 
 were invisible to the naked eye, and Kircher sought for similar 
 organisms in diseases which he considered might be due to their 
 agency. The microscope which he described obviously could not 
 
 1 
 
2 BACTERIOLOGY. 
 
 admit of the possibility of studying, or even detecting, the micro- 
 organisms which are now known to be associated with certain 
 diseases ; and it is not surprising that his teachings did not at the 
 time gain much attention. They were destined, however, to receive 
 a great impetus from the discoveries which emanated from "the 
 father of microscopy." 
 
 Antony van Leeuwenhoek had learned as a youth to grind and 
 polish lenses, and later in life employed his spare time in constructing 
 microscopes, and in conducting those researches which have made 
 for him a name which is familiar to all microscopists. His researches 
 were published in a series of letters to the Royal Society. In 1675 
 he described extremely minute organisms in rain-water, well-water, 
 infusions of pepper, hay, and other vegetable and animal substances,. 
 in saliva, and in scrapings from the teeth ; and, further, he was 
 able to differentiate these minute living things by their size, their 
 form, and the character of their movements. In 1683 these 
 discoveries were illustrated by means of woodcuts, and there can be 
 little doubt, from the drawings of these micro-organisms, that they 
 are intended to represent leptothrix filaments, vibrios, and spirilla. 
 Indeed, we can almost recognise these micro-organisms as bacteria 
 from Leeuwenhoek's graphic descriptions, apart from his figures. 
 They were described as moving in the most characteristic manner, 
 progressing with great rapidity, or spinning round like a top, and 
 so excessively minute that they were only perceived with great 
 difficulty. The smallest forms could hardly be examined individually ; 
 but, viewed en masse, they closely resembled a swarm of gnats or 
 flies. In another communication, published in 1692, he gives 
 some idea of the size of these animalcules by stating that the}' 
 were a thousand times smaller than a grain of sand. Others 
 which were, comparatively speaking, of considerable length, were 
 characterised by their peculiar mode of progression, bending and 
 rolling on themselves movements which, he adds, created both 
 delight and astonishment in the mind of the observer. Leeuwenhoek 
 himself was not disposed to believe in the possibility of such 
 organisms being found in the blood in disease; but as soon as he 
 had proved the actual existence of such minute creatures, theoretical 
 physicians were not wanting who at once attributed various maladies 
 to their agency. Among these, Nicholas Andry is made conspicuous 
 by his work published in 1701, Andry classed the minute organisms 
 discovered by Leeuwenhoek as worms. 
 
 In 1718 Lancisi believed that the deleterious effect of the air of 
 malarial districts depended upon animalcules, and others considered 
 
HISTORICAL INTRODUCTION. 
 
 that the plague in Toulon and Marseilles in 1721 arose from a 
 similar cause. In fact, by some, all diseases were attributed to 
 \vrmicules. and this led to the theory being ridiculed and discredited. 
 
 In spite of adverse criticism, the theory of contagium vivum 
 survived, and Linnaeus acknowledged it by placing the micro- 
 organisms discovered by Leeuwenhoek, the contagia of specific 
 fevers, and the causes of putrefaction and fermentation, into one 
 class " chaos." The theory was further supported by the writings 
 of Plenciz, who, in 1762, very ably discussed the nature of contagium r 
 as well as the relation of animalcules to putrefaction and disease. 
 However, no proofs in support of these theories were forthcoming, 
 and gradually the idea of contagium vivum fell into obscurity, and 
 indeed came to be regarded by some as an absurd hypothesis. 
 
 Though a causal relation of animalcules to diseases was for a 
 time discredited, the natural history of these micro-organisms was- 
 studied with increasing interest. In 1778 Baron Gleichen described 
 and figured a great number of micro-organisms which he had 
 discovered in various vegetable infusions. Joblot, Lesser, Reaumur, 
 Hill, and many others worked at the same subject. Hill remarked 
 that there was hardly the least portion of matter or the least drop 
 of fluid of any kind naturally found in the earth, which was not 
 inhabited by multitudes of animalcules. But these observers inclined 
 rather to searching for new forms than to studying more thoroughly 
 those which had been already discovered ; and, as a result, but little 
 scientific progress was made until the time of M tiller, of Copen- 
 hagen. Miiller, in 1786, criticised the work of previous writers, 
 and pointed out that they had been too much occupied with merely 
 finding new micro-organisms. Miiller took into account the form 
 of the micro-organism, its mode of progression, and other biological 
 characters, and on such data based a classification. Thus the 
 scientific knowledge of these minute beings was considerably advanced 
 ly his writings and illustrations. 
 
 The subject which now eclipsed all others in interest was the 
 origin of these micro-organisms. Two rival theories were widely 
 discussed spontaneous generation, and development from pre-exi-t 
 ing germs ; and the researches that were made in the course of 
 this di>cuion. and the discoveries which resulted, indirectly yet 
 materially advanced the germ theory of <li>tMM-. and explain many 
 of the phenomena in the life-history of the pathogenic microbes 
 which have been brought to light in recent years. 
 
 Spontaneous development of micro-organisms in putrescible 
 infusions was believed in by many, but was supported by no one 
 
4 BACTERIOLOGY. 
 
 with greater persistency than Needham. Needham found that 
 animalcules readily developed \vhen meat infusion was boiled an 
 transferred to a well-stoppered flask, and he could only explain this 
 by supposing that they originated spontaneously from the material of 
 the infusion. In 1768 Bonnet strenuously opposed these conclusions 
 on purely theoretical grounds, and maintained that it was far more 
 probable that the ova of the animalcules were present in the infusions 
 or were suspended in the air enclosed in the flask. 
 
 Spallanzani was the first to demonstrate by experiment the 
 correctness of Bonnet's arguments. It occurred to him to boil the 
 infusion in flasks, and to seal the vessels during the process of boiling. 
 As a result the flasks remained free from putrefaction, and 
 animalcules only developed when the infusion was exposed to the air 
 by making a hole in the flask. That Spallanzani's experiments 
 were reliable, and his conclusions correct, was evidenced by the fact 
 that his simple precaution led to great practical results, as Francois 
 Appert introduced, on this principle, the method of preserving meats, 
 vegetables, and other provisions. 
 
 The disciples of Needham nevertheless brought forward counter 
 objections. Treviranus urged that a certain quantity and quality of 
 air was necessary for the spontaneous development of these infusoria, 
 and that by sealing the flasks, too small a quantity of air was in 
 contact with the infusion, and, further, that this air had become 
 changed in quality by the process of boiling. 
 
 Spallanzani argued against these objections, but did not support 
 his opinions by further experiments, so that the question remained 
 for a time undecided. 
 
 In 1836 Francis Schulze devised an experiment which brought 
 still further evidence against Needham's theory. Schulze filled a 
 glass vessel half full with distilled water and different animal and 
 vegetable substances. This was plugged with a doubly-bored cork, 
 and through each perforation a glass tube was introduced, bent at 
 a right angle. On boiling the flask, steam issued freely from each 
 tube, and all parts were thoroughly sterilised. Each tube was then 
 connected with a bulbed tube, one bulb containing concentrated 
 sulphuric acid and the other a solution of potash. Fresh air was 
 drawn into the flask by aspiration, and this was deprived of any 
 germs which might be present by its passage through the sulphuric 
 acid. The result was that the infusion remained without any 
 development of micro-organisms. When, on the other hand, air was 
 admitted without first being drawn through the sulphuric acid, the 
 n asion in a short time teemed with animalcules. In other words 
 
HISTORICAL INTRODUCTION. 5 
 
 Scliulze demonstrated that in spite of free access to air, which Jiadnot 
 been heated, the infusions remained free from germs. 
 
 Schwann, in 1837, arrived at similar results. He found that 
 putrescible substances remained sterile if exposed to an abundant 
 supply of air which was heated by being passed through a melted mix 
 ture of metals. This convinced him that the cause of the decompo- 
 sition which would otherwise have occurred must exist in the air. 
 
 The objection remained that in the experiments of Schulze and 
 Schwann, the air which was admitted to the flasks had undergone 
 either a chemical or a thermal change, and therefore the theory of 
 Needham was not yet entirely disposed of. 
 
 In 1854 the final blow was dealt by Schroder and Van Dusch, 
 These investigators demonstrated that decomposition could be obviated 
 without resorting either to thermal or chemical treatment of the 
 air, as simple filtration of the air through cotton- wool was shown to- 
 be efficacious in excluding germs. Finally, Hoffman in 1860, and 
 independently, Chevreuil and Pasteur in 1861, showed that even 
 cotton-wool could be dispensed with, as a sterile solution would 
 remain sterile when the neck of the vessel was bent into an 
 S -shaped curve. Micro-organisms in the air entering the flask 
 were deposited by gravitation in the bend of the tube. 
 
 The advocates of spontaneous generation were ready with fresh 
 objections. They now urged that the medium lost its power of 
 undergoing decomposition by being boiled. This objection was at 
 once set aside by the fact that when unfiltered air w r as admitted to 
 the infusion, decomposition set in. Additional evidence was brought 
 against spontaneous generation by the experiments of Pasteur, 
 Burdon Sanderson, Lister, and others, in which it was shown that 
 blood, urine, and milk would remain without decomposition, when all 
 precautions were adopted to avoid contamination in filling the 
 sterilised flasks. 
 
 Even at this stage of this great scientific controversy fresh 
 difficulties arose, for it was found that in certain solutions which had 
 been boiled and hermetically sealed in flasks micro-organisms made 
 their appearance. In 1872 Charlton Bastian published a research 
 which was to prove that spontaneous generation actually took 
 place. Decoctions of turnip and cheese which had been filtered, 
 neutralised, and boiled for ten minutes, and hermetically sealed 
 during the boiling, were found after a time to contain micro 
 organisms. These results, however, were before long explained by 
 the fact that in milk, infusions of hay, and certain other decoctions, 
 the spores of bacilli are present, which are much more resistant 
 
6 BACTERIOLOGY. 
 
 than the bacilli themselves. In such cases mere scalding or boiling 
 for a few minutes will not sterilise the solution. The bacilli are 
 destroyed, but not their spores; and if the latter remain unhurt, 
 they will germinate, and rapidly multiply. But if, as Tyndall 
 found, the boiling be repeated a second and a third time, all the 
 spores will be destroyed ; for in the intervals between the boilings 
 the spores sprout into bacilli, and the bacilli at the next boiling 
 perish ; so that after three or four repeated boilings the infusion is 
 rendered perfectly free from germs. 
 
 While this discussion was occupying the attention of the whole 
 scientific world, some investigators had been again following up the 
 theory of a connection between micro-organisms and disease. 
 
 In 1837 Gagniard Latour and Schwaiin independently made the 
 discovery that the yeast plant was a living organism, and the true 
 cause of yeast fermentation. The close analogy between the pro- 
 cesses of fermentation and of certain diseases had long been held ; 
 and, therefore, when it was proved that fermentation was due to a 
 micro-organism, fresh advocates appeared in support of the theory 
 that diseases were produced by similar agencies. Boehm, in 1838, 
 described certain organisms in cholera, which was at that time 
 raging in Europe; but the researches of Bassi, who a year 
 previously had discovered the cause of a disease of silkworms, 
 attracted much greater attention. 
 
 Bassi discovered that in this disease extremely minute spores 
 existed on the bodies of the worms, which were conveyed from the 
 .sick to the healthy. They destroyed the healthy worms by 
 germinating in their skins and growing into their bodies. These 
 discoveries may be said to have brought the theory of contagium 
 vivum to life again ; and Henle, in reviewing the facts of the case in 
 1 840, came to the conclusion that the cause of all contagious diseases 
 must be of a living nature, and this he maintained, although he 
 had searched in vaccine and small-pox lymph, in the desquamation 
 of scarlet fever, and in other diseases without success. 
 
 Bassi's discovery and Henle's doctrine encouraged a number of 
 investigators, and remarkable results followed. In favus, in herpes 
 tonsurans, in pityriasis versicolor, fungus threads and spores were 
 found, and were regarded as being of etiological importance, 
 inasmuch as the morbid lesions corresponded with the growth of the 
 particular fungus. 
 
 Cholera became especially a subject for research. Swaine, 
 Brittan, and Budd found micro-organisms in choleraic dejecta, 
 Davaine described certain monads in the intestinal contents, but no 
 
HISTORICAL INTRODUCTION. 7 
 
 causal connection was established between these organisms and the 
 disease ; ami when the cholera disappeared the interest in contagium 
 i-ii-Hiii waned, and was eclipsed by the question of fermentation. 
 The discoveries which followed in this subject had a very important 
 1> 'aring on the micro- parasitic origin of communicable diseases. 
 
 Pasteur, following up the researches of Cagniard Latour and 
 Schwann, demonstrate 1 in 1857 that the lactic, acetic, and butyric 
 fermentations were produced by micro-organisms. 
 
 Previously to this, in 1850, Davaiue and Rayer had noted the 
 existence of little rod-like or filamentous bodies, about the size of a 
 blood corpuscle in the blood of a sheep that had died of splenic fever. 
 Pollender had seen similar bodies in the blood of cows. Davaine 
 did not at first pay much heed to this discovery; but in 1863 he 
 thoroughly rein vestiga ted the subject, and conducted a series of 
 experiments which led him to the conclusion that the actual cause 
 of splenic fever was an organised being whose presence and 
 multiplication in the blood produced changes in that fluid of the 
 nature of fermentation, resulting in the death of the animal. 
 
 These conclusions were not accepted by all, and indeed, the 
 evidence was so far incomplete that sceptics were justified in con- 
 sidering that these experiments afforded only a working hypothesis. 
 But Davaine's comparison between this disease and fermentation 
 attracted the attention of Pasteur, whose mind had been fully trained 
 for entering upon this investigation by the researches which he had 
 been carrying on in the interval between Davaine's publications of 
 1857 and 1863. 
 
 Pasteur, as already mentioned, had been working at fermentation, 
 and his attention was next directed to studying the so-called diseases 
 of wines, and subsequently to a contagious disease which committed 
 ravages among silkworms. Bv laborious researches Pasteur was 
 able to confirm the belief that this disease of silkworms was due to 
 the presence of micro-organisms discernible with the aid of the micro- 
 scope. These oval shining bodies in the moth, worm, and eggs had 
 been previously observed by Cornalia, and described by Nageli as 
 Nosema bombycis, and by Lebert as Paiihistophyton. But it was 
 reserved for Pasteur to introduce a means of combating the disease. 
 ur showed that when a silkworm, whose body contained these 
 micro- organism s. was pounded up with water in a mortar, and the 
 mixture painted with a brush on the leaves on which healthy worms 
 were fed, they would all without fail succumb to the disease. 
 
 As the contagious particles were transmitted to the eggs, a 
 method for ]>r -v.nting the spread of the disease suggested itself. 
 
8 BACTERIOLOGY. 
 
 Each female moth was kept separate from the others, and allowed to 
 deposit her eggs on a small linen cloth. The moth was then pinned 
 to the corner of the cloth, and left for future examination. When 
 the time for this arrived, the moth was crushed up with water in a 
 mortar, and a drop examined under the microscope. When any 
 trace of corpuscular matter was found to be present, the cloth with 
 its collection of eggs was burnt ; and if not, the eggs were set aside 
 for use. 
 
 Complete as this appears to be as a demonstration of a causal 
 connection between the micro-organisms and the disease, it could 
 obviously be objected that there was no distinct proof that the 
 corpuscular bodies constituted the actual contagium. There was no 
 isolation of the organisms, no artificial cultivation of them apart 
 from the diseased moth or worm, and subsequent production of the 
 disease by means of the isolated organisms. The same objection 
 was applicable to Davaine's investigations. Davaine found rods 
 in association with anthrax, and maintained that they were 
 causally related ; but others stated that it was possible to inoculate 
 animals with anthrax blood containing rods, and to produce the 
 disease without being able to detect the rods again in the blood 
 of the animal experimented upon. It was also urged that it was 
 possible to infect with anthrax blood after the rods had disappeared, 
 and to find a reappearance of the bacilli in the blood of the 
 inoculated animal. 
 
 The well-known fact that anthrax was especially prevalent in 
 certain seasons and certain localities appeared to lend great support 
 to these objections. The disease, in fact, was regarded by some 
 as originating from peculiar conditions of climate and soil. The 
 fallacies in these objections were, however, rapidly dispelled. 
 Bollinger, in 1872, pointed out that the blood, from which the rods 
 had disappeared, was still virulent owing to the presence of the 
 spores of the bacillus, and that it was owing to the soil being impreg- 
 nated with these spores that the disease broke out in certain 
 localities. Yet there still remained many who refused to regard 
 these particles as living bodies, some looking upon them simply as 
 crystals ; and the question of their importance remained undecided 
 for several years. 
 
 In 1877 Robert Koch published a memoir in which he fully 
 described the life-history of the anthrax or splenic fever bacillus, 
 ;tiid gave a complete demonstration of the life-history of the micro- 
 organism, and the definite proofs of its pathogenic properties. He 
 pointed out how the rods grew in the blood and tissues by lengthen- 
 
HISTORICAL INTRODUCTION. 9 
 
 ing and by cross division. Further, that in the blood or in serum 
 or in aqueous humour they not only grew into long leptothrix 
 filaments, but they produced enormous numbers of seeds or spores. 
 He traced, by continuous observation on the warm stage, the whole 
 life cycle, from the fission of the rods to the formation of spores and 
 the sprouting of the spores into fresh rods. Further, he carried 
 on the disease by inoculating from mouse to mouse for several 
 generations, and observed that in the blood of the animal and in 
 the swollen spleen the glass-like rods were always to be found. 
 
 Pasteur also studied the microbe of splenic fever, and amply 
 confirmed and extended the observations of Koch by his researches 
 on the attenuation of the anthrax virus. 
 
 Pasteur also met with adverse criticism. Paul Bert argued 
 that the bacilli were of no importance, because he could destroy 
 them by exposing material containing them to great pressure, and 
 yet the material produced the disease on inoculation. But such 
 measures did not destroy the spwes ; and finally, Paul Bert was 
 convinced of his error when Pasteur demonstrated cultures of the 
 anthrax bacillus in urine, from which successive generations were 
 started, and that with such cultivations the disease could always be 
 produced. 
 
 It was, however, principally the researches of Koch which 
 placed the doctrine of contagium vivum on a scientific basis. 
 
 Koch's improvements in the methods of cultivation, his recom- 
 mendation of the necessary microscopical apparatus, his histological 
 methods for examining these minute organisms, and his famous 
 postulates for proving beyond controversy the existence of specific 
 pathogenic micro-organisms, elevated the theory of contagium vivum 
 t a demonstrated and established fact. The chain of evidence 
 regarded by Koch as essential for proving the existence of a 
 pathogenic organism was as follows : 
 
 1. The micro-organism must be found in the blood, lymph, or 
 diseased tissue of man or animal suffering from or dead of the 
 disease. 
 
 2. The micro-organisms must be isolated from the blood, lymph, 
 or tissues, and cultivated in suitable media i.e., outside the animal 
 1 M >< 1 v. These pure cultivations must be carried on through successive 
 generations of the organism. 
 
 .'>. A pure cultivation thus obtained must, when introduced into 
 the body of a healthy animal, produce the disease in question. 
 
 4. In the inoculated animal the same micro-organism must 
 he found. 
 
1 BACTERIOLOGY. 
 
 The chain of evidence is still more complete if we can from 
 artificial cultures obtain a chemical substance which is capable 
 of producing the disease independently of living micro-organisms. 
 
 It is of very little value merely to detect or artificially to 
 cultivate a bacterium associated with disease. We must endeavour 
 to establish the exact relationship of the bacteria to disease processes, 
 and the determination of the true pathogenic microbe is beset 
 with fallacies. In. many diseases bacteria have been regarded as 
 the actual contagia, until a searching inquiry by other investigators 
 has shown that the evidence was most unsatisfactory or entirely 
 misleading. For example, in diseases with lesions of the external 
 or internal linings of the body, extraneous micro-organisms may 
 get into the circulation and be swept into the internal organs, 
 where they either perish in the battle with the healthy tissues 
 which are opposed to their existence, or they may gain the upper 
 hand, and set up destructive processes. Such organisms, when 
 found in association with these diseases, may be discovered in the 
 blood and internal organs ; and though only accidental epiphytes, 
 often associated with septic complication, they may too readily be 
 accepted by the enthusiast as the actual contagium of the disease 
 in question. 
 
 It is only when such fallacies are exposed that we are brought 
 once more face to face with the fact that the nature of the contagium 
 in hydrophobia, variola, vaccinia, scarlet fever, measles, and many 
 other diseases, is still undetermined. 
 
CHAPTER II. 
 
 MORPHOLOGY AND PHYSIOLOGY OF BACTERIA. 
 
 BACTERIA may be considered as minute vegetable cells destitute of 
 nuclei. They are distinguished from animal cells by being able to 
 <terive their nitrogen from ammonia compounds, and they differ 
 from the higher vegetable cells in being unable to split up carbonic 
 acid into its elements, owing to the absence of chlorophyll. Von 
 Eugelmami and Van Tieghem include among the bacteria certain 
 organisms, named by them Bacterium chlorinum, Bacterium viride, 
 and Bacillus virens, which are coloured green by this substance, but 
 it is quite possible that they may be Algse, and further researches 
 are required before any conclusions are definitely arrived at as to 
 the exact place these particular organisms occupy in the vegetable 
 kingdom. 
 
 Composition. For our knowledge of the chemical composition 
 of K-trteria we are chiefly indebted to Nencki. Their constituents 
 are found on analysis to vary slightly, according to whether the 
 bacteria are in zoolgcea or in the active state. In the latter condition 
 they are said to consist of 83-42 per cent, of water. In one hundred 
 parts of the dried constituents there are the following : 
 
 A nitrogenous body ..... 84*20 
 
 Fat . . 6-04 
 
 Ash 4-72 
 
 Undetermined substances . . . . 5 '04 
 
 This nitrogenous body is called myco-protein, and consists of 
 
 Carbon 52'32 
 
 Hydrogen ....... 7 '55 
 
 Nitrogen 14'75 
 
 but no sulphur or phosphorus. 
 
 The nitrogenous body appears to vary in different species, for 
 
 in Bacillus anthracis a substance has been obtained which does not 
 
 11 
 
12 BACTERIOLOGY. 
 
 give the reactions of myco-protein, and, therefore, is distinguished 
 as anthrax-protein. 
 
 Considering bacteria as cells, we may speak of the cell-wall and 
 the cell-contents. The cell-wall consists of cellulose, or, according 
 to Nencki, in the putrefactive bacteria of myco-protein. It may be 
 demonstrated by the action of iodine, which contracts the proto- 
 plasmic contents, and renders the cell-wall visible. By staining 
 cover-glass preparations of the anthrax bacillus by the method of 
 Gram, the rods are at first uniformly stained, by subjecting them to- 
 iodine solution the protoplasmic contents are contracted, and alcohol 
 decolorises the sheath, which may be then stained in contrast, with 
 eosin. 
 
 The cell-wall may be either pliable or rigid. Pliability is 
 observed in the long filaments, which are endowed with a slow 
 vermicular movement, while rigidity accounts for the maintenance 
 of the characteristic form of several species, such as spirilla. 
 
 The cell-protoplasm yields myco-protein. In some it is homogene- 
 ous, and in others granular. The action of the aniline dyes indicates 
 a close relation to nuclear protoplasm, though all nuclear stains are 
 not suitable for bacteria. In some cases also the bacteria remain 
 stained under the influence of a reagent, which removes the colour 
 from nuclei. The power of fixing the stain is not always present t 
 and indicates a difference in the protoplasm of different species. 
 Thus in staining phthisical sputum, the nitric acid removes the 
 stain from all bacteria and bacilli present, with the exception of the 
 tubercle bacillus. This difference in the protoplasm of different 
 species is also illustrated by the necessity, in many cases, of using 
 special processes, owing to the ordinary methods being unsatisfactory 
 or not producing any result. 
 
 The protoplasm of some bacteria contains starch granules ; thus 
 Clostridium butyricum gives the starch reaction with iodine. 
 Sulphur granules are present in some species of Beggiatoa which 
 thrive in sulphur springs. The colouring- matter of the pigmented 
 bacteria is probably external to the cell as a rule : for example, in 
 Micrococcus prodigiosus the pigment granules are distictly between 
 the cells; on the other hand, in Beggiatoa roseo-persicina, or the 
 peach-coloured bacterium, the special pigment bacterio-purpurin 
 appears to be dissolved in the cell protoplasm. In Bacillus 
 pyocyaneus the pigment is certainly not localised entirely in the 
 cell, for it becomes rapidly diffused in the surrounding medium, 
 considerably beyond the confines of the growth itself. 
 
 In several species, either as a result of a secretion from the cell or 
 
MORPHOLOGY AND PHYSIOLOGY OF BACTERIA. 13 
 
 of the absorption of moisture and consequent swelling of the outer 
 layer of the cell-wall, a mucinous or gelatinous envelope develops 
 around them. This envelope may form a capsule, such as we meet 
 with in certain bacteria found in the rusty sputum of pneumonia, and 
 in Micrococcus tetragenus ; or it may occur as a continuous sheath 
 around a chain of bacteria, which by its disappearance sets free 
 the individual links. The capsule is soluble in water, and under 
 some circumstances is difficult to demonstrate. In the pneumo- 
 coccus of Friedlander the capsule disappears on cultivation, but 
 reappears in preparations made from an inoculated animal. In the 
 pleuritic fluid of a mouse these cocci are often found with a parti- 
 cularly well-marked capsule, and in other encapsuled cocci the extent 
 of the envelope has been observed to vary considerably in the same 
 species of bacterium. 
 
 When this gelatinous material forms a matrix, in which 
 numbers of bacteria are congregated in an irregular mass, we have 
 what is termed a zoogloea. The zooglcean stage is a resting stage, 
 often preceded or followed by a motile stage. Thus bacteria may 
 be present in a solution in an active state, and after a time a scum 
 or pellicle forms on the surface of the liquid, which consists of 
 zooglcea. At the edges of the zooglcea, individuals may be seen to 
 again become motile, and after detaching themselves to swim off in 
 the surrounding fluid. 
 
 The zoogloean stage may be observed sometimes in cultivations in 
 broth, and also in nutrient gelatine which has become liquefied. 
 The inoculated bacteria grow and multiply, and after a time a film 
 appears on the surface of the liquefied layer. In cultivations on 
 potato the appearances in this stage are varied, and sometimes 
 extremely characteristic. In the case of a bacillus which readily 
 develops on unsterilised potatoes, the zoogloea may spread over the 
 cut surface, forming a pellicle which can be raised en masse like 
 a delicate veil. Another bacillus forms a zooglcea, consisting of a 
 tenacious layer w^hich can be drawn out in long stringy threads. 
 In Ascococcus Billrothii the gelatinous envelope develops to such 
 an enormous extent that it forms the characteristic feature of the 
 secies. (Fig. 1.) 
 
 Form. The individual cells vary in form, and may either 
 remain isolated or attached to each other. Hound cells and egg- 
 shaped cells are called cocci. The spherical form is the mast 
 common, but cocci are occasionally exclusively ovoid, as in Strepto- 
 coccus bombycis. The giant cocci of some species are spoken of as 
 j/iegacocci, to distinguish them from the ordinary cocci, or micrococci. 
 
14 
 
 BACTERIOLOGY. 
 
 The fission by which the cocci increase may take place in one 
 direction only, and if the two resulting cells remain attached to 
 each other they form a diplococcus. If these two cells again divide, 
 and the resulting cells remain linked together, we get a chain or 
 rosary, termed streptococcus. These chains may consist of a few 
 individuals linked together, or of several hundreds, in which case 
 the chains are generally curved or twisted. When the division 
 occurs in two directions, so that four cocci result, a tetrad or 
 merismopedia is formed ; when in three directions, one coccus divides 
 into eight, and the result is a packet form or sarcinacoccus. 
 Immediately after division, the daughter cells are not perfectly 
 circular, but are flattened or facetted where they are opposite to 
 
 FIG. 1. Ascococcus BILLROTHII, x 65. [After Cohn. 
 
 each other. They gradually become rounded off, and each daughter 
 cell is then ready to divide in its turn. In other cases the cocci 
 after division only form irregular heaps or collections like bunches 
 of grapes. This form is sometimes distinguished as staphylococcus, 
 but it cannot be considered an important feature. When we find 
 irregular masses of cocci united by intercellular substance and 
 embedded in a tough gelatinous matrix, the form is described as 
 ascococcus. 
 
 Another type is the rod, characteristic of bacterium and bacillus. 
 The rods may vary considerably in length. The very short rods 
 with rounded ends are difficult to distinguish from the oval cocci, 
 but differ in that a rod, however short it may be, must have 
 two sides parallel. The vibrio or bent rod may be considered as 
 the connecting link between the rods and the corkscrew forms or 
 
DESCRIPTION OF PLATE I. 
 Bacteria, Schizomycetes, or Fission Fungi. 
 
 1. Cocci singly and varying in size. 2. Cocci in chains or rosaries (strepto- 
 coccus). 3. Cocci in a mass (staphylococcus). 4 and 5. Cocci in pairs 
 (diplococcus). 6. Cocci in groups of four (merismopedia). 7. Cocci in packets 
 (sarcina). 8. Bacterium termo. 9. Bacterium ternio x 4000 (Dallinger and 
 Drysdale). 10. Bacterium gqriictgmia hamorrhagicce. 11. Bacterium pneu- 
 monia crouposce. 12. Bacillus subtilis. 13. Bacillus murisej)ticus. 14. 
 Bacillus diphtheria. 15. Bacillus typhosus (Eberth). 16. Spirillum undula 
 (Cohn). 17. Spirillum volutans (Cohn). 18. Spirillum cholerce Asiaticce. 
 19. Spirillum Obermeieri (Koch). 20. Spirochata plicatilis (Fliigge). 21. 
 Vibrio rugula (Prazmowski). 22. Cladothrix Forsteri (Cohn). 23. Cladothrix 
 dichotoma (Cohn). 24. Nonas Okenii (Cohn). 25. Nonas Warmingii (Cohn). 
 26. Rhabdomonas rosea (Cohn). 27. Spore-formation (Bacillus alvei). 28. 
 .>pore-formation (Bacillus anthracis). 29. Spore-formation in bacilli cultivated 
 from a rotten melon (Frankel and Pfeiffer). 30. Spore-formation in bacilli 
 cultivated from earth (Frankel and Pfeiffer). 31. Involution-form of Crenothrix 
 (Zopf). 32. Involution-forms of Vibrio serpens (Warming). 33. Involution- 
 forms of Vibrio rugula (Warming). 34. Involution-forms of Clostridium 
 polymyxa (after Prazmowski). 35. Involution-forms of Spirillum cholera 
 Axiaticte. 36. Involution-forms of Bacterium aceti (Zopf and Hansen). 
 37. Spirulina-form of Beggiatoa alba (Zopf). 38. Various thread-forms of 
 Bacterium merismopedi aides (Zopf). 39. False-branching of Cladothrix (Zopf) . 
 
: 
 
 i 
 
 V 
 
 
 
 : 
 
 1 
 
 
 
 t 
 
 I 
 
 
 
 i 
 
 
 
 
 s 
 
 \ 
 
 
 i 
 
 :S \l 
 
 : i \ 
 
 BACTERIA .srmZOMYCETES. OR FISSION FUNGI. 
 
MORPHOLOGY AND PHYSIOLOGY OF BACTERIA. 15 
 
 tf/iirillc. Lastly, we h;ive the filamentous forms, which may be 
 .straight, leptothrix, or wavy, spirochceta, or the wavy thread may 
 be looped and entwined on itself, spirulina. 
 
 The term involution form is applied to certain peculiar shapes, 
 wh idi result more especially in bacteria grown under abnormal 
 conditions. They are round, oval, pear-shaped, or club-formed 
 enlargements. 
 
 Movement. Many bacteria are devoid of movement through- 
 out the whole of their life history. Others, during certain stages of 
 their life cycle, and possibly some forms always, are endowed with 
 locomotive power. The character of the movement is very varied, 
 and ranges from a slow undulatory motion to one of extreme 
 rapidity. Many appear to progress in a definite direction. Others 
 move continuously, first in one direction and then in another, and 
 others again seem to hesitate before altering their course. They 
 may either glide along smoothly or progress with a tremulous motion. 
 
 FIG. 2. SPIROCH.ETA FROM SEWAGE WATER, x 1200. 
 
 They appear to be able to avoid obstacles, and to set themselves 
 free from objects with which they have accidentally come into 
 contact. Vibrios have a peculiar serpentine movement, but other 
 forms, such as the commonly known Bacterium termo and segments 
 of spirilla, such as comma -bacilli, revolve around their long axis 
 as well as make distinct progression. The complete spirilla are 
 characterised by the familiar corkscrew movement. With regard 
 to cocci there is some doubt as to whether they are endowed with 
 independent movement, any quivering or oscillation being generally 
 regarded as only Brownian or molecular. In some straight thread- 
 forins, which are motile, the movement is very slow and vermicular 
 in character, but in wavy threads, such as the Spirochaeta plicatilis, 
 there is not only an undulatory motion, with rapid progression acm 
 the field of the microscope, but if they are confined by more or 
 debris^ they give very peculiar and characteristic spasmodic 
 movements. (Fig. 2.) 
 
 The rod-forms of Proteus vulgaris exhibit very extraordinary 
 
16 
 
 BACTERIOLOGY. 
 
 movements on the surface of solid nutrient gelatine. Groups o 
 rods may be observed to pass each other in opposite directions 
 Single individuals meet and progress side by side, or one or mor< 
 individuals may part from a group and glide away independent!} 7 
 Occasionally a number of rods progress in single file. It is, however 
 difficult to believe that these movements can occur on a solid surface 
 
 FIG. 3. FLAGELLA. 
 
 1. Coccus with flagellum. 2. Similar coccus dividing, with two flagella. 3. Colony 
 of flagellated macrococci of Begcjiatoa roseo-persicina. 4. Short rod from the 
 same Beggiatoa with flagella [all after ZopfJ. 5. Bacillus with flagella [from 
 a photograph by Koch}. 6. Bacillus suUilis [after Brefeldj. 7, 8. Short rod- 
 forms of Begcjiatoa roseo-persicina with one flagellum [after Zopf]. 9. Very 
 long rod of the same, with flagellum at both ends [after Warming]. 10. Vibrio, 
 with double flagellum at each end [after Warming]. 11. Vibrio, with flagella 
 [from a photograph by the author]. 12. Spirillum with flagella [from a photo- 
 graph by Koch]. 13. Spirillum with flagella [after Zopf]. 14. Spirillum with 
 double flagella [after Zopf). 15. Beggiatoa roseo-persicina, with a triple 
 flagellum at one end ; and 16, with a double flagellum at both ends [after 
 Warming]. 
 
 The author is inclined to believe that there is an almost inappreciable 
 layer of liquid on the surface of the gelatine, which is expressed 
 after the gelatine sets. In tubes of nutrient agar-agar gelatinised 
 obliquely and then kept upright the liquid so expressed collects at 
 the bottom of the sloping surface. 
 
MnRPHOLoi.Y AM) PHYSIOLOGY OF BACTERIA. 
 
 17 
 
 The means by which bacteria are endowed with the power of 
 spontaneous movement and of progression may still be said, in 
 SOUK- rases, to be unsettled. The author has watched the move- 
 ment of long slender threads in. sewage-contaminated water, which 
 could only be explained by the inherent contractility of the proto- 
 plasmic contents ; for if any drawing or propelling organ existed 
 in proportion to the length of the organism, it would probably have 
 been visible. But in many cases the organism is provided with a 
 vibratile lash or flayellum at one end, or with one or more at both 
 ends, or with numerous lateral and terminal flagella. 
 
 Some observers believe that the movement of cocci is due to the 
 
 FIG. 4. BACILLUS MEGATHERIUM. 
 
 a. A chain of rods, x 250. The rest x 600. 
 6. Two active rods. 
 
 d to/. Successive stages of spore-formation. 
 h to 7/1. Successive stages of germination. 
 
 [After De Bary.j 
 
 existence of a nagellum. In Bacterium termo the existence of a 
 lash at either end was first determined by the researches of Dallinger 
 and l.)rysdale. In motile bacilli, such as the hay bacillus and 
 Bacillus ulna, and in vibrios and spirilla, the flagella can be readily 
 recognised by expert microscopists with the employment of the best 
 lenses, and, what is of equal importance, proper illumination. They 
 are objects of extreme delicacy and tenuity, and in stained prepara- 
 tions may be absent from retraction or injury. Koch succeeded 
 in photographing them aftfi- Maining with logwood, which turned 
 them a brown colour. The author ha> observed th'iu in vibrios in 
 preparations stained with gentian violet, from which also they have 
 been photographed, in spite of the violet colour, by th- use of 
 
 2 
 
18 
 
 BACTERIOLOGY. 
 
 isochromatic dry plates, and more recently special methods havt 
 been introduced, by Loffler arid others, by which they can be stainec 
 and photographed with comparative facility. 
 
 It is not certain whether the flagella are extensions of the cell- 
 wall, or derived from the internal protoplasm. Van Tieghem holds 
 the first view, and does not regard them as motile organs at all, 
 Zopf, on the other hand, adheres to the second view, and moreover 
 believes that they can be retracted within the cell-wall. 
 
 Reproduction. Bacteria multiply by fission and by processes 
 which may be considered as representing fructification. The 
 
 FIG. 5. CLOSTRIDIUM BUTYRICUM, x 1020. 
 
 B. Stages of spore-formation. 
 
 C. Stages of germination. 
 
 : [After Prazmowski.] 
 
 bacteria exhibiting the latter processes have been divided into two 
 groups, distinguished by the formation of endospores in the one and 
 of arthrospores in the other. In the process of fission the cell first 
 increases in size, and a transverse septum forms from the cell- wall, 
 dividing the internal protoplasm into two equal parts ; these may 
 separate and lead an independent existence, or remain linked 
 together. In chains of cocci the individual cells are easily visible 
 and distinct, but in the thread-forms resulting from the linking 
 together of rods, as in the anthrax bacillus, the composition of the 
 thread is only demonstrated by the action of reagents. 
 
 Endospore formation may be conveniently studied in Bacillus 
 anthracis, Bacillus megatherium, or Bacillus subtilis. The proto- 
 
MORPHOLOGY AND PHYSIOLOGY OF BACTERIA. . 19 
 
 plasm becomes granular, and at certain points in the thread a speck 
 appears, which gradually enlarges and develops into a circular 
 or egg-shaped, sharply defined, highly refractive body. The spore 
 grows at the expense of the protoplasm of the cell, which in time, 
 together with the cell-wall, entirely disappears, and the spore is set 
 free. These phenomena are best seen in an immotile bacillus in a 
 drop- cultivation on a warm stage ; the whole process may then be 
 observed continuously from beginning to end. Spores may form in 
 each link of the thread, so that a regular row results, or they may 
 occur at irregular intervals. Spore-formation also occurs in bacilli 
 which do not develop into leptothrix filaments. The spores may 
 develop in the centre or at one end of the rod. In the tetanus 
 bacillus a spore develops at the extreme end, producing the appear- 
 ance of a drum-stick. The spore may be considerably wider, but 
 is never longer than the parent cell. 
 
 FIG. 6. LEUCONOSTOC MKSENTEROIDES ; COCCI-CHAINS WITH ARTHROSPORKS 
 (after Van Tieghem and Cienkowski). 
 
 Arthrospore formation is illustrated in Leuconostoc mesenteroides. 
 Certain elements in the chain of cocci, apparently not differing from 
 the rest, become larger, with tougher walls, and more refractive. 
 The remaining cells die, and these cells having acquired the pro- 
 perties of spores are set free, and can reproduce a new growth in 
 jiny fresh nourishing soil. That this occurs in all species which 
 do riot form endospores is at present only a supposition. 
 
 Spores are invested by a thick membrane, which is believed to 
 consist of two layers. To this they probably owe the property they 
 possess of retaining vitality when desiccated, and of offering a 
 greater resistance to the action of chemical reagents and heat than 
 the parent cells. 
 
 Spore-formation has been regarded by some as occurring when 
 the nourishing soil is exhausted, thus providing for the perpetuation 
 
20 BACTERIOLOGY. 
 
 of the species. In anthrax the bacilli do not form spores in the 
 living body, but when the animal dies the development of spores 
 takes place, and hence the danger of contaminating the soil if the 
 body is disposed of by burial. Klein, however, has pointed out that 
 if mice and guinea-pigs which have died of anthrax are kept un- 
 opened, the bacilli simply degenerate and ultimately disappear. 
 Thus there is good reason for believing that spore-formation is 
 not due to exhaustion of the pabulum, but probably free access to 
 oxygen constitutes an important factor in inducing this condition. 
 If we inoculate a potato with anthrax, copious spore- formation 
 occurs, though we cannot consider that the nourishing soil has been 
 exhausted. But we have in this case the surface of the potato 
 freely exposed to the air in the damp chamber. In the same way, 
 in cultivations on agar-agar solidified obliquely (so as to get a large 
 surface), spore-formation readily takes place. Contamination of 
 
 Fit;. 7. SPORE-BEARING THREADS OF BACILLUS ANTHRACIS, DOUBLE-STAINED 
 
 WITH FUCHSINE AND METHYLENE BLUE, X 1200. 
 
 the ground results, therefore, from animals in which a post-mortem 
 examination has been made and the blood and organs freely exposed 
 to the air ; or from carcasses the hides of which have been soiled 
 with excretions, and with blood which issues from the mouth and 
 nostrils before death. 
 
 When spores are introduced into a suitable medium at a favour- 
 able temperature they develop again into rods. The spore loses its 
 sharp contour, and, at one pole or on one side, a pale process bursts 
 through the membrane, gradually growing into a rod from which 
 the empty capsule is thrown off. 
 
 Spores differ from the parent cells in their behaviour to staining 
 reagents. Like them, they can be stained with aniline dyes, but 
 not by the ordinary processes. They require to be specially treated. 
 This is probably due to the tough capsule, which must be altered 
 or softened by heat or strong acid, until it allows the stain to 
 penetrate. 
 
MORPHOLOGY AND PHYSIOLOGY OF BACTERIA. 21 
 
 Once stained, they again differ from the parent cells in resisting 
 decolonisation ; this fact is taken advantage of to double-stain spore- 
 Ivaring bacilli (Fig. 7). 
 
 In staining micro-organisms, the protoplasm is sometimes broken 
 iip into irregular segments or granules, as in many spirilla, and we 
 may add the bacilli of tuberculosis and leprosy. The beaded 
 appearance of the tubercle bacillus is well known. Some observers 
 have regarded the beads, others the bright spaces between them, 
 ;t- -[tores. But spores in unstained preparations appear as glistening 
 bodies with sharp contour. They do not stain at all, or very little, 
 by the ordinary processes. These considerations led the author to 
 .stain and examine tubercular sputum and pure-cultures under 
 careful illumination, and with such lenses as Powell and Lealand's 
 JL in. horn. imm. The tubercle bacillus in sputum (Fig. 8), as a rule, 
 
 FIG. 8. BACILLI OF TUBERCLE IN SPUTUM, x 2500 (from photographs). 
 
 r- insists of a very delicate sheath, holding together a number of 
 deeply stained granules, for the most part round or cylindrical, with 
 irregular contour, and differing considerably in size, while the light 
 interspaces are seen to vary in form according to the shape of the 
 granules. On the other hand, particularly in old cultures, more or 
 less spherical, sharply defined bodies are observed in the bacilli, and 
 also set free. These are the true spores of the tubercle bacillus, 
 and are quite distinct from the irregular granules. There can be 
 no doubt that a tubercle bacillus consists of a very delicate sheath, 
 with protoplasmic contents which have a great tendency to break 
 up or coagulate into little segments or roundish granules, partly 
 owing to their age and the conditions under which they are grown, 
 and partly to the treatment they are subjected to in making a 
 microscopical preparation. This does not always occur, for the 
 bacilli at times are not beaded, but are stained in their entirety. 
 Tn the leprosy bacilli a similar appearance occurs. In stained 
 
22 BACTERIOLOGY. 
 
 sections the rods have a beaded appearance, but the intervals between 
 the granules are sometimes very long, and occasionally the protoplasm 
 appears to have collected only at the extreme ends of the rod. 
 
 The appearances in the case of the bacillus of glanders and the 
 bacillus of hsemorrhagic septicaemia may be similarly explained. 
 
 The fact that tubercular sputum preserves its virulence for 
 several months, even after desiccation, is to be attributed to the 
 formation of spores. Babes claims to have succeeded in differen- 
 tiating them by double staining. 
 
 In his definition of spirilla, Zopf gives the spore-formation as 
 absent or unknown. In comma -bacilli in sewage water the author- 
 has often noted appearances suggestive of refractive spores ; and 
 the same also may be observed in vibrios, differing by their regular 
 contour from the irregular spaces occasionally observed in stained 
 preparations ; but they are only vacuoles. 
 
 FIG. 9. COMMA-BACILLI IN SEW- FIG. 10. VIBRIOS IN WATER CON- 
 
 AGE WATER, STAINED WITH TAMINATED WITH SEWAGE, 
 
 GENTIAN VIOLET, x 1200. x 1200. 
 
 Respiration and Nutrition. Like all a-chlorophyllous vegeta- 
 bles, bacteria require for their nutrition oxygen, nitrogen, carbon y 
 water, and certain mineral salts. Many require free access to oxygen, 
 others can derive it from the oxidised compounds in the medium 
 in which they grow. Pasteur divided bacteria into two great classes 
 the aerobic and anaerobic, and considered that the latter net 
 only had no need of oxygen, but that its presence was actually 
 deleterious. Though this view must be considerably modified, the 
 terms are convenient, and are still retained. They are well illus- 
 trated by the bacillus of anthrax, and the bacillus of malignant 
 oedema; and a simple plan of demonstration has been employed 
 by the author. A fragment of tissue from the spleen, for example, 
 known to contain anthrax bacilli, is deposited with a sterilised 
 inoculating needle, with the necessary precautions, on the surface of 
 nutrient agar-agar in a test-tube ; another tube of nutrient agar- 
 agar is liquefied, and when cooled down almost to the point of 
 
MORPHOLOGY AND PHYSIOLOGY OF BACTERIA. 23 
 
 p-latinisation, a part is poured into the first tube, so that when it 
 the piece of tissue is completely embedded. A piece of tissue 
 from an animal suffering from malignant oedema is treated in the 
 >am' way, and the tubes are placed in the incubator. If we 
 examine them after two or three days, we shall find no change in 
 the anthrax tube ; the bacillus being eminently aerobic, no growth 
 whatever has occurred. In the tube containing the bacilli of 
 malignant oedema there will be a more or less characteristic 
 cultivation. 
 
 The nitrogen which is essential for building up their protoplasm 
 can be obtained either from albumins, or from ammonia and its 
 derivatives. That the albumins can be dispensed with was shown 
 by Pasteur, who employed an artificial nourishing solution consti- 
 tuted upon a formula representing the essential food constituents. 
 
 Carbon is derived from such substances as cane sugar, milk 
 sugar, and glycerine, and, in some cases, by the splitting up of 
 complex proteid bodies. 
 
 Water is essential for their growth, but deprivation of water 
 does not kill all bacteria. Desiccation on potato is employed for 
 preserving some micro-organisms, as a new growth can be started, 
 when required, by transferring some of the dried potato to fresh 
 nourishing ground. Comma -bacilli, on the other hand, are 
 destroyed by drying. Sugar is used in making preserves, because 
 by abstracting water it prevents the development of micro- 
 organisms, 
 
 Mineral or inorganic substances, such as compounds of sodium 
 and pota>sium, and different phosphates and sulphates, are necessary 
 in small proportions. 
 
 CIRCUMSTANCES AFFECTING THE GROWTH OF BACTERIA. 
 
 X'tture of the Soil. Though we know the elements necessary, 
 wt- are, nevertheless, as yet unable to provide a pabulum suitable for 
 all kinds of bacteria. Thus we are quite unable to cultivate some 
 >|ccirs artificially. Others will only grow upon special media. 
 Many gnw U[M)II nutrient gelatine; but some species only if it be 
 acid or alkaline respectively. Whether in the latter case this is due 
 purely to the reaction or to the presence of the particular ingredients 
 i> an unsettled point. Though the comma-bacillus of Koch, like the 
 majority of organisms, grows best on an alkaline medium, yet it 
 i> wj'll known to flourish at the temperature of the blood on the 
 surface of potato, which is acid. 
 
24 BACTERIOLOGY. 
 
 Temperature. The influence of temperature on bacteria will l>e 
 found to vary according to the species, but still for the majority we 
 may distinguish a maximum, optimum, and minimum temperature. 
 
 Many grow best at the temperature of the blood, and hence the 
 value of nutrient agar-agar, which is not liquefied at 37 C. The 
 tubercle bacillus will only grow satisfactorily at a temperature 
 varying between 30 C. and 41 C. On the other hand, many forms 
 grow between the limits of 5 C. and 45 C. At these temperatures 
 their functional activity is paralysed, but they are not destroyed, 
 for by removal to favourable conditions they spring again into life. 
 Bacteria seem to have a special power of resisting the effects of cold. 
 It has been stated that comma-bacilli exposed to a temperature 
 of 10 C. for an hour, and bacilli of anthrax after exposure to a 
 temperature of 110 C., still retained their vitality. Temperatures 
 over 50 to 60 C. destroy most bacteria, but not their spores ; spores 
 of anthrax retain their vitality after immersion in boiling water, but 
 are destroyed by prolonged boiling. Roughly speaking, all patho- 
 genic bacteria grow best at the temperature of the blood, and 
 non-pathogenic bacteria at the ordinary temperature of the room. 
 
 Movement. Bacteria probably grow best when left undisturbed. 
 Violent agitation of a vessel in which they are growing certainly 
 retards their growth, but a steady movement is stated not to affect 
 it ; at any rate, anthrax bacilli grow with enormous rapidity in the 
 blood-vessels, in spite of the circulation. 
 
 Compressed Air. Paul Bert maintained that a pressure of 
 twenty-three to twenty-four atmospheres stopped all development 
 of putrefactive bacteria. Oxygen, under a pressure of five or six 
 atmospheres, is stated to stop their growth. Other observers have, 
 however, obtained different results. 
 
 Gases. Hydrogen and carbonic acid are stated to stop the 
 movements of the motile bacteria. Chloroform is believed to arrest 
 the changes brought about by the zymogenic species. 
 
 Electricity. Cohn and Mendelssohn found that a constant 
 galvanic current produced a deleterious effect owing to electrolysis. 
 At the positive pole the liquid became distinctly acid, and at the 
 negative pole distinctly alkaline. With a weak current there 
 appeared to be no effect, two powerful cells at the very least being 
 necessary. 
 
 Light. Dowries lias shown that sunlight is fatal to putrefactive 
 bacteria. This is believed to be due to a process of induced hyper- 
 oxidation, from which living organisms ordinarily are shielded by 
 protective developments of the cell-wall, or of colouring- matter, 
 
MORPHOLOGY AND PHYSIOLOGY OF BACTERIA. 25 
 
 which cut off injurious rays. Duclaux has investigated the same 
 subject, and observed that micrococci were more sensitive to sun- 
 light than the spore-bearing bacilli. Engelmann has described a 
 bacterium whose movements cease in the dark, and Zopf states that 
 in his cultures of Beggiatoa roseo-persicina the growth was much 
 more strongly developed on the side of the vessel facing the light. 
 Arloing, Marshall Ward, and Dieudonne have studied the effect 
 of the sun's rays on anthrax spores, and on chromogenic and 
 other bacteria, and maintain that they are bactericidal. The 
 effect is due chiefly, if not entirely, to the blue rays. 
 
 Chemical Reagents. Many substances, such as carbolic acid, 
 corrosive sublimate, chlorine, bromine, have a marked effect upon 
 the growth of bacteria. This will be more fully described in 
 another chapter. In several cases the bacteria themselves secrete 
 a substance which is injurious to their future development. 
 
 PRODUCTS OF GROWTH. 
 
 Bacteria may be grouped together according to the changes pro- 
 duced in the media in which they grow. Thus we have pigment- 
 forming, phosphorescent, fermentative, putrefactive, nitrifying, and 
 disease-producing bacteria. 
 
 Chromogenic or pigment-forming bacteria elaborate during their 
 growth definite colour stuffs. Such species are exemplified by Bacillus 
 violaceus, which produces a striking purple growth; Bacillus 
 pyocyaneus, which secretes pyocyanin, a substance which has been 
 isolated and obtained in a crystalline form ; Micrococcus prodigiosus, 
 which produces a pigment allied to fuchsine ; Beggiatoa roseo-per- 
 sicina, whidh is characterised by the presence, of bacterio-purpurin ; 
 Sarcina lutea, Bacillus cyanogenus, and many others. 
 
 Photogenic, or light-producing, bacteria are found more especially 
 in sea-water. There are several species of phosphorescent bacilli, 
 and according to Beyrinck the best medium for their cultivation is 
 fish-broth made with sea-water. Photographs can be obtained of 
 cultures by their own light. 
 
 Zymogenic or ferment bacteria produce their changes in non- 
 nitrogenised media. Bacterium aceti, by its growth produces the 
 acetic fermentation in wine by which alcohol taking up atmospheric 
 oxygen is converted into vinegar : 
 
 C 2 H 6 + O 2 = C 2 H 4 2 + H-'O. 
 
 The fermentation of urine, by which urea is converted into carbonate 
 of ammonia, can be brought about by several micro-organisms, but 
 
26 BACTERIOLOGY. 
 
 notably by the Bacterium urea?. The change produced is represented 
 by the following formula : 
 
 + 2H2 = (NH 2C 3 ' 
 
 Clostridium butyricum converts the salts of lactic acid into- 
 butyric acid, producing the butyric fermentation in solutions of 
 starch, dextrine, and sugar. These bacteria are agents in the 
 ripening of cheese, and the production of sauerkraut. Thus, in a 
 solution neutralised with calcium carbonate : 
 
 2[Ca(C 3 H 5 O s ) 2 ] + H 2 = Ca(C 4 H 7 0-) 2 + CaCO 3 + SCO 2 + H 8 . 
 
 In the so-called viscous fermentation of wines, Streptococcus viscosus- 
 produces a gummy substance. According to Pasteur, the change 
 may be thus represented : 
 
 25(C I2 H 22 11 ) + 25(H 2 0) = 12(C 12 H0 10 ) + 24(C 6 H 14 6 ) + 
 12(C0 2 ) + 12(H 2 0). 
 
 And as another example, the Bacillus acidi lactici may be mentioned f 
 through the agency of which sugar of milk is converted into lactic 
 acid : 
 
 Saprogenic or putrefactive bacteria play a most important part 
 in the economy of nature. They produce changes allied to fermenta- 
 tion in complex organic substances. Their action on proteids, 
 according to Hoppe-Seyler, may be compared to digestion ; bodies 
 like peptones are first produced, then leucin, tyrosin, and fatty 
 acids ; lastly indol, phenol, sulphuretted hydrogen, ammonia, carbonic 
 acid, and water. They abstract the elements they require, and the 
 remainder enter into new combinations. Associated with the forma - 
 tion of these substances are certain bodies which have a poisonous 
 effect when introduced into animals. These poisonous alkaloids, 
 ptomaines, produce a septic poisoning, which must be distinguished 
 from septic infection. The effects of septic poisoning depend on the 
 dose, whereas the effects of septic infection are, to a certain extent, 
 independent of the dose. A small quantity of a septic poison may 
 produce only transient effects, and a relatively large quantity may 
 be necessary to produce vomiting, rigors, arid death. Septic in- 
 fection, on the other hand, may result equally from a small dose,. 
 because the poison introduced is a living organism which is capable 
 of propagation and multiplication. Our knowledge of these 
 alkaloids is largely attributable to the researches of Selmi, Gautier, 
 and Brieger, and the result of their work will be referred to again. 
 
MORPHOLOGY AND PHYSIOLOGY OF BACTERIA. 27 
 
 .Vitrifying bacteria play a very important part by providing 
 plant life with a most necessary food. They occur in the soil, and 
 two kinds have been described the one kind converting ammonia 
 into nitrous acid, and the other changing nitrous into nitric acid. 
 To Winogradsky and Frankland we are principally indebted for our 
 knowledge of these bacteria. 
 
 J'rifhogenic bacteria are those which are genetically related to 
 disease. Many organisms have been supposed to be pathogenic, or 
 have been described in connection with diseases, which are only 
 saprophytic associates. By saprophytic we mean organisms which 
 feed upon dead organic matter. They include many forms which 
 are found on the skin, in the intestinal canal, and sometimes in the 
 internal organs, especially the liver and kidneys ; the tissues have 
 lost their vitality, and the organisms, through some lesion, have 
 been carried into the circulation. 
 
 That many organisms are causally related to disease, there is 
 strong evidence in proof. No organism can be considered to be pro- 
 ductive of disease unless it fulfils the conditions which have been 
 laid down by Koch. Great stress must be laid upon the importance 
 of successive cultivation through many generations, as the objection 
 that a chemical virus may be carried over from the original source 
 is thus overcome. Any hypothetical chemical poison carried over 
 from one tube to another would, after a great number of such 
 cultivations, be diluted to such an extent as to be inappreciable 
 and absolutely inert. 
 
 Though we may accept as a fact the existence of pathogenic 
 organisms, we are not in all cases in a position to assert the means 
 by which they produce their deleterious or fatal effects. Many 
 theories have been propounded. It has been suggested that the 
 pathogenic organisms may be compared to an invading army. 
 The cells or phagocytes arrayed against them endeavour to as- 
 similate and destroy them, but perish themselves in the attempt. 
 Thi> might explain the breaking down of tissue, and the for- 
 mation of local lesions, but does not assist us in understanding 
 the fatal ivsult in thirty-six to forty-eight hours produced by the 
 inoculation of the bacilli of anthrax. Another view is that the 
 invading army seizes upon the commissariat, appropriating the 
 general pabulum, which is so essential to the life of the tissues. 
 This would hardly account for so acute and fatal a result as in 
 anthrax, but would lead one to expect symptoms of inanition and 
 irradual exhaustion. Moreover, against this theory we have the 
 tact that death may result, in some cases, with the presence 
 
28 BACTERIOLOGY. 
 
 of comparatively few bacilli in the blood ; and, again, the 
 blood may teem with parasites such as the flagellated monads in 
 well-nourished, healthy- looking rats, without apparently causing 
 any symptoms whatever. In the same category may be placed the 
 theory that eminently aerobic organisms seize upon the oxygen of 
 the blood and produce death by asphyxia. Another explanation is 
 afforded by the suggestion of interference with the functions of the 
 lung and kidney by mechanical blocking of the capillaries. Here 
 the same objection is met with in the case of anthrax, the same 
 fatal result may occur with only a few bacilli, while other cases 
 yield very beautiful sections, looking like injected preparations from 
 the mapping out of the capillaries with the countless crowds of 
 bacilli. 
 
 Putrefactive bacteria derive their necessary elements from com- 
 plex organic substances, and accompanying the residue we find the 
 presence of poisonous substances. Pathogenic bacteria, in a similar 
 way, give rise to virulent poisons. Anthrax bacilli produce poisonous 
 principles in the blood which cause death, independently of the 
 number of bacilli, provided there are sufficient present to develop a 
 fatal dose. 
 
 It has been also suggested that possibly a special ferment is 
 .secreted by some organisms, and that by the changes ultimately 
 wrought by the action of this ferment the symptoms and phe- 
 nomena of disease arise. We have an analogy with this theory 
 in the alkaline fermentation of urine by means of the torula urese. 
 By the researches of Musculus, and later of Sheridan Lea, it has 
 been shown that a ferment is secreted by the cells which can be* 
 isolated in aqueous solution, and is capable of rapidly inducing an 
 active fermentation of urea. 
 
 We can now understand how it is that in anthrax or in tuber- 
 culosis we may find the presence of only a few bacilli, or that in 
 tetanus we can have such a violent disturbance of the system 
 produced by the presence of very few micro-organisms. We may 
 conceive that different species of bacilli may vary greatly in their 
 power of producing a toxin or secreting a ferment, just as the 
 elaboration of pigment is much more marked in some species than 
 in others ; thus it need not follow that the number of micro- 
 organisms bears any relation to the virulence or activity of the 
 .substance they produce. There is, however, yet another factor in 
 the production of disease. We know that in health we are proof 
 against most of these micro-organisms ; if it were not so, we should 
 all rapidly fall victims to the tubercle bacillus or others, which in 
 
MORPHOLOGY AND PHYSIOLOGY OF BACTERIA. 29 
 
 health we inhale with impunity. We know that a microbe may only 
 cause a local lesion in one animal, but death in another. It is still 
 more striking that the same micro-organism, as is the case with 
 anthrax, may have 110 effect whatever upon certain species of 
 animals, though it is deadly to others. Again, an animal naturally 
 susceptible to the effect of a pathogenic organism may be rendered 
 proof against it. These matters wiJJ be discussed in a future 
 chapter. 
 
 DISTRIBUTION OF BACTERIA. 
 
 Bacteria are commonly described as ubiquitous. They are ever 
 present in the air, though not in such exaggerated numbers as is 
 commonly supposed. In nutrient media exposed to the air one is often 
 astonished at times at the comparatively few bacteria which develop 
 in comparison to the amount of floating matter, such as mineral 
 particles, scales, spores of fungi and debris known to be present. 
 In water they are also present in considerable numbers, though of 
 course varying according to the character of the water. Wherever 
 there is putrefaction, they are present in vast numbers. In the soil, 
 in sewage, in the intestines and, in uncleanly persons especially, 
 011 the skin and between the teeth, various species may always be 
 found, but in the healthy blood and healthy tissues bacteria are 
 never present. In a previous chapter the method of examining the 
 blood of living persons has been described, and there is, by thin 
 means, ample opportunity for satisfying oneself that bacteria are 
 never to be found in the blood in health. The organs removed from a 
 perfectly healthy animal, with the necessary precautions, and placed 
 in sterilised media, can be kept indefinitely without undergoing 
 putrefaction, or giving any development of bacteria. This has been 
 established by many observers, notably Cheyne and Hauser ; and 
 the results of former observers to the contrary must be attributed 
 to imperfect methods admitting of accidental contamination. 
 
CHAPTER III. 
 
 EFFECT OF ANTISEPTICS AND DISINFECTANTS ON BACTERIA. 
 
 IN the previous chapter several conditions were alluded to whicl 
 affected the growth of bacteria, such as the nature of th* 
 nutrient soil, temperature, light, and electricity. The effect of 
 certain chemical substances, and of excessive heat and cold, w 
 also mentioned ; but this constitutes a subject of such practical 
 importance that it must be considered more fully. 
 
 Agents which retard the growth of bacteria are generally spoken 
 of as antiseptics, as distinguished from disinfectants which altogether 
 destroy their vitality. 
 
 Though chemical disinfectants, or germicides, when diluted, act 
 as efficient antiseptics, the converse, that an antiseptic in a suffi- 
 ciently concentrated form will act as a disinfectant, is not the case. 
 The term " antiseptic," indeed, should be restricted to those sub- 
 stances or agents which arrest the changes bacteria produce, but 
 which do not prevent their springing into activity when removed 
 to favourable conditions. Thus excessive heat, which destroys 
 bacteria and their spores, is a true disinfectant ; and excessive cold, 
 which only benumbs them, retarding their development without 
 killing them, is an antiseptic. 
 
 Spores have a greater power of resisting the action of these 
 various agents than the parent cells, and many species of micro- 
 organisms differ from each other in their resisting power. An 
 exact knowledge of the subject can, therefore, only be based upon 
 investigations which will determine the effect of these agents upon 
 pure cultivations of the different micro-organisms causally related to 
 putrefaction and disease. In the latter case, especially, this is not 
 possible in the present state of our knowledge. In some cases of 
 communicable disease there is considerable doubt as to the etiological 
 importance of the organisms which have been described; in other 
 cases no organisms have as yet been discovered, or the organisms 
 
 30 
 
EFFECT OF ANTISEPTICS AND DISINFECTANTS ON BACTERIA. 31 
 
 <Mimot be artificially cultivated, or the disease is not reproduced by 
 inoculation, so that there is no means of testing whether the agents 
 have had any effect. One can, therefore, only draw general 
 conclusions by selecting some well-known pathogenic and non- 
 pathogenic micro-organisms, and considering the influence of 
 c^ ^cals, of hot air and of steam upon them, as representing the 
 effect upon the various contagia of disease and the causes of 
 putrefaction. 
 
 Such knowledge must necessarily prove of the greatest im- 
 
 ce : to the sanitarian, who is concerned in preventing the 
 
 spreading of disease and in the disposal of putrefactive matter; to 
 
 the surgeon, who is anxious to exclude micro-organisms during 
 
 Mnrical operations, and to arrest the development of bacteria which 
 
 ive already gained an entrance in wounds; to the physician, in 
 
 c he treatment of micro-parasitic diseases. 
 
 The sanitarian and the surgeon must profit directly by such 
 experiments, for in the disinfection of clothes and the sick-room by 
 the one, and in the application of antiseptic dressings and lotions 
 liy the other, the micro-organisms are encountered, as in the experi- 
 ments, outside the living body. 
 
 The physician, on the other hand, is principally concerned in 
 dealing with micro- parasites when circulating in the blood, or 
 carrying on their destructive processes in the internal tissues. So 
 far as our knowledge at present goes, the physician can avail him- 
 self but little of the effect of the direct application of the substances 
 which have been found to retard or destroy the growth of the 
 organisms in artificial cultivations, for the concentrated form in 
 which they would have to be administered would prove as deleteri- 
 ous or as fatal to the host as to the parasites. Thus Koch has 
 stated that to check the growth of the anthrax bacillus in man it 
 would be necessary that there should be twelve grammes of iodine 
 constantly in circulation, and that the dose of quinine necessary 
 to destroy the spirilla of relapsing fever would be from twelve to 
 sixteen grammes. The retarding influence, however, of certain 
 .substances when diluted, and the fact that disinfectants are some- 
 times equally efficacious in a diluted form when their application is 
 prolonged, seem to indicate measures which may be adopted, in some 
 with chances of success, such as the inhalation of antiseptic 
 vapours in phthisis. For the most part the physician must look 
 rather to combating the effects of micro-organisms by restoring to 
 its normal standard the lowered vitality which enabled the bacteria 
 to get a footing. 
 
32 BACTERIOLOGY. 
 
 There is no wider field for research than the determination of 
 the real effect of disinfectants and antiseptics. Painstaking and 
 laborious as the researches are which have been hitherto made, the 
 subject is so beset with fallacies, leading, in some cases, to totally 
 erroneous conclusions, that it is not surprising that one meets on 
 all sides with conflicting statements. The author has no intention 
 of analysing these results, but a general idea will be given of the 
 methods which have been employed, and for further details reference 
 must be made to the original papers mentioned in the bibliography. 
 
 Chemical Substances. It was customary to judge of the power 
 of a disinfectant or antiseptic by adding it to some putrescent liquid. 
 A small portion of the latter was, after a time, transferred to 
 some suitable nourishing medium, and the enicacy of the substance 
 estimated by the absence of cloudiness, odour, or other sign of 
 development of bacteria in the inoculated fluid. Koch pointed out 
 the errors that might arise in these experiments from accidental 
 contamination, or from there being no evidence of the destruction 
 of spores, and we are indebted to him for a complete and careful 
 series of observations with more exact methods. 
 
 Instead of employing a mixture of bacteria, Koch's plan was to- 
 subject a pure cultivation of some well-known species with marked 
 characteristics to the reagent to be tested. A small quantity was 
 then transferred to fresh nourishing soil, under favourable con- 
 ditions, side by side with nutrient material inoculated from a 
 cultivation without treatment with the disinfectant. The latter 
 constituted a control test, which is most essential in all such 
 experiments. To test the resistant power of bacteria which are 
 easily destroyed, two species were selected, Micrococcus prodigiosus r 
 and the bacillus of blue pus. These were cultivated on potatoes r 
 the surfaces of which were sliced off and dried. A fragment trans- 
 ferred to freshly prepared potato gave rise to a growth of the 
 particular micro-organism ; but if after treatment with some reagent 
 no growth occurred, the conclusion was drawn that the reagent wa& 
 efficacious in destroying the vitality of the bacteria. 
 
 Anthrax bacilli in blood, withdrawn from an animal just killed, 
 were taken to represent sporeless bacteria, while silk threads steeped 
 in an artificial cultivation of the bacilli and dried, afforded a means 
 of testing the vitality of spores. 
 
 Even by employing pure cultivations on solid media, great 
 precautions were necessary to avoid mistakes. When, for instance, 
 .1 l;n-^c quantity of the growth which had been subjected to some 
 chemical solution was carried over to the fresh tube containing 
 
EFFECT OF ANTISEPTICS AND DISINFECTANTS ON BACTERIA. 33 
 
 the nutrient medium, or when a silk thread, which had been dipped 
 in a solution, was directly transferred to the new soil, enough of the 
 -upposed disinfectant might be mechanically carried over to retard 
 the development of the bacteria, though it was ineffectual in 
 drst roving them. From a growth not appearing, it was concluded 
 that the spores or the bacteria had been affected, and so ,-i 
 mistake occurred. To avoid this, Koch made a point of transfer- 
 ring a minimum, of the disinfected growth to as large a cultivation 
 area as possible, so that any chemical substance mechanically 
 carried over would be so diluted as to be inert. For the same 
 r<MM>n, threads, after withdrawal from the disinfecting solution, 
 were rinsed in sterilised water, or weak alcohol, and then trans- 
 planted; or, instead of judging from the development on nutrient 
 gelatine, the effect of inoculation in a healthy animal was made 
 the test. 
 
 A few examples may be quoted in illustration. Silk threads, 
 impregnated with anthrax spores, were placed in bottles containing 
 carbolic acid of various strengths. A thread was removed from each 
 011 successive days, and transferred to nutrient gelatine, and the 
 result noted. It was found that immersion of the thread in a 5 per 
 cent, solution of carbolic acid was sufficient in two days to effect 
 complete sterilisation, and seven days in a 3 per cent, solution was 
 equally efficacious. Since for practical purposes a strength should 
 be selected which would be effectual in twenty-four hours, Koch 
 recommended that for general use, allowing for deterioration by 
 keeping, a solution containing not less than 5 per cent, should be 
 employed, and for complex fluids probably a still higher percentage 
 would be necessary. In the case of sporeless bacilli the results were 
 very different. Blood containing the bacilli, from an animal just 
 killed, was dried on threads, and after exposure for two minutes to 
 a 1 per cent, solution, was completely sterilised; and fresh blood 
 mixed with a 1 per cent, carbolic solution produced no effect when 
 inoculated. On the other hand, when the blood was mixed with a 
 5 per cent, solution, the virulence was not destroyed. The facility 
 with which the bacilli are destroyed, compared with their spores, 
 illustrates how easily errors may occur, when mere arrest of growth 
 or loss of motility is regarded as a sign of the efficacy of disinfection. 
 
 To test vapours, Koch exposed anthrax spores or the sp<nv> 
 which occur in garden earth by suspending them over solutions, 
 siu-h as bromine or chlorine, in a closed vessel. After a time they 
 \\.-n- transferred to a nutrient medium to test their vitality. To 
 test the power of sulphurous acid gas, the spores were spread about 
 
 3 
 
34 BACTERIOLOGY. 
 
 in a room in which the gas was generated by burning sulphur in 
 the ordinary way for disinfecting a room. To test chemicals which 
 might be recommended for disinfecting vans and railway carriages, 
 spores were laid on boards, which were then washed or sprayed, and 
 the spores then transferred to the nutrient gelatine. 
 
 Sternberg has also made an elaborate series of experiments with 
 regard to the action of germicides. In this case cultivations of 
 well-known pathogenic organisms in liquid media were employed. 
 The supposed germicide was added to the liquid cultivation, and 
 after two hours a fresh flask of sterilised culture was inoculated from 
 the disinfected cultivation, and placed in the incubator. In twenty- 
 four to forty-eight hours, if the chemical proved inefficient, there 
 was evidence of a growth of bacteria. Blyth has investigated the 
 disinfection of cultivations of Bacterium termo, of sewage, and 
 typhoid excreta, and, in conjunction with Klein, the effect of well- 
 known disinfectant materials on anthrax spores. Miquel, Laws, 
 and others have also contributed to our knowledge of the effect 
 of antiseptics and disinfectants upon micro-organisms. In spite of 
 all that has been done there is room for many workers ; a great 
 deal of ground must be gone over again to rectify discrepancies, 
 examine conflicting results, arid thus determine what observations 
 may be relied upon for practical application. 
 
 This may be illustrated by referring in detail to some experiments 
 made with corrosive sublimate. Koch investigated a long list of 
 chemical reagents, and according to these experiments the salts of 
 mercury, and the chloride especially, proved most valuable. Where 
 heat is not admissible, these compounds were therefore highly 
 recommended, though their poisonous nature is a drawback to their 
 indiscriminate use. Koch stated that for disinfecting a ship's bilge, 
 where a 5 per cent, solution of carbolic acid must be left forty- eight 
 hours, a 1 in 1000 solution of mercuric chloride would only require 
 a few minutes. 
 
 But there was good reason for doubting the efficacy of very dilute 
 solutions; for, though according to Koch's experiments anthrax 
 spores subjected to a 1 in 20.000 solution of mercuric chloride for 
 ten minutes, and then trashed in alcohol, gave no growth in nutrient 
 gelatine, silk threads exposed for ten minutes to a 1 in 20,000 
 solution, or even 1 in 10,000, still proved' fatal to mice. 
 
 Herroun cultivated ordinary septic bacteria in albuminous 
 nitrates, containing 1 in 2000, and concluded that the value of 
 mercuric chloride as an antiseptic was much over-rated. It is pre- 
 cipitatfd by albumins though, as Lister has shown, the precipitate 
 
Kl-TKCr OK ANTISEPTICS ANI> DISINFECTANTS ON BACTERIA. 35 
 
 of albuniinnte of mercury is redissolved when there is an excess of 
 albumin present. 
 
 Geppert, and later Behring, recognised that the methods employed 
 for testing the efficacy of corrosive sublimate were unreliable. They 
 found, for example, that corrosive sublimate could not be removed 
 from silk threads by washing ; and therefore to study the effect of 
 this antiseptic acting for a given time, it was necessary to dip the 
 threads in ammonium sulphide solution after the treatment with 
 corrosive sublimate. 
 
 The author confirmed the results of Geppert and Behring, and 
 ma ile a scries of experiments to test the value respectively of carbolic 
 acid and corrosive sublimate in antiseptic surgery. The method 
 of dipping an infected thread into an antiseptic solution for a few 
 minutes, and then transferring it to the surface of a nutrient medium 
 to test its efficacy in a given time, was discarded as fallacious; the 
 thread being still wet with the solution when transferred to the 
 medium, it was obvious that the action of the antiseptic continued 
 for many days. To wash infected silk threads with alcohol after 
 exposure to the antiseptic to stop its further action, also proved to 
 be a fallacious method, for the author found in control experiments 
 that absolute alcohol will destroy Streptococcus pyogenes, erysipelatis, 
 and Staphylococcus pyogenes aureus, acting for only one minute. 
 < >ther methods were therefore resorted to, and cultures on the 
 sloping surface of nutrient agar were at first used. The antiseptic 
 was poured into the culture tube until the growth was covered, 
 and when it had acted for a definite time (one minute, five minutes r 
 or fifteen minutes) a solution was added which immediately stopped 
 further action. In the case of corrosive sublimate, ammonium sul- 
 phide was employed, which is quite inert as an antiseptic. The liquid 
 contents of the test tube were carefully poured off, and an inoculation 
 \\as made into a fresh tube of broth or agar from the culture still 
 adhering to the surface of the nutrient medium. As the results 
 disproved the efficacy of corrosive sublimate, it was thought possible 
 that the solution had not been able in the time to penetrate the 
 film of growth. Another plan was accordingly adopted. Cultures 
 were made in broth, and when fully developed the supernatant liquid 
 was carefully poured off. Corrosive sublimate solution was added to 
 the test tube, and agitated until any flocculent masses were dis- 
 integrated and the whole of the liquid became uniformly turbid. 
 Ammonium sulphide \\a> added when the time had expired, and 
 tubes of fresh broth were inoculated with the mixture. In the case 
 of carbolic acid the cultures, after its action, were thoroughly washed 
 
36 BACTERIOLOGY. 
 
 with water, and its efficacy tested by making inoculations from the 
 cultures in fresh media. The results were entirely in favour of 
 carbolic acid. Staphylococcus pyogenes aureus and Streptococcus 
 pyogenes were not destroyed, even when corrosive sublimate solution 
 of 1 in 1000 was allowed to act for an hour. In the case of the 
 cultures of streptococcus of erysipelas the results were different. 
 A solution of 1 in 10,000 had no effect, but 1 in 4,000, acting for 
 one minute, destroyed the culture. With carbolic acid the results 
 were very striking. Cultures were exposed to solutions of 1 in 20, 
 1 in 30, 1 in 40, 1 in 50, for one minute, five minutes, fifteen 
 minutes. The attempts to make subcultures in every case failed. 
 Carbolic acid I in 40, acting for only one minute, was sufficient to 
 destroy Streptococcus pyogenes and Streptococcus erysipelatis and 
 Staphylococcus pyogenes aureus. Further experiments were made 
 with tubercular sputum, the test being subsequent inoculation of 
 guinea-pigs. Corrosive sublimate solution as strong as 1 in 500 had 
 no effect, but 1 in 20 carbolic acid, shaken up with the sputum for 
 one minute, completely neutralised it. 
 
 Koch's statements with reference to the germicidal power of 
 coiTosive sublimate in extremely weak solutions had led Lister to 
 substitute it for carbolic acid as a detergent in surgery. The author's 
 experiments, which were undertaken in 1892, encouraged Lister to 
 revert to the use of carbolic acid, which, indeed, had always proved 
 efficacious in surgical practice. Lister pointed out that carbolic acid 
 has also the great advantage of combining eagerly with fats and 
 epidermis, so that the seat of operation can be effectually cleansed. 
 
 These experiments also point to the conclusion that carbolic acid 
 should be used in hospital wards for the disinfection of tubercular 
 .sputum instead of mercuric chloride and other less efficacious dis- 
 infectants commonly in use. 
 
 Hot Air and Steam. Koch, in conjunction with Wolfhiigel, also 
 made exhaustive experiments to test the value of hot air. A similar 
 plan was adopted to that employed in disinfection with chemicals. 
 Bacteria and spores were subjected for a certain time to a known 
 temperature in the hot-air chamber, and then were transferred to a 
 nourishing soil or inoculated in animals. 
 
 Paper parcels, blankets, bags, and pillows, containing samples of 
 micro-organisms wrapped up inside, were also placed in the hot-air 
 chamber, to test the power of penetration of heat. 
 
 The conclusions from these experiments were as follows : 
 
 Sporeless micro-organisms at a little over 100 C. are destroyed 
 in one hour and a half. 
 
K1KKCT OF ANTISEPTICS AND DISINFECTANTS ON BACTERIA. 37 
 
 S|M>IV> lit' bacilli require three hours at 140 C. 
 
 If enclosed in pilknvs and blankets, exposure from three to four 
 hours to 140 C. is necessary. 
 
 Sp.>iv- of fungi require one and a half hours at 110 C. to 
 ll.-i C. 
 
 Further experiments showed that at the temperature nece;iry 
 for the destruction of spores of bacilli almost all fabrics are more or 
 less injured. 
 
 Koch, in conjunction with Gaffky and Loffler, also investigated 
 the effect of steam under pressure and at the atmospheric pressure. 
 
 Rolls of flannel with anthrax spores or earth spores, and a 
 thermometer wrapped up inside, were subjected to steam, and 
 the results compared with the effect obtained with hot air. 
 
 Thus in hot air four hours' exposure to a temperature of 130 C. 
 to 140 C. brought the temperature inside the roll to 85 C., and the 
 spores were not injured ; on the other hand, exposure to steam 
 under pressure at 120 C. for one and a half hours, raised the 
 internal temperature to 117 C. and killed the spores. 
 
 By such experiments the superior penetrative power of steam- 
 heat was established. 
 
 To te>t >t -am-he:it at the atmospheric pressure, water was boiled 
 in a glass flask with its neck prolonged by means of a glass tube, the 
 temperature i;i which was found to be uniform throughout. Anthrax 
 and earth > pores placed in the tube were found to be unable to with- 
 stand -team at 100 (,'. even for a few minutes. It was, therefore, 
 concluded that disinfection by steam at atmospheric pressure wa> 
 superior to hot air from its greater efficiency, and to steam under 
 pressure from the simplicity of the necessary apparatus. 
 
 Parsons and Klein made some experiments which were more 
 in favour of dry heat than the above. These observers state that 
 anthrax bacilli are destroyed by an exposure of five minutes at 
 from 100 C. to 103 C. and that anthrax spores are destroyed 
 in four hours at 104 C., or in one hour at 118 C. Guinea-pigs 
 inoculated with tuberculous pus winch had been exposed for five 
 minutes to 104 C., remained unaffected. They concluded that as 
 none of the infectious diseases, for which disinfecting measure- 
 are in practice commonly applied, are known to depend upon the 
 pn-sence of bacilli in a spore-b3aring condition, their contagia 
 are not likely to retain their activity after being heated for an 
 hour to 105 0. (220 Fahr.) 
 
 In experiments with steam the results were in accordance 
 with those already -riven, and complete penetration of an object 
 
38 . BACTERIOLOGY. 
 
 by steam-heat for more than five minutes was deemed sufficient. 
 They also arrived at the same result as in Koch's experiments, 
 viz., that steam-chambers are preferable to those in which dry heat 
 is employed, though it must be borne in mind that some articles, 
 .such as leather, are injured by exposure to steam. 
 
 PRACTICAL APPLICATION. 
 
 Nurses and others attending infectious cases should freely use 
 1 in 40 carbolic for the hands and a weaker solution for the body 
 generally. The skin of patients after recovery should be sponged 
 with 1 in 40 carbolic. The dead should be wrapped up in a sheet 
 soaked in 1 in 20 carbolic acid or a strong solution of chloride of 
 lime. Infected clothing and bedding should be burnt unless in excep- 
 tional cases, when they may be disinfected by boiling, or by exposure 
 to dry heat at 105C. to 110C. for three hours, or by steaming 
 at 100C. for fifteen minutes. Leather and other articles which 
 would be destroyed by any of these processes should be sponged with 
 1 in 40 carbolic. The walls of the sick-room and furniture should 
 be exposed to the fumes of burning sulphur, and next day washed 
 down with 1 in 40 carbolic, and the room freely ventilated by 
 opening all windows and doors. Rags should be burnt, or dis- 
 infected by boiling or exposure to steam when supplied to manu- 
 facturers. The importation of rags from places where there are 
 cases of cholera or small-pox should be prohibited. Infected ships 
 must be fumigated with sulphur, and the bilge disinfected with 
 carbolic acid. Infected railway carriages should be disinfected in 
 the same way as a sick-room. 
 
 Tubercular sputum, cholera and typhoid evacuations and other 
 excreta should be disinfected by 1 in 20 carbolic acid, or by a strong 
 solution of chloride of lime. 
 
CHAPTER IV. 
 
 ( HKMICAL PRODUCTS OF BACTERIA. 
 
 THK products of the metabolism induced by bacteria may l)e divided 
 into three classes : (1) ptomaines or alkaloids; (2) albumoses or tox- 
 albumins ; and (3) enzymes. Alkaloids and albumoses are directly 
 poisonous ; enzymes or ferments are harmless except in the presence 
 of proteids, which they are capable of transforming into poisonous 
 album 
 
 PTOMAINES AND Tox- ALBUMINS. 
 
 The study of these products may be said to date back to 1822, 
 when Gaspard and Stick found an intensely poisonous principle 
 in cadaverous extracts. In 1856 Panum discovered a poisonous 
 Mibstunce in putrid flesh; and in 1863 Bergmann and Smiedeberg 
 found a nitrogenous crystallisable substance in putrid beer which 
 they named sepsin. In 1872 Gautier found that the decomposition 
 of fibrine led to the formation of various complex alkaloidal 'sub- 
 stances, and in 1875 Richardson obtained in pyaemia an alkaloid, 
 septin. This subject, however, received most attention from the 
 classical researches of Selmi, the Italian toxicologist. Selnri. in a 
 celebrated poisoning case, demonstrated the presence of an alkaloid 
 as the result of post-mortem changes. Similar substances were 
 found in alcohol in which morbid specimens had been preserved. 
 Thus the researches of Gautier and Selmi established the fact that 
 albuminoid material undergoing decomposition leads to the forma- 
 tion of cadaveric alkaloids. These animal alkaloids Selmi named 
 ptomaines. Brieger, finding the bases derived from the products of 
 putrefaction less poisonous than those obtained from the pathogenic 
 bacteria, suggested the term toxins for the latter. Ptomaines have 
 been divided into two classes those which are non-oxygenous, liquid, 
 and volatile, and those which are oxygenous, >olid. and ci \>tallisable. 
 They are, for the most part, precipitated by tlu* ordinary rfa<r'iits 
 
 39 
 
40 BACTERIOLOGY. 
 
 for alkaloids, such as chloride of gold, double iodide of mercury 
 and potassium, picric acid, arid tannin. Phospho-molybdic acid 
 precipitates them without exception. They are powerful reducing 
 agents. Ferro-cyanide of potassium is converted into ferri-cyanide in 
 their presence, and the addition of ferric chloride gives the Prussian 
 blue test. Selmi discovered this test, and Brouardel and Boutmy 
 regarded it as absolutely characteristic of ptomaines ; but this is 
 not the case ; some vegetable alkaloids, for example, behave in the 
 same way. 
 
 As examples of the non-oxygenous ptomaines there are : 
 
 Parvolin (C 9 H 13 N) an oily base of an amber colour prepared 
 from putrid mackerel and horse-flesh. 
 
 Hydrocollidin (C 8 H 13 N), from the same source. It is highly toxic, 
 being compared by Gautier to the venom of the cobra di capello. 
 
 Collidin (C 8 !! 11 !^), from putrid gelatine and the pancreas of a 
 bullock, also highly toxic. 
 
 NcMTulm (C 5 H 14 N 2 ), from fish, flesh, and decaying cheese. 
 
 Saprin (C 5 H 14 N 2 ), isoineric with neuridin. 
 
 Cadaverin (C 5 H 14 ]Sr 2 ), a third isomeride, from ordinary putrefac- 
 tion and herring brine. 
 
 Putrescin (C 4 H 12 N 2 ) from putrefaction. 
 
 The oxygenous ptomaines are in some instances found also in 
 healthy tissues. They include the following : 
 
 Xeurin (C 5 H 13 NO), found in cadaveric putrefaction. 
 
 Cholin (C S H 5 N0 2 ), in bile. 
 
 Muscarin (C 5 H 13 N0 2 ), in a poisonous mushroom, Agaricus mus- 
 carius, and in putrid fish. These are all highly poisonous. 
 
 Gadinin (C 7 H 16 NO 2 ), in putrefying codfish. 
 
 Mytilotoxin (C 6 H 15 N0 2 ), in poisonous mussels. 
 
 Poisonous alkaloids are of great importance in connection with 
 those cases of meat poisoning produced by sausages, hams, poultry, 
 and cheese. Tyrotoxicon is a poisonous alkaloid obtained from cheese. 
 
 The toxic substances of most interest to the bacteriologist are 
 those isolated from pure cultivations of pathogenic bacteria, such as 
 typhotoxin, isolated by Brieger from cultivations of the bacillus of 
 typhoid fever, and tetanin, from cultivations of the tetanus bacillus ; 
 and the poisons known as albumoses or tox-albumiiis, which are 
 allied to the albumose of snake poison. 
 
 Pasteur, in 1885, suggested that in anti-rabic inoculations the 
 immunity resulted from the action of a substance secreted by a 
 microbe, though the microbe has not as yet been discovered in 
 rabies. Salmon produced immunity from hog cholera by the injec- 
 
CHKMICAL PRODUCTS OF BACTERIA. 41 
 
 tiou of tlic toxic products in filtered culture fluids. Wooldridge, 
 Ilankin, and Martin studied the products of Bacillus anthraci>. 
 ( 'h.-irrin. and later Woodhead, Wood, and Blagovestchensky, investi- 
 gated on these lines Bacillus pyocyaneus. Roux and Chamberland 
 experimented with the bacillus of malignant oedema; Roux with 
 >vmptomatic anthrax; Ohantemesse and Widal with the typhoid 
 bacillus. Roux, Yersiii, Brieger, Fraiikel, Martin, and Behring 
 worked on the same lines with diphtheria. Koch introduced 
 tuberculin, Kalning mallein, while others have utilised the products 
 of streptococci and pneumococci. Anrep found an albumose in the 
 medulla of rabid animals, and Babes claims to have found an 
 albumose in both rabies and glanders. 
 
 Cholera. Brieger found several ptomaines, including putresciii 
 and cadaverin, in pure cultures of the spirillum of Asiatic cholera, 
 and Petri found in addition to poisonous bases a proteid body which 
 produces in guinea-pigs muscular tremors, paralysis, and a rapidly 
 fatal result. Roux and Yersiii obtained from cultures a tox-albumin 
 insoluble in water, which kills guinea-pigs in two or three days, 
 but has no effect on rabbits. Pfeiffer also investigated the toxic 
 substances in cultures. Chloroform, thymol, and drying destroyed 
 comma -bacilli, leaving their toxic products unaffected. Concentrated 
 solutions of neutral salts and boiling produced secondary toxic 
 substance^, but the original toxic substances were ten or twenty 
 tirnes more virulent. 
 
 Typhoid Fever. Typhotoxin (C 7 H 17 NO 2 ), the alkaloid ob- 
 tained by Brieger from cultures of the typhoid fever bacillus, produces 
 in mice and guinea-pigs salivation, rapid breathing, dilatation of the 
 pupil, diarrhoea, and death in twenty-four to forty-eight hours. At 
 the po>t- mortem examination the heart is found in a state of systolic 
 contraction, and the condition of the heart after death and the 
 absence of convulsions during life serve to distinguish typhotoxin 
 from an isomeric base obtained by Brieger from putrid horse -flesh. 
 Roux and Yersin have obtained a tox-albumin. It is soluble with 
 ditiiculty in water, and more toxic to rabbits than guinea-pigs. 
 
 Tetanus. Brieger obtained the alkaloid tetanin from impure 
 cultures of the tetanus bacillus. It is a base having the formula 
 ; 1!--X-O 4 . The hydrochloride is a very deliquescent salt, and 
 soluble in alcohol. Tetanin injected into guinea-pigs produces 
 rapid breathing, followed by tetanic convulsions. Another toxic 
 product, tetfinotrj.i:in (C^H^N), produces the same effects as tetanin. 
 The formula of a third base, apasiiwtoxin, has not been determined. 
 C'adaverin and putre>cin are also present in cultures. Kitasato and 
 
42 BACTERIOLOGY. 
 
 Weyl analysed the products of pure-cultures, and obtained the same 
 substances, tetanin and tetanotoxin ; and subsequently Brieger and 
 Frankel found that in pure-cultures a tox- albumin could be 
 obtained which is soluble in water, and infinitely more active than 
 the toxic ptomaines. 
 
 Anthrax. In 1887 Woolclridge succeeded in protecting rabbits 
 from anthrax by a new method. A proteid body obtained from the 
 testis and from the thymus gland was used as the culture fluid. 
 This proteid substance was dissolved in dilute alkali, and the solution 
 .sterilised by repeated boiling. This was inoculated with the anthrax 
 bacillus, and kept at 37 C. for two or three days. A small quantity 
 of the filtered culture fluid injected into the circulation in. rabbits 
 produced immunity from anthrax. Subcutaneous inoculation of 
 extremely virulent anthrax blood, made simultaneously with the injec- 
 tion of the protecting fluid, produced no effect. Wooldridge showed 
 that the growth of the anthrax bacillus in special culture fluids 
 gave rise to a substance which, when injected into the organism, 
 protected not only against an immediate but also subsequent attacks. 
 
 In 1889 Hankin worked under the guidance of Koch in the 
 Hygienic Institute of Berlin. The acquired tolerance of the effect 
 of ordinary albuinoses, and the experiments of Sewall, who pro- 
 duced immunity against lethal doses of the albumose of snake 
 poison by the injection of minute doses, led Hankin to expect that 
 an albumose developed in anthrax cultures, and that the anthrax 
 albumose would probably confer immunity fromthedisea.se. Hankin 
 succeeded in isolating it from culture fluids. It was precipitated by 
 excess of absolute alcohol, well washed in alcohol to free it from 
 addition of ptomaines, filtered, dried, then redissolved and filtered 
 through a Chamberland filter. With this substance Hankin suc- 
 ceeded in producing immunity in mice and rabbits. 
 
 Sidney Martin, working quite independently, grew anthrax bacilli 
 in a solution of pure alkali albumin made from serum proteids. After 
 ten or fifteen days the organisms were removed by filtration through 
 a Chamberland filter. The filtrate contained proto-albumose and 
 deutero-albumose, a trace of peptone, an alkaloid, and small quantities 
 of leucin and tyrosin. The mixture of albuinoses proved poisonous 
 to mice. The anthrax alkaloid produced symptoms and lesions 
 similar to the albumoses, but much more rapidly and severely. It 
 is an amorphous yellow body, soluble in alcohol and alkaline in 
 reaction. Martin concluded that the anthrax bacillus formed the 
 albumoses and the alkaloid by digesting the alkali albumin; and 
 suggested that the alkalinity of the albumoses explained their toxic 
 
CI1KMICAL PRODUCTS OF BACTERIA. 43 
 
 properties, the alkaloid probably being in ;i nascent condition in the 
 .-iDmmose molecule. 
 
 Tuberculosis. Koch prepared a glycerine extract of the 
 product of the tubercle bacillus in pure cultivations, and found 
 that the injection of small doses produced a remarkable reaction, 
 l>otli local and general, in tubercular cases, and especially lupus. 
 This extract, called tuberculin, came to be extensively used a.- a 
 therapeutic agent, but with disappointing results. Very shortly 
 after the first announcement of Koch's discovery, the author, in 
 conjunction with Herroun, investigated the chemical properties and 
 physiological effects of the products of the tubercle bacillus. 
 <'ultures in glycerine- broth were filtered through porcelain, and 
 a clear amber-coloured liquid was obtained, which gave important 
 and suggestive chemical reactions. As this filtrate contained the 
 products of the growth of the bacillus most probably in minute 
 quantities, it was evaporated at a low temperature over sulphuric 
 acid. The viscous residue was dissolved in distilled water and 
 tested on the healthy guinea-pig. The result was a marked fall of 
 temperature, staring coat, extreme irregularity of the heart's action, 
 muscular spasms, loss of control over the extremities, and death. 
 
 A preliminary examination of glycerine -broth cultivations having 
 .shown the presence of iion-coagulable proteid bodies of the nature 
 of albumose and peptone, and a crystallisable precipitate of a 
 remarkable character resulting on the addition of iodine, the idea 
 naturally suggested itself that the tubercle bacillus might form 
 albumoses and an alkaloid or ptomaine similar to the substances 
 isolated by Martin from pure cultivations of the Bacillus anthracis. 
 
 Koch pointed out that the effective substance in his extract 
 could be precipitated by absolute alcohol; the author and Herroun 
 determined to investigate the properties and physiological effects 
 of the separated products. They accordingly set to work to isolate 
 the ptomaine, of the existence of which they had some qualitative 
 indication, and at the same time to examine the properties of the 
 albuminous bodi->. 
 
 In this endeavour the general method they found satisfactory was 
 as follows. The clear filtrate from the culture was evaporated at 
 40 C. to a very small bulk, and the residue thus obtained was 
 mixed with an excess of absolute alcohol, which precipitated the 
 albumoses and peptone. It was found that by adding the alcohol 
 by degrees a partial separation of the albumose from the peptone 
 could be effected, the latter being only precipitated when the alcohol 
 \\a- nearly absolute. The precipitated albumose was collected on 
 
44 BACTERIOLOGY. 
 
 M filter and redissolved in distilled water. In another experiment 
 the albumose underwent a second precipitation, and after washing 
 \vas again dissolved. 
 
 The alcoholic nitrate from the precipitated albuminous bodies was 
 then concentrated at a very gentle heat until a viscous residue was 
 left containing the glycerine originally present in the cultivating 
 medium and the extractives and products of the bacillus soluble 
 in alcohol. With this residue definite reactions of an alkaloidal 
 substance or ptomaine were obtained. 
 
 Careful experiments, however, led to the belief that the whole 
 of the ptomaine was not separated from the albuminous precipitate 
 by simple addition of alcohol, and the above method was therefore 
 slightly modified. 
 
 The ptomaine is soluble in water and alcohol, and sparingly 
 soluble in amyl-alcohol, but insoluble in benzine, ether, or chloro- 
 form, which liquids therefore fail to extract it from aqueous 
 solutions. In its aqueous solutions it is distinctly but not strongly 
 alkaline to test-paper. Phospho-tungstic acid gives with it a white 
 flocculent precipitate. Phospho-molybdic acid gives a pale yellow 
 precipitate, soluble in ammonia to a blue solution which becomes 
 colourless on boiling. In this respect it resembles the vegetable alka- 
 loids, aconitiri and atropiii. It must be remembered, however, that 
 albuminous bodies are precipitated by both this and the preceding 
 reagents, and in the case of the former a reduction of the phospho- 
 molybdate giving the blue solution with ammonia is obtained. 
 
 The reducing power of the ptomaine is shown by the conversion 
 after a short time of ferri-cyanide of potassium to ferro-eyanide, 
 giving the Prussian blue test with ferric chloride, to which much 
 undue importance was attached by Brouardel and Boutmy. The 
 solution of albumose and solution of peptone are both capable of 
 giving this reaction as well as many vegetable alkaloids. A solution 
 of the ptomaine is not precipitated by ferro- cyanide of potassium or 
 pot issici bichromate. 
 
 In strong solutions it yields precipitates with platinic chloride 
 (yellow), gold chloride (pale yellow), arid mercuric chloride (white). 
 That yielded by the first of these reagents is granular in character, 
 and quite insoluble in alcohol, though apparently soluble in water. 
 The precipitation by gold chloride excludes amides and ammonium 
 salts. 
 
 With iodine in hydriodic acid or potassic iodide a precipitate is 
 obtained which is occasionally crystalline, more often granular or 
 amorphous. 
 
CHEMICAL PRODUCTS OF BACTERIA. 45 
 
 This precipitate is soluble in alcohol, and is redeposited when the 
 alcohol is evaporated. On heating it is redissolved into oily drops of 
 a dark colour. With picric acid a granular precipitate is obtained, 
 which under the microscope is seen to consist of minute crystal*. 
 This precipitate, on standing, is converted into rounded crystalline 
 H with numerous small crystals admixed. 
 
 The ptomaine appears to be easily broken up by heating, 
 '>p'cially in the presence of mineral acids or of baryta. The actual 
 quantity obtained from a considerable amount of culture fluid vnu 
 v-ry small, and as it was possible that when the bacilli were grown 
 in a medium richer in albumin, such as the animal body, more of 
 the>e products might be formed, the liquid obtained by extracting 
 large masses of tubercular growths from cattle was examined in a 
 >imilar manner. In this extract, after nitration through porce- 
 lain, an albumose, and minute quantities of a ptomaine were 
 obtained which in reactions was identical with that obtained from 
 the artificial cultivation of the bacillus, but present in even smaller 
 amount. The probable explanation of this is, that in the living 
 animal the ptomaine is constantly being removed ; or it may indicate 
 that it is only formed in minute quantity under those conditions. 
 
 1 1 jiving succeeded in obtaining the albumose and the ptomaine 
 in separate solutions, we next proceeded to ascertain the effects of 
 these substances upon healthy and tubercular guinea-pigs. 
 
 The effect of the ptomaine isolated from different series of 
 cultures was as follows. A rise of temperature occurred in tuber- 
 cular animals, and distinct enlargement of tubercular glands. Theie 
 was a slight indication of a depression of temperature or hypothermic 
 effect on healthy animals. The albumose, whether obtained from 
 pure cultivations of the bacillus or from tubercular tissue, pro- 
 duced a marked rise of temperature in tubercular guinea-pigs. On 
 the other hand, in a control experiment on a healthy guinea-pig 
 there was an equally well-marked fall of temperature. The effect 
 upon the tubercular glands in the cases associated with marked rise 
 of temperature was to render them well-defined, indurated, and 
 painful, rather than any considerable increase in volume. 
 
 Hunter made a chemical examination of Koch's crude extract, 
 and confirmed the presence of albumoses and alkaloidal 8ubetanoe& 
 The albumoses consisted chiefly of proto-albumose and deutero-albu- 
 mose with hetero-albumose, and occasionally a trace of dys-albuinose. 
 Two alkaloidal sultances were obtained in the form of platinum 
 Compounds of their hydrochlorate salts. In addition there were 
 extractives, iniicin. inorganic .salts, glycerine, and colouring-matter. 
 
46 BACTERIOLOGY. 
 
 Swine Fever. Schweinitz applied Brieger's methods in the 
 investigation of the products of the swine fever, or hog cholera 
 bacillus. Broth- cultures were neutralised with dilute hydrochloric 
 acid, and evaporated in the water bath. The residue was treated 
 with 96 per cent, alcohol, and the filtered solution with mercuric 
 chloride. A heavy crystalline precipitate was separated by filtra- 
 tion, treated with water, and decomposed with sulphuretted hydrogen, 
 and cadaverin and methylamine were isolated. The filtrate from 
 the mercuric chloride precipitate was freed from excess of mercury 
 by sulphuretted hydrogen, and the mercury sulphide filtered off. 
 The residue, after concentration of the filtrate, was extracted with 
 absolute alcohol, and the solution showed the presence of an alka- 
 loidal salt. The double salt obtained with platinum chloride was 
 submitted, after crystallisation, to an analysis, and the results gave 
 the formula (C 14 H 34 N 2 PtCl 6 ). The hydro- chloride is soluble in abso- 
 lute alcohol as well as in water, and produces needle-like crystals. 
 
 On treating the culture fluids with excess of absolute alcohol a 
 white flocculent precipitate was obtained partly soluble in water, and 
 re-precipitated by alcohol. It was obtained in the form of white 
 crystalline plates. A watery solution gives almost insoluble needle- 
 crystals on the addition of platinum chloride. These products were 
 respectively termed sucholo-toxin and sucholo-albuinin. Small doses 
 of these substances produce in guinea-pigs a. slight rise in tempera- 
 ture, and ulcsration at the seat of injection. Large doses produce 
 a fatal result in six to twenty-four hours. Schweinitz asserts that 
 he has produced immunity in guinea-pigs. An attempt to produce 
 immunity in swine by injection of the albumose gave unsatisfactory 
 results. 
 
 Diphtheria. Roux and Yersin finding that filtered cultures of 
 the diphtheria bacillus produced paralysis, affecting chiefly the hind 
 legs, and a fatal result in rabbits and guinea-pigs, proceeded to 
 investigate the chemical products. They succeeded in obtaining a 
 white amorphous substance which was extremely active when 
 injected into guinea-pigs. It was precipitated by alcohol from an, 
 aqueous solution, and it was calculated that '0004 gram would 
 destroy eight guinea-pigs of 400 grams, or two rabbits of 3 kilos, 
 each. They concluded that the poison was an enzyme or ferment, 
 as it not only acted in extremely small doses, but it was attenuated 
 by heat and destroyed by boiling. 
 
 Brieger and Frankel confirmed these experiments, and asserted 
 that the poison was a tox-albumin ; but according to Martin their 
 chemical analysis and reactions were vitiated by the fact that they 
 
< I1KMKAL PRODUCTS OF BACTERIA. 47 
 
 had peptone in their cultivating medium. Martin examined the 
 products l>y using as a culture medium a 1 to 2 per cent, solution 
 of alkali-albumin in broth made from beef, omitting the peptone. 
 Afier about thirty days the bacillus had converted the alkali-albumin 
 into albumoses, which gave the reactions of proto- and deutero- 
 albumose, \vith small quantities of an organic acid. A single dose 
 of these albumoses produced weakness of the hind limbs, which after 
 a time passed off. The animal was killed, and the nerves which 
 were examined showed degeneration. Repeated intravenous in- 
 jection on successive days, amounting in all to a dose of 1'69 grams 
 per kilo, of body weight, produced high fever, followed by depres- 
 >ion of temperature, severe watery diarrhoea, and emaciation. The 
 i n Ion reflexes began to diminish after the ninth day, 011 the 
 eleventh or twelfth day theie was definite paralysis of the hind legs, 
 and on the seventeenth day reflexes could scarcely be obtained. 
 
 Martin thus gives his method of abstracting the poisonous pro- 
 ducts either from cultures or from diphtheritic tissues. In dealing 
 with tiu'>. the spleen and other organs are first finely minced and 
 placed in rectified spirit, and the blood is also placed in spirit, and 
 allowed to stand till the proteids are coagulated; they are then 
 filtered, and the residue extracted with cold water, all the extracts 
 are mixe.l together, and evaporated at 35 C. to a small bulk, and 
 thrown into absolute alcohol. Mast of the albumoses are precipi- 
 tated, the alcohol is poured off, evaporated to dryness at a low 
 temperature, and extracted by absolute alcohol until nothing more 
 dissolves. The residue is deutero-albumose and mineral salts. All 
 the proteid is mixed together, dissolved in water, and precipitated by 
 alcohol, the proces> being repeated to remove any traces of bodie> 
 soluble in alcohol and the excess of mineral salts. At the last 
 precipitation the precipitate is allowed to stand under alcohol for 
 alxjtit two months. The alcohol is then poured off, and the pre- 
 cipitate dried 'm vacua, 
 
 The resulting product is a light yellowish -brown powder soluble 
 in water, cold or boiling, giving a yellowish and faintly acid or 
 nearly neutral icaction. It is composed of deutero-albumose with 
 a slight amount of proto-albumose but no peptone. It gives the 
 ordinary actions of proteids and a well-marked biuret reaction. It 
 i> precipitated from solution by ammonium sulphate, and slightly 
 by nitric acid. The reactions are similar to those of peptic deutero- 
 albumose. The alcoholic extract of the tissues is strongly acid, and 
 contains free fatty acid and an organic acid insoluble in chloroform. 
 The organic acid is ivadily soluble in water and absolute alcohol. 
 
48 BACTERIOLOGY. 
 
 and insoluble in ether, chloroform, and benzine. It is a yellowish 
 amorphous body, becoming a deep brown when made alkaline. 
 
 Martin concludes that whereas the Bacillus anthracis produces 
 albumoses and an organic base, in diphtheria we find albumoses and 
 an organic acid. 
 
 Glanders. Kalming has obtained from cultures of the glanders 
 bacillus an extract similar to tuberculin. This crude extract is 
 known as mullein, and is extensively used for the diagnosis of 
 glanders. In a glandered horse it causes a rise of temperature 
 and swelling at the seat of the injection, and the glandered nodules 
 become swollen and painful. Finger claims to have produced 
 immunity from glanders by inoculation of the products contained 
 in sterilised cultures. Schweiiiitz extracted from cultures a noii- 
 poisonous albumose, and obtained only traces of a ptomaine. 
 
 Suppuration and Pneumonia. Brieger obtained a ptomaine 
 from cultures of Staphylococcus pyogenes aureus, and Roux and 
 Yersin a tox-albumin fatal to rabbits and guinea-pigs in a few 
 days. There was pus-formation at the seat of inoculation, with 
 redness and swelling of the surrounding parts. 
 
 From pure-cultures of the micrococcus of pneumonia Klemperer 
 obtained a tox-albumiii, for which the name pneumo-toxin has been 
 suggested. 
 
 ENZYMES OR FERMENTS. 
 
 Many bacteria liquefy the nutrient gelatine m which they are 
 cultivated. This is due to the development of a ferment or enzyme, 
 which dissolves the albumin and gelatine. 
 
 Enzymes are products of the vital activity of living bacteria. 
 Bitter, and independently Sternberg, showed that when a liquefying 
 bacterium is removed by filtration or destroyed by heat, the culture 
 fluid retains the power of liquefying gelatine. As this occurs 
 usually when the reaction is alkaline, bacterial enzymes resemble 
 trypsin and papain rather than pepsin. They can be extracted with 
 glycerine, and are quite harmless. If injected into animals 110 effect 
 is produced, and after a few hours no trace of them can be found. 
 According to Fermi, the influence of temperature on the enzymes 
 produced by different bacteria will be found to vary very consider- 
 ably. The enzyme of Staphylococcus pyogenes aareus is destroyed 
 at 55 C., while the enzyme of Bacillus anthracis succumbs at a 
 temperature of 65 C. to 70 C. 
 
 Some bacteria produce both enzymes and toxins, but many pro- 
 duce enzymes and not toxins, and others toxins but not enzymes. 
 
CHAPTER V. 
 
 IMMUNITY. 
 
 THE condition of being insusceptible to an infective disease may be 
 either natural or acquired. In studying the pathogenic organisms 
 several examples of natural immunity will be encountered. The 
 bacillus of septicaemia, so fatal to house mice, has been shown to 
 have 110 effect upon field mice. The bacillus of anthrax is innocuous 
 to cats and white rats. The bacterium of rabbit septicaemia is 
 e< iiially inert in dogs, rats, and guinea-pigs. The immunity may 
 be as in these cases complete, or only partial. Ordinary sheep are 
 very easily affected with anthrax, but Algerian sheep succumb only 
 to large doses of the virus. Natural immunity may not only be 
 characteristic of certain species, but it may occur in certain indi- 
 viduals of a susceptible species. The same immunity occurs in man, 
 for certain individuals, though equally exposed during an epidemic 
 of small-pox, may escape, whereas others readily fall victims to the 
 disease. 
 
 Acquired immunity is illustrated by the protection afforded by 
 one attack of the exanthemata against subsequent attacks. Thus 
 one attack of measles or small-pox, as a rule, affords complete 
 protection. A knowledge of the immunity resulting in the latter 
 case led to the introduction of inoculation of small- pox as a 
 protection against natural small-pox. 
 
 Immunity may be acquired by acclimatization, for the inhabit- 
 ants of tropical climates are less susceptible to the diseases of the 
 country, malarial fevers, for instance, than strangers. 
 
 In civilised communities also, there appears to be a degree of 
 acquired immunity, for infectious diseases like measles introduced 
 among savages or isolated communities have assumed the most 
 malignant type. 
 
 The immunity acquired by protective inoculation constitutes, in 
 connection with the study of pathogenic micro-organisms, a subject 
 of pre-eminent interest and importance. Pasteur, in his researches 
 
 49 4 
 
50 BACTERIOLOGY. 
 
 upon fowl-cholera, observed that after non-fatal cases the disease 
 either did not recur, or the severity of a subsequent attack was in 
 inverse proportion to the severity of the first attack. It occurred 
 to him to endeavour to obtain the virus of this disease in a form 
 which would provoke a mild attack of the disease, and thus give 
 protection against the virulent form. This attenuation or miti- 
 gation of the virus was successfully attained by allowing cultiva- 
 tions of the microbe in chicken-broth to remain with a lapse 
 of several months between the carrying on of successive cultiva- 
 tions in fresh media. The new generations which were then 
 obtained were found to have diminished in virulence, and ultimately 
 a virus was obtained which produced only a slight disorder ; on 
 recovery the animal was found to be proof against inoculation with 
 virulent matter. The explanation given by Pasteur of this change 
 was, that prolonged contact with the oxygen of the air was the 
 influence which diminished the virulence, and he endeavoured to 
 prove this by showing that when broth was inoculated in tubes 
 which could be sealed up, so that only a small quantity of air 
 was accessible to the microbe, the virulence of the cultures was 
 retained. 
 
 Toussaint investigated the possibility of attenuating the virus of 
 anthrax. Sheep injected with 3 cc. of defibrinated blood, con- 
 taining anthrax bacilli, which had been exposed to 55 C. for ten 
 minutes, recovered, and were afterwards insusceptible. Pasteur 
 subsequently argued that this method did not admit of practical 
 application, because difficulties would arise in dealing with infective 
 blood in quantity, and artificial cultivations started from this blood 
 could not be relied upon, as they proved sometimes as virulent as 
 ever. 
 
 Pasteur endeavoured to apply the same method for obtaining 
 an attenuated virus of anthrax, as he had successfully employed 
 in fowl-cholera. A difficulty was soon encountered, for in culti- 
 vations of this bacillus, with free access of air, spore-formation 
 readily takes place, and the spores are well known to have an 
 extraordinary power of retaining their virulence. Pasteur found 
 that the bacilli ceased to develop at 45 C., and he believed that 
 spore-formation ceased at 42 to 43 C C., the bacilli continuing to 
 develop by fission only,. The cultivations were, therefore, kept at 
 this temperature, and at the end of eight days the bacilli were 
 found to have lost their virulence, and were quite inert when 
 inoculated in guinea-pigs, sheep, or rabbits. This total destruction 
 was, however, preceded by a gradual mitigation, so that a virus 
 
IMMUNITY. 51 
 
 could be obtained, by taking it at the right time, which gave only a 
 mild disease, and afforded subsequent protection. 
 
 At Melun, in 1881, the protective inoculation against anthrax 
 was put to a practical test. Sheep and oxen were inoculated with 
 the mitigated virus, and then with a virulent form ; at the same 
 time other sheep and oxen were inoculated with the virulent form 
 without previous vaccination, as a control experiment. The unpro- 
 tected sheep died without exception ; the unprotected oxen suffered 
 from cedematous swellings at the seat of inoculation, and a rise of 
 temperature ; but all the protected animals remained healthy. 
 
 As a result of these experiments an idea arose that by preventive 
 inoculation with attenuated virus all communicable diseases would 
 in time be eradicated ; but this does not follow, for all communi- 
 cable diseases do not confer immunity after a first attack ; in 
 influenza the very reverse is believed to occur, and erysipelas of the 
 face leads to an increased liability to subsequent attacks. Even 
 with regard to the prevention of anthrax, Pasteur's researches were 
 opposed and criticised. Koch investigated the subject, and came to 
 the conclusion that the process did not admit of practical applica- 
 tion, chieflv on the ground that as immunity lasted only a year, the 
 losses from the vaccination process would be as great or even 
 greater than from the spontaneous disease ; further, there was 
 danger in disseminating a vaccine of the strength required to be 
 effectual. 
 
 Chauveau proved that the attenuation was due to the tempera- 
 ture, and not to the prolonged effect of oxygen. By keeping 
 cultivations at 42 to 43 C. in vacuo, the virulence was found 
 to disappear in twenty-four hours, and by keeping cultivations 
 at a low temperature with free access of air, the virulence was 
 retained. Chauveau considered, therefore, not only that oxygen \\a> 
 not the agent, but that the mitigation was much more easily effected 
 in its absence. In spite of these adverse criticisms, these researches 
 nevertheless confirmed the principle of Pasteur's conclusion, that 
 immunity could be induced by experimental measures, and further 
 showed that he had considerably advanced the methods by which this 
 could be effected. 
 
 Chauveau succeeded also in attenuating the virus by a modifica- 
 tion of Toussaiut's method. Sterilised broth was inoculated with 
 the bacilli, and placed in the incubator at 42 to 43 C. After the 
 lapse of twenty hours it was removed to another incubator at 47 C. 
 According to the time of exposure to this increased temperature, the 
 mitigation varied in degree. Thus inoculation with the virus, l^efore 
 
52 BACTERIOLOGY. 
 
 it was exposed to 47 C., was fatal to guinea-pigs; but after one 
 hour at 47 C. the virulence was diminished, and, though ultimately 
 fatal, life was prolonged ; after two hours' exposure at 47 C. only 
 half the animals died; and after three hours' exposure they 
 recovered, and were rendered refractory to subsequent inoculation. 
 
 Attenuation of the virus of anthrax has also -been induced by 
 chemical means. Chamberland and Roux stated that a fresh growth 
 started from a cultivation of bacilli which had been subjected for 
 twenty- nine days to ^J^ of carbolic acid was found to be inert in 
 guinea-pigs and rabbits. Bichromate of potash added to a cultiva- 
 tion in the proportion of y^-g^- to ^^^ gave, after three days, a 
 new growth, which killed rabbits, guinea-pigs, and half the sheep 
 inoculated ; after ten days, rabbits and guinea-pigs, but not sheep ; 
 and after a longer time even guinea-pigs were unaffected. 
 
 In other diseases similar results have been obtained. Arloing, 
 Cornevin, and Thomas found that by inoculating a small quantity 
 of the virus of symptomatic anthrax anywhere in the subcutaneous 
 connective tissue, or a moderate quantity at the root of the tail, 
 and even by intravenous injection, immunity was obtained from a 
 virulent dose. 
 
 In swine -erysipelas, Pasteur and Thuillier obtained attenuated 
 virus upon quite another principle. They discovered that by 
 passing the virus through pigeons the virulence was increased, but 
 by passing it through rabbits it was progressively diminished. Thus 
 a virus was obtained from the rabbit, which produced only a mild 
 disease in pigs, and after recovery complete immunity. Similarly 
 in rabies, Pasteur found that passage of the virus through various 
 animals considerably modified its properties. By inoculating a 
 monkey from a rabid dog, and then passing the virus through other 
 monkeys, the virulence was diminished ; but by inoculating a rabbit 
 from the dog, and passing the virus from rabbit to rabbit, the 
 virulence increased. 
 
 In rabies, Pasteur has employed another method of attenuating 
 the virus. The spinal cord of inoculated rabbits is removed with 
 all possible precautions, and portions a few centimetres in length 
 are suspended in flasks in which the air is dried by fragments of 
 potash. By this process the virulence is found to gradually diminish 
 and finally disappear. Animals inoculated with portions of these 
 cords, after suspension for a certain time, are rendered refractory to 
 inoculation with virulent cords. Having rendered dogs, which had 
 been previously bitten, free from the svipervention of symptoms of 
 hydrophobia by means of protective inoculation, Pasteur proceeded 
 
IMMUNITY. 53 
 
 to apply the same treatment to persons bitten by rabid animals, with 
 results which tend to the belief that a real prophylactic for rabies 
 lias been discovered. 
 
 Immunity may also be produced by injecting the toxic products 
 existing in pure cultivations after removal of the bacilli. Salmon 
 w;is the first to produce immunity in this way, by utilising the toxic 
 products of the bacterium of hog-cholera, which were separated by 
 filtration from the living micro-organisms; and shortly afterwards 
 Wooldridge demonstrated that filtered anthrax cultures contained 
 a substance which conferred immunity. Behiing and Kitasato 
 produced immunity by mixing cultures with terchloride of iodine. 
 Yaillard filtered the cultures through porcelain, and attenuated the 
 products by heating at different temperatures. 
 
 Lastly, in the course of Behring's and Kitasato's experiments, it 
 was found that the blood serum of animals rendered immune was 
 capable of conferring immunity on other animals. The injection 
 of the toxic products of pathogenic bacteria leads to the development 
 of substances in the blood to which the term " antitoxin" has been 
 applied. These protective substances neutralise or destroy the 
 injected poison, and blood serum which has thus been rendered 
 antitoxic can be utilised to confer immunity on other animals. 
 
 Haffkine's system of vaccination as a protection against Asiatic 
 cholera is supposed to be based upon the principle of inducing the 
 formation of antitoxins or defensive proteids. 
 
 MECHANISM OF IMMUNITY. 
 
 Raulin has shown that Aspergillus niger develops a substance 
 which is prejudicial to its own growth, in the absence of iron salts 
 in the nutrient soil, and Pasteur suggested that in rabies, side by 
 side with a living microbe, there is possibly some chemical product 
 or anti-microbe which has, as in Haulm's experiment, the power of 
 arresting the growth of the microbe. If we accept the theory of 
 arrest by some chemical product, we must suppose that in the 
 acquired immunity afforded by one attack of an infectious disease 
 this chemical substance is secreted, and, remaining in the system, 
 opposes the onset of the micro-organism at a future time. In the 
 natural immunity of certain species and individuals we must suppose 
 that this chemical substance is normally present. 
 
 Another theory is, that the micro-organisms assimilate the 
 elements which they require for their nutrition from the blood and 
 tissues, and render the soil impoverished or otherwise unsuitable for 
 
54 BACTERIOLOGY. 
 
 the development of the same species of micro-organisms hereafter ; 
 this condition may be permanent, or the chemical constitution of 
 the tissues may be restored to normal, when immunity ceases. If, 
 however, we explain acquired immunity by the result of the growth 
 of a previous invasion of micro-organisms, we are still confronted 
 with the difficulty of explaining natural immunity. 
 
 A third theory is that the tissues are endowed with some 
 power of vital resistance to the development of micro-organisms, 
 similar to the vital resistance to coagulation of the blood, which is 
 supposed to exist in the lining membrane of the healthy blood- 
 vessel; that in some species arid individuals this exists to a high 
 degree, and hence their natural immunity. But this does not 
 explain how one attack confers immunity from a subsequent one. 
 One would expect that the vital resistance would invariably be 
 lowered by a previous attack, and increased liability be the constant 
 result. 
 
 A fourth theory was propounded by Metchnikoff, who maintains 
 that immunity depends upon phagocytosis. If anthrax bacilli are 
 inoculated in the frog, white blood-cells, or phagocytes, are observed 
 to incorporate and destroy them until they entirely disappear, and 
 the animal is not affected. But if the animal, after inoculation, 
 is kept at a high temperature, the bacilli increase so rapidly that 
 they gain the upper hand over the phagocytes, and the animal 
 succumbs. 
 
 It has also been suggested that bacteria may attract or repel 
 the phagocytes, exercising either a positive or a negative chemio- 
 taxis. This power is supposed to depend upon some special product 
 of the bacteria or possibly upon their toxins, as suggested by Roux. 
 We must suppose that the negative chemio-taxis has become changed 
 to a positive chemio-taxis in an immunised animal, so that the 
 phagocytes, instead of withdrawing and leaving the bacteria to 
 multiply, are readily drawn into the contest and destroy the 
 invaders. 
 
 In septicaemia of mice, the white blood-cells are attacked and 
 disintegrated by the bacilli in a remarkable way. It is difficult, 
 however, to accept these observations as affording a complete ex- 
 planation of immunity. It is difficult to conceive that the leucocytes 
 in the blood and tissues in the field mouse are differently constituted 
 from those in the house mouse, so that they form an effectual 
 barrier to the onset of bacteria in the one case, though so readily 
 destroyed in the other, or that in acquired immunity the result is 
 due to educating the phagocytes to respond to a positive chemio-taxis. 
 
IMMUNITY. 55 
 
 Phagocytosis cannot explain the immunity which results from 
 the injection of filtered cultures, or of antitoxins, but when 
 blood serum of immunised animals was shown to possess antitoxic 
 properties, a new explanation of immunity was at once forthcoming. 
 In the light of these discoveries immunity, whether natural or 
 ac |iiireil, was regarded as due to the accumulation in the blood 
 and tissues of substances which have the property of counteracting 
 partially or entirely the products by which pathogenic bacteria 
 produce their poisonous effects. These antitoxins, or protecting 
 proteids, can be obtained not only from the blood but also from 
 the spleen and the lymphatic and other glands. They result 
 from the metabolism of the cells of the tissues of the body. 
 Phagocytes in their conflict with bacteria may play a small 
 part, but it is more than probable that immunity is altogether 
 independent of phagocytosis. 
 
CHAPTER VI. 
 
 ANTITOXINS AND SERUM THERAPY. 
 
 IT has been clearly shown by the experiments of Fodor and Nuttall 
 that some species of bacteria are killed by a mixture with fresh 
 blood. Fodor pointed this out in the case of the anthrax bacillus, 
 and Nnttall confirmed the experiments, and repeated them with 
 a number of different species of bacteria. 
 
 Behring and Nissen followed up this line of inquiry, and found 
 that there was a great difference in the behaviour of freshly drawn 
 blood to different bacteria. In some cases the bacteria were 
 destroyed, in others their growth was only retarded, and in others 
 again they were not affected at all. Bouchard pointed out that 
 although the normal blood serum of a rabbit may be used for the 
 cultivation of Bacillus pyocyaneus, the blood serum of a rabbit, which 
 has been rendered immune, will attenuate or entirely nullify the 
 pathogenic properties of the bacillus. 
 
 Ogata and Jasuhara obtained similar results by cultivating 
 anthrax bacilli in the blood of immune animals. Buchner demon- 
 strated that this property of fresh blood belonged to the serum 
 and not to the cellular elements, and strongly advocated the theory 
 that the force opposed to invading bacteria was to be found in the 
 serum rather than in phagocytes. 
 
 Similar experiments were made with the bacteria of swine-fever, 
 and Emmerich and Mastbaum discovered that the blood serum of 
 immune rabbits could be used as a therapeutic agent to prevent 
 the progress of the disease in animals already showing symptoms 
 of infection. 
 
 A new light was thrown upon this question by the experiments 
 of Behring, Kitasato, Tizzoni and Cattani, and others in connection 
 with tetanus and diphtheria. In these diseases the bacteria do not 
 invade the body, but the poisonous principles elaborated at the 
 seat of inoculation are absorbed into the system and produce 
 deleterious effects. It was obvious that attention must be turned 
 towards counteracting or destroying these poisonous products. 
 
 56 
 
ANTITOXINS AND SERUM THERAPY. 
 
 57 
 
 It was in this direction that the experiments of Behring and 
 Kitusato, in 1890, proved to be of profound importance. It was 
 >lm\vn that the blood serum of a rabbit rendered immune against 
 t -t;i uus or diphtheria had no destructive or retarding effect on the 
 irnnvth of the bacilli, but it possessed the power of neutralising the 
 poison developed by the agency of the bacilli. In short, the serum 
 was shown to possess an antitoxic instead of a bactericidal power. 
 
 Hankin conceived the idea that this property is due to substances 
 of the nature of defensive proteids, and the blood serum of the 
 naturally immune rat was found to contain a proteid body with well- 
 marked alkaline reaction, possessing the power of destroying anthrax 
 bacilli. Injection of this proteid into mice, together with fully 
 virulent anthrax spores, prevented the development of the disease. 
 Young rats are susceptible to anthrax, and, according to Hankin, 
 they can be protected from anthrax by injection of the blood serum 
 of the parent. Tizzoni and Cattani expressed the opinion that the 
 antitoxic substance in the blood serum of animals rendered immune 
 against tetanus is a globulin to which they gave the name tetanus 
 antitoxin. Buchner proposed the term alexins (dAeo>, I defend), 
 to signify these substances. Hankin subdivided them into sozins 
 and phylaxins. Sozins are defensive proteids occurring in normal 
 animals ; phylaxins are only found in animals artificially immune ; 
 and each of these are sub -classed by Hankin according to their power 
 of attacking the bacteria themselves or the products they generate. 
 
 Defensive proteids 
 
 (Hankin) 
 Alexins (Buchner) 
 
 Sozins : 
 
 Defensive proteids y 
 present in the nor- 
 mal animal. 
 
 Phylaxins : 
 Defensive 
 present 
 
 proteids 
 in the 
 
 animal after it has 
 artificially been 
 made immune. 
 
 'Myco-sozins : 
 
 Alkaline globulins from rat 
 (Hankin). destroying an- 
 thrax bacillus. 
 
 Toxo-sozins : 
 
 Of rabbit, destroying poison 
 of Vibrio Metchnikovi 
 (Gamaleia). 
 
 Myco-phylaxins : 
 
 Of rabbit, destroying pig 
 typhoid bacillus (Em- 
 merich). 
 
 Toxo-phylaxins : 
 
 Of rabbit, etc., destroying 
 diphtheria and tetanus 
 poisons (Behring and 
 Kitasato, anti-toxin of 
 Tizzoni and Cattani;. 
 
 Tizzoni and Cattani immunised dogs and other animals against 
 tetanus, and employed the antitoxin as a therapeutic agent. Its 
 
58 BACTERIOLOGY. 
 
 active substance was precipitated by alcohol. Behring, Kitasato, and 
 Schiitz experimented with a view to conferring immunity upon 
 horses. The cultures were mixed with terchloride of iodine, and 
 injected at intervals of eight days, and the antitoxic power tested 
 on mice. By using increasingly virulent cultures, the blood became 
 increasingly antitoxic. 
 
 Vaillard filtered tetanus cultures through porcelain, and heated 
 the filtrate at gradually diminishing temperatures. The first in- 
 jections were made with 10 cc., which had been raised to 60 C. for 
 an hour, then a filtrate was used which had been heated to 55 C., 
 and lastly, a filtrate which had been heated to 50 C. The blood 
 became antitoxic, and by injecting increasing quantities of virulent 
 filtrates the antitoxic power was rapidly intensified, and animals 
 which were injected with antitoxin of full strength possessed 
 immunity many months afterwards. 
 
 Roux and Vaillard introduced another method. Virulent cultures 
 were filtered through porcelain, and the filtrate mixed with Gram's 
 solution of iodine in iodide of potassium. To give immunity to a 
 rabbit, 3 cc. of toxin with 1 cc. of Gram's solution were injected on 
 the first day, and increasing doses of toxin mixed with increasing 
 doses of Gram's solution on the following days. The same method 
 was applied to horses, sheep, and cattle. The antitoxin was found 
 not only in the blood, but in the urine and saliva, and in the 
 milk in cows. With cows and goats it is necessary to proceed 
 with the utmost care ; while horses, on the other hand, bear the 
 injections well, and are therefore more suitable for this purpose. 
 It is also very easy to obtain large quantities of blood from the 
 horse by inserting a trocar and cannula into the jugular vein. 
 
 Frankel was the first to produce immunity against diphtheria 
 by injecting guinea-pigs with toxin which had been heated to 70 C. 
 Behring mixed the toxin with terchloride of iodine, or employed small 
 doses of pure toxin. Horses, sheep, goats, and dogs were rendered 
 immune. 
 
 PREPARATION OF DIPHTHERIA ANTITOXIN. 
 
 For the preparation of diphtheria antitoxin Roux cultivates the 
 diphtheria bacillus in alkaline broth with 2 per cent, of peptone, and 
 by preference, in flasks in which the cultivating liquid can be exposed 
 to a current of moist air at 37 C. After about three weeks the 
 culture is filtered through a Chamberland filter, and if tested on 
 a guinea-pig it will be found that y 1 ^ of a cc. will kill an animal 
 weighing five hundred grammes in forty-eight hours. The diphtheria 
 
ANTITOXINS Ai\D SERUM THERAPY. 59 
 
 toxin immediately before the injection is mixed with ^ of its 
 volume of Gram's solution. This is used for several weeks, and 
 afterwards only pure toxin is injected. 
 
 The horses employed for this purpose are animals no longer fit 
 tor work, and it is necessary to inject them first of all w^ith mallein 
 to l>e sure that they are not suffering from glanders. 
 
 In a horse inoculated by Roux, the injection began with j cc. 
 of iodised toxin, increased to 1 cc. by the thirteenth day, and the 
 injection continued daily. On the seventeenth day | cc. of pure 
 toxin was injected, and this was increased by the forty-first day 
 to 10 cc. ; and on the forty-third day 30 cc. of pure toxin were 
 injected, causing pronounced oedema. The doses were still further 
 increased, until on the eightieth day 250 cc. were injected. In 
 two months and twenty days the horse had received 800 cc. of 
 toxin. 
 
 On the eighty-seventh day the serum obtained had an immunising 
 power of over 50,000. Bv this is meant that a guinea-pig resisted 
 inoculation of | cc. of virulent diphtheria culture when injected 
 twelve hours beforehand with serum in quantity equal to the -5337577 
 part of its body weight. 
 
 There are two tests which can be applied to the serum. First, 
 the antitoxic serum added to diphtheria toxin renders it inert ; and r 
 secondly, if serum is injected into a guinea-pig and toxin injected 
 several hours afterwards, no result follows. 
 
 Several ways have been suggested for estimating the immunising 
 power of the serum. 
 
 In Ehrlich's system, the unit is represented by *1 cc. of anti- 
 toxic serum, which, added to '8 cc. toxin, will neutralise it so that 
 the w r hole may be injected subcutaneously in a guinea-pig without 
 producing oedema. The standard toxin is a toxin of which *3 cc. 
 is fatal to 1 kilo, of guinea-pig. 
 
 But the preventive power of the serum is best expressed by the 
 result of a subsequent injection of toxin. The immunising power is 
 estimated by the number of grammes of guinea-pig which can be 
 protected against the minimum fatal dose of toxin by 1 cc. of anti- 
 toxic serum. 
 
 The antitoxic serum can be kept in sterilised flasks in the dark, 
 with the addition of a small piece of camphor, or it may be dried 
 191 I-///-//0. powdered, and thus supplied in a convenient form for trans - 
 I -"it. It has merely to be dissolved in water before use. 
 
 Klein employed a modified plan by which he claimed to have 
 obtained antitoxin in a far shorter time than is possible by Roux's 
 
60 BACTERIOLOGY. 
 
 method. Unfiltered attenuated cultures were injected into the 
 horse. Later, large quantities of living diphtheria bacilli from 
 the surface of solid cultures, of gradually increasing virulence, were 
 repeatedly injected so as to allow the bacilli to grow and multiply. 
 In twenty-three days an antitoxic serum was obtained, one part 
 of which was found capable of protecting 20,000 to 40,000 
 grammes of guinea-pig against more than a fatal dose of both living 
 bacilli and the resulting toxin. 
 
 Serum Treatment of Diphtheria. The results obtained by 
 Behring, Ehrlich, Kossel, and Wasserman, in the treatment of 
 diphtheria in children in Germany by means of the curative serum, 
 and by Roux and others in France, led to the adoption of the treat- 
 ment in this country. It is best to use an especially constructed 
 hypodermic syringe, which can be easily taken to pieces, and placed 
 in boiling water to sterilise it. The skin surface of the flank is 
 washed, and disinfected with 1 in 20 carbolic, and the antitoxin is 
 then injected. The syringe is taken to pieces, placed again in boiling 
 water, and thoroughly cleaned. 
 
 The dose will depend upon the age of the patient and the 
 strength of the serum. From 10 cc. to 20 cc. are injected in children 
 under fifteen, and 30 cc. to 40 cc. in older patients, and the 
 injection may be repeated in 12 hours. The best results are said 
 to be obtained by injecting every 12 hours, for the first 12, 36 or 
 48 hours, according to the nature of the case, 1,000 Behring's units, 
 this being the dose calculated according to the immunising power of 
 the serum. The result of the injection is to lower the temperature 
 and pulse, but frequently the reverse occurs, and in about half the 
 cases an urticarial and sometimes a scarlatiniform rash is produced. 
 Pains in the joints, in rare cases effusion, may also result from the 
 injection. 
 
 The beneficial results of the treatment are, according to the 
 Report of the Medical Superintendents of the hospitals of the 
 Metropolitan Asylums Board, as follows : 
 
 (1) Diminution of the faucial swelling and of the consequent 
 distress ; 
 
 (2) Lessening or entire cessation of the irritating and offensive 
 discharge from the nose ; 
 
 (3) Limitation of the extension of membrane ; 
 
 (4) Earlier separation of the exudation ; 
 
 (5) Limitation and earlier separation of membrane in laryngeal 
 cases ; 
 
 (6) Improvement in general condition and aspect of patients ; 
 
ANTITOXINS AND SERUM THERAPY. 61 
 
 (7) Prolongation of life, in cases which terminate fatally, to an 
 not obtained with former methods of treatment. 
 
 Statistics have also been brought forward which show, assuming 
 them to be reliable, a great reduction in the mortality after the 
 antitoxin treatment. A few instances may be quoted to illustrate 
 the statistical evidence. 
 
 According to Behring, in the four years prior to the employment 
 of antitoxin, there were in Berlin 15,958 cases of diphtheria, with a 
 mortality of 35'2 per cent. In 1894-5 there was an epidemic of 
 5,578 cases. Behring asserts that if the mortality had not been 
 reduced by the antitoxin treatment 1,963 would have died instead 
 of 1,056. Behring also states that in the Charite Hospital there 
 were 299 patients, with 53 deaths, or 16-7 per cent. In the Bethania 
 Hospital, where antitoxin was not employed, there were 249 
 patients, with 112 deaths, or 43 per cent. 
 
 At Vienna, at the Anna Hospital for children, the mortality 
 in 760 cases was 50'65 per cent., but after the introduction of anti- 
 toxin there were 40 deaths in 159 cases, giving a mortality of 
 2 5 '5 per cent. 
 
 In New York, it is said that before the introduction of anti- 
 toxin the mortality ranged from 30 '67 to 37 '34, while in 1895, under 
 treatment with antitoxin, the mortality fell to 19*43; but it was 
 also pointed out that since the introduction of antitoxin many 
 children with trifling attacks had been treated, and reported as 
 suffering from actual diphtheria, and that they would have recovered 
 without antitoxin, and therefore these cases have given the remedy 
 some credit which it does not deserve. 
 
 In London, according to the Report of the Medical Superin- 
 tendents of the hospitals of the Metropolitan Asylums Board there 
 were in 1894, before antitoxin was employed, 3,042 cases of 
 diphtheria with 902 deaths or 2 9 '6 per cent., and in 1895, when 
 antitoxin was used, 3,529 cases with 796 deaths or 22'5 per cent. : 
 a reduction of 7 ! 1 per cent, below that of 1894. The conclusions 
 drawn from the statistical and clinical observations are summed up 
 in the Report thus : 
 
 The improved results in the diphtheria cases treated during the year 
 1 895, are : 
 
 (I.) A great reduction in the mortality of cases brought under treat- 
 ment on the first and second day of illness. 
 
 (II.) The lowering of the combined general mortality to a point 
 below that of any former year. 
 
 (III.) The still more remarkable reduction in the mortality of the 
 laryngeal cases. 
 
62 BACTERIOLOGY. 
 
 (IV.) The uniform improvement in the results of tracheotomy at 
 each separate hospital. 
 
 (V.) The beneficial effect produced on the clinical course of the 
 disease. 
 
 A consideration of the statistical tables and clinical observations, 
 covering a period of 12 months and embracing a large number of cases, 
 in our opinion sufficiently demonstrates the value of antitoxin in the 
 treatment of diphtheria. 
 
 It must be clearly understood, however, that to obtain the largest 
 measure of success with antitoxin it is essential that the patient be 
 brought under its influence at a comparatively early date if possible not 
 later than the second day of disease. From this time onwards the chance 
 of a successful issue will diminish in proportion to the length of time 
 which has elapsed before treatment is commenced. This, though 
 doubtless true of other methods, is of still greater moment in the case 
 of treatment by antitoxin. 
 
 Certain secondary effects not infrequently arise as a direct result of 
 the injection of antitoxin in the form in which it has at present to be 
 administered, and, even assuming that the incidence of the normal com- 
 plications of diphtheria is greater than can be accounted for by the 
 increased number of recoveries, we have no hesitation in expressing the 
 opinion that these drawbacks are insignificant when taken in conjunction 
 with the lessened fatality which has been associated with the use of this 
 remedy. 
 
 We are further of the opinion that in antitoxin serum we possess 
 a remedy of distinctly greater value in the treatment of diphtheria than 
 any other with which we are acquainted. 
 
 On the other hand it has been urged that the decline in the 
 mortality in 1895 in London, which has been attributed entirely 
 to the antitoxin treatment, may possibly be partly due to the pre- 
 valence of a mild type of the disease, and that the fall in the 
 mortality during the seven previous years from 59 per cent, in 
 1888 to 29 per cent, in 1894, continued in 1895. 
 
 It is obvious that the whole subject requires to be very carefully 
 considered, and before any final conclusion can be arrived at as to 
 the therapeutic value of antitoxin, the evidence of others who have 
 had great experience in the treatment of diphtheria by the old and 
 the new methods must be taken into account, and reliable statistics 
 allowed to speak for themselves. 
 
 PREPARATION OF TETANUS ANTITOXIN. 
 
 Antitoxin for use in the serum treatment of tetanus is obtained 
 from the horse. The tetanus bacillus is cultivated in an atmosphere 
 of hydrogen, in flasks specially constructed for the purpose. In 
 
ANTITOXINS AND SERUM THERAPY. 63 
 
 a K>ut a fortnight the cultures are extremely toxic. The toxin is 
 obtained free from bacilli by nitration through porcelain. Injec- 
 tions may be given daily, subcutaneously or intravenously, beginning 
 with 1 cc. of iodised toxin, and gradually increasing the dose until 
 the pure toxin may be injected without danger. 
 
 Roux and Yaillard produced immunity in about three months. 
 \Vht-ii a few days have elapsed after the last injection, the blood is 
 drawn, by means of a trocar and cannula, from the jugular vein into 
 a sterilised glass vessel, and set aside to coagulate ; next day the 
 serum is drawn off with a pipette, and used in the liquid state, or 
 dried in a vacuum over sulphuric acid, and subsequently powdered. 
 When required for use the powder is dissolved in cold water. About 
 5 grammes are used for a dose. 
 
 Serum Treatment of Tetanus. The result, so far, of the 
 employment of tetanus antitoxin in animals suffering from tetanus 
 is disappointing, and the serum treatment is not likely to be of much 
 value in veterinary practice. Nocard infected sheep with tetanus 
 by inserting splinters of wood infected with spores into the muscles 
 of the leg. Tetanus supervened in eleven days, and the splinters 
 were removed, the tissues excised, and the wounds dressed with 
 iodoform. About twelve hours after the symptoms had shown 
 themselves, the sheep were inoculated with antitoxic serum at 
 intervals of one hour, but they all succumbed to tetanus. In one 
 case the total amount injected was 160 cc. of highly antitoxic serum. 
 
 The antitoxin has been employed in tetanus in man. Kanthack 
 has collected the history of a number of cases, and they indicate 
 that the treatment is useless in acute cases in man with a short 
 incubation period, while chronic cases with a long incubation period 
 often recover after the treatment. At the same time it must be 
 remembered that recovery often took place in chronic cases before 
 the introduction of the antitoxin treatment. 
 
 The question must still be considered to be sub judice, and a 
 trustworthy conclusion can only be based upon a more extended use 
 of the antitoxin and impartial reports of every individual case. 
 
 ANTITOXIN OF SEPTIC INFECTIONS. 
 
 An anti-streptococcic serum has been prepared by Marmorek. 
 Culture of streptococcus was intensified in virulence by inoculation 
 from rabbit to rabbit, and highly virulent cultures gave rise to a 
 powerful toxin. Roget and Charrin also, found that the serum of 
 immunised rabbits and of a horse conferred immunity. A patient 
 
64 BACTERIOLOGY. 
 
 with puerperal fever was injected with 8 cc., on the follow- 
 ing day with 16 cc., and on the third day with 25 cc. On the 
 fourth day the temperature had fallen, and the patient recovered. 
 Favourable results are said to have followed the use of the serum 
 in 46 cases of erysipelas. 
 
 Bokenham, working independently, cultivated the streptococcus 
 in a mixture of broth and serum. Horses and asses were inocu- 
 lated, and a considerable degree of immunity established. The 
 serum of an inoculated ass possessed antitoxic power. 
 
 Ruffer and Bullock succeeded in immunising four horses against 
 the toxin of Streptococcus pyogenes ; two had been previously immu- 
 nised against the toxin of the diphtheria bacillus. The streptococcus 
 was cultivated by Marmorek's methods in a mixture of two parts 
 of blood-serum and one part of peptonised broth, and the virulence 
 of cultures maintained by inoculation of rabbits. On testing the 
 immunising power of the antitoxic serum on rabbits, the effect 
 appeared to be slight in comparison with the antitoxins of the 
 bacilli of diphtheria and tetanus. In treating cases of septic infection 
 in the human subject, it has been recommended to commence with 
 two injections of 10 cc., and it is said that no unfavourable results 
 have been met with which could be attributed to the effect of the 
 serum. 
 
 ANTITOXIN OF TYPHOID FEVER AND OTHER DISEASES. 
 
 An antitoxic serum has been obtained by Chantemesse for use in 
 cases of typhoid fever, and it is probable that attempts will be made 
 to extend the principle of the antitoxic treatment to other infective 
 diseases. 
 
CHAPTER VII. 
 
 THE BACTERIOLOGICAL MICROSCOPE. 
 
 THE instruments sometimes in use in biological and pathological 
 laboratories are not sufficient for the study of bacteria. It is 
 absolutely essential for the examination of such minute objects that 
 the microscope should be equipped with an objective of sufficiently 
 high magnifying power and with a special illuminating apparatus, 
 while the mechanical arrangements of the stage must admit of the 
 examination of plate-cultivations. It would not be within the scope 
 of this work to give a detailed account of the mechanical arrange- 
 ments and optical principles of the microscope. These matters are 
 fully dealt with in special works on the subject,* but sufficient will 
 be said to afford assistance in the selection of a suitable instrument, 
 and to explain the improvements in the microscope which have been 
 such an aid in bacteriological investigations. 
 
 A magnified image of an object is the result of the change 
 produced in the direction of rays of light which are made to pass 
 through lenses. This alteration in the course of the rays is known 
 as refraction. A ray of light passing from a larer into a denser 
 medium is refracted towards a line drawn perpendicularly to the 
 surface of the latter. A ray of light passing through air and 
 impinging on water will not pass on in the same direction, but will 
 be refracted towards a line drawn perpendicularly towards the 
 MirfiK-e of the water. If the ray pass into glass instead of water 
 a greater refraction will take place, and if it pass into diamond the 
 bending in its course will be still greater (Fig. 11). 
 
 The sines of the angle of incidence and refraction of different 
 
 substances have a constant ratio to each other, which is known 
 
 he index of refraction, and this is determined for different 
 
 substances by the refraction produced by the passage of rays from 
 
 ;i vacuum. Thus the index of refraction for flint glass is about 1*6, 
 
 * Carpenter : The Microscope. Nageli and Schwenderer : The Microscope 
 in Theory and Practice. 
 
 65 n 
 
66 BACTERIOLOGY. 
 
 the sine of the angle of incidence of a ray passing from a vacuum 
 into glass being to the sine of the index of refraction as 1'6 to 1. 
 
 If we study the course of a pencil of rays we find that some 
 of the rays are reflected instead of entering the medium and being 
 refracted. When, for example, a pencil of rays falls upon water 
 or glass, after passing through air, some of the rays are lost by 
 reflection, and the proportion of the lost rays will increase with 
 their obliquity. The diminution of the brightness of the image 
 when pencils of rays have to pass through lenses is thus accounted 
 for, and this loss of light increases when the number of surfaces 
 
 -B 
 
 FIG. 11. THE REFRACTION OF LIGHT. 
 
 through which the rays pass are, as in high-power objectives,. 
 increased. There is an additional loss when there is an increase in 
 the difference between the refractive power of the different media 
 through which light passes. When pencils of rays pass from glass 
 into air, and then into glass again, the loss is much greater than 
 when the air is replaced by a medium with a refractive index mor& 
 nearly approaching that of glass. This explains the value of tha 
 immersion system, which will be referred to more fully later on, 
 and also the advantage of cementing pairs of lenses with Canada 
 balsam or glass paste. The lenses used in the optical arrangements 
 
THE BACTERIOLOGICAL MICROSCOPE. 
 
 67 
 
 of a microscope are principally convex, and the imperfections which 
 result must, if possible, be entirely overcome. These imperfections 
 are spherical and chromatic aberration. 
 
 Spherical aberration results from the unequal refraction of 
 rays passing through lenses with equal curvatures. The rays passing 
 through an ordinary convex lens do not all come to the same focus. 
 The rays passing through the marginal portion come to a focus at a 
 
 7?' 
 
 r F F 
 
 FIG. 12. SPHERICAL ABERRATION. 
 
 point much nearer to the lens than the focus of the rays passing 
 through the more central portion of the lens (Fig. 12), If the whole 
 aperture of the lens is used there must of necessity be blurring, for 
 at the point at which the marginal rays form a distinct image the 
 central rays will be out of focus, and at the point at which the 
 central rays form a distinct image the marginal rays will have 
 diverged, causing indistinctness. 
 
 This is partially remedied by using a diaphragm 
 and shutting out the marginal rays ; but this is 
 at the cost of loss of light and diminution of the 
 angle of aperture. The difficulty is approximately 
 overcome in practice by using a combination of 
 lenses. The aberration of a convex lens is the 
 opposite of that of a concave lens (Fig. 13). The 
 makers of the best lenses endeavour to obtain this 
 correction as perfect as possible to get the sharpness 
 of the image, so essential in studying the mor- 
 phology of bacteria. 
 
 Chromatic aberration is the result of the 
 unequal refrangibility of the coloured rays which 
 compose white light. If parallel rays of light pass through a 
 convex lens the violet rays, which are the most refrangible, will 
 come to a focus at a point much nearer to the lens than the 
 focus of the red rays, which are the. least refrangible; and the 
 intermediate rays of the spectrum will be focussed at points between 
 the red and the violet. A screen held at either of these foci 
 will show an image with prismatic fringes (Fig. 14). 
 
 FIG. 13. COM- 
 BINATION O F 
 LENSES IN 
 ABBE'S HOMO- 
 GENEOUS IM - 
 
 MERSION. 
 
68 BACTERIOLOGY. 
 
 The chromatic aberration may be reduced by stopping out the 
 marginal rays; but as it is necessary to get the most perfect 
 correction possible, advantage is taken of the different relations 
 which the refractive and dispersive powers bear to each other in 
 different glasses. By combining a double convex lens of crown 
 glass with a plano-convex lens of flint glass, correction is obtained 
 for the violet and red rays. An achromatic objective is constructed 
 on this principle, but the result is not perfect, as the intermediate 
 coloured rays remain uncorrected, and what is termed a secondary 
 spectrum gives rise to images with coloured fringes, especially at the 
 margin of the field. Abbe and Schott, after a great number of 
 experiments, succeeded in discovering a glass with optical properties 
 which removed the secondary spectrum, and objectives made with 
 the new glass are termed apo-chromatic. There is much more 
 
 FIG. 14. CHROMATIC ABERRATION. 
 
 perfect concentration of the component rays than in the ordinary 
 achromatic objectives, and the advantages thus obtained are very 
 great. The objectives can be made of higher angle and admit of 
 higher eye-pieces being used without materially diminishing the 
 brilliancy and definition of the image. There is a complete absence 
 of coloured fringes, and the perfect definition is invaluable in 
 micro-photography. 
 
 Another fault which has to be corrected is the aberration caused 
 by covering a microscopical preparation with a cover-glass. Ross 
 was the first to point out the difference in the image w-hen the 
 object was examined under a cover-glass, and that by altering the 
 position of the front pair of lenses, in an objective corrected for an 
 uncovered object, the objective could be corrected for the covered 
 object (Fig. 15). 
 
 Objectives are generally corrected for a standard thickness of 
 cover-glass, but H. Lister devised a screw-collar adjustment by 
 which the position of the front pair of lenses could be altered at 
 will ; and as it is almost impossible to obtain cover-glasses which 
 
THE BACTERIOLOGICAL MICROSCOPE. 69 
 
 do not vary slightly in thickness, the most perfect definition 
 can only be obtained by adjusting for each separate cover-glass 
 preparation. 
 
 Immersion system. All objectives were formerly used dry 
 that is to say, with an air space between the objective and the 
 specimen to be examined but high-power objectives are now almost 
 entirely made on the immersion system, a drop of liquid being 
 interposed between the objective and the cover-glass. 
 
 About fifty years ago Amici observed that if a drop of water 
 intervened between the cover-glass or an uncovered object and the 
 lens the image was more brilliant. The passage of rays from the 
 object or the cover-glass into air, 
 and again from air into glass, caused 
 considerable loss of light. With 
 objectives of wide angle of aperture 
 the advantages were counteracted 
 by the reflection of rays falling ob- 
 liquely upon the lens. By inter- -. 
 
 posing water more rays are bent 
 in. or refracted, and enter the lens 
 instead of being reflected and lost. 
 
 Hartnack, Nachet, and others 
 
 adopted the immersion svstem, and 
 
 , . , . , FIG. 15. OBJECTIVE WITH COLLAR 
 
 high-power water immersion lenses CORRECTION (6). 
 
 were constructed with high angle 
 
 of aperture.* It was found that there was less necessity for 
 correcting for covers of different thickness, as the aberration from 
 this cause was diminished. The lenses were corrected for an average 
 thickness of cover, and slight deviations produced hardly any 
 appreciable effect. 
 
 Wenham, Stephenson, Abbe, and Zeiss carried the system to 
 perfection. They argued that the advantages obtained by water 
 immersion would be intensified if a liquid could be found of the 
 same refractive and dispersive power as crown glass. The media 
 would be optically uniform, and the residt a homogeneous immersion 
 system. 
 
 * The angle of aperture is " the angle made by the most diverging of the 
 rays of the pencil issuing from any point of an object that can enter the lens, 
 and take part in the formation of an image of it." 
 
 The numerical aperture is defined by Abbe as equal to " the sine of the 
 angle of aperture multiplied by the refractive index of the medium between 
 the object and the objective." 
 
70 BACTERIOLOGY. 
 
 After experimenting with different liquids solutions of salts, 
 and various essential oils Abbe recommended cedar oil as most 
 suitable for the purpose. In its optical properties it very closely 
 resembles crown glass, and it is far more convenient for use than 
 any watery solutions of salts, especially when it is necessary to 
 make a more or less prolonged examination of an object. 
 
 The difference between the dry, water, and oil immersion systems 
 may be illustrated, as Frankel has pointed out, by a very simple 
 experiment. If a -glass rod is inserted into an empty test-tube, it 
 is easily visible owing to the difference in refraction between the 
 glass and the surrounding air. If the tube is filled up with 
 water the rod is seen with difficulty, and if, instead of w r ater, cedar 
 oil is used, the part of the rod immersed in the oil will entirely 
 disappear from view. The rays of light pass through an optically 
 uniform medium in the experiment with cedar oil, and no refraction 
 or reflection of rays of light can occur. 
 
 To use an oil immersion objective, a minute drop of cedar oil 
 is placed on the centre of the cover-glass, and the lens lowered 
 by means of the coarse adjustment until it touches the oil. The 
 specimen is then carefully brought into focus with the fine adjust- 
 ment. If the slide is held between the finger and thumb of one 
 hand, and moved from side to side while the other hand is working 
 the fine adjustment, there can be no danger of injuring either the 
 objective or the specimen. 
 
 Microscopes are made upon either the Ross or the Jackson 
 model. In the Ross model the body of the microscope is fixed 
 at its base to a transverse arm, which is raised or lowered with 
 it by the rack arid pinion. In the Jackson model the body is 
 supported for a great part of its length on a solid " limb." 
 
 In the Ross model, unless the body and transverse arm are very 
 solid as in Powell and Lealand's microscopes (Fig. 23), there will be 
 vibration at the ocular end ; but in the Jackson model vibration is 
 practically prevented, and this is most essential, especially in working 
 with very high powers. 
 
 The steadiness of the microscope also largely depends upon the 
 form of stand. There are four different types of stands. The 
 tripod (Fig. 23) ; the plate, with double columns ; the single column, 
 ending in a plate or a bent claw; and the horse-shoe (Fig. 18). 
 
 The tripod stand with cork feet is the steadiest form of stand, 
 but it is cumbrous and expensive, and these objections also apply to 
 the model made by Ross. 
 
 The single upright should be unquestionably condemned, as it 
 
THE BACTERIOLOGICAL MICROSCOPE. 
 
 71 
 
 FIG. 16. ENGLISH MODEL. 
 
72 BACTERIOLOGY. 
 
 freely admits of vibration, and is most inconvenient for laboratory 
 work, The heavy horse-shoe form is compact and firm, and the 
 weight of it can hardly be considered an objection. 
 
 The tubular body is from eight to ten inches in length, and within 
 it is a draw-tube with engraved scale. By extending the draw-tube 
 greater magnification is obtained ; but as this is at the cost of 
 definition it should hardly ever be used in the examination of 
 bacteria. 
 
 A triple nose-piece is a great convenience, saving the time which 
 is otherwise spent in replacing objectives of different magnifying 
 power, and there is less risk of injuring them. 
 
 Focus should be obtained by means of a rack and pinion coarse 
 
 FIG. 17. REMOVABLE MECHANICAL STAGE. 
 
 adjustment. The sliding tube is not to be recommended, as the 
 motion may be stiff, encouraging the use of force, which in turn may 
 result in the objective being brought violently into contact with the 
 specimen, injuring the lens or damaging the preparation ; or it may 
 get too loose and readily slip out of focus. 
 
 The stage should be flat and rigid, either rectangular or circular, 
 so long as it is sufficiently large to accommodate a plate- cultivation. 
 A removable mechanical stage is of great advantage for working 
 with high powers, as a motile bacterium can be constantly kept in 
 view while one hand is engaged in working the fine adjustment 
 (Fig. 17). It may also be employed as a finder if it is engraved 
 with a longitudinal and vertical scale, and provided with a stop. 
 The mechanical stage must be removable, so that the stage proper 
 
THE BACTERIOLOGICAL MICROSCOPE. 
 
 73 
 
 FIG. 18. COXTIXEXTAL MODEL. 
 
74 BACTERIOLOGY. 
 
 may be free from any attachments when required for the examina- 
 tion of cultures. 
 
 Diaphragms are necessary for regulating the amount of light. 
 The plan of using a series of discs, with apertures of different sizes, 
 should be avoided, as they are easily lost, and bacteriological investi- 
 gations may have to be made under conditions in which it is difficult 
 to replace them. A better plan is a revolving plate with apertures 
 of different sizes, but the most convenient form is the iris diaphragm 
 (Fig. 19). 
 
 The sub-stage condenser is quite as necessary in bacteriological 
 work as a high-power objective. In fact, the condenser and the 
 objective should be considered as forming one optical apparatus, and 
 
 the microscope regarded quite as 
 incomplete without a condenser as 
 'it would be without an objective. 
 
 By means of the sub- stage con- 
 denser (Fig. 20) the rays of light 
 are concentrated at one point or 
 on one particular bacterium ; and 
 for the best definition it is essential 
 that there should be mechanical 
 
 FIG. 19,-lRis DIAPHRAGM. arrangements for accurately cen- 
 
 tring and focussing the condenser. 
 It may even with advantage be provided with a fine adjustment. 
 
 To sum up, a microscope for bacteriological investigation should 
 be provided with (1) a steady stand of either the tripod or horse- 
 shoe form ; (2) a tubular body on the Jackson model ; (3) a wide- 
 angled sub-stage condenser, such as Abbe's; (4) objectives of an inch, 
 ^th of an inch, and a yVfch homogeneous immersion ; (5) a removable 
 mechanical stage ; and for the most accurate work there should be 
 centring arrangements and a coarse and fine adjustment to an oil- 
 immersion sub-stage condenser such as Powell and Lea-land's, and 
 a j\th homogeneous oil-immersion apo- chromatic objective. 
 
 With regard to the choice of a microscope, it is chiefly a 
 question of price. The most perfect instrument is the large model 
 by Powell and Lealand, but it is most expensive, and quite unsuit- 
 able for laboratory work. For general use excellent instruments 
 are made by Zeiss, Leitz, Reichert, or Swift. The bacteriological 
 microscopes of these makers are in the necessary equipment 
 practically identical. The Zeiss microscope is the most finished, and 
 costs about twenty pounds. A similar microscope by Leitz and 
 by Swift costs about eighteen, and both make an excellent students' 
 
THE BACTERIOLOGICAL MICROSCOPE. 75 
 
 bacteriological microscope, with a cheap form of adjustment to the 
 suit-stage condenser, at a total cost of about fifteen pounds. 
 
 Method of Illumination. Good daylight is the best for 
 general work. The microscope should be placed near a window 
 with a northern aspect. Direct sunlight should never be utilised, and 
 the best light is that reflected from a white cloud. When daylight 
 is not available good results can be obtained with either gas or a 
 
 FIG. 20. ABBE'S CONDENSER CONSTRUCTED BY ZEISS. 
 
 paraffin lamp. In the author's laboratory the microscope lamps 
 are fitted with Welsbach incandescent mantles. These have many 
 advantages over an Argand burner or a paraffin lamp. A steady 
 and beautifully white light is obtained, and the lamps are quickly 
 lit, and require comparatively little attention. In using high powers 
 and carefully focussing the sub-stage condenser, the image of the 
 fabric of the mantle is embarrassing, and is an objection to this 
 light for the most accurate observations, but in other respects, and 
 
76 
 
 BACTERIOLOGY. 
 
 for general use, it is the best form of artificial illumination for the 
 microscope. 
 
 An ordinary paraffin lamp of the cheapest form may be used, 
 but there are many objections to it, such as the shape of the 
 chimney, and the striae and defects in the glass. The best form of 
 paraffin lamp is constructed by Baker and by Swift from sug- 
 
 FIG. 21. MICROSCOPE LAMP. 
 
 gestions by Nelson and Dallinger (Fig. 21) ; there is also a similar 
 but much larger pattern which is made by Swift (Fig. 22). This 
 form of lamp has a large flat bowl for the oil: It is attached to 
 a standard, and can be raised or lowered to the desired position. 
 The chimney is of metal and blackened, so that there is no reflected 
 light, and it may also with advantage be provided with a shade, 
 so that no light reaches the eye except through the microscope. 
 The burner may be made to revolve, so that either the edge 
 
 or 
 
THE BACTERIOLOGICAL MICROSCOPE. 
 
 77 
 
 the flat of the flame may be utilised. Great care should be taken 
 to have the wick evenly trimmed. The best paraffin oil should 
 be burnt, and it is as well to add a small lump of camphor. The 
 iiu-tal chimney has an aperture in front, giving exit to the rays of 
 light, which is closed in by a slip of glass. The glass is very liable 
 to crack when exposed to the full force of the flame, and it is as well, 
 therefore, to be provided with a stock of glass slips, which have 
 
 FIG. 22. LARGE MICROSCOPE LAMP. 
 
 been annealed by being enveloped in a cloth and boiled for two or 
 three hours. 
 
 The flat of the flame is used with low powers. The image of 
 the flame is reflected by a plane mirror, and a bull's-eye condenser 
 interposed between the lamp and the mirror to give an equal 
 illumination of the whole field. In working with high powers the 
 lamp is turned with the flame edgewise, and the mirror is dispensed 
 with. By working, as it is termed, directly on the edge of the 
 name, the illumination is greatly increased, and a band of light can 
 
78 BACTERIOLOGY. 
 
 be concentrated on that part of the microscopical preparation which 
 requires most careful study (Fig. 23). 
 
 To obtain the best definition considerable time must be spent in 
 the arrangement of the illumination. The lamp and microscope 
 having been placed in position, a low power is first used and the 
 smallest diaphragm. On looking through the microscope it will 
 probably be observed that the image of the diaphragm is not in 
 the centre of the field. By moving the centring screw of the con- 
 denser this may be adjusted. The image of the edge of the flame 
 may not be central, and this must be adjusted by moving the lamp 
 into position. The low power is then replaced by a high power, 
 the largest diaphragm used, and the bacteria brought into focus. 
 The diaphragm must now be replaced by one of medium size, and 
 by racking the condenser up and down, a point will be arrived at 
 when the image of the edge of the flame appears af an intensely 
 bright band of light. If this is not exactly in the centre of the 
 field the centring screws of the condenser must again be adjusted. 
 Lastly, by trying different sizes of diaphragms, and focussing with 
 the fine adjustment, and using the correction collar, we arrive at the 
 sharpest possible image of the bacteria. 
 
 When the condenser has been accurately centred, it will .still be 
 necessary to focus it for each individual specimen, so as to correct 
 for difference in the thickness of slides and the layers of mounting 
 medium. Correction for different thickness of cover-glasses must 
 in each case be made by means of the collar adjustment in the follow- 
 ing way. A high-power eye-piece is substituted for the ordinary 
 eye- piece, and the fault in the image will thereby be intensified. By 
 moving the collar completely round, first in one direction and then 
 the other, while carefully observing the effect on the image, it will 
 be seen to become obviously worse whichever way the collar is turned. 
 The collar must then be turned through gradually diminishing dis- 
 tances until an intermediate point is reached at which the best image 
 results with the high-power eye-piece, and on replacing this by the 
 low-power eye-piece the sharpest possible image will be obtained. 
 
 Effect of the sub-stage condenser. The sub-stage condenser 
 gives the most powerful illumination when it has been racked up 
 until it almost touches the specimen. It produces a cone of rays of 
 very short focus, and the apex of the cone should correspond with the 
 particular bacterium or group of bacteria under observation. The 
 effect of the condenser without a diaphragm is to obliterate what 
 Koch has termed the structure picture. If the component parts of a 
 tissue section were colourless and of the same refractive power as 
 
THE BACTERIOLOGICAL MICROSCOPE. 
 
 79 
 
 5 
 
 
 1 1 
 
 IS 
 
 rH O 
 
 'r v 
 
80 BACTERIOLOGY. 
 
 the medium in which the section is mounted, nothing would be 
 visible under the microscope. As, however, the cells and their nuclei, 
 and the tissue fibres do differ in this respect, the rays which pass 
 through them are diffracted, and an image of lines and shadows is 
 developed. If in such a tissue there were minute coloured objects, 
 and if it were possible to mount the tissue in a medium of exactly 
 the same refractive power, the tissue being then invisible, the 
 detection of the coloured objects would be much more easy. This 
 is exactly what is required in dealing with bacteria which have been 
 stained with aniline dyes, and the desired result can be obtained 
 by the use of the sub-stage condenser. 
 
 If we use the full aperture of the condenser the greatly converged 
 rays play on the component parts of the tissue, light enters from 
 
 FIG. 24. RAMSDEX MICROMETER EYE-PIECE. 
 
 nil sides, the shadows disappear, and the structure picture is lost. 
 If now a diaphragm is inserted, so that we are practically only 
 dealing with parallel rays, the structure picture reappears. As the 
 diaphragm is gradually increased in size the structure picture 
 gradually becomes less and less distinct, while the colour picture, 
 the image of the stained bacteria, becomes more and more intense. 
 When, therefore, bacteria in the living condition and unstained tissues 
 are examined a diaphragm must be used, and when attention is 
 to be concentrated upon the stained bacteria in a section or in a 
 cover-glass preparation, the diaphragm must be removed and the 
 field flooded with light. 
 
 Micrometer. For the measurement of bacteria a stage micro- 
 meter may be used with a camera lucida. The stage micrometer 
 consists of a slip of thin glass ruled with a scale consisting of tenths 
 and hundredths of a millimetre. The image of this can be projected 
 
THE BACTERIOLOGICAL MICROSCOPE. 
 
 81 
 
 on a piece of paper, and a drawing made, and the object to be 
 measured can then be projected on the paper and compared with the 
 scale. 
 
 In the Ramsden micrometer eye- piece (Fig. 24) two fine wires 
 are stretched across the field of an eye-piece, one of which can be 
 moved by a micrometer screw. In the field there is also a scale 
 with teeth, and the interval between them corresponds to that of the 
 threads of the screw. The circumference of the brass head is usually 
 divided into one hundred parts, and a screw with one hundred threads 
 to the inch is used. The bacterium to be measured is brought into a 
 
 FIG. 25. MICROMETER EYE-PIECE BY ZEISS. 
 
 position in which one edge appears to be in contact with the fixed 
 wire, and the micrometer screw is turned until the travelling wire 
 appears to be in contact with the other edge. The scale in the 
 field and the scale on the milled head together give the number of 
 complete turns of the screw and the value of a fraction of a turn in 
 separating the wires. 
 
 In the micrometer eye-piece constructed by Zeiss, the eye- piece 
 with a glass plate with crossed lines is carried across the field by 
 means of a micrometer screw (Fig. 25). Each division on the edge 
 of the drum corresponds to '01 mm. Complete revolutions of the 
 drum are counted by means of a figured scale in the visual field. 
 Another method of measuring bacteria will be referred to in the 
 
82 BACTERIOLOGY. 
 
 chapter on micro-photography. The unit of measurement is one 
 thousandth of a millimetre or a micro-millimetre or micron, and is 
 expressed by the Greek letter /JL. 
 
 CARE OF THE MICROSCOPE. 
 
 After use the objectives, sub-stage condenser, and eye-piece 
 should be carefully wiped with soft linen, an old silk handkerchief, 
 or chamois leather, and the microscope covered with a bell-glass to 
 protect it from dust. If a lens comes into contact with Canada 
 balsam it must be very carefully wiped with a soft rag moistened 
 with alcohol, and then cleaned with a soft leather. Microscopes 
 should not be exposed to the fumes of sulphuretted hydrogen^ 
 chlorine, or volatile acids. 
 
CHAPTER VIII. 
 
 MICROSCOPICAL EXAMINATION OF BACTERIA. 
 
 (A) BACTERIA ix LIQUIDS, CULTURES, AND FRESH TISSUES. 
 
 Ix conducting bacteriological researches the importance of absolute 
 cleanliness cannot be too strongly insisted upon. All instru- 
 ments, glass vessels, slides, and cover-glasses should be thoroughly 
 cleansed before use. A wide-mouthed glass jar should always be 
 close at hand, containing refuse alcohol for the reception of re- 
 jected slide preparations or dirty cover-glasses. When required 
 again for use, slides can be easily wiped clean with a soft rag. Cover- 
 glasses require further treatment, for, unless they are perfectly 
 clean, it is difficult to avoid the presence of air bubbles when 
 mounting specimens. They should be left in strong acid (hydro- 
 chloric, sulphuric, or nitric) for some hours ; they are then washed, 
 first with water and then with alcohol, and carefully wiped with a 
 soft rag. The same principle applies in the preparation and 
 employment of culture media ; any laxity in the processes of 
 sterilisation, or insufficient attention to minute technical details, 
 will surely be followed with disappointing results by contamination 
 of the cultures, resulting in the lass of much time. 
 
 For the preparation of microscopical specimens it will be found 
 convenient to use a platinum inoculating needle. This consists of 
 two or three inches of platinum wire fused into the end of a gla>> 
 rod about eight inches in length. Platinum is employed as it 
 rapidly cools after being raised to a white heat in the flame of a 
 Bunsen burner. It is thus completely sterilised, and in a few 
 moments is cool enough not to destroy the bacteria with which it is 
 brought into contact. 
 
 When using platinum needles, either for inoculating fresh tubes 
 in carrying on a series of pure cultures, or in transferring a small 
 portion of a cultivation to a cover-glass for examination under the 
 microscope, the careful sterilisation of the needle by heating the 
 
 83 
 
84 BACTERIOLOGY. 
 
 platinum wire till it is white hot in every part, and heating also 
 as much of the glass rod as is made to enter the test-tube, must 
 be carried out with scrupulous care. Indeed it is a good plan to 
 
 FIG. 26. INOCULATING NEEDLES. 
 
 let it become & force of habit to sterilise the needle before and after 
 use on every occasion, whatever may be the purposes for which it 
 is employed. 
 
 UNSTAINED BACTERIA. 
 
 The bacteria in liquids, such as pus, blood, and culture- fluids, can 
 be investigated in the unstained condition by transferring a drop with 
 a looped platinum needle or a capillary pipette to a slide, covering 
 it with a clean cover-glass, and examining without further treat- 
 ment. If it is desirable to keep the specimen under prolonged 
 observation, a drop of sterilised water or salt solution must be run in 
 at the margin of the cover-glass to counteract the tendency to dry. 
 
 Cultures on solid media can be examined by transferring a small 
 portion with a sterilised needle to a drop of sterilised water on a 
 slide, thinning it out, and covering with a cover-glass as already 
 described. 
 
 Tissues in the fresh state may be teased out with needles in 
 sterilised salt solution, and pressed out into a sufficiently thin layer 
 between the slide and cover-glass. Glycerine may in many cases 
 be substituted for salt solution, especially for the examination of 
 micro-organisms such as Actinomyces and mould fungi. 
 
 There is, as a rule, no difficulty in recognising the larger micro- 
 organisms such as those just mentioned; but when we have to 
 deal with very small bacilli and micrococci, they may possibly be 
 mistaken for granular detritus or fat-crystals, or vice versa. They 
 are distinguished by the fact that fatty and albuminous granules 
 are altered or dispersed by acetic acid, and changed by solution 
 of potash; alcohol, chloroform, and ether dissolve out fat-crystals 
 
MICROSCOPICAL EXAMINATION OF BACTERIA. 85 
 
 or fatty particles ; 011 the other hand, micro-organisms remain 
 unaffected by these reagents. Baumgarten demonstrated tubercle 
 bacilli in sections by treating them with potash, which clarified 
 the ti>su's and brought the bacilli clearly into view. Actinomyces 
 and other vegetable structures will not disappear when sections are 
 immersed in weak hydrochloric acid and mounted in glycerine. 
 
 In examining unstained bacteria, it is necessary, in order to 
 obtain the structure picture, that the light entering the microscope 
 should be reduced by employing a small diaphragm, and the sub-stage 
 condenser carefully centred and focussed. To focus an unstained 
 specimen in which only bacteria are present, is often difficult. The 
 slide may be gently raised towards the objective, and the stage 
 may be constructed to enable this to be done with the index finger 
 (Fig. 16). If on tilting the slide the organisms come into focus it 
 will serve as a guide in working the fine adjustment. Another plan 
 when bacteria are examined in water, is to look for an air-bubble, 
 and then to focus its edge until the bacteria appear in view. 
 
 The simple method of covering the liquid with a cover-glass will 
 not answer for a prolonged examination, as the liquid evaporates and 
 the specimen dries up. To keep living bacteria under observation 
 for any length of time, in order to study their movements or spore- 
 formation, a special slide must be employed (p. 120). 
 
 STAINED BACTERIA. 
 
 Yv'eigert first pointed out the value of the aniline dyes for 
 staining bacteria, and we are principally indebted to Koch, Ehrlich, 
 Gram, and Loftier for many valuable processes. 
 
 The staining of fresh preparations, especially those with no 
 coagulable albumen to fix them, may be carried out by the method 
 of His. A slide is prepared as already described for the exami- 
 nation of micro-organisms in the fresh state. The reagents are 
 then applied by placing them with a pipette drop by drop at 
 one margin of the cover-glass, and causing them to flow through 
 the preparation by means of a strip of filter-paper placed at the 
 opposite margin. 
 
 Babes recommends another rapid means of examining cultivations. 
 A little of the growth, removed by means of a sterilised platinum 
 hook or small loop, is spread out on a cover-glass into as thin a 
 film as possible : when almost dry, a drop or two of a weak aqueous 
 solution of methyl violet is allowed to fall from a pipette upon the 
 film. The cover-glass with the drop of stain is, after a minute, 
 
86 BACTERIOLOGY. 
 
 carefully turned over on to a slide, and the excess of stain gently 
 and gradually removed by pressure with a strip of filter-paper. 
 This affords a rapid means of demonstration for example, of a 
 cultivation of Koch's comma bacilli in nutrient gelatine enabling 
 the microbes to be seen in some parts of the preparation both 
 stained and in active movement. 
 
 COVER- GLASS PREPARATIONS. 
 
 Bacteria may be spread out into a thin layer on a cover-glass, 
 and then treated with a dye, or sections of tissues containing bacteria 
 can be stained and then mounted in the usual way. 
 
 The method of making a cover-glass preparation is one which is 
 very commonly employed. In addition to its value as a means of 
 examining bacteria in liquids and solid culture media, it affords 
 the additional advantage of enabling, if necessary, a large number 
 of preparations to be made, which, when dried, can be preserved, 
 stained or unstained, in ordinary cover-glass boxes ; they are 
 then in a convenient form for transport, and can be mounted 
 permanently at leisure. 
 
 The method is as follows : A cover-glass is smeared with the 
 cut surface of an organ or pathological growth, or with sputum ; 
 or a drop of blood, pus, or culture- fluid is conveyed to it with a 
 looped platinum needle. It is absolutely necessary to spread out 
 the micro-organisms into a sufficiently thin layer, so that the 
 individual bacteria may be as much as possible in the same plane, 
 otherwise some in the field will be in focus and others out of 
 focus, and it would be impossible to obtain a satisfactory photograph 
 of such a specimen. To overcome this it will be necessary, in the 
 case of cultures on solid media, to diffuse the bacteria in a little 
 sterilised water ; and even cultures in liquids may sometimes with 
 advantage be diluted in the same way. By means of another 
 cover-glass the juice or fluid is squeezed out between them into a 
 thin layer, and on sliding them apart each cover-glass bears on one 
 .side a thin film of the material to be examined ; or a culture is 
 spread out into a thin film by means of a hooked platinum needle. 
 The cover-glass is then placed with the prepared side upwards, and 
 allowed to dry. After a few minutes, it is taken up with a pair of 
 flat-bladed or spring forceps, with the prepared side uppermost, -and 
 passed rapidly from above downwards three times through the 
 flame of a spirit lamp or Bunsen burner. Two or three drops of 
 an aqueous solution of fuchsine or methyl violet will be sufficient to 
 cover the film, arid after a minute or two the surplus stain is washed 
 
MICROSCOPICAL EXAMINATION OF BACTERIA. 87 
 
 off with distilled water by means of a siphon apparatus or a wash- 
 bottle. The cover-glass may be allowed to dry, and then mounted 
 in Canada balsam, or it may, while still wet, be turned over on to a 
 slide, the excess of water removed with filter-paper, and the exposed 
 surface wiped dry. It may first be examined with a power of about 
 250 diam*. ; and if a high magnification is required, which is usually 
 the case, a droplet of cedar oil is placed on the cover-glass, and the 
 specimen examined with an immersion lens. 
 
 If the specimen is to be made permanent, fix the cover- glass at 
 one corner with the thumb, and with a soft rag carefully wipe off 
 the cedar oil ; then float off the cover-glass by running in distilled 
 water at its margin, and having made a little ledge with a strip of 
 filter-paper, place the cover-glass up against it upon one of its 
 edges and leave it to dry. When perfectly dry mount in Canada 
 balsam, or put it away in a cover-glass box provided with a label of 
 contents. 
 
 In many cases it is necessary or preferable to apply the stain 
 for a much longer period. This may best be effected by pouring 
 some of the staining solution into a watch-glass, and allowing the 
 cover-glasses to swim on the surface, with their prepared side, of 
 course, downwards. Throughout all these manipulations it is 
 necessary to bear in mind which is the prepared surface of the 
 cover-glass. 
 
 Instead of using the watery solutions of the aniline dyes the 
 author prefers in many cases to use stronger solutions, and to reduce 
 the staining by a momentary immersion in alcohol. Yery beautiful 
 preparations of streptococci, sarcinse and other bacteria can be 
 obtained by this method, which is as follows : Cover-glass prepara- 
 tions are stained with carbolised fuchsine (Neelsen's solution) for 
 about two minutes, rinsed in alcohol for a few seconds, quickly 
 washed in water, and either examined in water or dried and 
 mounted in the usual way. The extent of decolorisation is a 
 matter of practice : a momentary immersion in alcohol is sometimes 
 sufficient ; too long immersion will remove too much of the colour ; 
 too short immersion will leave the delicate outlines indistinct. This 
 method is especially valuable for sarcinse and streptococci, the 
 divisions between the elements being sharply defined, and as any 
 albuminous particles or debris in the preparation are decolorised, 
 much cleaner and sharper preparations are obtained than with the 
 watery solutions. Loffler's and other concentrated solutions may 
 also be used, but Neelsen's solution may be regarded as the standard 
 one for this method. 
 
88 BACTERIOLOGY. 
 
 Aniline oil, carbolic acid, and some other chemicals, when added 
 to the aniline dyes, have the property of acting in the manner 
 of mordants, in some way fixing the colour in the bacteria, so that 
 they are not so readily acted upon by decolorising agents. 
 
 Loffler's Solution. Potash intensifies the staining power, and 
 Koch and Lomer have both used it with methylene blue. Loftier' s 
 solution consists of 30 grammes of methylene blue in 100 grammes 
 of 1 in 10,000 solution of potash. It may be used with advantage 
 for almost all kinds of bacteria. 
 
 Gram's Method. With a solution of gentian-violet the whole 
 film on the cover-glass is at first stained violet. By immersing the 
 cover-glass in a solution of iodine in iodide of potassium the stain 
 is fixed in the bacilli, but not in any debits, pus cells, or tissue 
 elements present in the film. Consequently by transferring the 
 cover-glass to alcohol the bacilli alone remain stained, the violet 
 colour being merely changed to blue. By employing a contrast 
 colour, such as eosin, a double staining is obtained. In some 
 bacteria the sheath is by this method differentiated from the 
 protoplasmic contents. 
 
 The stock solution of gentian-violet is prepared by shaking up 
 1 cc. of pure aniline with twenty parts of distilled water, and 
 filtering the emulsion. Half a gramme of the best finely powdered 
 gentian-violet is dissolved in the clear filtrate, and the solution filtered 
 before use. 
 
 The details of the method will now be described. In the first 
 place, it is much better to employ the aniline-gentian-violet solution 
 quite freshly prepared, and the following useful method is invariably 
 used by the author : Place four or five drops of pure aniline in 
 a test-tube, fill it three-quarters full with distilled water, close the 
 mouth of the tube with the thumb, and shake it up thoroughly. 
 Filter the emulsion twice, and pour the filtrate into a watch-glass 
 or glass capsule. To the perfectly clear aniline water thus obtained 
 add drop by drop a concentrated alcoholic solution of gentian-violet 
 till precipitation commences. Cover-glasses must be left in this 
 solution about ten minutes, transferred to iodine-potassic-iodide 
 solution until in itwo or three minutes the film becomes uniformly 
 brown, and then rinsed in alcohol. The process of decolorisation may 
 be hastened by dipping the cover-glass in clove-oil and returning it 
 again to alcohol. The cover-glass is once more immersed in clove-oil, 
 then dried by gently pressing between two layers of filter-paper, 
 and finally mounted in Canada balsam. 
 
MICROSCOPICAL EXAMINATION OF BACTERIA. 89 
 
 DOUBLE STAINING OF COVER-GLASS PREPARATIONS. 
 
 To double stain cover-glass preparations they can be treated by 
 Ehrlich's method for staining tubercular sputum, or by Neelsen's 
 modification, or by staining with eosin after treatment by the 
 nit -thod of Gram. 
 
 Ehrlich's method is as follows : Five parts of aniline oil are 
 shaken up with one hundred parts of distilled water, and the 
 emulsion filtered through moistened filter-paper. A saturated 
 alcoholic solution of fuchsine, methyl- violet, or gentian-violet, is 
 added to the filtrate in a watch-glass, drop by drop, until precipitation 
 commences. Weigert recommended that exactly eleven parts of the 
 dy j should be used to one hundred parts of the aniline solution. 
 
 Cover-glass preparations are floated in this mixture for fifteen 
 minutes to half an hour, then washed for a few seconds in dilute 
 nitric acid (one part nitric acid to two of water), and then rinsed 
 in distilled water. The stain, is removed from everything except 
 the bacilli; but the ground substance can be after-stained brown 
 if the bacilli are violet, or blue if they have been stained red. 
 
 Xeelsen's Solution and Methylene Blue. Ziehl suggested the use of 
 carbolic acid as a substitute for aniline oil, and Neelsen recommended 
 a solution composed of 100 cc. of a 5 per cent, watery solution 
 of carbolic acid, 10 cc. of absolute alcohol, and 1 gramme of 
 fuchsine. This stain is commonly known as the Neelsen or Ziehl- 
 Neelsen solution. Cover-glass preparations are floated on the hot 
 dye for two minutes, they are then rinsed in dilute sulphuric acid 
 25 per cent., washed in water, immersed in watery solution of 
 methylene blue for three minutes, again washed in water, dried, 
 and mounted in balsam. 
 
 (Irani s Solution and Eosin. Double staining of cover-glasses can 
 be obtained by combining Gram's method with eosin. The method 
 is very useful for differentiating the sheath of Streptococcus 
 pyogenes and Bacillus anthracis, from the protoplasmic contents, 
 and for staining preparations of pneumonic sputum, or of micrococci 
 and other micro-organisms in pus. After decolorising the prepara- 
 tion in alcohol, the cover-glass is transferred to a weak solution 
 of eosin for two or three minutes, then washed again in alcohol, 
 immersed in clove-oil, dried between filter-paper, and mounted in 
 balsam. 
 
 STAINING OF SPORES. 
 
 A slight modification of the ordinary process employed in making 
 cover-glass preparations has to be adopted to stain the spores of 
 
90 BACTERIOLOGY. 
 
 bacilli. Under ordinary circumstances the stain will not penetrate 
 the sheath, but if it can be made to penetrate, it is not readily 
 removed. The cover-glass preparation must be heated to a tem- 
 perature of 210 C., for half an hour, or passed as many as twelve 
 times through the flame of a Bunsen burner, or exposed to the 
 action of strong sulphuric acid for several seconds, and then a few 
 drops of a watery solution, of an aniline dye may be applied in the 
 usual way. 
 
 To double stain spore- bearing bacilli the cover-glass preparations 
 may be floated, for from twenty minutes to an hour, on Ehrlich's 
 fuchsine-amline-water, or on the Ziehl-IsTeelsen solution. The stain 
 must be heated by preference in a capsule placed in a sand-bath 
 until steam rises. The fuchsine is removed from the bacilli by 
 rinsing in water and washing in weak hydrochloric acid, and then 
 the preparations are washed again in water, and floated for a few 
 minutes on a watery solution of methylene blue. They are again 
 rinsed in water, dried, and mounted. ISTeisser's decolorising solution 
 consists of 25 parts of hydrochloric acid to 75 parts of alcohol. 
 
 STAINING OF FLAGELLA. 
 
 Koch first stained flagella by floating the cover-glasses 011 a 
 watery solution of hsematoxylin. From this they were transferred 
 to a 5 per cent, solution of chromic acid, or to M tiller's fluid, by 
 which the flagella obtain a brownish-black coloration. The author 
 succeeded in demonstrating and photographing flagella in prepara- 
 tions stained with a saturated solution of gentian violet in absolute 
 alcohol ; but these methods are now superseded owing to the much 
 more satisfactory method introduced by Loffler. 
 
 Loffler's method depends upon the employment of a mordant. 
 Loffler tried tannate of iron, and after a number of experiments 
 the following method was introduced. An aqueous solution of 
 ferrous sulphate is added to an aqueous solution of tannin (20 per 
 cent.), until the mixture turns a violet-black colour, then 3 or 4 cc. 
 of a 1 in 8 aqueous solution of logwood are added. This constitutes 
 the mordant, and a few drops of carbolic acid may be added, and the 
 solution kept in well-stoppered bottles. The dye consists of 1 cc. of 
 a 1 per cent, solution of caustic-soda, added to 100 cc. of aniline 
 water, in which 4 or 5 grammes of either methyl violet, methylene blue, 
 or fuchsine, are dissolved. A cover-glass preparation is made in 
 the ordinary way, the bacteria being diffused in water, and then 
 .spread out in a very thin film. After drying and very carefully 
 fixing, the film is covered with the mordant, and the cover-glass 
 
MICROSCOPICAL EXAMINATION OF BACTERIA. 91 
 
 held over the flame until steam rises. The mordant is then washed 
 off with distilled water, and all traces removed from the edge of 
 the cover-glass with alcohol. The stain is filtered, and a few drops 
 allowed to fall on the film, and after a few minutes the cover-glass 
 i> again very carefully warmed until steam rises. The stain is then 
 washed off with distilled water, and is ready to be examined and 
 subsequently mounted. For some bacteria it is necessary to modify 
 the solutions, either by the addition of acetic or sulphuric acid, or 
 l)v varying the quantity of soda solution. 
 
 Trenkmann introduced a modification of Loffler's system. Cover- 
 glasses are floated for from two to twelve hours on a solution 
 consisting of 1 per cent, tannin and ^ per cent, hydrochloric acid. 
 After washing in water the preparation is stained with a saturated 
 alcoholic solution of any of the aniline dyes diluted in the propor- 
 tion of 2 drops of the dye to 20 of water. The cover-glasses 
 which remain in the solution for from two to four hours are then 
 washed in water, and examined. The best results are obtained with 
 carbolised fuchsine, diluted in the proportion of 2 drops to 20 drops 
 of 1 per cent, carbolic. Trenkmann also recommended the use of 
 <-ateclm and logwood as mordants, with the addition of very dilute 
 acid, and subsequent staining with fuchsine. 
 
 Lutesch suggested the use of ferric acetate. To avoid any 
 deposit on the surface of the preparation, freshly prepared saturated 
 ferric acetate is used, and 5 to 10 drops of acetic acid are added to 
 16 cc. of the mordant. After warming the solution the preparation 
 i> washed in water, followed by 20 per cent, acetic acid, again 
 thoroughly washed, and then stained with hot solution of fuchsine or 
 gentian -violet in aniline water. 
 
 Van Ermengem used a mordant composed of 1 part of 
 2 per cent, solution of osmic acid, 2 parts of 10 to 25 per cent, 
 solution of tannin, with to every 100 cc. of this mixture 4 or 5 drops 
 of acetic acid. A black ink is thus formed, and the solution is 
 applied for from five to thirty minutes. After washing in water and 
 alcohol the cover-glasses are placed in a solution of nitrate of silver 
 and transferred to another solution composed of 5 grammes of gallic 
 acid, 3 grammes of tannin, 10 grammes of acetate of soda, and 330 
 grammes of distilled water. In a few moments they are again placed 
 in nitrate of silver, and then washed and mounted in balsam. 
 
 Sclavo's method answers well for certain micro-organisms. The 
 preparations are left for one minute in solution of tannin, washed in 
 distilled water, transferred for a minute to 50 per cent, phospho- 
 molybdic acid, again washed and stained from three to five minutes 
 
92 BACTERIOLOGY. 
 
 in hot saturated solution of f uchsine in aniline water, washed in water y 
 dried on filter paper, and mounted in balsam. The tannin solution 
 consists of 1 part of tannin to 100 cc. of 50 per cent, alcohol. 
 
 Nicolle and Morax also, have modified Loffler's method. Per- 
 fectly clean cover-glasses are used, and the film is dried without 
 fixing in the flame. Cover-glasses are covered with the mordant, 
 and heated for about ten seconds, and when steam rises the mordant 
 is shaken off and the film rinsed with water. The same process is 
 repeated three or four times, and finally the cover-glass is stained 
 with Neelsen's solution, holding it over the flame once or twice 
 for a quarter of a minute ; it is then washed and examined. 
 
 Bunge prefers as a mordant a mixture of aqueous solution of 
 tannin with 1 in 20 aqueous solution of sesquichloride of iron in the 
 proportion of 3 parts of the tannin solution, 1 part of the iron 
 solution, with the addition of 1 cc. of a saturated watery solution of 
 f uchsine added to 10 cc. of the mixture. The mordant is kept before 
 use, and applied for five minutes. The preparation is then washed and 
 stained with Neelsen's solution. In another plan the cover-glasses 
 are immersed for one half to one minute in 5 per cent, solution of 
 acetic acid, washed and dried. The mordant is then applied three or 
 four times, and the cover-glasses washed, dried, and then stained with 
 gentian-violet, dipped in 1 per cent, acetic acid, washed, dried, and 
 mounted. Peroxide of hydrogen may be added to the mordant, 
 drop by drop; it becomes reddish-brown in colour, and must be 
 shaken up and filtered before use. Cover-glasses are exposed to its 
 action for about a minute, and Neelsen's solution is used for 
 staining. 
 
 Hessert dispenses with the mordant. The film is fixed by 
 treating cover-glasses with a saturated alcoholic solution of corrosive 
 sublimate. After washing, the cover-glass is stained for thirty to 
 forty minutes in a hot dye, by preference a 10 per cent, watery 
 solution of saturated alcoholic solution of fuchsine. 
 
 COVER-GLASS IMPRESSIONS. 
 
 One of the most instructive methods for examining micro- 
 organisms is to make an impression-preparation. This enables us, 
 in many cases, to study the relative position of individual micro- 
 organisms one to another in their growth on solid cultivating media, 
 and in some cases produces the most exquisite preparations for the 
 microscope. A perfectly clean, usually small-sized, cover-glass is 
 carefully deposited on a plate-cultivation, and gently and evenly 
 pressed down. One edge is then carefully levered up, with a needle, 
 
MICROSCOPICAL EXAMINATION OF BACTERIA. 93 
 
 and the cover-glass lifted off by means of forceps. It is then 
 allowed to dry, passed through the flame three times, and stained 
 a> already described. In some cases of plate-cultures, especially 
 where no liquefaction has taken place, the growth is bodily trans- 
 ferred to the cover-glass, and a vacant area left on the gelatine 
 or agar-agar, corresponding exactly with the form and size of the 
 cover-glass employed. 
 
 PRESERVATION OF PREPARATIONS. 
 
 After examining a cover-glass preparation with an oil immersion 
 objective the cedar oil must be carefully wiped off, and the slide 
 get aside for the Canada balsam to set. At a convenient time all 
 preparations should be sealed with a ring of Hollis' glue ; the 
 cedar oil used at subsequent examinations of the specimen will 
 not be able to work its way under the cover-glass, and prevent 
 the balsam from hardening. When it is ringed cedar oil can be 
 readily wiped off, and the specimen cleaned without danger of 
 moving the cover-glass and injuring the preparation. 
 
 (B) BACTERIA IN SECTIONS OF TISSUES. 
 
 Methods of Hardening and Decalcifying Tissiies. To harden small 
 organs, such as the viscera of a mouse, they should be placed on 
 a piece of filter-paper at the bottom of a small wide-mouthed glass 
 jar, and covered with about twenty times their volume of absolute 
 alcohol. Larger organs, pathological growths, etc., are treated in 
 the same way, but must first be cut into small pieces, or cubes, 
 varying from a quarter of an inch to an inch in size. M tiller's 
 fluid may also be employed, and methylated spirit may be sub- 
 stituted for alcohol, from motives of economy. Tissues hardened in 
 absolute alcohol are ready for cutting in two or three days, and 
 those hardened in M tiller's fluid in as many weeks. 
 
 Teeth, or osseous structures, must first be placed in a decalcifying 
 solution, such as Kleinenberg's. When sufficiently softened, they 
 are allowed to soak in water, to wash out the picric acid, and then 
 transferred through weak spirit to absolute alcohol. Ebner's solu- 
 tion also gives excellent results, especially when the structures to 
 be decalcified are placed in fresh solution from time to time. 
 
 Methods of Embedding, Fixing, and Cutting. The author 
 tii ids that freezing with ether combined with the method of em- 
 bedding in celloidin gives excellent results. The pieces of tissue 
 to be embedded are placed, after the process of hardening is com- 
 
94 
 
 BACTERIOLOGY. 
 
 pleted, in a mixture of ether and alcohol for an hour or more. 
 They are then transferred to a solution of celloidin in equal parts 
 of ether and alcohol, and left there, usually for several hours. 
 
 The piece of tissue is then placed in a glass capsule, and some 
 of the celloidin solution poured over it. The capsule can be placed 
 
 FIG. 27. SWIFT'S FREEZING MICROTOME. 
 
 >x)dily in 60 to 80 per cent, alcohol, and left until the following 
 morning. The celloidin will then be of the consistency of wax. 
 The piece of tissue is next cut out, and after trimming off superfluous 
 celloidin is put in water until it sinks. It is then transferred to 
 gum, and frozen and cut with a freezing microtome. 
 
 For cutting with Jung's microtome, the tissues are embedded 
 
MICROSCOPICAL EXAMINATION OF JiACTERIA. 95 
 
 in paraffine or celloidin, and mounted on cork; or, if* firm enough, 
 they may be fixed upon cork without any embedding material at 
 all. Paraffiiie, dissolved in chloroform, will be found very service- 
 able as an embedding material. 
 
 < 1 orks ready cut for the clamp of the microtome are smeared 
 over with the solution of celloidin. This can be applied with a 
 glass rod to the surface which is to receive the piece of tissue- 
 The corks are then set aside for the film of celloidin to harden. 
 In the rise of lung, or degenerated broken-down tissue, the 
 specimen should be left for a much longer time than is found to 
 }>e sufficient for firmer structures. When ready, it is removed 
 from the celloidin solution with forceps ^and placed upon the pre- 
 
 FIG. 28. JUNG'S MICROTOMK. 
 
 pared cork. Enough of the solution, which is of syrupy consistence, 
 is allowed to fall on the piece of tissue to cover it completely, and 
 the mounted specimen is placed in the alcohol to harden. The 
 specimen will be ready for cutting next day. 
 
 The specimen may be more neatly embedded by fixing it with 
 a pin in a small paper tray, pouring the celloidin solution over it, 
 and then placing the tray in alcohol to harden the celloidin. The 
 embedded specimen is then fixed on a cork, which has been cut for 
 the clamp of the microtome. The celloidin in the section disappears 
 in the process of clearing with clove-oil. 
 
 In the case of specimens embedded in celloidin, or mounted 
 directly on a cork, the tiue. u well as the blade of the knife, should 
 be kept constantly bathed with alcohol, and the sections transferred 
 from the blade with a camel's-hair brush, and floated in alcohol. 
 
96 BACTERIOLOGY. 
 
 For fixing directly on cork, small organs and pieces of firm tissue 
 such as the kidneys of a mouse, or liver, we may employ gelatine or 
 glycerine gelatine, liquefied over a Bunsen burner in a porcelain 
 capsule. Glycerine gelatine may be used with advantage for fixing 
 irregular pieces of tissue, as it does not become of a consistency 
 that would inj are the edge of the knife. The cork, with specimen 
 affixed, is placed in alcohol, and is ready for cutting sections next 
 clay. 
 
 Material infiltrated with paraffine must be cut perfectly dry, 
 and the sections prevented from rolling up by gentle manipulation 
 with a camel's-hair brush. They must then be picked off the blade 
 of the knife with a clean needle, and dropped into a watch-glass 
 containing xylol. This dissolves out the paraffine. The sections are 
 then transferred to alcohol to get rid of the xylol, and then to the 
 staining solution. 
 
 Staining Bacteria in Tissue Sections. Sections of fresh tissues 
 made with the freezing microtome are to be floated in '8 per cent, 
 salt solution, and then carefully transferred, well spread out on a 
 platinum lifter, to a watch-glass containing absolute alcohol. Simi- 
 larly, sections selected from those cut with Jung's microtome may 
 be transferred from the spirit to absolute alcohol. The sections 
 may be then stained by any of the methods to be described. 
 
 It is often advisable to employ some method which will enable 
 one to study the structure of the tissue itself ; and sections, however 
 stained, should always be first examined with low powers, to enable 
 one to recognise the tissue under examination, and to examine in 
 many cases the topographical distribution of masses of bacteria. 
 With a power of about 250 diams. (one-sixth), very many bacteria 
 can be distinguished ; and with the oil immersion lenses the minutest 
 bacilli and micrococci can be recognised, and the exact form of 
 individual bacteria accurately determined. As most good modern 
 instruments are provided with a triple nose-piece, there is no loss 
 of time in examining a preparation successively with these different 
 powers. 
 
 Weigert's Method. A very useful method for staining both 
 the tissue and the bacteria is as follows : Place the sections for 
 from six to eighteen hours in a 1 per cent, watery solution of any of 
 the basic aniline dyes (methyl violet, gentian violet, fuchsine, Bis- 
 marck brown). To hasten the process, place the capsule containing 
 the solution in the incubator, or heat it to 45 C. A stronger 
 solution may also be employed, in which case the sections are far 
 more rapidly stained, and are easily over-stained. In the latter case 
 
MICROSCOPICAL EXAMINATION OF BACTERIA. 97 
 
 they must be treated with a half-saturated solution of carbonate of 
 potash. In either case the sections are next washed with distilled 
 water, and pas>e I through 60 per cent, alcohol into absolute alcohol. 
 When almost decolorised, spread out the section carefully on a 
 platinum lifter and transfer it to clove-oil, or stain with picro-carmine 
 solution (Weigert's) for half an hour, wash in water, alcohol, and 
 then treat with clove-oil. After the final treatment with clove-oil, 
 transfer with the platinum lifter to a clean glass slide. Dry the 
 preparation by pressure with a piece of filter-paper folded several 
 times, and preserve in Canada balsam, dissolved in xylol. 
 
 Gram's Method. In the method of Gram sections are stained 
 for ten minutes in a capsule containing aniline-gentian- violet solution. 
 <M<at care must be taken not to injure the sections. If there is 
 any difficulty in finding them, it is best to carefully pour off the 
 stain and fill up the capsule with water. The sections are then readily 
 visible, and can be taken up on the end of a glass rod and placed 
 in the iodine and iodide of potassium solution, where they remain for 
 two or three minutes, until stained uniformly brown and resembling 
 in appearance a boiled tea-leaf. They are then placed in absolute 
 alcohol, and washed by carefully moving the sections in the liquid 
 with a glass rod. When completely decolorised they are spread out 
 on a lifter, and transferred to clove-oil until completely clarified. 
 Each is transferred with a lifter to a slide, and the clove-oil is 
 run off and then completely removed by gently pressing two or 
 three layers of filter-paper upon the section. Finally, the section 
 is mounted in Canada balsam. 
 
 The process of decoWisation. may be hastened by transferring the 
 section from alcohol to clove-oil, and back again to alcohol, repeating 
 this two or three time>. 
 
 On examination the tissue appears colourless, or >lightly tinged 
 yellow from too long immersion in the iodine solution, while the 
 micro-organisms are stained blue or blue-black. 
 
 Double -taining is obtained by transferring the >ei-ti<ns after 
 d.colorisution to fiin. Ili-marck brown, or vesuvin. They are left 
 in a waterv solution for two or three minutes, then again washed in 
 alcohol, before clarifying in clove-oil and mounting in balsam. 
 
 Another instructive method is to place the decolorised sections 
 in pioro-rarminate of ammonia for three or four minutes, and then 
 treat with alcohol and clove-oil. 
 
 A >'miilar result is obtained by placing the sections in Orth's 
 >olutioii (picro-lithiuin carmine), transferring to acidulated alcohol, 
 and then passing through clove-oil and mounting in balsam. 
 
 7 
 
98 BACTERIOLOGY. 
 
 In Ehrlich's method delicate sections are liable to be injured by 
 immersion in the nitric acid, and therefore Watson-Oheyne suggested 
 the use of formic acid. 
 
 The Ziehl-Neelsen. method, in which sulphuric acid is used instead 
 of nitric acid, is much to be preferred to Ehrlich's method. 
 
 Ziehl-Neelsen Method. The solution is warmed, and sections 
 left in it for ten minutes. The red colour, which disappears when 
 the section is placed in weak sulphuric acid (25 per cent.), may 
 partly return when the section is placed in water. In this case the 
 section must be again immersed in acid and passed backwards and 
 forwards from acid to water until the red colour has completely, or 
 almost completely, disappeared. It must be thoroughly washed in 
 water to remove all traces of the acid, and then placed in a watery 
 solution of methylene blue for two or three minutes, washed again 
 in water, immersed in alcohol, clarified in clove-oil, and mounted in 
 the usual way. Sections are brilliantly stained, and the results are 
 very permanent. 
 
 Many special methods of staining have been introduced, and will 
 be given in subsequent chapters with the description of the bacteria 
 to which they apply. The methods already described are those 
 which are more or less in constant use in studying bacteria and in 
 conducting original researches. 
 
CHAPTER IX. 
 
 PREPARATION OF NUTRIENT MEDIA AND METHODS OF 
 CULTIVATION. 
 
 To cultivate micro-organisms artificially, and, in the case of the 
 pathogenic bacteria, to fulfil the second of Koch's postulates, they 
 must be supplied "with nutrient material free from pre-existing 
 micro-organisms. Hitherto various kinds of nutrient liquids have 
 been employed, and in many cases they still continue to be 
 iiM'.l with advantage, but for general use they have been, in a 
 great measure, supplanted by the methods of cultivation on sterile 
 solid media about to be described. The advantages of the latter 
 methods are numerous. In the first place, in the case of liquid 
 media, in spite of elaborate precautions and the expenditure of much 
 labour and time, it was almost impossible or extremely difficult to 
 obtain a pure culture. When a drop of liquid containing several kinds 
 of bacteria is introduced into a liquid medium, we have a mixed 
 cultivation from the very first. If in the struggle for existence 
 some bacteria were unable to develop in the presence of others, or 
 a change of temperature and soil allowed one form to predominate 
 over another, then we might be led to the conclusion that many 
 bacteria were but developmental forms of one and the same micro- 
 organism ; while possibly the contamination of such cultures might 
 lead to the belief in the transformation of a harmless into a patho- 
 genic bacterium. The secret of the success of Koch's methods greatly 
 depends upon the possibility, in the case of starting with a mixture 
 of micro-organisms, of being able to isolate them completely one 
 from another, and to obtain an absolutely pure growth of each 
 cultivable specie-. When sterile nutrient gelatine has been liquefied 
 in a tube and inoculated with a mixture of bacteria in such a way 
 that the individual micro-organismf! arc distributed throughout it, 
 and the liquid i> poured out on a plate of glass and allowed to solidity, 
 the individual bacteria, instead of moving about freely as in a liquid 
 medium, are fixed in one sjM>t. where they develop individual.-, of 
 
 99 
 
100 
 
 BACTERIOLOGY. 
 
 their own species. In this way colonies are formed each possessing 
 its own biological characteristics and morphological appearances. 
 When an adventitious germ from the air falls upon the culture, it also 
 grows exactly upon the spot upon which it fell, and can be easily 
 recognised as a stranger. To maintain the individuals isolated from 
 one another daring their growth, and free from contamination, it is 
 only necessary to thin out the cultivation, and to protect the plates 
 from the air. The slower growth of the micro-organisms in solid 
 media, affording much greater facility for examining them at various 
 intervals and stages of development, is an additional point in favour 
 of these methods ; and the characteristic macroscopical appearances 
 so frequently assumed are, more especially in the case of morpho- 
 logical resemblance or identity, of the greatest importance. The 
 colonies on nutrient gelatine (examined with a low power) of micro- 
 organisms such as Bacillus anthracis and Proteus mirabilis, the 
 naked eye appearances in test-tubes of the growth of the bacilli 
 of anthrax and tubercle, and the brilliant growth of Micrococcus 
 prodigiosus, may be quoted as examples in which the appearances are 
 often very striking and sometimes quite characteristic. 
 
 SOLID MEDIA. 
 
 (A) PREPARATION OF NUTRIENT GELATINE AND NUTRIENT 
 AGAR-AGAR. 
 
 Nutrient Gelatine is prepared as follows: Take half a kilo- 
 gramme of beef (one pound), as free as possible from fat. Chop it 
 up finely, transfer it to a flask or cylindrical 
 vessel, and shake it up well with a litre of 
 distilled water. Place the vessel in an ice- 
 pail, ice-cupboard, or in winter in a cold 
 cellar, and leave for the night. Next morn- 
 ing commenca with the preparation of all 
 requisite apparatus. Thoroughly wash and 
 rinse with alcohol about 100 test-tubes, and 
 allow them to dry. Plug the mouths of the 
 test-tubes with cotton-wool, taking care that 
 the plugs fit firmly but not too tightly. 
 Place them in their wire cages in the hot-air 
 steriliser, to be heated for an hour at a temperature of 150 C. In 
 the same manner cleanse and sterilise several flasks and a small 
 glass funnel. In the meantime the meat infusion must be again 
 well shaken, and the liquid portion separated by filtering and 
 
 FIG. 29. WIRE CAGE 
 FOR TEST-TUBES. 
 
DESCRIPTION OF PLATE II. 
 Pure-cultivations of Bacteria. 
 
 FIG. 1. In tJie depth of Nutrient Gelatine. A pure-cultivation of Kochs 
 comma-bacillus (Spirillum cholerae Asiaticae) showing in the track of 
 the needle a funnel-shaped area of liquefaction enclosing an air-bubble, 
 and a white thread. Similar appearances are produced in cultivations of 
 the comma-bacillus of Metchnikoff. 
 
 FIG. 2. On the surface of Nutrient Gelatine. A pure-cultivation of Bacillus 
 typhosus on the surface of obliquely solidified nutrient gelatine. 
 
 FIG. 3. On the surface of Nutrient Agar-agar. Pure-cultivation of Bacillus 
 indicus on the surface of obliquely solidified nutrient agar-agar. The 
 growth has the colour of red sealing-wax, and a peculiar crinkled 
 appearance. After some days it loses its bright colour and becomes 
 purplish, like an old cultivation of Micrococcus prodigiosus. 
 
 FIG. 4. On tlie surface of Nutrient Agar-agar. A pure-cultivation obtained 
 from an abscess (Staphylococcus pyogenes aureus). 
 
 FIG. 5. On the surface of Nutrient Agar-agar. A pure-cultivation obtained 
 from green pus (Bacillus pyocyaneus). The growth forms a whitish, 
 transparent layer, composed of slender bacilli, and the green pigment 
 is diffused throughout the nutrient jelly. The growth appears green by 
 transmitted light, owing to the colour of the jelly behind it. 
 
 FIG. (>. On the surface of Potato. A pure-cultivation of the bacillus of 
 glanders on the surface of sterilised potato. 
 
^pi 
 
 
 op 
 
 fi 
 
MTH1ENT MEDIA AM> MKTHODS OF CULTIVATION. 
 
 101 
 
 ><[uee/.iiii: through a linen cloth or a meat press. The red juice thus 
 obtained must be brought up to a litre by transferring it to a large 
 measuring glass and adding distilled water. It is then poured 
 into a sufficiently large and strong beaker, and set aside after the 
 addition of 10 grammes of peptone, 5 grammes of common salt and 
 KM) grammes of best gelatine. 
 
 In about half an hour the gelatine is sufficiently softened, and 
 heating in a water-bath causes it to be completely 
 
 
 FIG. 30. HOT AIR STERILISEH. 
 
 diolved. The danger of breaking the beaker may be avoided by 
 placing a cloth, several times folded, at the bottom of the water-bath. 
 
 The next process requires the greatest care and attention. Some 
 micro-organisms grow best in a slightly acid, others in a neutral 
 or slightly alkaline, medium. For example, for the growth and 
 characteristic' appearances of the comma bacillus of Asiatic cholera 
 a faintly alkaline soil is absolutely essential. This slightly alkaline 
 me limn will be found to answer best for most micro-organisms, and 
 may be obtained as follows : - 
 
 With a clean glass rod dipped in the mixture, the- reaction 
 upon litmus-paper may be ascertained, and a concentrated solution 
 of carbonate of soda must h- added dmp by drop, until red litmus- 
 
102 
 
 BACTERIOLOGY. 
 
 paper becomes faintly blue. If it has been made too alkaline, it can 
 be neutralised by the addition of lactic acid. 
 
 Finally, the mixture is heated for an hour in the water-bath. 
 Ten minutes before the boiling is completed, the white of an egg 
 beaten up with the shell is added, and the liquid is then filtered 
 while hot. For the filtration, the hot-water apparatus (Fig. 31) 
 can be used with advantage, furnished with a filter of Swedish 
 
 paper, which may be conveniently 
 made in the following way : 
 
 About- eighteen inches square 
 of the best and stoutest filter paper 
 is first folded in the middle, and 
 then creased into sixteen folds. The 
 filter is made to fit the glass funnel 
 by gathering up the folds like a fan, 
 and cutting off the superfluous part. 
 The creasing of each fold should be 
 made firmly to within half an inch 
 of the apex of the filter, which part 
 is to be gently inserted into the 
 tube of the funnel. To avoid 
 bursting the filter at the point, the 
 broth, w r hen poured out from the 
 flask, should be clhected against 
 the side of the filter with a glass 
 rod. During filtration the funnel 
 should be covered over with a 
 circular plate of glass, and the pro- 
 cess of filtration must be repeated, 
 if necessary, until a pale, straw- 
 coloured, perfectly transparent 
 filtrate results. 
 
 The sterilised test-tubes are filled to about a third of their depth 
 by pouring in the gelatine carefully and steadily, or by employing a 
 small sterilised glass funnel. The object of this care is to prevent 
 the mixture touching the part of the tube with w T hich the plug 
 comes into contact ; otherwise, when the gelatine sets, the cotton- 
 wool adheres to the tube and becomes a source of embarrassment in 
 subsequent procedures. As the tubes are filled they are placed in 
 the test-tube basket, and must then be sterilised. They are either 
 lowered into the steam steriliser, when the thermometer indicates 
 100 0., for twelve minutes for four or five successive days, or they 
 
 FIG. 31. HOT-WATER FILTERING 
 APPARATUS. 
 
NUTRIENT MEDIA AND METHODS OF CULTIVATION. 
 
 103 
 
 may be transferred to the test-tube water-bath, and heated for an 
 hour a day for three successive days. 
 
 If the gelatine shows any turbidity after these processes it must 
 
 FK;. 32. METHOD OF MAKING A FOLDED FILTKK. 
 
 be poured back from the test-tubes into a flask, boiled up for ten 
 minutes, and filtered once more, and the processes of sterilisation just 
 described must be repeated. 
 
 Fn;. :. STEAM STEKIMSEI:. 
 
 Nutrient Agar-agar.- Aptr-agar is .1 >ultauce prepared 
 from seaweed which grows on the coasts of Japan and India, 
 and is supplied in long crinkled strips. It boils at 90 C., and 
 
104 
 
 BACTERIOLOGY. 
 
 remains solid up to a temperature of about 45 C. It is there- 
 fore substituted for gelatine in the preparation of a jelly for the 
 cultivation of those bacteria which will only grow, or grow best, in 
 the incubator at the temperature of the blood. It may also be 
 employed at ordinary temperatures for bacteria which liquefy 
 gelatine. The preparation is conducted on much the same principles 
 as those already described. Instead, however, of 100 grammes of 
 gelatine, only about 20 grammes of agar-agar are employed (1*5 to 
 2 per cent.), and to facilitate its solution it must be allowed to soak 
 in salt water overnight. For the nitration, flannel is substituted 
 
 for filter- paper, or may 
 be used in combination 
 with the latter. The 
 hot -water apparatus 
 is invariably employed, 
 unless, to accelerate 
 the process, the glass 
 funnel and receiver are 
 bodily transferred to 
 the steam steriliser. If 
 the conical cap cannot 
 be replaced, cloths laid 
 over the mouth of the 
 steriliser must be em- 
 ployed instead. It may 
 be necessary to repeat 
 the process of nitra- 
 tion, but it must not be 
 expected that such a 
 brilliant transparency 
 can be obtained as with 
 gelatine. The final 
 
 result, when solid, should be colourless and clear; but if slightly 
 milky, it may still be employed. 
 
 A little liquid gradually collects in the tubes, being expressed by 
 the contraction of the agar-agar. 
 
 Wort-gelatine is used in studying the bacteria of fermentation. 
 It is made by adding from 5 to 10 per cent, of gelatine to beer- wort. 
 Glycerine Agar-agar. This is prepared by adding 5 per 
 cent, of glycerine to nutrient agar-agar, after the boiling and before 
 the filtration, and other modifications can be made for special 
 purposes by the addition of grape-sugar or of gelatine. 
 
NUTRIENT MEDIA AND METHODS OF CULTIVATION. 
 
 105 
 
 After the final treatment in the steam steriliser some of the 
 tubes of gelatine and agar-agar are placed upright and allowed to 
 set. and others are placed on an inclined plane or in the blood- serum 
 inspissator, and left to gelatinise with an oblique surface. 
 
 (B) METHODS OF KMIM.OVIXG NUTRIENT JELLY IN TEST-TUBES 
 AND ON GLASS PLATES. 
 
 Test-tube-cultivations. To inoculate test-tub:-* containing 
 nutrient jelly, the cotton- wool plug is first twisted round in case 
 there are any adhesions between the plug and the test-tube. It is 
 then removed with the thumb and 
 index finger of the right hand, and 
 placed between the fourth and fifth 
 fingers of the. left hand, instead of 
 being put down on the laboratory table 
 and thereby probably contaminated 
 with bacteria or the spores of mould 
 fungi. A sterilised needle charged, 
 for example, with blood or pus con- 
 taining bacteria, or with a colony from 
 a plate -culture, is thrust once in the 
 middle line into the nutrient jelly, 
 and steadily withdrawn. The tube 
 should be held horizontally or with 
 its mouth downward, to avoid, as far 
 as possible, accidental contamination 
 from the gravitation of germs in the 
 air ; and the plug replaced as quickly 
 as possible. The cotton- wool project- 
 ing beyond the mouth of the tube is 
 
 then thoroughly burnt in the flame of a Bunsen burner or blow- 
 pipe, and an india-rubber cap fitted over the mouth of the tube 
 
 The clianct > of rrr<>r arising from contamination of the culti- 
 vations are reduced by avoiding draughts at the time of inoculation, 
 and it is best that these manipulations should be carried on in a 
 quiet room in which the tables and floor are wiped with damp cloths, 
 rather than in a laboratory in which the air becomes charged with 
 germs through constant sweeping and dusting, and the entrance 
 and exit of clae> of studMit>. In conducting any investigation 
 a dozen or more tubes slioul 1 be inoculated, and if by chance an 
 adventitious germ, in spite of all precautions, gains ., entrance, 
 
 FIG. 35. METHOD OF INOCULAT- 
 ING A TKST-TI HI-: COXTAIXIXO 
 STKKILK XITKIKNT JKLLY. 
 
106 BACTERIOLOGY. 
 
 the contaminated tube can be rejected, and the experiments con- 
 tinued with the remaining pure cultivations. 
 
 When, however, one tube containing a liquid medium is in- 
 oculated from another, as in the process of preparing plate-cultures, 
 or when a culture is made from a tube in which the growth has 
 liquefied the gelatine, it is obvious that the tubes cannot be inverted 
 or held horizontally, and they must then be held and inoculated as 
 in Fig. 38. To inoculate those tubes of nutrient media which have 
 been solidified obliquely, the point of a straight sterilised needle 
 charged with the material to be cultivated is traced over the surface 
 ,of the jelly from below upwards, or the inoculated material may be 
 spread out with a hooked or looped needle. 
 
 Examination of Test- tube- cultivations. The appearances pro- 
 duced by the growths in test-tubes can be in most cases sufficiently 
 examined with the naked eye. In some cases the jelly is partially 
 or completely liquefied, while in others it remains solid. The 
 growths may be abundant or scanty, coloured or colourless. The 
 nutrient jelly may itself be tinged or stained with products resulting 
 from the growth of the organisms. When liquefaction slowly takes 
 place in the nesdle track, or the organism grows without producing 
 this change, the appearances which result are often very delicate, 
 and in some cases very characteristic. The appearance of a simple 
 white thread, of a central thread with branching lateral filaments, 
 of a cloudiness, or of a string of beads in the track of the needle, 
 may be given as examples. 
 
 In some cases much may be learnt by examining the growth with 
 a magnifying glass. Here, however, a difficulty may be encountered, 
 for the cylindrical form of the tube so distorts the appearance of its 
 contents, that the examination is rendered somewhat difficult. To 
 obviate this, a very simple contrivance may be employed with 
 advantage. This consists of a rectangular vessel, about four inches 
 in height and two inches in width, which may be easily constructed 
 by cementing together two slips of glass to form the back and front, 
 with three slips of stout glass with ground edges forming the sides 
 and base (Cheshire). The front may be constructed of thin glass, 
 and the base of the vessel made to slope so that the test-tube when 
 placed in the vessel has a tendency to be near the front. The 
 vessel is filled with a mixture of the same refractive index as the 
 nutrient gelatine. The latter has a refractive index rather higher 
 than water, which is about 1-333 ; alcohol has a refractive index of 
 1'374. The vessel is filled with water, and alcohol is then added 
 until the proper density is reached. The test-tube is placed in the 
 
NTTRIKXT MEDIA AND METHODS OF CULTIVATION. 
 
 107 
 
 1, and held in position by means of a clip. The vessel can be 
 fixed on the inclined stage of the microscope, and the contents of the 
 tube conveniently examined with low-power objectives. 
 
 Plate -cultivations. By this method, as already mentioned, a 
 mixture of bacteria, whether in fluids, excreta, or in cultivations on 
 solid media, can be so treated that the different species are isolated 
 one from the other, and perfectly pure cultivations of each of the 
 cultivable bacteria in the original mixture established in various 
 nutrient media. We are enabled also to examine under a low 
 power of the microscope the individual colonies of bacteria, and to 
 distinguish by their characteristic appearances, micro-organisms 
 which, in their individual form, closely resemble one another, or 
 are even identical. The same process, with slight modification, is 
 also employed in the examination of air, soil, and water, which will 
 be referred to later. 
 
 The preparation 
 of plate-cultivations, 
 therefore, must be 
 described in every de- 
 tail : and to take an 
 example, we will sup- 
 pose that a series of 
 plates is to be pre- 
 pared from a test- 
 cube-cultivation. 
 
 Arrangement of 
 
 Levelling Apparatus. In order to spread out the liquid jelly 
 evenly on the surface of a glass plate, and hasten its solidifica- 
 tion, it is necessary to place the glass plate upon a level and 
 cool surface. This is obtained in the following manner : Place a 
 large shallow glass dish upon a tripod stand, and fill it to the brim 
 with cold water ; carefully cover the dish with a slab of plate-glass, 
 or a pane of window-glass, and level it by placing the spirit-level in 
 the centre and adjusting the screws of the tripod. Substitute for 
 the spirit-level a piece of filter-paper the size of the glass plates to 
 be employed, and cover it with a shallow bell-glass. 
 
 >'/ r'dlsation of Glass Plates. The glass plates are sterilised in 
 an iron box placed in the hot-air steriliser, at 150 C., from one to 
 two hours. As these plates are used also for other purposes, a 
 quantity ready sterilised should always be kept in the box. 
 
 Preparation of Damp Chambers. The damp chambers for the 
 reception of the inoculated plates are prepared thus : Thoroughly 
 
 FIG. 36. LEVELLING APPARATUS. 
 
108 
 
 BACTERIOLOGY. 
 
 cleanse and wash out with 1 in 20 carbolic acid a shallow glass dish 
 and bell. Cut a piece of filter- paper to line the bottom of the glass 
 dish, and moisten it with the same solution. 
 Method of Inoculating the Test-tubes. In 
 a glass beaker or an ordinary glass tumbler, 
 with a pad of cotton-wool at the bottom, 
 place the tube containing the cultivation, 
 the three tubes to be inoculated, three glass 
 rods which have been sterilised by heating 
 in the flame of a Bunsen burner, and a 
 thermometer. Provide a strip of paper, a 
 large label, a pencil, a pair of forceps, and 
 inoculating needles. All is now ready at 
 hand to commence the inoculation of the tubes. 
 
 Liquefy the gelatine in the three tubes by placing them in a 
 beaker containing water at 30 C., or by gently warming them in 
 the name of the Bunsen burner. Keep the tubes, both before and 
 after the inoculation, in the warm water, to maintain the gelatine 
 in a state of liquefaction. Hold the tube containing the cultivation 
 
 FIG. 37. IRON Box FOR 
 GLASS PLATES. 
 
 FIG. 38. METHOD OF INOCULATING TEST-TUBES IN THE PREPARATION 
 OF PLATE-CULTIVATIONS. 
 
 and a tube of the liquefied gelatine as nearly horizontal as possible 
 between the thumb arid index finger of the left hand. With the index 
 finger and thumb of the right hand loosen the plugs of the tubes. 
 Take the looped platinum needle in the right hand and hold it like 
 a pen. Remove the plug from the culture-tube by using the fourth 
 and fifth fingers of the right hand as forceps, and place it between 
 
DESCRIPTION OF PLATE III. 
 Plate-cultivation. 
 
 This represents the appearance of a plate-cultivation of the comma- bacillus 
 of Cholera nostras, when it is examined over a slab of blackened plate-glass. 
 The drawing was made from a typical result of thinning out the colonies by 
 the process of plate-cultivation. At this stage they were completely isolated 
 one from the other ; but later they became confluent, and produced complete 
 liquefaction of the gelatine. 
 
NUTRIENT MEDIA AM) METHODS OF CULTIVATION. 109 
 
 the fourth and fifth fingers of the left. Remove the plug of the 
 other tube in the same way, placing it between the third and fourth 
 fingers of the left hand. With the needle take up a droplet of the 
 cultivation and stir it round in the liquefied jelly. Replace the plugs, 
 and set aside the cultivation. Hold the freshly inoculated tube be- 
 tween the index finger and thumb of either hand, almost horizontally, 
 then raise it to the vertical, so that the liquid gelatine gently flows 
 back. By repeating this motion and rolling the tube between the 
 fingers and thumbs the micro-organisms which have been introduced 
 are distributed throughout the gelatine. Any violent shaking, and 
 consequent formation of bubbles, must be carefully avoided. From 
 the inoculated tube, in the same manner inoculate a fresh tube of 
 liquefied gelatine, introducing into it three droplets with a sterilised 
 needle. After tilting and rolling this tube, as in the previous case, 
 the same process is repeated with a third tube, which is inoculated 
 from the second tube. This last tube must be inoculated in different 
 ways, according to experience, for different micro-organisms. Some- 
 times a sufficient separation of the micro-organisms is attained by 
 inoculating the last tube with a straight, instead of a looped, needle, 
 dipping it from the one into the other from three to five times. 
 
 The next process consists in pouring out the gelatine on glass 
 plates and allowing it to solidify. 
 
 Preparation of the Gelatine- Plates. The directions to be observed 
 in pouring out the gelatine are as follows : 
 
 Place the box containing sterilised plates horizontally, and so 
 that the cover projects beyond the edge of the table ; remove the 
 cover, and withdraw a plate with sterilised forceps ; hold it between 
 the finger and thumb by opposite margins, rapidly transfer it to 
 the filter-paper under the bell-glass, and quickly replace the cover 
 of the box. On removing the plug from the tube which was first 
 inoculated, an assistant raises the bell-glass, and the contents of the 
 tube are poured on to the plate ; with a glass rod the gelatine must 
 be then rapidly spread out in an even layer within about half an 
 inch of the margin of the plate. The assistant replaces the bell- 
 glass, and the gelatine is left to set. Meanwhile a glass bench or 
 metallic shelf is placed in the damp chamber, ready for the reception 
 of the plate-cultivation, and when the gelatine is quite solid the 
 plate is quickly transferred from under the bell-glass to the damp 
 chamber; precisely the same process is repeated with the other 
 tubes, and the damp chamber, labelled with the details of the 
 experiment, Is set aside for the colonies to develop. Not only plate- 
 cultures should be carefully labelled with date and description, but 
 
HO BACTERIOLOGY . 
 
 the same remark applies equally to all preparations tube-cultures, 
 potato-cultures, drop-cultures, etc. 
 
 Corresponding with the fractional cultivation of the micro- 
 organisms obtained in this manner, the colonies will be found to 
 develop in the course of a day or two, the time varying with the 
 temperature of the room. The lower plate will contain a countless 
 number of colonies which, if the micro-organism liquefies gelatine, 
 speedily commingle, and produce, in a very short time, a complete 
 liquefaction of the whole of the gelatine. On the middle plate the 
 colonies will also be very numerous, but retain their isolated position 
 for a longer time ; while on the uppermost plate the colonies are 
 completely isolated from one another, with an appreciable surface of 
 gelatine intervening. 
 
 Examination of Plate- cultivations. The macroscopical appear- 
 ances of the colonies are best studied by placing the plate on a 
 
 FIG. 39. DAMP CHAMBER CONTAINING PLATE-CULTIVATIONS. 
 
 slab of blackened glass, or on a porcelain slab if the colonies are 
 coloured. 
 
 To examine the microscopical appearances, a selected plate is 
 placed upon the stage of the microscope. The smallest diaphragm 
 is employed, and the appearances studied principally with a low 
 power. These appearances should be carefully noted, and a 
 sketch or photograph of the colony made. The morphological 
 characteristics of the micro-organisms of which the colony is formed 
 can be examined in the following way : A small looped or 
 hooked platinum needle is held like a pen, and the hand steadied 
 by resting the little finger on the stage of the microscope. The 
 extremity of the needle is steadily directed to the space between 
 the lens and the gelatine without touching the latter, until, on 
 looking through the microscope, it can be seen in the field, above 
 or by the side of the colony under examination. The needle 
 is then dipped into the colony, steadily raised, arid withdrawn. 
 Without removing the eye from the microscope this manipulation 
 can be seen to be successful by the colony being disorganised or 
 
NUTKIKNT MEDIA AND METHODS OF CULTIVATION. 
 
 Ill 
 
 completely removed from the gelatine. It is, however, not easy to 
 lit* successful at first, but with practice this can be accomplished 
 with rapidity and precision. A preparation is then made by rubbing 
 
 FIG. 40. PASTEUR'S LARGE INCUBATOR. 
 
 the extremity of the needle in a droplet of water on a slide, covering 
 with a cover-gla. and examining in the fresh Mate, in- l>y spreading 
 nut th- droplet on a co\vr-gl:t>>. drying, );issing three times through 
 the riame. and staining witli a drop of fuchsine or gentian violet. 
 Inoculation.- should lie made in te>t-tube> of nutrient gelatine 
 
112 BACTERIOLOGY . 
 
 and agar-agar, from the micro-organisms transferred to the cover- 
 glass before it is dried and stained, from any remnants of the colony 
 which was examined, or from other colonies bearing exactly similar 
 appearances. In this way pure cultivations are established, and 
 the macroscopical appearances of the growth in test-tubes can be 
 obtained. The plates should be replaced in the damp chamber 
 as soon as possible ; drying of the gelatine, or contamination with 
 micro-organisms gravitating from the air during their exposure, 
 
 may spoil them for subsequent examination. 
 A much simpler method of plate-cultivation 
 
 is to dispense with the levelling apparatus, and 
 
 pour the liquefied ielly into shallow, flat dishes. 
 FIG. 41. PETRI'S 
 
 j) ISH They take up much less room, and in many 
 
 ways are more convenient (Fig. 41). 
 
 Nutrient agar-agar can also be employed for the preparation 
 of plate-cultivations, but it is much more difficult to obtain satis- 
 factory results. The test-tubes of nutrient agar-agar must be 
 placed in a beaker with water and heated until the agar-agar 
 is completely liquefied. The gas is then turned down, and the 
 temperature of the water allowed to fall until the thermometer 
 stands just above 50 C. The water must be maintained at this 
 temperature, and the test-tubes must be in turn rapidly inoculated 
 and poured out upon the glass plates, or better still, into glass 
 dishes, as already described. 
 
 A very much simpler plan is to liquefy the agar, pour it into 
 
 FIG. 42. GLASS BENCHES AND SLIDES. 
 
 a shallow dish, and allow it to solidify. The culture material is 
 thinned out in sterilised broth, and a few drops are spread out over 
 the surface of the agar. The dishes are then placed in the incubator 
 at 37 C. 
 
 Glass plates may also be employed in a much simpler way. 
 The nutrient jelly is liquefied, poured out, and allowed to set. A 
 needle charged with the material to be inoculated is then drawn 
 in lines over the surface of the jelly. This method is of use for 
 inoculating different organisms side by side, and watching the effect 
 of one upon the other, or a micro-organism in this way may be 
 
NUTRIENT MEDIA AND METHODS OF CULTIVATION. 113 
 
 sown upon the gelatine which has been already altered by the 
 growth of another micro-organism; the change produced in the 
 jiflatine, as in the case of the Bacillus pyocyaneus, extending far 
 beyond the limits of the growth itself. 
 
 Nutrient jelly may also be spread out on sterilised glass slides, 
 which after inoculation are placed in damp chambers for the growths 
 to develop. 
 
 Esmarch's Roll-cultures. Esmarch introduced a modification 
 of the method of plate- cultivation which may sometimes be used with 
 advantage. The ordinary test-tubes may be employed, or tubes 
 considerably larger in size. 
 
 After the liquid jelly has been inoculated in the tube, instead 
 of pouring it out on to a glass plate or into a dish, the cotton- wool 
 plug is replaced, and an india-rubber cap fitted over the mouth of 
 the tube. 
 
 The tube is then placed horizontally on a block of ice or 
 in a vessel containing iced water. The neck of the tube is steadied 
 with the left hand, and the tube turned round and round with the 
 right hand. In a very short time the gelatine sets, and the tube 
 is lined inside with a thin coating. There is far less danger of 
 contamination, and the cultures are in a much more convenient 
 form when circumstances render it necessary to move them. 
 
 (c) PREPARATION AND EMPLOYMENT OF SOLIDIFIED BLOOD SERUM. 
 
 Solid Blood Serum. The tubercle-bacillus, the bacilli of 
 glanders and of diphtheria, and many other micro-organisms, thrive 
 well when cultivated on solid blood serum. This medium has' the 
 additional advantage of remaining solid at all temperatures. The 
 technique required for its preparation and sterilisation is as follows : 
 Several cylindrical vessels, about 20 cm. high, are thoroughly washed 
 with carbolic acid (1 in 20), and then with alcohol, and finally 
 rinsed out with ether. The ether is allowed to evaporate, and the 
 vessels are then ready for use. The skin of the animal selected 
 calf, sheep, or horse is washed with carbolic at the seat of operation, 
 and the bleeding performed with a sterilised knife or a trocar and 
 rannula. The first jet of blood from the vein is rejected, and that 
 which follows is allowed to flow into the vessels until they are almost 
 full. The ground-glass stoppers, greased with vaseline, are replaced, 
 and the vessels set aside in ice, as quickly as possible, for from 
 t \\vnty-f our to thirty hours. By that time the separation of the 
 clot is completed, and the clear serum can then be transferred to 
 
 8 
 
114 
 
 BACTERIOLOGY. 
 
 plugged sterilised test-tubes. These should be filled, with a sterilised 
 pipette, to about one-third of their capacity. 
 
 Formerly the tubes were sterilised by Tyiidall's process of dis- 
 continuous sterilisation. The tubes were placed in Koch's serum 
 steriliser, with the temperature maintained for an hour or more at 
 56 C., and this was repeated for six successive days, the temperature 
 on the last day being gradually raised to 60 C. This completed the 
 sterilisation, and to solidify the serum the tubes were arranged in 
 the inspissator at the angle required, and the temperature was kept 
 
 between 65 C. and 68 C. 
 Directly solidification took 
 place the tubes were removed. 
 The new process is much 
 less tedious, and consists in 
 taking every possible pre- 
 caution to obtain the blood 
 without contamination by bac- 
 teria in the air or in the 
 vessels employed. There is 
 then no need to sterilise the 
 serum, and it can be coagu- 
 lated immediately. The tubes 
 are tested by placing them 
 in an incubator at 37 C. for 
 a week, and if any show 
 signs of contamination they 
 are discarded, and the rest 
 can be used or kept in stock. 
 
 The serum should then 
 
 present the character of being hard, solid, of a pale straw colour, 
 and transparent. A little liquid collects at the lowest point, and 
 the serum is sometimes milky in appearance at its thickest part. 
 
 Loffler's Blood Serwm is prepared by mixing two-thirds of fresh 
 serum with one-third of broth, prepared in the usual way but with 
 the addition of 1 per cent, grape-sugar. The mixture is decanted 
 into test-tubes, avoiding the formation of air-bubbles, and it is then 
 coagulated in the usual way. The serum may be employed not only 
 in test-tubes, but also in small flasks, glass capsules, or other vessels, 
 all of which must be cleansed and sterilised. 
 
 Hydrocele fluid and other serous effusions] may be prepared in 
 the same manner. Gelatine may be added to the serum in the pro- 
 portion of 5 per cent. 
 
 FIG. 43. KOCH'S SERUM STERILISER. 
 
MTKIKNT MKI'IA AND METHODS OF CULTIVATION. 115 
 
 Inoculation of the Tubes. A small portion of a culture or of the 
 material to be inoculated is taken up with a sterilised platinum 
 needle, and traced over the sloping surface of the serum ; or a 
 fragment of tissue, such as diphtheritic membrane or tubercle, may 
 be introduced into the tube and rubbed gently over the serum so as. 
 not to bivak the surface. 
 
 FK;. 44. HUKPPK'S SEKUM INSPISSATOR. 
 
 (D) PREPARATION AND EMPLOYMENT OF STERILISED POTATO. 
 
 Potato-cultivations. -Sterilised potatoes form an excellent 
 medium for the cultivation of many micro-organisms, more especially 
 the chromogenic species. Potato-cultivations al>o give in some cases 
 very oharactrri.-tic appearam-f>, which are of value in distinguishing 
 bacteria which possess morphological resemblances. 
 
 Prepn-'itiim <>f SbrHiMd Potatoes. Potatoes, preferably smooth- 
 skinned, which are free from "eyes" and rotten sput>. should be 
 selected. If they cannot be obtained without eyes ami spots, these 
 must }^' cart-fully picked out with the point of a knife. The potatoes 
 are well x-rubbed with a stiff brush, and allowed to >oak in 1 in _< 
 carbolic for a few minutes. They are then transferred to the potato- 
 
116 BACTERIOLOGY. 
 
 receiver, and steamed in the steam steriliser for twenty minutes to 
 half an hour, the time varying according to the size of the potatoes. 
 When cooked, the potato -receiver is withdrawn and left to cool, the 
 potatoes being retained in it until required for use. 
 
 Damp chambers are prepared ready for the potatoes, the vessels 
 being cleansed and washed with carbolic as described for plate- 
 cultivations. Small glass dishes of the same pattern as the large 
 ones may be employed for single halves of potatoes. Potato-knives 
 and scalpels, which have been sterilised in an iron box by heating 
 them in the hot-air steriliser at 151 C. for one hour, should 
 be ready to hand. Knives sterilised by heating them in the name 
 of a Bunsen burner should afterwards be placed upon a sterilised 
 glass plate and covered with a bell-glass. It must not be forgotten, 
 
 FIG. 45. Box FOR STERILISING INSTRUMENTS. 
 
 however, that heating the blades in the flame destroys the temper 
 of the steel, and therefore knives and other instruments should 
 preferably be sterilised in the hot-air steriliser, enclosed in an iron 
 box, or simply enveloped in cotton-wool. 
 
 Inoculation of Potatoes. The coat-sleeves should be turned back, 
 and the hands, after thorough washing with good lathering soap, 
 be dipped in 1 in 40 carbolic. An assistant opens the potato- 
 receiver, and a potato is selected and held between the thumb and 
 index ringer of the left hand. With the knife held in the right 
 hand, the potato is almost completely divided in the direction 
 which will give the largest surface. The assistant raises the cover 
 of the damp chamber, and the potato is introduced, and while the 
 knife is withdrawn, allowed to fall apart. The cover is quickly 
 replaced, and another potato treated in the same way, is placed 
 in the same damp chamber. The four halves are then quite ready 
 for inoculation. As an extra precaution, the left hand is again 
 dipped in carbolic, and one half of a potato is taken up between the 
 tips of the thumb and index finger, care being taken to avoid 
 touching the cut surface. Holding it with its cut surface vertical, 
 
MTRIENT MEDIA AND METHODS OF CULTIVATION. 117 
 
 a small portion of the substance to be inoculated is placed on the 
 centre with a sterilised platinum needle. With a sterilised scalpel 
 the inoculated substance is rapidly spread over the surface of the 
 potato with the flat of the blade, to within a quarter of an inch 
 of the margin, and the potato is then as quickly as possible replaced 
 in the damp chamber. With another sterilised scalpel a small 
 portion of the potato from the inoculated surface of the first half 
 is in the same way spread over the surface of the second half, thus 
 thinning out the bacteria as in plate-cultivations. Exactly the 
 same is repeated with a third potato, and even a fourth, so that 
 a still further thinning out or fractional cultivation of the micro- 
 organisms may be obtained. In some cases it is necessary to place 
 the cultures in an incubator (Fig. 40) ; others grow very well at the 
 
 Ki':. 4('>. DAMP CHAMBER FOR POTATO-CULTIVATIONS. 
 
 temperature of the room. As in plate-cultivations, the potato may 
 also be inoculated by simply streaking it in lines with a needle 
 charged with the material to be cultivated. 
 
 Potato in Test-tubes. Large surfaces of potato are employed when 
 we wish to obtain cultures of micro-organisms in considerable quanti- 
 ties, as in the examination of the products of chromogenic bacteria ; 
 but under ordinary circumstances potato is employed in test-tubes. 
 The central portions of raw potatoes are cut out in cylindrical pieces 
 with a cork-borer. These are divided obliquely in their whole 
 length, and each half is placed in a test -tube. The test-tubes are 
 plugged with cotton-wool, and then steam in the steam- steriliser 
 for twenty minutes. The sloping surface is inoculated in the same 
 way as obliquely solidified jelly, and the advantages are great. The 
 culture.-, an- obtained in a more convenient form, and there is le>^ 
 danger of contamination. 
 
 Potato-paste may be employed when it is desirable to obtain an 
 extensive growth of certain bacteria. The potatoes are boiled for 
 an hour, and the floury centre squeezed out of the skins. This 
 is then mashed up with sufficient sterilised water to produce a thick 
 
118 BACTERIOLOGY. 
 
 paste, and is heated in the steam steriliser for half an hour for three 
 successive days. 
 
 (E) PREPARATION AND EMPLOYMENT OF BREAD-PASTE, VEGETABLES, 
 FRUIT, WHITE OF EGG. 
 
 Some micro-organisms, more especially mould fungi, grow very 
 well on bread-paste. This is prepared by removing the crust from 
 slices of bread and drying them in the oven. They are then 
 broken up, and reduced to a fine powder with a pestle and mortar. 
 Small, carefully cleansed, conical, or globe-shaped flasks are plugged 
 with cotton-wool and sterilised in the oven. When cool, a small 
 quantity of the powder is placed in them, and sterilised water added 
 in the proportion of one part to every four of the powder. The 
 paste is sterilised by steaming in the steriliser at 100 C. for half an 
 hour for three successive days. The flasks can be reversed, and may 
 be inoculated with a platinum needle. 
 
 Boiled carrots and other vegetables, and various kinds of stewed 
 fruit, are also occasionally employed for the cultivation of bacteria. 
 The sterilisation of these media must be carried out on the principles 
 already explained. 
 
 White of egg may be solidified in shallow glass dishes, in the 
 steam steriliser. After inoculation the dishes should be placed 
 in a damp chamber. 
 
 LIQUID MEDIA. 
 
 (F) PREPARATION OF STERILISED BROTH, LIQUID BLOOD SERUM, 
 URINE, MILK, VEGETABLE INFUSIONS, AND ARTIFICIAL NOURISH- 
 ING LIQUIDS. 
 
 Nutrient liquids are still largely employed. For inoculation ex- 
 periments when the presence of gelatine is undesirable, for studying 
 the physiology and chemistry of bacteria, and when for any object 
 a rapid growth of micro-organisms is necessary, the employment 
 of liquid media is not only advisable, but absolutely necessary. 
 Liquid media comprise two distinct groups natural and artificial. 
 Natural media include meat broth, blood, urine, milk, and vegetable 
 infusions ; artificial media are solutions composed from a chemical 
 formula representing essential food constituents. 
 
 Broth may be made from beef, pork, chicken, or fish in the 
 manner which has been described for the preparation of nutrient 
 gelatine, simply with omission of the gelatine. After the process 
 
MTKIENT MEDIA AM) METHODS OF CULTIVATION. 
 
 119 
 
 of neutralisation with carbonate of soda solution, the flask of broth 
 is placed m the steam steriliser for half an hour at 100 C. A 
 clear liquid results on nitration which is transferred to plugged 
 sterilised flasks or test-tubes, and sterilisation effected by exposing 
 them in the steam steriliser for half an hour at 100 0. for two 
 
 FIG. 47. APPARATUS FOR STERILISATION BY STEAM UXDKK PKE>- 
 
 or three successive days, or by-lusing the apparatus for sterilising 
 by steam under pressure. For some bacteria a more suitable 
 cultivating medium is obtained by the addition of glycerine or 
 grape-sugar. 
 
 Liquid Blood Serum. The preparation of blood serum has 
 already been described. ' It may be required for cultivations before 
 the final treatment by which it is solidified, for example, in the 
 method of drop-cultivation, and may be used with the addition of 
 glycerine or grape-sugar. Hydrocele fluid, peritonitic and pleuritic 
 
120 BACTERIOLOGY. 
 
 effusions, can also be employed after sterilisation in the steam 
 steriliser. The fluid should be withdrawn with a sterilised trocar 
 and cannula, and received into plugged sterilised flasks. 
 
 Urine. In order to obtain urine free from micro-organisms the 
 following precautions must be observed : The orifice of the urethra 
 must be thoroughly cleansed with weak carbolic. The first jet of 
 urine should be rejected, and the rest received into sterilised vessels, 
 which must be quickly closed with sterilised plugs. If these pre- 
 cautions be not attended to, the urine must be rendered sterile by 
 the means described for the sterilisation of broth. 
 
 Milk. If milk has been drawn into sterile flasks, after thoroughly 
 cleansing and disinfecting the teats and hands, it may be kept 
 without change. If procured without these precautions, it must 
 be steamed in the steriliser for half an hour for five successive 
 days. 
 
 Vegetable and other Infusions. Infusions of hay, cucumber, 
 and turnip are used for special purposes, and more rarely decoctions 
 of plums, raisins, malt, and horse-dung. They are mostly prepared 
 by boiling with distilled water, after maceration for several hours. 
 The filtrate is received into sterile flasks and sterilised in the usual 
 way in the steam steriliser. 
 
 Artificial Fluids. Pasteur's solution is prepared by mixing the 
 ingredients in the following proportions : 
 
 Distilled water 100 
 
 Pure cane-sugar 10 
 
 Ammonium tartrate 1 
 
 Ash of yeast -075 
 
 Mayer's modification of the nourishing fluid employed by Colin is 
 as follows : 
 
 Distilled water . . . . . 20 
 
 Ammonium tartrate .... *2 
 
 Phosphate of potassium . . . . -1 
 
 Sulphate of magnesium ... -1 
 
 Tribasic calcium phosphate .... -01 
 
 Drop-cultures. This method of cultivation is a particularly 
 instructive one. It enables us to study many of the changes which 
 take place during the life history of micro-organisins. This is 
 illustrated, for example, in a drop-culture of the anthrax bacillus, 
 in which we can watch the gradual growth of a single bacillus into 
 
NUTRIENT MEDIA AND METHODS OF CULTIVATION. 
 
 121 
 
 a long filament, and the subsequent development of bright oval 
 sport's. It is necessary carefully to observe the minutest details 
 in order to maintain the cultivation pure. An excavated slide is 
 thoroughly cleaned, and then sterilised by being held with the 
 cupped side downwards in the flame of the Bunsen burner. A ring 
 of vaseline is painted round the excavation, and the slide is then 
 placed under a glass bell. Meanwhile a carefully cleansed cover- 
 glass i> al>o sterilised by passing it through the flame, and should 
 be deposited on a sterilised glass plate. With a sterilised looped 
 needle, a drop of sterile broth is transferred to the cover-glass, 
 and this is inoculated by touching it with another sterilised needle 
 charged with the material to be examined, without disturbing 
 the form of the drop. It is quite sufficient just to touch the drop 
 
 1*5 
 
 
 FIG. 48. DROP CULTIVATION'. 
 () Drop of broth ; (6) layer of vaseline. 
 
 instead of transferring a visible quantity of blood, juice, or growth, 
 as the case may be. The slide is then inverted and placed over the 
 cover-glass, so that the drop will come exactly in the centre of the 
 excavation, and is gently pressed down. On turning the slide over 
 airaiii the cover-glass adheres, and an additional layer of vaseline 
 i- painted round the edges of the cover-glass itself. The slide must 
 be labelled, and if necessary, placed in the incubator, and the results 
 \\atehed from time to time. Instead of broth, liquid blood serum 
 may be employed in this form of cultivation. If it is required 
 to preserve the drop-cultivation as a microscopic preparation, the 
 cover-glass Ls gently lifted off and allowed to dry. Any vaseline 
 adhering to the cover-glass should be wiped off, and the cover-glass 
 can then be passed through the flame and stained in the usual manner. 
 Moist Cells. Unless drop-cultures are very carefully prepared 
 
122 BACTERIOLOGY. 
 
 they are liable to dry up, if kept for examination for several days. 
 Many therefore prefer employing a moist cell, of which there are 
 several different forms in use. 
 
 The drop-culture slide may be converted into a moist cell 
 by having a deep groove cut round the circumference of the con- 
 cavity. This groove is filled with sterilised water by means of a 
 pipette. A ring of vaseline is painted with the camel's-hair brush 
 outside the groove, and the cover-glass, with the drop-cultivation, 
 is inverted and placed over the concavity. This form is very useful, 
 as the slide can be easily cleansed and effectually sterilised by 
 holding it in the flame of the Bunsen burner. 
 
 A very simple form of moist cell recommended by Schafer 
 
 FIG. 49. SIMPLE METHOD OF FORMING A MOIST CELL. 
 
 may be used in some cases, but possesses the disadvantage of not 
 admitting of sterilisation by heat. A small piece of putty or 
 modelling wax is rolled into a cord about two inches long and J inch 
 thick. By uniting the ends a ring is formed, which is placed on the 
 middle of a clean glass slide. A drop of water is placed in the 
 centre of the ring, and the cell roofed in by applying a cover-glass. 
 
 A cell somewhat similar in form, which has the advantage of 
 permitting of thorough cleansing, may be constructed by cementing 
 a glass ring with flat surfaces to an ordinary slide. Vaseline is 
 applied with a camel's-hair brush to the upper surface of the ring, 
 and one or two drops of water placed with a pipette at the bottom 
 of the cell. The cover-glass, with the preparation, is then inverted 
 over the cell and gently pressed down upon the glass ring. The 
 vaseline renders the cell air-tight, and, to a certain extent, fixes 
 the cover-glass to the ring. 
 
NUTRIENT MEDIA AND METHODS OF CULTIVATION. 
 
 123 
 
 Stages. To apply warmth while a preparation is under 
 continuous observation, we must either place the microscope bodily 
 
 FIG. 50. WARM STAGE. 
 
 within a special incubator, with the eye-piece protruding through an 
 opening, or we must employ some means of applying heat directly 
 to the preparation. 
 
 A simple warm stage may be made of an oblong copper plate, 
 
 FIG. 51. WARM STACK SHOWN IN. OPERATION. 
 
 two inches long by one inch wide, from one side of which a rod of 
 the same material projects. The plate has a round aperture in the 
 
124 
 
 BACTERIOLOGY. 
 
 middle, half an inch in diameter, and is fastened to an ordinary 
 slide with sealing-wax. The drop to be examined is placed on a 
 large-sized cover-glass and covered with a smaller one. Olive oil 
 or vaseline is painted round the edge of the smaller cover-glass to 
 prevent evaporation, and the preparation is placed over the aperture 
 in the plate. 
 
 The slide bearing the copper plate is clamped to the stage of 
 
 FIG. 52. ISRAEL'S WARMING APPARATUS IN OPERATION. 
 
 the microscope. The flame of a spirit-lamp is applied to the 
 extremity of the rod, and the heat is conducted to the plate and 
 thence transmitted to the specimen. In order that the temperature 
 of the copper plate may be approximately that of the body, the lamp 
 is so adjusted that a fragment of cacao butter and wax, placed close 
 to the preparation, is melted. 
 
 Israel's Warming Apparatus. It is obvious that in employing 
 very high powers a difficulty will be presented by the warm stages 
 
NUTRIENT MEDIA AND METHODS OF CULTIVATION. 
 
 125 
 
 commonly used for accurate observations, such as Schafer's or 
 Strieker's, owing to their interference with the illumination. To 
 
 FIG. 53. SECTION OF ISRAEL'S WARMING APPARATUS AND DROP-CULTURE 
 
 SLIDE. 
 
 overcome this an apparatus has been constructed by which the 
 -liile is warmed from above (Figs. 52, 53). 
 
 The drop-culture slides are provided with a shallow groove, '1 mm. 
 deep and 1 mm. broad, cut round the concavity. Into this the 
 cover-glass fits, so that its upper surface is level with that of 
 
 FIG. 54. ISRAEL'S WARMING APPARATUS 
 
 the slide. The heating apparatus consists of a flat disk-sliapcd box 
 with a central conical aperture. 
 
 The entrance and exit pipes are fixed on at a right angle to the 
 
126 
 
 BACTERIOLOGY. 
 
 side (Fig. 54). The former, , is of metal, ancLthe latter, a, of glass 
 fitted with a thermometer, the bulb of which, k, is contained within 
 the box. A partition, s, keeps up, a current between the openings 
 
 FIG. 55. GAS CHAMBER IN USE WITH APPARATUS FOR GENERATING 
 CARBONIC ACID. 
 
 of the pipes, which are supported on a stand and connected by 
 tubing with the hot-water supply. 
 
 A mixture of paraffine and vaseline is recommended for indicating 
 the temperature of the chamber, and experience has shown that if 
 a temperature of 37 C. is required the temperature of the water 
 in the box must range between 42 and 47 C. 
 
 FIG. 56. GAS CHAMBER. 
 
 Gas Chambers. To investigate the action of gases or vapours 
 upon micro-organisms, a modification of the moist cell may be 
 employed. 
 
 A piece of glass tubing is first fixed to the slide by means of 
 
NUTRIENT MEDIA AND METHODS OF CULTIVATION. 
 
 127 
 
 sealing-wax, and the ring of putty is so placed as to include one end 
 of it, leaving a small interval at the side, or a little notch is made 
 iii the putty opposite, so as to afford an exit for the gas or vapour. 
 
 FIG. 57. MOIST CELL ADAPTED FOR TRANSMISSION OF ELECTRICITY. 
 
 Application of Electricity. To study the effect of electricity 
 we may prepare a drop-culture in the moist cell. The cover- 
 glass to be used is provided with two strips of tinfoil, which are 
 
 Fii.. 58. APPARATUS ARRANGED FOR TRANSMITTING ELECTRICITY. 
 
 isolated from the brass of the microscope, and so arranged that a 
 current of electricity may be passed through them. 
 
 A much simpler plan, which may also be employed, is to tak- 
 an ordinary irlax slide an. I coat the surface with gold-size. The 
 
128 
 
 BACTERIOLOGY. 
 
 slide is then pressed firmly down on gold-leaf or tinfoil and allowed 
 to dry. When dry, the metal is scraped away, leaving two triangles 
 with a small interval between them. 
 
 FIG. 59. SLIDE WITH GOLD-LEAF ELECTRODES. 
 
 The liquid containing the micro-organisms is placed between the 
 electrodes, covered with a cover-glass, and then subjected to the 
 electric current. 
 
 (G) METHODS OF EMPLOYING AND STORING LIQUID MEDIA. 
 
 Cultivations in liquid media can be carried on in test-tubes, but 
 it is more satisfactory to employ special forms of flasks, bulbs, and 
 U tubes, such as those employed by Pasteur and his school, and by 
 Lister, Sternberg, and Aitken. 
 
 Lister's flasks. These flasks were especially introduced by Lister 
 as a means of storing liquid nutrient media. 
 They are so constructed that after removal 
 of a portion of the contents, on restoring 
 the vessel to the vertical position, a drop of 
 liquid always remains in the extremity of 
 the nozzle, which prevents regurgitation of 
 unflltered air. 
 
 Sternberg' s Bulbs. The method of intro- 
 ducing liquid into the bulb employed by 
 Sternberg, and of sterilising and inoculating 
 it, is as follows : The bulb is heated slightly 
 over the flame, and the extremity of the neck, after the sealed point 
 has been broken off, is plunged beneath the surface of the liquid. 
 As the air cools the liquid is drawn into the bulb, 
 usually filling it to about one-third of its capacity. ~^ = ^^====== 
 
 The neck of the flask is again sealed up, and the 
 
 liquid which has been introduced is sterilised by IG ' ,'~ TERN- 
 BERG'S BULB. 
 repeatedly boiling the flasks in the water- bath. 
 
 They should then be placed in the incubator for two or three days ; 
 
 FIG. 60. LISTER'S 
 FLASK. 
 
NTTRIKXT MEDIA AND METHODS OF CULTIVATION. 
 
 129 
 
 FIG. 62. ATTKEN'S 
 TUBE. 
 
 and if the contents remain transparent and free from film, they 
 may be set aside as stock-bulbs, to be used when required. 
 
 To inoculate the liquid in the bulb the end of the neck is heated 
 to sterilise the exterior, the bulb is gently warmed, and the extremity 
 of the neck nipped off with a pair of sterilised forceps. The open 
 extremity is plunged into the liquid containing the micro-organisms, 
 and a minute quantity enters the tube and mingles with the fluid in 
 the bulb without fear of contamination by atmospheric germs. The 
 extremity of the neck is once more sealed up in the flame of a 
 Li i nst MI burner. 
 
 A itken's Tubes. These tubes are plugged and sterilised, and the 
 nutrient medium introduced as into ordinary test-tubes. Instead 
 of withdrawing the cotton-wool plug, they are 
 inoculated through a lateral arm. The sealed 
 extremity of the arm is nipped off with 
 sterilised forceps, and the inoculating needle is 
 carefully introduced through the opening thus 
 made. It is directed along the arm until it 
 touches the opposite side of the test-tube, where 
 it deposits the material with which it was charged. 
 The needle is withdrawn, and the end of the 
 lateral arm apiin sealed up in the flame ; the test-tube is then 
 tilted until the liquid touches the deposited material ; on restoring 
 the tube to the vertical, the material is washed down with the 
 nutrient liquid. 
 
 Uiquel's Butts. The tube a 
 hfnde of Miquel is also a very 
 useful form. It consists of a 
 lulb of 50 cc. capacity, blown 
 in the middle of a glass tube. 
 The part of the tube above the 
 bulb is contracted in two places, 
 and can either be left quite 
 >traight or made to curve 
 slightly. Between the contrac- 
 tions the tube is plugged with 
 asbestos. The portion of the 
 tube below the bulb is $ shaped, 
 and drawn out at its extremity 
 
 into a fine point. The bulb is charged with nutrient liquid and 
 inoculated by aspiration, and the point of the $ tube sealed in the 
 Maine of a Bunsen burner. 
 
 9 
 
 Ki... i;::. . 
 
 lin.u. 
 
130 
 
 BACTERIOLOGY. 
 
 FIG. 64. PASTEUK'S FLASK. 
 
 Pasteur's Apparatus. Special forms of tubes, bulbs, and pipettes 
 are employed by the school of Pasteur. The tubes are provided 
 
 with lateral or with 
 curved arms drawn 
 out to a fine point, 
 and with slender 
 necks plugged with 
 cotton-wool. A 
 double form (Fig. 65) 
 shaped like a tuning- 
 fork, each limb with 
 a bent arm, is con- 
 venient for storing 
 sterilised broth. The 
 
 sealed end of an arm is nipped off with sterilised forceps, the sterile 
 broth aspirated into each limb, and the arm again sealed in the 
 flame ; a series of such tubes can be 
 arranged upon a rack on the working 
 table. 
 
 Bulbs with a vertical neck drawn out 
 to a fine point, others with a neck bent 
 at an obtuse angle, plugged with cotton- 
 wool, and a lateral curved arm drawn 
 out to a fine point, are also employed. 
 For a description of these various vessels 
 and their special advantages, the works 
 
 of Pasteur and Duclaux must be con- 
 
 FIG. G5. PASTEUR'S DOUBLE 
 suited. TUM . 
 
 (H) CULTIVATION OF ANAEROBIC BACTERIA. 
 
 To cultivate anaerobic organisms the same media are employed 
 as for aerobic organisms, but the methods must be modified, or special 
 apparatus used, so that the oxygen in the air may be excluded. 
 
 In the preparation of plate -cultivations, before the film of gelatine 
 has completely hardened it is covered with a sheet of mica, and the 
 edges are sealed with melted paraffine. By this process the air is not 
 completely excluded, so that only those organisms which are not 
 strictly anaerobic can be grown by this method. Liborius recom- 
 mends boiling a considerable volume of gelatine in a tube, cooling it f 
 ;i n<l after thoroughly distributing the organisms in the still liquid jelly, 
 rapidly solidifying it by placing the tube in iced water. By this 
 
NUTRIENT MEDIA AND METHODS OF CULTIVATION. 
 
 131 
 
 process very little air re-enters the jelly, and colonies of even strictly 
 anaerobic bacteria will develop in the lower part of the tube. The 
 drawback is the difficulty encountered in examining the colonies, 
 and in preparing sub-cultures. For this purpose the tube 
 must be broken, or carefully warmed until the jelly can be 
 shaken out. 
 
 Esmarch first prepares a roll culture, and when the gelatine film 
 has set, the tube is completely filled with liquefied gelatine which has 
 been cooled down almost to the temperature at which it solidifies. The 
 same difficulty arises as 
 in the previous method, in 
 the examination of the 
 colonies. 
 
 Buchner places the 
 culture tube inside a much 
 larger tube containing a 
 small quantity of pyro- 
 gallic acid and closed with 
 a gutta-percha cap. The 
 pyrogallic acid absorbs the 
 oxygen, but the method 
 i> not altogether suc< 
 ful. 
 
 The most satisfactory 
 plan is to exhaust the air 
 with an air pump, or to 
 substitute an atmosphere 
 of hydrogen which does 
 not affect the growth of 
 the bacteria. 
 
 Various forms of flasks 
 and tube* for cultivating 
 bacteria have been devised, 
 which can be easily con- 
 nected with an exhausting 
 aj para t us. and readily 
 sealed by the flame of the blowpipe when the air has been removed. 
 
 If hydrogen is employed the most convenient plan is to use 
 a Kipp's apparatus, from which the hydrogen is passed through 
 two bottles, one containing a solution of lead, to remove any 
 sulphuretted hydrogen, and the other pyrogallic acid, to intercept 
 any oxygen. 
 
 ANAKROBIC TUBE-CULTURE" 
 Glag9 tube through which h y drogen is 
 
 passed; b, exit tube; c, india-ruMx-i 
 stopper coated externally with paraffin 
 
 (FKANKI.AXI)). 
 
132 
 
 BACTERIOLOGY. 
 
 In the method recommended by Frankel a tube of gelatine is 
 liquefied, and inoculated. A gutta-percha stopper is substituted 
 for the cotton-wool plug (Fig. 66). It is perforated by two holes, 
 through which two tubes pass which are bent at a right angle. 
 One tube only just passes through the stopper, the other reaches 
 down to the bottom of the test-tube. The 
 horizontal part of each tube has a narrow 
 neck. The long tube has a plug of steril- 
 ised cotton-wool, and is connected with 
 a short piece of india-rubber tubing by 
 which it can be connected with Kipp's 
 apparatus. The hydrogen drives the air 
 out of the liquefied jelly and out of the 
 test-tube, and after about half an hour 
 the horizontal tubes are sealed up, arid 
 the test-tube is made into a roll culture. 
 
 Liborius employs a tube with a narrow 
 neck and a lateral arm (Fig. 67). The 
 tube is filled up to the height of the arm 
 with either nutrient agar or a mixture of 
 nutrient agar with 2 per cent, of grape- 
 sugar. The liquefied jelly is inoculated 
 in the usual way, and hydrogen passed 
 through the lateral arm. When the air 
 has been completely driven out, the tube 
 is sealed up. 
 
 To cultivate anaerobic organisms in 
 broth, such as the tetanus bacillus, a flask 
 is inoculated with the bacillus, and a 
 stream of hydrogen is passed through the 
 broth by means of a tube passing down to the bottom of the 
 flask. The air in the flask escapes by a lateral arm which is bent 
 down at a right angle, and immersed in a capsule of mercury. 
 When the air has been completely expelled the entrance tube is 
 hermetically sealed, and the mercury in the capsule prevents any 
 air from re-entering the flask by the lateral arm (Fig. 68). 
 
 FIG. 67. ANAEROBIC CUL- 
 TURE-TUBE (LlBORIUS). 
 
 METHOD OF FIXING CULTURES. 
 
 The colonies in plate-cultivations arid the growths of bacteria 
 in test-tubes may be stopped at any stage of their growth, and 
 permanently fixed by exposing the culture to the fumes of formic 
 
NUTRIENT MEDIA AND METHODS OF CULTIVATION. 
 
 133 
 
 aldehyde. The test-tubes, dishes, or capsules are placed in a cylin- 
 drical irlass vessel containing a pledget of cotton- wool moistened with 
 
 FIG. 68. APPARATUS FOR ANAEROBIC CULTURES. 
 
 (ROSCOE AND LU.NT.) 
 
 formic aldehyde. The vessel is fitted with a ground glass stopper 
 and >-t aside. The growth almost immediately ceases. Any 
 liqueliM gelatine is hardened, so that the exact appearances of 
 cultures are obtained in a permanent form. 
 
CHAPTER X. 
 
 EXPERIMENTS UPON THE LIVING ANIMAL. 
 
 To carry out the last of Koch's postulates, and so complete the 
 chain of evidence in favour of the causal relation of micro-organisms 
 to disease, and to study the mode of action of a pathogenic bacterium, 
 it is necessary to introduce into a living animal a pure cultivation 
 of the micro-organism or its chemical products. For this purpose 
 various animals are employed, such as mice, rabbits, guinea-pigs, 
 pigeons, and fowls. 
 
 Inhalation. The animals may be made to inhale an atmosphere 
 impregnated with micro-organisms by means of a spray. In this 
 way Friedlander succeeded in administering the bacteria of pneu- 
 monia to mice ; and the production of tuberculosis by experimental 
 inhalation has thrown light upon the clinical records of cases 
 reported as instances of the infectiousness of phthisis. 
 
 Ingestion. A sheep fed upon potatoes which have been the 
 medium for the cultivation of the anthrax bacillus dies in a few 
 days. Eabbits fed on cabbage sprinkled with a culture of the 
 bacillus of fowl cholera, rapidly succumb to the disease. Animals 
 fed upon the nodules of bovine tuberculosis or upon tubercular 
 flesh and milk will be readily infected. 
 
 Milk, or bread soaked in milk, is a very convenient medium, 
 and from a public health point of view, a most instructive way of 
 administering and testing the effect of pathogenic bacteria. 
 
 Vaccination and Subcutaneous Inoculation. Vaccination may be 
 performed by making a few superficial scratches and inoculating the 
 wound with a sterilised platinum needle charged with the micro- 
 organisms. Another simple method is to take a sterilised scalpel, 
 infect the point with the material to be inoculated, and then make 
 a minute puncture or incision. In either case a situation should be 
 selected, such as the root of the ear, which cannot be licked by the 
 animal after the operation. 
 
 134 
 
EXPERIMENTS UPON THE LIVING ANIMAL. 
 
 135 
 
 Subcutaneous inoculation is very simple and effectual, and con- 
 sequently the method most frequently employed. The animal 
 selected for example, a guinea-pig is held by an assistant, who 
 covers it with a towel, leaving only the hind extremities exposed. 
 By so doing, and gently laying it upon its back, with its head low, 
 a guinea-pig passes apparently into a state of hypnotism, and the 
 
 FIG. 69. KOCH'S SYRINGE. 
 
 trivial operation can be performed with little or no movement on the 
 part of the animal. From a spot on the inner side of the thigh the 
 hair is cut close with a small pair of scissors curved on the flat, and 
 the skin must be thoroughly purified with 1 in 20 carbolic acid. A 
 small fold of skin is then pinched up with a pair of sterilised forceps, 
 and with a pair of sharp sterilised scissors, or with a tenotomy knife, 
 a minute incision is made. A sterilised platinum loop is charged 
 with the material to be inoculated, and the loop is gently inserted 
 under the skin, forming a small pocket in the subcutaneous tissue. 
 The needle is then withdrawn, and the sides of the wound gently 
 pressed into apposition and painted over with collodion. 
 
 FIG. 70. SYRINGE WITH ASBESTOS Pi.n;. 
 
 In inoculating a mouse the same process is adopted, with the 
 exception that the root of the tail is the usual site of the 
 operation. 
 
 In some cases it may be necessary to inoculate cultures diffused 
 in sterilised salt solution, or blood or lymph containing bacteria, 
 or a culture in broth, or a filtrate containing the toxic products, 
 
136 BACTERIOLOGY. 
 
 and then a hypodermic syringe may be required. One of the ordinary 
 pattern may be used, but it is very much better to employ a syringe 
 which has been especially constructed to admit of thorough dis- 
 infection. Koch's syringe is a convenient form, the liquid being 
 expressed by pressure on a rubber ball. 
 
 The author has generally preferred to improvise a substitute for 
 the hypodermic syringe which can be quickly made, and is destroyed 
 after use, so that there can be no possible risk of accidentally 
 infecting other animals. A short length of ordinary glass-tubing is 
 sterilised, and plugged at one end with sterilised cotton-wool ; about 
 three inches from the plug a bulb is blown about the size of a 
 marble, and two inches below this the glass is drawn out into a long 
 capillary tube. A sufficient quantity of the liquid to be injected rises 
 up into the tube by capillary attraction, or can be drawn up by 
 means of an india-rubber ball, until the bulb is full. The point of 
 the capillary tube is inserted through the opening in the skin, and 
 gently pushed into the subcutaneous tissue, and then withdrawn for 
 a short distance. By pressure on the bulb the contents of the tube 
 are injected. In dealing with chemical products there is no risk in 
 applying the lips and blowing out the contents of the tube, or indeed 
 of filling it by suction, for if too much force were applied the liquid 
 which might enter the mouth would be stopped by the cotton-wool 
 plug. 
 
 A number of these capillary tubes can be placed in a small case, 
 and when it is necessary to go to a distance to investigate an outbreak, 
 they will be found most convenient to bring back lymph or blood to 
 the laboratory for further study. 
 
 Sternberg takes a piece of glass-tubing, blows a bulb at one end, 
 and draws out the other end into a thin tube. By heating the bulb 
 and then ^dipping the tube into the liquid to be inoculated the latter 
 rises in the tube as the bulb cools. After inserting the point of 
 the tube subcutaneously the bulb is again heated, and the liquid is 
 forced out into the tissues. 
 
 Intravenous Injection. r-A cultivation of micro-organisms may be 
 mixed with sterilised water, and then injected with a syringe directly 
 into the circulation. This may be performed without much difficulty 
 by injecting, with a hypodermic syringe, the large vein at the base 
 of the ear in rabbits, or the jugular vein in large animals. 
 
 Special Operations. In many cases it is absolutely necessary to 
 perform an operation of greater severity. After the administration 
 of an anaesthetic, infective material may be inserted, or injected, into 
 the peritoneal cavity, or injected into the duodenum in the manner 
 
EXPERIMENTS UPON THE LIVING ANIMAL. 137 
 
 employed in the case of Koch's comma bacilli by Nicati and Rietsch. 
 In such oases antiseptic precautions must be rigidly followed, and 
 use made of iodoform and other antiseptic dressings. The disinfec- 
 tion of the skin of the animal, of the instruments employed, and of 
 the hands of the operator, are details essential to secure success. 
 
 To inoculate tubercular matter, sputum may be rubbed up with 
 distilled water, and some of the mixture injected into a tracheal 
 fistula ; or the first steps of the operation of iridectomy may be 
 performed and tubercular material inserted into the anterior chamber 
 of the eye, but this method is only justifiable when it is absolutely 
 necessary for the results and changes to be observed from day 
 to day. 
 
 To inoculate rabbits or other animals with the virus of rabies,, 
 the skull is trephined, and an emulsion prepared from the spinal cord 
 of a rabid animal is injected beneath the dura mater. 
 
 Before every inoculation the instruments must be sterilised in a 
 hot-air steriliser or by immersion in boiling water in a flat dish or 
 enamel tray heated by a spirit-lamp, and after each operation all 
 instruments should be placed in carbolic acid (1 in 20) or in boiling 
 water, wiped dry, and again sterilised in the hot-air steriliser, before 
 they are put away. If these precautions are not observed, instances 
 of accidental infection are sure to occur. 
 
 After the inoculation is completed a careful record must be made 
 of the date and details of the experiment. The form in which, the 
 virus was used, the quantity employed, and the seat of inoculation,, 
 must be taken into account. The animals must be kept under close 
 observation, the temperature taken, and any signs of illness, such as 
 ceasing to feed, difficulty in breathing, staring coat, and any local 
 signs, >uch as the development of a tumour or an enlargement of the 
 lymphatir glands, must be carefully noted. 
 
 It is perhaps hardly necessary to add that in this country no 
 experiments of any kind may be performed on living animals without 
 a licen-e. 
 
 METHOD OF DISSECTION AND EXAMINATION. 
 
 All animals that die after an experimental inoculation should 
 be examined immediately after death. Every precaution must be 
 taken in conducting the dissection, to exclude extraneous micro- 
 ori:ani-iii>. and all instruments employed must have been sterilised 
 in the hot-air steriliser, or by immersion in boiling water. If a mouse r 
 for example, has died after inoculation with anthrax, it should be at 
 
138 BACTERIOLOGY. 
 
 once pinned out by its feet 011 a slab of wood or in a gutta-percha 
 tray, and bathed with 1 in 40 carbolic. In the same way, before 
 examining a dead rabbit, a stream of carbolic should be directed 
 over it to lay the fur, which otherwise interferes with the dissection. 
 The hair should be cut away with sterilised scissors from the seat 
 of inoculation, which is the first part to be examined, and any 
 suppuration, haemorrhage, oedema, or other pathological change should 
 be carefully noted. From any pus or exudation that may be 
 present, material for inoculations should at once be taken, and 
 cover- glass- preparations made for microscopical examination. 
 
 To examine the internal organs and to make inoculations from 
 the blood of the heart or spleen, the skin is cut through from below 
 upwards in the median line of the abdominal and thoracic regions. 
 The abdominal cavity is then opened, and the walls pinned back 
 on either side of the animal. Any abnormal appearances in the 
 peritoneum should be noted, and the state of the spleen should be 
 carefully examined by turning the intestines aside. After noting 
 its appearances, it should be removed with sterilised forceps and 
 scissors, and deposited upon a sterilised glass slide, and incised with 
 sterilised scissors. The cut surface is then touched with the point 
 of a sterilised inoculating needle, and cultures are made in. test-tubes 
 of nutrient gelatine and agar-agar, and also on potato, and in broth 
 in the form of drop-cultivations. Precisely the same care must be 
 taken in examining lymphatic glands, tubercles, or pathological 
 nodules ; any chance putrefactive micro-organisms on the surface 
 should be destroyed by carbolic acid or the actual cautery; an 
 incision is then made, and a minute fragment snipped out of the 
 centre of the nodule, which can be inoculated in the living animal or 
 transferred to a cultivating medium. 
 
 The examination of the thorax is made by cutting through the 
 ribs on either side of the sternum with sterilised scissors, and 
 turning the sternum up where it will be out of the way. The 
 pericardium is then opened, and the right auricle or ventricle pierced 
 with the point of a sterilised scalpel, and inoculations and cover- 
 glass-preparations are made from the blood which escapes. 
 
 The lungs also require to be especially studied. They should be 
 incised with a sterilised scalpel, and inoculations and cover-glass- 
 preparations made from the cut surface. It may be necessary to 
 embed a piece of lung or fragment of spleen, so that it shall be free 
 from air. This may be done by isolating a fragment with the 
 precautions just described, and depositing it upon the surface of a 
 test-tube of nutrient agar-agar. The contents of another tube, 
 
KXIKKIMI.NTS UPON THE LIVING ANIMAL. 139 
 
 which have been liquefied, and allowed to cool almost to the point 
 of gelatinisation, must then be poured over it. From a potato a 
 little cube must be cut, the tissue deposited in the trough thus 
 formed, and the cube replaced, or cultures may be prepared by any 
 of the methods which have been described for dealing with anaerobic 
 bacteria. Blood may also be taken directly from a vein by laying it 
 bare by dissection, making a small opening with sterilised scissors, 
 and inserting a looped platinum needle, the needle of a hypodermic 
 syringe, a capillary tube, or the extremity of the capillary neck of 
 a Sternberg's bulb. If the cultivation, in spite of these precautions, 
 is contaminated, or if there was more than one organism present 
 in the blood or tissues under examination, it will be necessary to 
 separate the different kinds by plate-cultivation. 
 
 Having completed the dissection, the organs of such a small 
 animal as a mouse may be removed en masse, and transferred to 
 absolute alcohol for subsequent examination. In other cases it may 
 be only necessary to reserve portions of each organ, in experimenting 
 with a virulent micro-organism like anthrax, any remaining part of 
 the animal should be cremated, and the hands and all instruments 
 should be thoroughly disinfected. 
 
 Isolation of Micro-organisms during Life. Micro-organisms in 
 the living subject may be isolated from the pus of abscesses, or 
 other discharges, and from the blood and tissues. Abscesses should 
 be opened, and other operations performed, when practicable, with 
 Listerian precautions, and a drop of the discharge taken up with 
 ;i looped needle or capillary pipette, as already explained. 
 
 To make a cultivation from the blood of a living person, the tip 
 of a finger must be well washed with soap and water and sponged 
 with 1 in 20 carbolic. Venous congestion is produced by applying 
 an elastic band or ligature to the finger, which is pricked with a 
 sterilised sewing needle. From the drop of blood which exudes the 
 necessary inoculations and examinations can be made. Another 
 way of extracting blood from the living patient is to apply a leech. 
 This method has been found of considerable value in experimenting 
 upon the blood of patients suffering from malaria, and may be 
 useful in other diseases, if the blood is required for further 
 examination, or in quantity. 
 
CHAPTER XL 
 
 EXAMINATION OF AIR, SOIL, AND WATER. 
 
 AlR. 
 
 THE air, as is well known, contains in suspension, mineral, aninial r 
 and vegetable substances. The mineral world is represented 
 by such substances as silica, silicate of aluminium, carbonate and 
 phospate of calcium, which may be raised from the soil by the 
 wind, and particles of carbon, etc., which gain access from acci- 
 dental sources. Belonging to the animal kingdom we find the 
 debris of perished creatures, as well as, sometimes, living animals. 
 The vegetable world supplies micrococci, bacilli, and other forms 
 of the great family of bacteria, spores of other fungi, pollen seeds, 
 parts of flowers, and so forth. The air of hospitals and sick rooms 
 has been found to be especially rich in vegetable forms; fungi 
 and spores have been stated to be present in particularly large 
 numbers in cholera wards; spores of tricophyton have been dis- 
 covered in the air of hospitals for diseases of the skin, and of achorioii 
 in wards with cases of favus. The tubercle bacillus is said to have 
 been detected in the breath of patients suffering from phthisis. 
 
 These points indicate that, in addition to the interest for the 
 micro-biologist, considerable importance, from a hygienic point of 
 view, must be attached to the systematic examination of the air. 
 A knowledge of the microbes which are found in the air of marshy 
 and other unhealthy districts, and in the air of towns, dwellings, 
 hospitals, workshops, factories, and mines, will be of practical value. 
 
 Miquel, who has particularly studied the bacteria in the air, 
 has found that their number varies considerably. The average 
 number per cubic metre of air for the autumn quarter at Mont- 
 souris is given as 142, winter quarter 49, spring quarter 85, and 
 summer quarter 105. In air collected 2,000 to 4,000 metres above 
 the sea-level, not a single bacterium or fungus spore was found, 
 while in 10 cubic metres of air from the Rue de Rivoli (Paris) the 
 number was computed at 55,000. 
 
 140 
 
EXAMINATION OF AIR, SOIL, AND WATER. 141 
 
 The simplest method for examining the organisms in air eon>i>t> 
 in exposing plates of glass or microscopic slides coated with 
 irlycerine, or with a mixture of glycerine and grape sugar, which is 
 stable, colourless, and transparent. Nutrient gelatine spread out on 
 glass plates may be exposed to the air for a certain time, and then 
 put aside in clamp chambers for the colonies to develop. Sterilised 
 potatoes, prepared in the usual way, may be similarly exposed. In 
 both the last -mentioned methods separate colonies develop, which 
 may be isolated, and pure cultivations carried on in various other 
 nutrient media. Nutrient gelatine has also been employed in the 
 special methods of Koch and Hesse. 
 
 Koch's Apparatus. This consists of a glass jar, about six inches 
 high, the neck of which is plugged with cotton-wool. In the 
 interior is a shallow glass capsule, which can be removed by means 
 of a brass lifter. The whole is sterilised by exposure to 150 C. 
 for an hour in the hot-air steriliser. The nutrient gelatine in a 
 stock-tube is liquefied, and the contents emptied into the glass 
 capsule. The jar is exposed to the air to be examined for a definite 
 time, the cotton -wool plug replaced, and the apparatus set aside for 
 the colonies to develop. 
 
 Hesse's Apparatus. The advantage of this apparatus is that it 
 enables the experimenter to examine a known volume of air. A 
 glass cylinder, 70 cm. long and 3*5 cm. in diameter, is closed at one 
 end by an india-rubber cap, perforated in the centre. Over this iit> 
 another cap, which is not perforated. The opposite end of the 
 cylinder is closed with a caoutchouc stopper, perforated to admit 
 a glass tube plugged with cotton- wool. The tube can be connected 
 by means of india-rubber tubing with an aspirating apparatus, 
 which consists of a couple of litre-flasks, suspended by hooks from 
 the tripod-stand which supports the whole apparatus. The cylinder, 
 <;ij. and plug are washed with solution of carbolic acid, and 
 then with alcohol. After being thus cleansed, 50 cc. of nutrient 
 gelatine are introduced, and the whole sterilised by steaming for 
 half an hour for three successive days. After the final sterilisation, 
 tlit cylinder is rotated on its long axis, so that the nutrient medium 
 solidities in the form of a coating over the whole of the interior. 
 When required for use, the cotton- wool plug is removed from the 
 Mnall glass tube, and the latter connected with the upper flask by 
 n:-ni> of the india-rubber tubing. 
 
 The apparatus is placed in the air which is to be examined, the 
 outer india-rubber cap removed from the glass cylinder, and the 
 upper flask tilted until the water begins to flow into the lower one. 
 
142 
 
 BACTERIOLOGY. 
 
 The emptying continues by siphon action, and air is drawn in along" 
 the cylinder to replace the water. When the upper flask is empty r 
 the position of the two is reversed, and the flow again started. 
 When a sufficient volume has been drawn through the cylinder, the 
 outer cap and the cotton- wool plug are replaced, and it is set aside 
 for the colonies to develop. As an example, twenty-five litres of air 
 from an open square in Berlin gave rise to three colonies of bacteria 
 and sixteen moulds ; on the other hand, two litres from a school- 
 
 FIG. 71. HESSE'S APPARATUS. 
 
 room just vacated by the scholars gave thirty-seven colonies of 
 bacteria and thirty-three moulds. 
 
 Porous substances, such as sand, powdered glass, or sugar, may 
 be used for the filtration of samples of air; and an apparatus is 
 employed in a convenient form to be conveyed to the laboratory for 
 the subsequent examination. 
 
 Pefri's Apparatus consists of a glass-tube 9 cm. long, containing 
 two sand-filters separated from each other. A known volume of air- 
 is aspirated through the tube. The bacteria are arrested and can 
 
i:\AMINATIOX OF AIR, SOIL, AND WATER. 
 
 143 
 
 be examined by spreading the sand out in a dish and covering it 
 with nutrient gelatine ; or it may be shaken up with sterilised water 
 and plate- cultivations prepared. The sand-filter nearest to the 
 aspirator should remain free from bacteria. 
 
 Sedgwick and Tucker employ a glass cylinder which is drawn out 
 at one end into a narrow tube to contain sterilised powdered cane 
 sugar. Both ends of the apparatus are plugged with sterilised 
 cotton-wool. By means of an exhausting apparatus a known volume 
 of air is drawn through the tube. The cotton-wool plug is re- 
 
 FIG. 72. SEDGWICK AND TUCKKR'S TUBE. 
 
 moved, and liquid gelatine is introduced into the cylinder, the 
 plug is replaced, and the sugar is shaken into and quickly dissolves 
 in the jelly. The cylinder is then treated in the same way as a 
 roll-culture, and set aside for the colonies to develop (Fig. 72). 
 
 Various forms of " aeroscopes " and " aeroniscopes " have from 
 
 Fi'.. 73. POUCHET'S AEKOSCOPK. 
 
 time to time been employed. Pouchet's aeroscope c<>n-i>t- of a small 
 funnel, drawn out to a point below which is a glass slip coated with 
 
144 BACTERIOLOGY. 
 
 glycerine. The end of the funnel and the glass slip are enclosed 
 in an air-tight chamber, from which a small glass tube passes out 
 and is connected by india-rubber tubing with an aspirator (Fig. 73). 
 The air passing down the funnel strikes upon the glycerine, which 
 arrests any solid particles. For a full description of the apparatus 
 employed by Maddox, Cunningham, and Miquel, reference should 
 be made to the writings of these authors, and particularly to the 
 treatise published by the last-named. 
 
 SOIL. 
 
 Surface soil is exceedingly rich in bacteria. Miquel has com- 
 puted that there exists in a gramme of soil an average of 750,000 
 germs at Montsouris, 1,300,000 in the Rue de Remies, and 2,100,000 
 in the Rue de Monge. As agents in putrefaction and fermenta- 
 tion they play a very important role in the economy of nature ; 
 but there exist in addition, bacteria in the soil which are patho- 
 genic in character. Pasteur has succeeded in isolating the bacillus 
 of anthrax from the earth. Sheep, sojourning upon a plot of 
 ground where animals with anthrax have been buried, may succumb 
 to the disease. Pasteur considered that the spores were conveyed 
 by worms from buried carcasses to the surface soil. The bacilli 
 of malignant oedema and tetanus are also present in soil. Nicolaier 
 produced tetanus in mice and rabbits by inoculating a little garden 
 earth under the skin. 
 
 To obtain a cultivation of the microbes in soil a sample of the 
 latter must be first dried and then triturated. It may then be 
 .shaken up with distilled water, and from this a drop transferred to 
 .sterilised broth. The employment of solid media is, however, 
 much more satisfactory : a sample of earth is collected, dried, and 
 triturated, and a small quantity sprinkled over the surface of 
 nutrient gelatine prepared for a plate-cultivation. In another 
 method the gelatine is liquefied in a test-tube, the powder added, 
 and distributed, in the usual way, throughout the medium, which 
 is then poured out upon a glass plate or made into a roll-culture. 
 In the same way the dust which settles from the air in houses and 
 hospitals, or food substances in powder, may be distributed in 
 nutrient gelatine, arid examined both for aerobic and anaerobic 
 bacteria. The different kinds which develop, must be thoroughly 
 investigated n,s regards their morphological and biological charac- 
 ters, and pathogenic properties. 
 
EXAMINATION OF AIR, SOIL, AND WATER. 145 
 
 WATER. 
 
 In the case of water, as in that of air, a knowledge of the 
 micro-organisms which may be present is not only of interest 
 to the bacteriologist, but of the greatest importance in practical 
 hygiene. Common putrefactive bacteria and vibrios may not be 
 hurtful in themselves, but they indicate the probability of the 
 presence of organic matter in which there may be danger. The 
 detection of Bacillus coli communis may be taken to indicate a 
 probable contamination with human excreta. 
 
 The Microzyme Test which was introduced for the detection 
 of putrefactive bacteria, consisted in adding three or four drops of 
 the sample of water to 1 or 2 cc. of Pasteur's fluid, the nourishing 
 fluid having been previously boiled in a sterilised test-tube. If the 
 microzymes or their germs existed in the water, the liquid in a few 
 days became turbid from the presence of countless bacteria. This 
 test is of no real value, for it does little more than indicate that 
 bacteria are present, which we know to be the case in all ordinary 
 water, and even in ice. On the other hand, the bacteriological test 
 of Koch is a most valuable addition to the usual methods of water 
 analysis. It enables us not only to detect the presence of bacteria, 
 but to ascertain approximately their number, and to study very 
 minutely their morphological and biological characteristics. The 
 importance of a thorough acquaintance with the life-history of the 
 individual micro-organisms cannot be too strongly insisted upon. 
 For example, by such means the spirillum of Asiatic cholera can 
 be distinguished from most other comma-shaped organisms, and 
 inasmuch as its presence may be an indication of contamination 
 with choleraic discharges, such water should be condemned for 
 drinking purposes, even though we are not yet in a position to 
 affirm that the microbe is the cause of the disease. The detection 
 of the bacillus of typhoid fever or of the Bacillus coli communis 
 in suspected water or milk would be evidence of considerable 
 importance. 
 
 Koch's test, in short, consists in making plate-cultivations of a 
 known volume of water, counting the colonies which develop, 
 isolating the micro-organisms, and studying the characters of each 
 individual form. 
 
 Collection and Transport of Water Sa?nples.Sternberg's bulbs, 
 or Erlenmeyer's conical flasks of about 100 cc. capacity, may be 
 employed with advantage for collecting the samples of water. The 
 latter are cleansed, plugged, and sterilised in the hot-air steriliser. 
 
 10 
 
146 BACTERIOLOGY. 
 
 When required for use, the plug is removed and held between the 
 fingers, which must not touch the part which enters the neck of 
 the flask. About 30 cc. of the water to be examined are intro- 
 duced into the flask, and the plug must be quickly replaced and 
 covered with a caoutchouc cap. If collected from a tap, the water 
 should first be allowed to run for a few minutes, and the sample 
 should -be received into the flask without the neck coming into contact 
 with the tap. From a reservoir or stream, the flasks may be filled 
 by employing a sterilised pipette. During transport contact between 
 the water and cotton-wool plug must be avoided, and if likely to 
 occur the sample must be collected and forwarded in a Sternberg's 
 bulb. 
 
 Examination by Plate-cultivation. The apparatus for plate- 
 cultivation should be arranged as already described. Crushed ice 
 
 FIG. 74. APPARATUS FOR ESTIMATING THE NUMBER OF COLONIES IN A 
 PLATE-CULTIVATION. 
 
 may be added to the water in the glass dish to expedite the setting 
 of the gelatine, so that the plate may be transferred as quickly 
 as possible to the damp chamber. The caoutchouc cap is removed 
 from the flask, and the cotton- wool plug singed in the flame to 
 prevent contamination from adventitious germs on the outside of 
 the plug. The flask is then held slantingly in the hand, and the 
 plug twisted out and retained between the fingers. With a 
 graduated pipette a measured quantity (y 1 ^ or -^ cc.) of the sample is 
 transferred to a tube of liquefied nutrient gelatine, and the plugs of 
 the flask and of the tube quickly replaced. If the water is very 
 impure, it may be necessary to first dilute the sample with sterilised 
 water. The inoculated tube must be gently inclined backwards and 
 forwards, and rolled as already explained, to distribute the germs 
 throughout the gelatine, and the gelatine finally poured 011 a plate. 
 When the gelatine has set, the plate is transferred to a damp 
 chamber, which should be carefully labelled and set aside in a place 
 
EXAMINATION OF AIR, SOIL, AND WATER. 147 
 
 of moderate temperature. In about two or three days the cultivation 
 may be examined. In some cases the colonies may be counted at 
 once ; more frequently they are so numerous that the plate must 
 be placed on a dark background, and a special process resorted to. 
 A glass plate, ruled by horizontal and vertical lines into centimetre 
 squares, some of which are again subdivided into ninths, is so 
 arranged on a wooden frame that it can cover the nutrient -gelatine 
 plate without touching it (Fig. 74). A lens is used to assist in dis- 
 covering minute colonies. If then the colonies are very numerous, 
 the number in some small division is counted, if less in some large 
 one, and an average is obtained from which the number of colonies 
 on the entire surface is calculated. A separate calculation of the 
 liquefied colonies should be also made, and their number, as well 
 as the total number of colonies present in 1 cc. of the sample, 
 recorded. Any peculiar macroscopical appearances, colours, etc., 
 should be noted, and then the microscopical appearances of the 
 colonies studied. Lastly, examination of the individual organisms 
 should be made by cover-glass preparations, and by inoculation of 
 nutrient gelatine, potatoes, and other media. 
 
 Instead of plates, Petri's dishes may be used both for gelatine 
 and agar-agar cultivations. 
 
 FIG. 75. ESM ARCH'S ROLL-CULTURE. 
 
 a., India-rubber caps; b b b, longitudinal line drawn on the tube; c,c,c, transverse 
 lines for counting colonies (FRANKLAXD). 
 
 Another plan is to take a measured quantity of the sample of 
 water and prepare a roll-culture, using a large-sized test-tube 
 (Fig. 75). The colonies can be counted with the aid of a lens (Fig. 76). 
 Microscopical preparations and sub-cultures can be made from the 
 colonies, and the anaerobic bacteria can be examined by Frankel's 
 modification of this method (p. 131). 
 
 A drop of the sample of water may also be added to liquefied 
 nutrient gelatine in a test-tube, the organisms distributed, and the 
 gelatine allowed to solidify in the tube. A rough comparison of 
 water samples may be made in this way. 
 
 Microscopic Examination. A drop of the water may be mounted 
 and examined without staining; or allowed to evaporate on a cover- 
 
148 BACTERIOLOGY. 
 
 glass, which is then passed through the flame, and stained in the 
 usual manner. 
 
 Parietti's Method. As typhoid fever bacilli are apt to be 
 crowded out by more rapidly growing micro-organisms, some method 
 had to be devised for restraining the growth of the latter, and 
 Chantemesse and Widal suggested the use of carbolic acid. Parietti 
 put this into practice by the method he introduced. This consists in 
 adding to tubes of broth about five drops of a mixture composed of 
 sterilised water (100 parts), hydrochloric acid (4 parts), and carbolic 
 acid (5 parts). The tubes are first tested by incubation, and are 
 then ready for use. A few drops of the suspected water are added 
 
 FIG. 76. APPARATUS FOR COUNTING COLONIES IN A KOLL CULTURE. 
 
 to the broth, and if it becomes turbid in a day or two the typhoid 
 fever bacillus is present in the form of a pure- culture. 
 
 An excess of bacteria in a fresh sample indicates an excess of 
 organic matter, and points to possible contamination with sewage. 
 Where there is such contamination we are very likely to find 
 pathogenic bacteria ; and moreover impure water is a constant 
 source of danger, for if the contagia of infectious diseases are 
 introduced they will retain their vitality in such water for a long 
 period, and will in some cases even multiply, whereas the same 
 organisms introduced into pure water would in a short time perish. 
 
 The actual number of bacteria in water is not of very great 
 importance, and it must be remembered that if a sample is set aside 
 for a few days there will be an enormous increase in the number 
 of bacteria present ; but in dealing with perfectly fresh samples it 
 
EXAMINATION OF AIR, SOIL, AND WATER. 149 
 
 may be said that water containing less than 100 bacteria to the 
 cubic centimetre is very pure water. Water containing 1,000 or 
 more should be filtered. Water containing 100,000 to 1,000,000 
 is contaminated with surface water or sewage. It is necessary to 
 bear in mind that in typhoid fever and Asiatic cholera the excreta 
 contain the bacteria in great numbers, and wells and streams 
 receiving surface water may be contaminated in various ways. 
 The cholera bacillus dies as a rule quickly in distilled water, while 
 it preserves its vitality for a long time in water of a bad quality. 
 
 It is necessary to lay stress upon the fact that a bacterio- 
 logical analysis may show the presence of pathogenic bacteria when 
 their detection is not possible by any other means. They may 
 be present in water in such small numbers that no chemical 
 analysis would detect any contamination, but as they are living 
 organisms capable of increasing in a suitable environment, they can 
 readily be discovered by bacteriological methods. 
 
 The examination of rain water, drinking water, tap water, sea 
 water, various liquids and infusions, by these methods, opens up 
 a wide field for research. Pettenkofer has shown that impregna- 
 tion of water containing many bacteria with carbonic acid diminishes 
 the number of the latter. The examination of waters before and 
 after filtration, or after addition of chemical substances, are matters 
 which require further investigation, though a great deal of work has 
 already been accomplished. The reader will find in Micro- organisms 
 in Water by P. and G. Frankland, a very complete account of this 
 subject with valuable analytical tables. 
 
CHAPTER XII. 
 
 PHOTOGRAPHY OF BACTERIA. 
 
 THE production of pictures of microscopic objects by photographic 
 means was attempted at an early date. Some authorities regard 
 the very earliest recorded experiments as being the first experi- 
 ments alike in photography and micro-photography. The experi- 
 ments of Wedgwood and Sir Humphry Davy were embodied in 
 a paper read before the Royal Institution in 1802. They obtained 
 with the solar microscope impressions upon paper, and with greater 
 success upon white leather, though the results were transitory when 
 exposed to daylight. 
 
 In 1816 Nicephore Niepce described his experiments in con- 
 nection with fixing the image obtained by the camera. He was 
 at first only able to obtain negatives, and these were transitory. 
 But, after joining with Daguerre, who had been experimenting 
 in the same direction, a process was invented which was published 
 in 1839 under the name of daguerreotype. 
 
 This invention, and the rapid improvements which followed, 
 were taken advantage of by Reade, Donne, Hodgson, Kingsley, and 
 Talbot, who were early workers in the field of micro-photography. 
 
 So early as 1845 it is stated that Donne produced a work 
 illustrated with engravings copied from daguerreotypes. 
 
 Subsequently this interesting branch of photography was taken 
 up by many in France and Germany, in America, and in England. 
 Of those to whom we are indebted for the literature of the subject, 
 and for many improvements, the names of Wenham, Dancer, 
 Draper, Maddox, Shadbolt, Redmayne, Woodward, Highley, Deecke; 
 Moitessier, Gerlach, Koch, Sternberg, Frankel, Pfeiffer, and Pringle 
 may especially be mentioned. 
 
 Of these workers the name of Woodward stands pre-eminently 
 foremost. His skill in microscopical manipulations, combined 
 with access to the very best apparatus and objectives, placed at 
 
 150 
 
PHOTOGRAPHY OF BACTERIA. 151 
 
 his disposal in the Museum at Washington, enabled him to obtain 
 photographs of diatoms which probably have never been surpassed. 
 
 To Koch belongs the credit of being the first to extend the 
 application of micro-photography to the delineation of bacteria. 
 A .-erifs of instructive photographs was first published by him 
 in 1877. These were photographs of cover-glass preparations, 
 and all admirably illustrated the subjects from which they were 
 taken ; while two, showing the flagella of bacilli and spirilla, 
 were triumphs in this new departure. 
 
 Lewis, in India, was one of the first to illustrate his writings on 
 the subject of micro-organisms by means of photographs. 
 
 About the same time Sternberg, in America, took some excellent 
 photographs of bacteria. Heliotype reproductions of these were 
 published in 1884. 
 
 Ha user and Van Ermengem and many other bacteriologists 
 successfully resorted to photography for illustrating their researches, 
 and Frankei and Pfeiffer's, and Itzerott and Niemann's atlases of 
 photographs of bacteria, in microscopical specimens and cultivations, 
 are especially worthy of mention. 
 
 Opinions have differed widely as to the merits of photographic 
 illustrations. Many, taking the standpoint solely of a comparison 
 with drawings, have decried their use. By judging from such a 
 comparison alone the real value of photographs may be lost sight 
 of. On the other hand, many who have looked at the question 
 from all sides, have been led to value even a defective photograph 
 more than an ordinary drawing. 
 
 In his first publication on this subject, Koch strongly advocated 
 photography on the ground that illustrations would then be as true 
 to nature as possible. The photographs which accompanied his 
 paper were all taken from preparations of bacteria which had 
 been made from blood, cultivations, or infusions, by drying a 
 thin layer on a cover-glass and staining, or from specimens prepared 
 in the same way but left unstained. But when, having committed 
 himself to this opinion, Koch attempted, later, to photograph the 
 bacteria in animal tissues, he was led to modify his previous 
 conclusion. For though no trouble was spared, yet disappointing 
 results were met with. This was owing, he explains, to the fact 
 that the smallest 'and most interesting bacteria can only be made 
 visible in animal tissues by staining them, and thus obtaining the 
 advantage of colour. 
 
 This introduced the same difficulties which are met with in 
 photographing coloured objects, such as tapestry and oil paintings. 
 
152 BACTERIOLOGY. 
 
 As these difficulties had been to a certain extent obviated by the 
 use of eosin-collodion, Koch adopted the same method for photo- 
 graphing stained bacteria. By the use of eosin-collodion, and by 
 shutting off portions of the spectrum by coloured glasses, he 
 succeeded in obtaining photographs of bacteria which had been 
 stained with blue and red aniline dyes. But, owing to the long 
 exposure which was necessary, and the unavoidable vibrations of 
 the apparatus, the results were so wanting in definition that they 
 not only proved unsatisfactory as substitutes for drawings, but did 
 not in some cases give any evidence of what was to be seen in the 
 preparations. 
 
 Koch, in consequence, stated that he would abstain from 
 publishing photographic illustrations until he had the advantage 
 of improved methods. 
 
 We find, however, in spite of this, that in 1881 Koch published 
 a, series of reproductions from his negatives in illustration of what 
 could be accomplished by photography. 
 
 Here again we find that many of the photographs of cover- 
 glass preparations were admirable, but those of tissue-sections gave 
 evidence of the difficulties Koch encountered, and were undoubtedly 
 unsatisfactory from the want of flatness of field, some of the 
 illustrations recalling rather a map of a mountainous country than 
 a microscopical preparation. 
 
 In consequence of the difficulties met with in attempting to 
 photograph bacteria stained with the aniline dyes most commonly 
 used, Koch recommended that the preparations should be stained 
 brown, pointing out as his reason that, though the bright and 
 concentrated colour of the red and blue aniline dyes catches the 
 eye far more readily than the somewhat sombre brown colours, 
 yet no one up to the time of his publication had succeeded in 
 obtaining good photographs of bacteria which had been stained 
 either blue or red, and mounted in Canada balsam, while there was 
 no difficulty in obtaining photographic representations of prepara- 
 tions stained yellow or brown. 
 
 Though this stain could be easily employed in most cover-glass 
 preparations, it was by no means easy to obtain a good differential 
 stain of bacteria in the tissues by employing Bismarck brown. 
 An attempt was, therefore, made to photograph preparations 
 stained blue and red by the aid of the dry-plate process, and by 
 interposing glasses of suitable tints. After many fruitless experi- 
 ments this method had to be abandoned, and the method of staining 
 the object brown was adopted. In many cases this gave excellent 
 
PHOTOGRAPHY OF BACTERIA. 153 
 
 results ; in others again, compared with the results of staining 
 with blue or red stains, there was much to be desired, and further 
 improvement was needed. 
 
 That a stain, such as yellow or brown, must be employed which 
 absorbs the blue rays, and acts on the sensitive plate like black, 
 which absorbs all the light, constituted the first condition laid 
 down by Koch as an essential for success. It was further pointed 
 out that the suitability of the stain could be ascertained by first 
 passing the light, to illuminate the preparation, through a solution 
 of ammonio-sulphate of copper, under which condition the bacteria 
 would appear black on a blue ground. 
 
 The second condition was, that sunlight must be employed, but 
 that direct projection upon the object was disadvantageous, and it 
 must, therefore, be diffused by the interposition of one or more plates 
 of ground glass. 
 
 Lastly, an illuminating condenser was recommended, of such 
 construction that the diffused sunlight brightly illuminates the object 
 from all sides. 
 
 Sternberg encountered the same difficulty in photographing red, 
 blue or violet preparations, while he produced excellent pictures of 
 preparations stained with aniline brown, or a weak solution of iodine 
 (iodine grs. iij, potassic iodide grs. v, distilled water grs. 200). Thus 
 the results of a large number of attempts to photograph the tubercle 
 bacillus in sputum, only ended in producing such extremely faint 
 impressions, that any one unacquainted with the object as seen under 
 the microscope could form scarcely any idea of its form or minute 
 structure with even an accompanying explanation and the closest 
 inspection of the photograph. 
 
 Dufrenne, in attempting to photograph the same object by the 
 ordinary method, found the plates were uniformly acted on, or the 
 image was so faint, or so lacking in contrast, that they were useless 
 for obtaining proofs on paper or glass. By interposing green glass 
 between the objective and the sensitive plate, so that the red rays 
 were absorbed, while the green rays passed through and acted on 
 the plate, he states that better results were obtained. 
 
 The work of Hauser illustrated the great value of photography 
 in the production of pictures of impression-preparations and colonies 
 in nutrient gelatine. To give the general effect, as well as faithfully 
 reproduce the minute details in these difficult subjects would in most 
 cases create insurmountable difficulties, except to the most accom- 
 plished draughtsman. 
 
 Hauser employed Gerlach's apparatus and Schleussner's dry 
 
154 BACTERIOLOGY. 
 
 plates, and obtained the illumination by means of a small incan- 
 descent lamp, which gave a strong, white light, with three or 
 four Bunsen elements. In another respect Hauser's results were 
 of practical value. The preparations to be photographed were 
 stained brown as recommended by Koch, but they were mounted in 
 the ordinary way in Canada balsam. The objection to the mounting 
 medium most commonly employed was thus set aside. The prevalent 
 idea, however, that the preparations must be stained brown was still 
 a formidable obstacle, and the way out of this difficulty was clearly 
 shown by Yan Ermengem's photographs. These were pictures of 
 comma-bacilli which had been stained with fuchsine and methyl 
 violet. These photographs afforded the first practical illustration 
 of the value of isochromatic plates in micro-photography which had 
 been previously noted by Yan Ermengem in 1884, and their intro- 
 duction marks a distinct era in the progress of micro- photography. 
 
 A short explanation may be given of what is meant by isochro- 
 matic, or what have been more properly termed orthochromatic dry 
 plates. The difficulties encountered in photographing certain stained 
 preparations have been mentioned. It is a familiar fact that in 
 portraits, blue or violet comes out almost or quite white, while 
 other colours, such as yellow, are represented by a sombre shade 
 or perhaps black. This failure in correctly translating colours is 
 explained by the want of equality between the strength of the 
 chemical and luminous rays. If the rays of the spectrum are pro- 
 jected upon a photographically sensitive surface, the greatest effect 
 is found to take place at the violet end. In other words, the violet 
 and blue rays are more actinic or chemically powerful, while the 
 yellow and orange have scarcely any effect. The dyes employed in 
 staining give corresponding results : blue and violet give but a faint 
 impression, yellow and orange a black picture. These results are 
 most clearly demonstrated in a photograph of an oil painting taken 
 in the ordinary way ; and they led to experiments being made which 
 have resulted in orthochromatic photography. 
 
 The effect of interposing coloured glasses has already been 
 referred to. It was found later that, if plates were coloured yellow, 
 e.g., with turmeric, the blue and violet rays were intercepted, and 
 their actinism reduced. In 1881, Tailfer and Clayton produced the 
 so-called isochromatic plates. The emulsion of bromide of silver 
 and gelatine was stained with eosin, and it was claimed that colours 
 would be represented with their true relative intensity. Chlorophyll 
 and other stains have been tried, and by such methods the ordinary 
 gelatine dry plates can be so treated that they will reproduce 
 
PHOTOGRAPHY OF BACTERIA. 155 
 
 various colours, according to their relative light intensity, and thus 
 be rendered iso- or, what is now more commonly known as, ortho- 
 chroma ti-.-. 
 
 APPARATUS AND MATERIAL. 
 
 Micro-photographic Apparatus. As is well known, various forms 
 of apparatus have from time to time been recommended and em- 
 ployed by different workers. 
 
 Many use the microscope in a vertical position, with the camera 
 superposed or fitted to the eye-piece end of the microscope tube ; 
 or the microscope tube may be screwed off from the body of the 
 microscope, and a pyramidal camera, adjusted in its place, the base of 
 the pyramid being represented by the ground glass screen. 
 
 Others again prefer that the microscope and camera should be 
 arranged horizontally. 
 
 In another form the ordinary microscope is dispensed with, the 
 objective, stage, and mirror are adapted to the front of the camera, 
 and provided with suitable arrangements for holding the object, 
 supporting the mirror, and adjusting the different parts. 
 
 Lastly, the camera may be dispensed with, the operating-room, 
 which must be rendered impervious to light, taking its place, while 
 the image is projected and focussed upon a ground glass screen, 
 winch has a separate support.* 
 
 The horizontal position affords greater stability than the vertical, 
 so that the former is to be preferred. The vertical model with the 
 camera fixed to the microscope is particularly to be avoided, as the 
 weight of the camera bears directly upon the microscope, and must 
 affect the fine adjustment, and any vibration in one part of the 
 apparatus is communicated throughout. 
 
 The simplest apparatus consists of a camera fixed upon a base- 
 board four or five feet in length, upon which the microscope can be 
 clamped, and which carries also a lamp and a bull's-eye condenser 
 (Fig. 77). 
 
 Simplicity and economy must always be borne in mind in 
 recommending any apparatus of this kind, for to insist upon the 
 necessity of a very elaborate apparatus, or a specially fitted- up room, 
 or that a special room should be built with windows facing in a 
 definite direction, will in most cases at once place photography beyond 
 the reach of those who might otherwise employ it. Yet to fulfil 
 
 * For an excellent account of the forms of apparatus which have been 
 employed by different workers the reader is referred to the section on Micro- 
 photography in Beale's Horn tn icork with the Microx< 
 
156 
 
 BACTERIOLOGY. 
 
 all the purposes for which the apparatus may be required, including 
 the employment of the highest powers, and also that one may be 
 enabled to work for long intervals of time with due comfort, an 
 accurate and complete apparatus will be found to be most desirable. 
 Though most preparations will admit of being photographed 
 when the stage of the microscope is vertical, yet if we require to 
 photograph micro-organisms in liquids, or colonies upon partially 
 liquefied gelatine, the apparatus must admit of being placed so that 
 the stage of the microscope becomes horizontal. In addition, the 
 apparatus is rendered somewhat complex if we employ powerful 
 
 FIG. 77. HORIZONTAL MICRO-PHOTOGRAPHIC APPARATUS. 
 
 artificial light. Sunlight, no doubt, is the best and cheapest, but 
 it is not always available, especially in a city like London ; and, 
 moreover, evenings and dull days will probably be the very time 
 which can be best spared for this work. We must, therefore, fall 
 back upon the paraffine lamp, or the magnesium, oxyhydrogen, or 
 electric light. 
 
 To fulfil all these conditions Swift has constructed an apparatus 
 under the author's directions (Fig. 78). It is merely a modification 
 of the ordinary horizontal model, which admits of being readily placed 
 
PHOTOGRAPHY OF BACTERIA. 
 
 157 
 
 in the vertical position, while the illumination is supplied from an 
 oxyhydrogen lantern. 
 
 To place the apparatus in the vertical position two small hinged 
 
 brackets, at the end distant from the camera, are forced up with 
 a smart blow of the hand. The corresponding ends of the stretcher 
 bars are dislodged from their fittings, and allowed to descend ; when 
 
158 
 
 BACTERIOLOGY. 
 
 horizontal, the opposite extremities of the bars are easily released 
 from their sockets. The leg or support at this end can then be 
 
 FIG. 79. REVERSIBLE MICRO-PHOTOGRAPHIC APPARATUS ARRANGED IN THE 
 VERTICAL POSITION. 
 
 turned up and fixed underneath the apparatus by a button, and 
 the end of the apparatus itself gently lowered to the ground. 
 
PHOTOGRAPHY OF BACTERIA. 159 
 
 A hinged end- piece is also to be turned out to increase the base upon 
 which the whole apparatus will stand when raised to the vertical. 
 The two-legged support at the opposite end of the apparatus is 
 next worked down by a quick thread screw, and on raising the 
 apparatus to the vertical, the two-legged support drops to the 
 ground, and assists in maintaining the stability of the whole. If 
 it is thought necessary, a simple means can be readily devised for 
 clamping the apparatus, in either position, to the wall of the room, 
 so as to eliminate as much as possible all chances of vibration. A 
 second quick thread screw moves the base-board upon which the 
 camera and central sliding-board are mounted, so that the camera, 
 microscope and lantern can be raised to a convenient height from 
 the ground. 
 
 The various parts of this apparatus may be described in 
 detail. 
 
 The Microscope and its Attachments. It is most essential that 
 the microscope should be perfectly steady. The microscope was made 
 by Zeiss, and to ensure steadiness, the horse- shoe foot piece fits under 
 a projecting ledge, and is then clamped by a cross-piece, so that 
 it is firmly fixed. 
 
 The microscope with the means for clamping it and the oxy- 
 hydrogen lantern are carried upon an independent sliding-board, 
 which admits of movement to or from the camera. The sliding-board 
 also moves upon a centre, which enables the microscope to be turned 
 out from the median line ; in fact, to be turned at a right angle to the 
 position it occupies when ready for the exposure. The object of this 
 contrivance is to enable the operator to sit down by the side of the 
 apparatus, and with comfort to arrange the object in the field of the 
 microscope. On turning the microscope back into the median line, it 
 is fixed in the optical axis of the apparatus by means of a suitable 
 stop. The sliding-board is provided w r ith a small grooved wheel 
 receiving an endless, cord, made of silk or fishing-line, which passes 
 'round the grooved, milled head of the fine adjustment of the 
 microscope. When the sliding-board is returned to the median 
 line of the apparatus, the milled wheel connected with the fine 
 adjustment impinges upon the wheel of the long focussing rod. 
 The latter is provided with an india-rubber tire, which grips the 
 teeth of the milled wheel, and thus the long focussing rod is placed 
 in connection with the fine adjustment of the microscope. 
 
 Illumination. The oxy-hydrogen lamp has been more frequently 
 employed by the author than the paraffine lamp, partly on account 
 of the diminished time in exposure, especially when employing very 
 
160 
 
 BACTERIOLOGY. 
 
PHOTOGRAPHY OF BACTERIA. 161 
 
 high powers ; this is of great importance where there is likely to be 
 vibration from passing traffic. With rapid plates and the highest 
 powers, the exposure has only been two or three seconds, whereas 
 with the parafline lamp it may vary from three to ten minutes, or 
 even longer. 
 
 Walmsley gives the following table for exposures with the 
 par a (fine lamp : 
 
 1| inch objective . . . . 3 to 45 seconds. 
 | - 7 90 
 
 iV . \ 3 minutes. 
 
 i 97 
 
 5- ., & ., < , 
 
 i 4-10 
 
 TO" " " " 
 
 The illuminating apparatus represented in the accompanying 
 engraving (Fig. 78) consists of a lantern which not only moves 
 together with the microscope on the central sliding-board, but 
 can be moved independently to or from the microscope, and be 
 clamped with screws at the requisite distance for obtaining the best 
 illumination. It is provided with two 3-inch condensing lenses of 
 long focus, constructed of optical glass, which is much whiter than 
 that used for ordinary lantern condensers. The lime-cylinders should 
 be of the hardest and best quality, as they give a more actinic light 
 than those made of soft lime. The " Excelsior " lime-cylinders are 
 strongly recommended. They are supplied in hermetically sealed 
 tins which can be easily opened and re-sealed, so that a cylinder can 
 be taken out and used, and the rest preserved for a future occasion. 
 The hydrogen can be obtained by using the coal-gas supplied to the 
 house, and the oxygen should be supplied preferably in a compressed 
 state in iron bottles. Not only are the bottles much less cumbrous 
 than the bags, but a small quantity of gas can be used, and the 
 residue left for an indefinite time ; moreover, the gas is always 
 at hand to be turned on when required. On the other hand, the 
 ivtrntion of unused gas in bags is liable to cause their corrosion, 
 owing, it is believed, to impurities which are carried over in the 
 manufacture of the oxygen. If gas is not laid on in the house, then 
 it also must be procured in a compressed state in bottles. As the 
 blow-over jet is recommended on account of its safety, the bottles 
 should be supplied in this case with a supplementary valve. It is 
 then just as easy and free from danger to employ the compressed 
 ua it is to make use of the house-supply. 
 
 The Camera. A long-focus, half-plate camera is mounted upon 
 a sliding platform. This admits of the camera being pushed up to 
 
 11 
 
162 
 
 BACTERIOLOGY. 
 
 the microscope when it is in the long axis of the apparatus, so as to 
 make a light-tight combination. The opening which is filled in an 
 ordinary camera by the lens can be shut off by means of an internal 
 shutter, which is opened and closed by turning a screw at the side 
 of the camera. The dark-back is provided with plate-carriers, so 
 that either half, quarter, or lantern-size plates can be employed. It 
 will be found convenient to have two or more dark- backs, so that 
 several plates may be exposed without rearranging the light for 
 each exposure. 
 
 Much more elaborate and expensive micro-photographic cameras 
 have been constructed by Zeiss, and also by Swift. The latter has 
 
 FIG. 81. PHOTOGRAPH OF AN IMPRESSION PREPARATION. 
 
 carried out a suggestion made by Pringle for a support at the 
 ocular end (Fig. 80). 
 
 The Dark-room. In every bacteriological laboratory there should 
 be a developing room provided with shelves, gas, water-tap, and sink, 
 but these arrangements are not absolutely indispensable. All that 
 is essential is a room impervious to light ; and a closet or cupboard, 
 if it can be ventilated, will answer perfectly well, with a jug and 
 basin instead of the tap and sink. The steam -steriliser employed 
 in the preparation of nutrient media for cultivating bacteria, if not 
 required at the time for such purposes, may be filled to the brim 
 with water, and will form an excellent cistern and tap, while a pail, 
 or small sanitary bin, may be utilised as a sink. 
 
 Various kinds of lamps are made for the dark-room, burning 
 
PHOTOGRAPHY OF BACTERIA. 163 
 
 either candles, oil, or gas. In any case, the light must pass through 
 t\vo thicknesses of ruby gla>-. 
 
 />/>/ Plates. A small supply of any of the ordinary plates in 
 the market may be procured for preliminary trials in acquiring a 
 knowledge of the processes ; but to overcome the difficulties of certain 
 stained preparations, the isochromatic or orthochromatic plates should 
 be used. The J plate will be found to be the most suitable size. 
 
 There are numerous formulae for the requisite solutions for 
 developing and fixing the negatives, and instructions are usually 
 enclosed in the boxes of dry plates, but it is best to abstain from 
 trying a number of different formulae, as it leads to a great expendi- 
 ture of time. There is a temptation to do this, it being supposed 
 that there is probably some great advantage in one formula over 
 another. It is much better to get accustomed to the behaviour 
 under different exposures of one, or perhaps two methods. 
 
 In France the iron developer is much in vogue, and is recom- 
 mended by Tailfer and Clayton for use with their isochromatic 
 plates. It has the advantage of great simplicity in the mode of 
 employment, and, therefore, is very suitable for a beginner. In 
 England, on the other hand, the alkaline developer is very 
 much used, as it gives more command over the plate, enabling the 
 photographer more fully to compensate for incorrect exposure. 
 
 It is very desirable before attempting to take photographs with 
 the microscope to learn how to take photographs with an ordinary 
 landscape camera, and to get thoroughly accustomed to the use of 
 some good developer, so that mistakes may be corrected and the 
 clearest and sharpest negatives obtained. 
 
 PRACTICAL MANIPULATION. 
 
 Arrangement of Apparatus. For working with the paraffine 
 lamp, the mode of procedure is, as regards the illumination, briefly 
 ,i- follows. The sub-stage condenser is dispensed with when a 
 low power is employed, as well as the mirror, and the lamp is 
 so placed that the image of the flat of the flame appears accurately 
 in the centre of the field of the microscope. A bull's-eye condenser 
 i> then interposed, ><> that the image of the flame disappears, and 
 the whole field is equally illuminated. With high powers the 
 sub-star*' achromatic condenser is necessary, and a more intense 
 illumination is obtained by using the flame edgewise. In using a 
 low power with the oxyhydrogen light, the lantern is withdrawn 
 -cine little distance from the microscope, and the top combination 
 of the achromatic condenser removed. 
 
1 64 BACTERIOLOGY. 
 
 It is best to begin with the use of a low power, and a trial 
 object, such as the blow-fly's tongue, spine of Echinus, or trachea 
 of silkworm. 
 
 In order to explain the management of the apparatus (as 
 represented in Fig. 78) the steps in the arrangement of the 
 apparatus and exposure of the plate will be described in detail 
 for the employment of a high power and the oxyhydrogen light. 
 The solutions being ready for use, it is proposed to take a photograph 
 of tubercle bacilli in sputum, with a % apochromatic oil-immersion 
 objective. The first point to claim attention is the arrangement 
 of the light. Having lighted the gas at the hydrogen jet, the 
 lime- cylinder should be revolved until heated equally all round. 
 The oxygen is then carefully turned on until only a small spot 
 of incandescence is produced. The central sliding-board is turned 
 out, a low power screwed on to the microscope, and the image of 
 the bright spot focussed and accurately centered. To protect the 
 sight, an eye-piece provided with a smoked glass shade is used. 
 The immersion objective is then substituted for the low power, 
 and th% oxygen turned on until the right admixture of gas is 
 obtained to produce a brilliant illumination. It is well at this 
 stage to sit down to focus the selected object, and to spend some 
 little time in searching for the most characteristic part of the 
 specimen to be photographed. This being decided upon, the eye- 
 piece is carefully withdrawn, and the central sliding-board rotated 
 back into the median line. To make a light-tight connection 
 between the camera and the microscope, the camera is pushed up 
 until a velvet-lined tube, which occupies the position of the lenses 
 of an ordinary camera, is enclosed within a short wide tube which 
 is adapted to the eye-piec3 end of the microscope. 
 
 On opening the camera-shutter the image will be projected upon 
 the ground glass screen of the camera. It is necessary, however, 
 to obtain the exact focus, and to effect this the ground-glass 
 screen is turned away, and the dark-back with a piece of plain 
 glass is substituted. Here again time may be well spent in 
 getting the sharpest image, with the aid of a focussing glass of 
 proper focal length. 
 
 The greatest delicacy in manipulation is necessary, as in working 
 with such high powers a turn too much of the fine adjustment will 
 cause the image to vanish. Having determined the best visual 
 focus, which will be found with the high-power objectives of most 
 makers to correspond with the chemical focus, the dark-back must 
 be cautiously removed, to prevent any vibration, and the plain 
 
PHOTOGRAPHY OF BACTERIA. 165 
 
 i. r la>s replaced by a sensitive plate. To effect this change, the 
 operator retires to the dark-room, and opens a box of plates with 
 as little exposure to the red light as possible. Having removed 
 a plate, it is netv>sary to ascertain which is the sensitive side. 
 This may be done by momentarily exposing it to the red light, 
 and seeing which is the sensitive side by the dull appearance of 
 the film. A less satisfactory way is to moisten the tip of a 
 finger, and press it at one corner of the plate. The film side will 
 be recognised by imparting a sticky sensation. The film must be 
 lusted with a camel's-hair brush, as well as the dark-back, and 
 the plate is placed film-side downwards in the dark-back, which 
 is then securely closed. 
 
 Care should be taken that the plates then remaining in the box 
 are packed away before light is admitted to the dark-room. 
 
 Exposure of the Plate. On returning to the apparatus, the 
 camera-shutter is closed. Then the dark-back is gently slid into 
 its place, and its slide withdrawn. A few moments are allowed 
 to elapse, so that the least possible vibration, which might be 
 caused by inserting the dark-back, has had time to cease, and all 
 is ready for the exposure. 
 
 In the case of the object we have selected, three seconds will 
 probably be the exposure required. This is done by opening and 
 closing the camera -shutter with one hand, while a watch can be 
 held in the other. The slide of the dark- back is then carefully 
 closed, and the plate is ready to be carried off to the developing 
 room. 
 
 As the light will not be again required until the next exposure, 
 the oxygen must be turned off, while the coal-gas may be allowed 
 to play over the lime. 
 
 Development and Fixation of the Image. It is well to be 
 >\>tematic, and therefore, before the plate is taken out of the 
 dark-back, light is admitted to the dark-room, and everything 
 arranged so that the position of the trays and bottles may be 
 remembered in the dark. First, let the ruby lamp be lit, place 
 two dishes or trays close by, and a row of four dishes within easy 
 reach. Pour out some fixing solution in the first porcelain dish, 
 alum in No. 2, and water in Nos. 3 and 4. Put the necessary 
 quantity of " pyro " solution into the glass measure, and place 
 it with the ammonia drop-bottle in front of the ruby light. 
 Then, when all light except that from the ruby lantern has been 
 excluded, everything is ready to commence the development of the 
 plate. 
 
166 BACTERIOLOGY. 
 
 Opening the dark-back, the plate may be turned out on to the 
 palm of the hand. The film side is then uppermost, and the plate is 
 to be transferred in the same position to a tray, and covered with 
 water. This is to soak the film and obtain an equal action of 
 the developer ; or the solution of fresh pyro may be poured on to 
 the plate without previous soaking, if the flow is uniform, and the 
 formation of bubbles avoided. In the first case the water is run off 
 and the pyro allowed to flow evenly over the plate. To protect the 
 plate from prolonged exposure to the ruby light, a second tray may 
 be inverted over it, or the developing tray covered with a piece 
 of card-board. Gently rock the tray for a minute or so, then 
 to a few drops of ammonia in a measuring glass add the pyro 
 from the developing tray, and pour the mixture back again 
 over the plate. After again gently rocking the tray for a few 
 minutes, more ammonia is added by drops in the same way. 
 If the exposure has been properly timed, and the time necessary 
 must be ascertained by trial for each preparation, the image will 
 gradually begin to appear, and the action must be allowed to 
 continue until sufficient density has been obtained. To determine 
 this requires some experience. It is generally recommended to take 
 the plate out of the tray and hold it for a moment, film-side towards 
 the operator, in front of the ruby light. Though the plate is not 
 nearly so sensitive when the image has commenced to develop, and 
 there is, therefore, not the same danger of fogging, a safer plan is 
 to occasionally turn the plate film downwards in the tray, and when 
 the image appears on the back the development will be found to be 
 completed. 
 
 With such a preparation as tubercle bacilli in sputum it is 
 not easy to trace the gradual formation of the image, and hence the 
 advantage of commencing with a well-marked object such as the 
 blow- fly's tongue. It is then easy to watch the gradual progress of 
 the image. The. bright parts or high-lights appear first, then 
 gradually the half-tones, or less brightly-lighted parts, and lastly 
 every shade except the deepest shadows is represented. When, 
 however, all action seems to have ceased, we must still wait until 
 we have judged, in the manner already described, that the density 
 is sufficient. This being determined, we pour off the developing 
 solution and thoroughly wash the plate with water. It is then 
 ready to be placed in dish No. 1, containing " hypo," and here it must 
 be left for some minutes after all appearance of creaminess has 
 disappeared from the back. White light may now be admitted, the 
 plate removed from the hypo and thoroughly washed under the tap, 
 
PHOTOGRAPHY OF BACTERIA. 167 
 
 and then placed in dish No. 2. When another plate is ready to 
 take its place, transfer it to dish No. 3, and then to No. 4, and, 
 after a good final washing under the tap, place it upon a rack 
 to dry. If there is any tendency for the film to detach itself from 
 the plate, " to frill," the alum bath must be used before fixing, as 
 well as after. 
 
 Frilli/Hj or blistering may be due to an error in manufacture, 
 and is liable to occur in hot weather, or when using a developer too 
 strong in ammonia. If it occurs during washing or fixing, the alum 
 bath must be employed before the hypo. Fogging, or the appear- 
 ance of a veil over the plate, may arise from error in the manu- 
 facture, from admission of extraneous light, from over-exposure, 
 or from prolonged exposure to the ruby light during development. 
 Care must be taken that the camera and dark-room are light-tight. 
 Crystallisation, or powdery deposit, upon the negative when dry, is 
 due to insufficient washing out of the hyposulphite of soda. Thin- 
 ness of the image, or ivant of density, may be due to insufficient 
 development, too weak a developer, or too short or too long an 
 exposure. Too great density results from too long immersion in the 
 developer. 
 
 Spots may sometimes occur upon the negatives. They may be 
 caused by dust upon the plate or by air bubbles in the developer. 
 
 In the text-books of photography full accounts of failures will be 
 found, their causes and prevention ; but it will be advantageous 
 when these difficulties are encountered to take the negatives to a 
 skilled photographer and get advice upon them. It is necessary 
 to persevere, and not be disheartened if several negatives have to 
 be made of a preparation before a successful result is obtained. 
 
 It may here be remarked that the beginner will far more 
 rapidly learn the technique if he avail himself of a practical demon- 
 stration from a photographer. When he has learnt to obtain suc- 
 < ful negatives, if he prefer silver prints, and time is an object, it 
 will be found to be true economy to get the printing and mounting 
 done by a professional photographer. The credit of a successful 
 photograph of bacteria is due to the bacteriologist who prepares 
 the microscopical specimen and obtains the negative. 
 
 Determination of the Amplification. The amplification varies 
 not only with the objective employed, but with the distance of 
 the focussing screen from the object. In order to ascertain the 
 amplification afforded by a certain objective at a certain distance, a 
 photograph should be taken, under the same conditions, of the lines of 
 a micrometer slide. It is easy then to calculate the amplification 
 
168 
 
 BACTERIOLOGY. 
 
 obtained in the micro-photograph ; supposing, for example, in 
 the micro- photograph the lines which are 1Ty Vo" " 1C ^ a P ar * are 
 delineated 1 inch apart, the magnifying power must be 1,000 
 diameters. Moreover, having thus ascertained the amplification, 
 we can accurately compute from the photograph the size of the 
 objects taken. 
 
 Value of Photographs. It is not necessary to compare the 
 relative merits of diagrams and photographs. Diagrams which do 
 not purport to be accurate representations, but are 
 intentionally the means for simplifying instruction, 
 will always be valuable, even if we have the 
 original preparations under the microscope before 
 us. We must consider the relative merits of 
 photographs and of drawings which purport to be 
 exact representations of what is seen under the 
 microscope. Thus in the case of micro-organisms, 
 when their biological characters are studied under 
 low powers of the microscope, photographs are 
 preferable because they give a more faithful re- 
 presentation. At the same time, apart from this 
 comparative value, we must not lose sight of the 
 actual value of photography in placing within the 
 reach of the student or investigator, who may not 
 be a draughtsman, a most valuable means for 
 illustrating all kinds of preparations. 
 
 For double-stained or triple-stained tissue pre- 
 parations an accurately coloured drawing leaves 
 OF little to be desired ; but if we reproduce the same 
 N " by a wood engraving, and so lose the advantage 
 of the coloured picture, which is instructive in 
 indicating the method of staining, a photograph will, in many cases, 
 be far more satisfactory. 
 
 When we have to deal with the growth of bacteria en masse, 
 as in test-tube and plate- cultivations, with colonies as seen under 
 a low power of the microscope, and with impression-prepara- 
 tions both under low and high powers, unless the bacteriologist 
 is a most accomplished draughtsman as well as an accurate and 
 reliable observer, photography undoubtedly affords the best mode 
 of illustration. The apparatus being ready and at hand, a negative 
 can be produced in a few minutes of a preparation which, from the 
 amount of detail it contains, would take perhaps several hours to 
 draw and colour. From that negative any number of facsimiles can 
 
 FIG. 82. PHOTO- 
 GRAPH OF 
 CULTIVATION 
 BACILLUS 
 
 THRACIS. 
 
PHOTOGRAPHY OF BACTERIA. 169 
 
 be obtained, whereas an original drawing, even in the best hands, 
 if cut on wood or lithographed, is almost certain to fall short of 
 lieing an exact copy. 
 
 With regard to individual bacteria, the result is more satisfactory 
 in many cases than a drawing ; for there is the advantage of being 
 absolutely certain that any particular structure, form, or shape 
 win rli may be represented is actually what exists, and not what 
 may have been evolved by unconscious bias in the mind of the 
 observer. Many illustrations might be given of this. Thus Lewis, 
 who was a most conscientious observer, published an account of 
 organisms in the blood of rats in India, and illustrated it with a 
 wood engraving and with micro-photographs. The identity of the 
 organisms which were found in the common brown rat of this 
 country was established much more readily from these photographs 
 than from the wood engraving or the description in the letterpress. 
 
 A micro-organism, even under the highest powers, appears as 
 so minute an object that to represent it in a drawing requires a 
 very delicate touch, and it is only too easy to make a picture which 
 gives an erroneous impression to those who have not seen the 
 original. If, on the other hand, to represent the object more 
 clearly we draw an enlarged picture, we can only do so by repre- 
 senting what we think the object would be like if it could be 
 amplified to the size represented. In such cases a photographic 
 enlargement is certainly more valuable. 
 
 Photography enables us also to record rapid changes, and it 
 is possible that as the art advances we may find that the film is 
 more sensitive than the human retina, and brings out details in 
 bacteria which would be otherwise unseen. 
 
 Photographs can be readily transmitted by post, and when we 
 can neither make a great number of preparations to illustrate 
 .-ome object, nor perhaps be able to go to the expense of having a 
 drawing reproduced, this method will be of value in enabling others 
 to benefit by our observations. 
 
 The author is convinced that if the employment of photography 
 is encouraged in bacteriological and other research laboratories for 
 depicting microscopical preparations and cultivations of bacteria, the 
 i -Milts of increasing experience and practice will lead to its being 
 made more general use of as a faithful and graphic method, valuable 
 alike for class demonstrations and for illustrating publications. 
 
PAET II. 
 
 ETIOLOGY AND PREVENTION OF INFECTIVE 
 DISEASES. 
 
 171 
 
CHAPTER XIII. 
 
 SUPPURATION, PY^MIA, SEPTIOEMIA, ERYSIPELAS. 
 
 ABSCESS. 
 
 WHEN inflammation is followed by an accumulation of leucocytes 
 and of plasma which does not coagulate, the result is a white or 
 creamy liquid called pus, and when the surrounding tissues are 
 involved so that a cavity develops containing pus, we have what 
 is termed an abscess. The almost constant association of bacteria 
 with the production of pus has created a belief that they are 
 the direct cause of suppuration. Ogston found micrococci present 
 in all acute abscesses, and concluded that acute inflammation was 
 invariably due to their presence. The fact that inflammation 
 occurs more frequently in the external tissues of the body is 
 accordingly explained by the ready entrance of bacteria which 
 are in the air; and suppuration following pericarditis, pleurisy, 
 and other conditions in which air is excluded is attributed to the 
 presence of pyogenic cocci, which have gained access by the blood 
 stream. There is no pyogenic organism constantly present, but 
 several different species of bacteria have been isolated from pus 
 and carefully studied, and the antiseptic treatment is based upon the 
 principle of excluding bacteria in surgical operations, and destroying 
 any which may have previously obtained access to wounds and 
 broken surfaces. Inflamed tissue and pus form a most suitable 
 medium for the growth of bacteria, which in some cases are 
 unquestionably only accidental epiphytes. 
 
 In tuberculosis, actinomycosis, and glanders, pus formation may 
 take place without the presence of pyogenic cocci ; and it is gener.illy 
 believed that chemical irritants, such as croton oil, turpentine, iodine, 
 rsidu \ ci-iii, and tuberculin, will excite the formation of pus in the 
 absence of bacteria, although Klemperer, after a number of very 
 careful experiments, maintains that no genuine pus will be produced 
 if the chemical irritants are first carefully sterilised. 
 
 i::; 
 
174 
 
 INFECTIVE DISEASES. 
 
 The bacteria which have been isolated from pus include : 
 Staphylococcus pyogenes aureus, albus, and citreus, Staphylo- 
 coccus cereus flavus and albus, Streptococcus pyogenes, Micrococcus 
 pyogenes tennis, Micrococcus pneumonias crouposse, Bacillus pyo- 
 cyaneus, Bacillus pyogenes foetidus, Micrococcus tetragenus, Bacillus 
 intracellularis meningitidis, Gonococcus, Bacillus septicus vesicae, 
 Urobacillus liquefacieiis septicus, Bacillus typhosus, Bacillus coli 
 communis, Bacillus anthracis, Bacillus tuberculosis, Bacillus mallei, 
 and Actinomyces. 
 
 FIG. 83. SUPPURATION OF SUBCUTANEOUS TISSUE. 
 
 d, Leucocyte containing micrococci ; d', leucocyte with pale nucleus showing 
 necrosis ; c, fixed connective tissue cells, much enlarged, containing several 
 nuclei, of which some (') are pale and necrotic ; numerous cocci, diplococci, 
 and short chains. (CoRNiL and RANVI KK. ) 
 
 Some idea of the distribution of the bacteria most commonly 
 occurring in pus may be gathered from the records made by Passet 
 and by Karlinski. 
 
 Passet examined acute abscesses, and found Staphylococcus 
 pyogenes aureus and albus in 11 cases, Staphylococcus pyogenes 
 albus alone in 4, Staphylococcus pyogenes albus and citreus in 2, 
 Streptococcus pyogenes alone in 8, Staphylococcus pyogenes albus 
 and Streptococcus pyogenes in 1, and Staphylococcus pyogenes albus 
 and citreus, and Streptococcus pyogenes in 1. 
 
SUPPURATION, PY^MIA, SEPTICAEMIA, ERYSIPELAS. 
 Karlin>ki tabulated his cases thus : 
 
 175 
 
 
 
 
 
 
 
 
 i 
 
 
 
 
 
 
 ! 
 
 r S 
 
 II 
 
 1 
 
 /. 
 
 1 
 
 || 
 
 | 
 
 
 f 
 
 e< 
 
 .- .- 
 
 -:< 
 
 g | 
 
 $3 
 
 ~,: 
 
 r~ 
 
 1 
 
 DISEASE. 
 
 J 
 
 
 
 
 II 
 
 C ,. 
 
 ! i 
 
 
 Bacillus A 
 
 Mastitis 
 Subcutaneous Abscess 
 
 36 
 
 90 
 
 22 
 10 
 
 4 
 2 
 
 4 
 
 8 
 
 6 
 6 
 
 2 
 
 2 
 
 
 
 
 
 Phlegmon ..... 
 
 24 
 
 
 
 
 
 
 24 
 
 
 
 
 
 
 
 
 
 B'uruncle . . . 
 
 20 
 
 9 
 
 
 
 10 
 
 
 
 1 
 
 
 
 
 
 
 
 Bubo 
 
 17 
 
 - 
 
 1 
 
 1 
 
 7 
 
 
 
 
 
 
 
 
 Subperiosteal Abscess 
 
 16 
 
 6 
 
 
 
 10 
 
 
 
 
 
 
 
 
 
 
 
 i P 
 
 1 7 
 
 
 
 
 
 
 
 
 
 1 n 
 
 7 
 
 
 
 
 
 
 
 
 Dental Abscess .... 
 
 10 
 1 A " 
 
 1 
 
 
 
 4 
 
 1 
 
 3 
 
 1 
 
 
 
 
 
 Abscess of the Middle Ear 
 
 1U 
 
 4 
 
 2 
 
 _ 
 
 
 _ 
 
 
 
 
 
 2 
 
 
 
 Carbuncle 
 
 4 
 
 2 
 
 
 
 1 
 
 1 
 
 
 
 
 
 - 
 
 4 
 
 
 
 
 
 
 
 
 
 
 
 Total . 
 
 
 82 
 
 7 
 
 55 
 
 45 
 
 
 
 8 
 
 2 
 
 4 
 
 PYAEMIA AND SEPTKLEMIA. 
 
 When pyogenic micrococci get access to the blood stream they 
 may be carried into distant parts, and by multiplying produce meta- 
 >tatic abscesses in the lymphatic glands, bones, joints, and internal 
 organs, a condition which is recognised as pyaemia. 
 
 If there is a general invasion of the blood stream by micrococci, 
 and absorption of their poisonous products, septicaemia results, and 
 death may occur before the development of any secondary lesions. 
 When septic micro-organisms multiply locally, and their chemical 
 products are absorbed, or their products are separated from putrid 
 materisil and injected into the circulation, the result may be called 
 .il,,;i ,',. The blood in septicaemia contains living or#ani>ins. and is 
 infective. The blood in saprsemia contains only the toxic chemical 
 products, and is not infective. The one is septic infection ami t lie- 
 other septic intoxication. Pyaemia may follow accidental wounds, 
 surjrical operations, parturition, acute suppuration of bones, scarlet 
 te\er. typhoid fever, and other di>-a---. 
 
 To avoid pyuMiiia in surgery and midwifery, the greatest care 
 must be taken to prevent micro-organisms from being conveyed by 
 
 nim.'nts, sponges, bandars, and by the hand> of tin- >urgeon 
 
 the obstetric physician. By the use of anti>-j,tic> and absolute 
 cleanliness the chances of infection are reduced to a minimum. 
 
176 INFECTIVE DISEASES. 
 
 Rosenbach examined six cases of metastatic pyaemia : Strepto- 
 coccus pyogenes was found five times, partly in the blood and partly 
 in the metastatic deposits, and twice in company with Staphylococcus 
 pyogenes aureus. 
 
 Baumgarten, also, found Streptococcus pyogenes in the internal 
 organs in pysemic cases, and Eiselsberg found Streptococcus pyogenes 
 in company with Staphylococcus pyogenes aureus in the blood of 
 cases of septicaemia. 
 
 Frankel isolated a streptococcus from puerperal fever, which 
 he at first called Streptococcus puerperalis, but subsequently 
 identified with Streptococcus pyogenes. These researches have 
 been confirmed by others. Winkel obtained a pure cultivation of 
 a streptococcus from the blood of the heart in a case of puerperal 
 peritonitis. It produced erysipelatous redness when inoculated 
 in the rabbit's ear, and in form and in cultivation was similar to 
 the streptococcus in erysipelas. Gushing also found Streptococcus 
 pyogenes associated with puerperal infection. The cocci were 
 found in endometritis diphtheritica as well as in secondary puerperal 
 inflammation. These observations were still further confirmed by 
 Baumgarten, and Bumm isolated the same organism in puerperal 
 mastitis. 
 
 DESCRIPTION OF BACTERIA IN Pus. 
 
 A description may now be given of the cocci most frequently 
 found. Staphylococcus pyogenes aureus and albus and Strepto- 
 coccus pyogenes and Gonococcus are the most important of these. 
 Staphylococcus pyogenes citreus, cereus albus and flavus, are pro- 
 bably merely epiphytic. Micrococcus tetragenus, Micrococcus 
 pyogenes tenuis, Ba'cillus pyogenes foetid us, Bacillus pyocyaneus, 
 Bacillus coli communis, Bacillus septicus vesicae, Urobacillus lique- 
 faciens septicus, and Bacillus intracellularis meningitidis will be 
 described fully in Part III. The description of Actinomyces, of 
 Micrococcus pneumoniae crouposae and of the bacilli of anthrax, 
 tuberculosis, glanders, and typhoid fever, will be found in other 
 chapters in Part II. 
 
 Staphylococcus pyogenes aureus (Rosenbach). Yellow 
 coccus in pus (Ogston). Cocci singly, in pairs, very short chains, and 
 irregular masses. Cultivated on nutrient agar-agar, an orange- 
 yellow culture develops, looking like a streak made with oil paint. 
 One variety grows on nutrient gelatine without liquefying it ; 
 another produces rapid liquefaction, and the growth subsides as 
 
SUPPl'H. \IION, PY/KMIA, SEFHCEMIA, ERYSIPELAS. 177 
 
 
 FIG. 84. Pus WITH STAPHYLO- 
 cocci, x 800 (FLUGGE). 
 
 an orange-yellow sediment, On potatoes and blood serum a similar 
 orange-yellow culture grows luxuriantly. They may also form 
 colourless growths in sub-cultures, and are then indistinguishable 
 from Staphylococcus pyogeiies albus. The cocci do not cause any 
 septic odour in pus. nor doe> any i:a> develop. Albumin is con- 
 verted I iy their action into peptoii' 
 They produce rapid anmioniacal fer- 
 mentation in urine (Shattock). 
 
 The micro-orirani>iii> injected into 
 the pleura or knee of a rabbit produce, 
 as a rule, a fatal result on the following 
 day ; but if it survives longer, it eventu- 
 ally dies of severe phlegmon. If injected 
 into the knee of a dog, suppuration 
 occurs, followed by disintegration of the 
 joint. Injected into the peritoneal 
 cavity of animals, they set up perito- 
 nitis, and introduced into the jugular 
 
 vein they produce septicaemia and death. When a small quantity of 
 a cultivation is introduced into the jugular vein after previous fracture 
 or contusion of the bones of the leg, the animal dies in about ten days, 
 and abscesses are found in and around the bones, and in some cases in 
 
 the lungs and kidneys, 
 and the cocci are found 
 in the blood and pu>. 
 
 Garre caused sup- 
 puration by inoculating 
 a pure-culture in a 
 wound near his finger 
 nail. Bockhart suffered 
 from several pustule- 
 
 "ft,.,- v, !.,!,* In- 
 
 arm wit ha pure-culture 
 suspended in >alt solu- 
 tion, and Burnm gave 
 hiiiiM-lf a hypodermic 
 injection of a pure- 
 culture and produced 
 an absoeae. Thi> micro- 
 orpuiiMM is practically iibiMiitou>. It has hren cultivated from 
 the >kin and mucous membrane.- and secretions of healthy per>on>. 
 and it occurs in the air. in soil, in du>t. and in water, and in 
 
 12 
 
 - Tii i <>t A KABBIT 48 
 
 !( - ran kX IN.IKCTION >K S-iAi'HVi.'x-occi, x 
 '.I.~H i P,AIM<;AKTKN). 
 
178 INFECTIVE DISEASES. 
 
 association with suppuration, pyaemia, puerperal fever, and acute 
 osteomyelitis. 
 
 Staphylococcus pyogenes albus (Rosenbach). Cocci micro- 
 scopically indistinguishable from the above. In cultivations also 
 they resemble Staphylococcus pyogenes aureus, but the growth 
 consists of opaque white masses. They, as a rule, liquefy nutrient 
 gelatine very rapidly, and subside to the bottom as a white sediment ; 
 more rarely they liquefy very slowly ; and a variety has also been 
 described which does not produce any liquefaction. They are similar 
 to the above-mentioned in their pathogenic action. Pure-cultivations 
 of the organism were obtained from a case of acute suppuration 
 of the knee-joint. 
 
 Staphylococcus pyogenes citreus (Passet). Cocci singly, in 
 pairs, very short chains, and irregular masses. If cultivated on 
 nutrient gelatine or nutrient agar-agar, a sulphur or lemon-yellow 
 growth develops. When inoculated under the skin of mice, guinea- 
 pigs, or rabbits, an abscess forms after a few days, from which a 
 fresh cultivation of the micro-organism can be obtained. 
 
 Staphylococcus cereus albus (Passet). Cocci, morphologic- 
 ally similar to the above, but distinguished by forming on nutrient 
 gelatine a white, slightly shining layer, like drops of stearine or wax, 
 with somewhat thickened, irregular edges. In the depth of gelatine 
 they form a greyish -white, granular thread. In plate -cultiva- 
 tions, on the first day, white points are observed, which spread 
 themselves out on the surface to spots of 1 to 2 mm. When culti- 
 vated on blood serum a greyish-white, slightly shining streak 
 develops, and on potatoes the cocci form a layer which is similarly 
 coloured. 
 
 Staphylococcus cereus flavus (Passet). Cocci which produce 
 in nutrient jelly a growth which, at first white, becomes lemon - 
 yellow, somewhat darker in colour than Staphylococcus pyogenes 
 citreus. Microscopically Staphylococsus cereus flavus corresponds 
 with Staphylococcus cereus albus. Inoculation experiments with 
 both kinds give negative results. 
 
 Streptococcus pyogenes (Rosenbach).- Cocci occurring singly, 
 in masses, and in chains. The individual cocci are small spherical 
 cells, with a special tendency after fission for the resulting elements 
 to remain attached to each other, forming chains or rosaries. In 
 cultures on solid media they often occur in the form of staphylococci, 
 but in liquid cultures there may be a few, three or more elements, 
 linked together ; or a great number, forming long chains which may 
 be straight, serpentine, or twisted. 
 
DESCRIPTION OF PLATE IV. 
 Streptococcus Pyogenes. 
 
 FIG. 1. From a cover-glass preparation of pus from a pysernic abscess. 
 Stained with gentian-violet by the method of Gram, and contrast-stained 
 with eosin. x 1200. Powell and Lealand's apochromatic T \ Horn. imm. 
 E. P. 10. 
 
 FIG. 2. From cover-glass preparations of artificial cultivations of the strepto- 
 coccus in broth and in milk at different stages of growth, x 1200. Powell 
 and Lealand's apochromatic ^ Horn. imm. E. P. 10. 
 
 In these preparations there is a great diversity in size and form of the 
 chains and their component elements. In the drawing examples are 
 figured of the following: 
 
 (a) Branched chains. 
 
 (b) Simple chains composed of elements much smaller than the 
 average size. 
 
 (c) Chains with spherical and spindle-shaped elements at irregular 
 intervals. These are conspicuous by their size, and are sometimes 
 terminal. 
 
 (d e) Chains in which the elements are more or less uniform in size. 
 (/) Complex chains with elements dividing both longitudinally and 
 
 transversely, and varying considerably in size in different lengths 
 
 of the same chain. 
 
Rgl. 
 
 , "x.-O '*'""'* 
 
 ? v .... **, 
 
 
 ./ 
 
 ,, ./- f / 
 
 4% ,. % 
 
 x-f^X? 
 
 
 
 X V 
 
 STREPTOCOCCUS PYOGENES 
 
SUPPURATION, PY^MIA, SEPTICAEMIA, ERYSIPELAS. 179 
 
 The individual elements composing the chains will be found to 
 vary considerably in size : here and there in a preparation will 
 be found a chain composed of excessively small cocci, in another 
 part the elements will be all on a larger scale, and again in 
 another part they will be peculiarly conspicuous on account of their 
 >i/.e. So great is the diversity in the size of the cocci in some of the 
 chains, that one might imagine that there was more than one kind 
 of streptococcus present in a preparation, until on examining some 
 of the longest chains it is observed that various sizes are repre- 
 sented in different lengths of the same chain. Very characteristic 
 appearances result from the fact that the cocci enlarge and divide 
 both longitudinally and transversely ; and, indeed, the largest, for 
 the mast part, clearly show a division in two directions, resulting 
 in the formation of tetrads. In addition to the forms resulting 
 from the fission of the cocci, there are here and there in a chain, 
 and sometimes terminally, larger elements, which are spherical, 
 spindle-shaped, or in the form of a lemon. In the length of the 
 chains, as in the size of the individual cocci, there is usually great 
 diversity. In some cases they are composed of only a few, three 
 or four cocci; in others eight, ten, or twenty. Here and there 
 an exquisite rosary will extend in a straight line across the field 
 of the microscope, or be twisted, curved, or serpentine ; in some 
 preparations twisted or entangled strands are observed which are 
 composed of several hundred elements. Such chains will be found 
 to be much, thicker in one part than another. Another char- 
 acteristic appearance is produced by separation of the elements 
 resulting from fission in the long direction of the chain, by which 
 lateral twigs or branches are formed. Another character, which 
 is very striking, may be seen when the individuals in a chain 
 have become separated ; an unstained or faintly stained membrane 
 may be found bridging across the interval. This will become still 
 more visible in preparations contrast-stained with eosin. 
 
 In plate-cultivations the appearances of the colonies are not very 
 striking. They appear to the naked eye after three or four days 
 as t-xtremely minute, greyish- white, translucent dots, which under 
 the microscope have a slightly yellowish -brown colour. They are 
 finely granular and well defined. They do not liquefy the gelatii.e. 
 and after several weeks may not exceed the size of a pin's head. 
 
 If the surface of nutrient gelatine solidified obliquely be traced 
 over once or twice with a platinum needle bent at the extremity 
 into a little hook charged with the cocci, a ribbon-shaped film 
 develops in two or three days. This film is composed of minute, 
 
180 INFECTIVE DISEASES. 
 
 greyish-white, translucent dots or droplets, which can be more easily 
 recognised with the aid of a pocket lens (Fig. 87). According to 
 the number of organisms sown on the jelly, the dots or colonies 
 may be completely isolated, or form a more or less continuous 
 film. The film by reflected light has an iridescent appearance like 
 mother-of-pearl, but has a bluish or bluish-grey tint by transmitted 
 light, and with a pocket lens appears distinctly brownish. The 
 gelatine is not liquefied, and even after several weeks the cultivation 
 is limited to the inoculated area, and the individual colonies are, as 
 a rule, not larger than pins' heads. In gelatine- cultivations of the 
 same age, but kept in the incubator at 18 C., the colonies get 
 irregular in form, especially at the margin of the film, arid give the 
 growth an arborescent, fringed, or serrated appearance. Cultivated 
 on the oblique surface of nutrient agar-agar at 37 C. the growth is 
 very similar, forming a film composed of minute white colonies 
 like grains of sand ; but the film appears less transparent, is 
 whiter, and the colonies have a greater tendency to get irregular 
 in form. If inoculated with. one tracing of the needle the growth 
 is scanty, but tends to get thicker in the centre than towards the 
 margins, which may have a terraced appearance. Inoculated in 
 the depth of gelatine, there appears after a day or two at 18 C. a 
 thread-like growth along the track of the inoculating needle. This 
 delicate growth is found on examination with a pocket lens to consist 
 of a linear series of extremely minute granules. In a few days 
 more, the beads or granules become more marked, but even after 
 weeks, the cultivation only appears like a string of minute, white, 
 compact, globular masses or grains. In broth at 37 C. the cocci in 
 twenty-four hours create a turbidity, and gradually develop beauti- 
 ful chains varying in length according to the age of the cultivations. 
 Even in forty-eight hours there may be chains of eight, ten, twenty, 
 or a hundred elements. After a few davs the growth settles down 
 at the bottom of the tube in the form of a white deposit, while the 
 supernatant liquid becomes again clear. 
 
 Inoculated subcutaiieously in the ear of rabbits, they produce in 
 two days an inflammatory thickening with erysipelatous redness, 
 or sometimes suppuration. 
 
 They may occur in vaccine lymph, as the result of con- 
 tamination, and Pfeiffer suggested that before calf lymph is 
 employed for vaccination it should be tested on a rabbit's ear. 
 If in two days no rash has been produced, the possibility of the 
 presence in the lymph of Streptococcus pyogenes or erysipelatis 
 is excluded. 
 
SUPPURATION, PY^MIA, SEPTICAEMIA, ERYSIPELAS. 181 
 
 A.vording to Fliigge and others, after subc-utaneous inoculation 
 of mice with a small quantity of a cultivation, there is no result in 
 80 per cent, of the animals experimented upon. Sometimes there 
 is limited pus formation at the seat of inoculation, sometimas the 
 animals die without any very striking pathological appearances. 
 
 They occur in abscessas, pyaemia, and septicaemia, and are often 
 found in diseases such as scarlet fever and typhoid, associated with 
 septic complications. They have been isolated from air, soil, and 
 water. 
 
 The streptococcus found in erysipelas agrees in description, and 
 is merely a variety of Streptococcus pyogenes. It has been definitely 
 established by the researches of Frankel and Freudenberg, and later 
 by those of the author, Raskin, Prudden, and Bayard Holmes, 
 that Streptococcus pyogenes is frequently found in scarlet fever and 
 diphtheria, and in other diseases associated with septic complica- 
 tions. The author has isolated Streptococcus pyogenes from acute 
 abscesses, from suppuration after surgical operations, from pyaemia, 
 from pyaemia after scarlet fever, and from purulent peritonitis. 
 Some of these cultures have been kept up for very long periods, 
 extending over some years, so that opportunities occurred for a 
 complete investigation into the life history of this micro-organism. 
 Variations in the appearances of cultures have been observed when 
 obtained from the same source. A number of cultures from pus 
 were prepared on gelatine and agar, made according to the usual 
 formula, but at different dates, and, therefore, varying slightly in 
 composition and quality. Sub-cultures were also started in nutrient 
 gelatine of precisely the same composition, but from primary cul- 
 tures of the same micro-organism in different media agar-agar, 
 milk, and broth. The descriptions of the streptococcus hitherto 
 published were then found to be inadequate. The different cultures 
 and sub-cultures presented striking variations in the microscopical and 
 macroscopical appearances. Some sub-cultures on gelatine, for exam- 
 ple, exhibited a finely dotted appearance, others showed every variety 
 in the size, and degree of opacity of the colonies (Fig. 89). Cultures 
 in broth al>n. variril in appearance, owing to slight variation in the 
 composition of the medium, to slight differences of temperature, and 
 other conditions difficult to determine. The addition of glycerine" to 
 broth materially alters the appearance of the culture. It was con- 
 rlusi v'ly proved that minute differences arise from different conditions 
 of the cultivating media. The author was led to study exhaustively 
 tlit- streptococci^ .t' acute suppuration in bovines. Primary cultures 
 of Streptococcus pyogenes from man, and primary cultures from 
 
182 INFECTIVE DISEASES. 
 
 a case of purulent peritonitis in a cow, were carried through 
 sub-cultures under exactly similar conditions. -Cultivations of the 
 Streptococcus pyogenes bovis exhibited variations in microscopical 
 and cultural characters which were even more marked than in the 
 case of the Streptococcus pyogenes hominis. By selecting certain 
 cultures from both sources there was a striking similarity if not 
 identity between them, but, when compared under exactly identical 
 conditions, there was more difference in cultural characters between 
 the Streptococcus pyogenes bovis and the Streptococcus pyogenes 
 hominis than between the Streptococcus pyogenes hominis and the 
 Streptococcus erysipelatis, and they may therefore be regarded as 
 distinct varieties (Figs. 89, 90). 
 
 Some of the diseases and conditions in which Streptococcus 
 pyogenes has been found may be alluded to more in detail. 
 
 Spreading Gangrene. From a case of spreading gangrene, which was 
 identical with Ogston's erysipelatoid wound gangrene, and regarded by 
 him as the most intense and dangerous form of erysipelas, Rosenbach 
 obtained pure-cultivations of a streptococcus by incising the skin of the 
 limb, and inoculating tubes from the turbid reddish fluid which escaped. 
 That the streptococcus was identical with Streptococcus pyogenes was 
 ascertained by comparison with a cultivation derived from pus, of the 
 mode of growth, and of the effect on animals. 
 
 Surgical Fever. Eiselsberg proved the presence of a streptococcus 
 in the blood of several cases of surgical fever in Billroth's clinic. The 
 organism was identified by cultivation with Streptococcus pyogenes. 
 
 Diphtheria. In three cases of typical diphtheria, Loffler found a 
 streptococcus. He isolated it by cultivation, found that it was similar 
 in form, characters on cultivation, and effects after inoculation, to 
 Fehleisen's streptococcus of erysipelas. Loffler was not inclined to 
 regard them as identical, because Fehleisen never found his cocci in the 
 blood-vessels. Flligge named the organism Streptococcus articulorum, 
 and states, that after subcutaneous inoculation or injection of a cultiva- 
 tion in mice, a large proportion of the animals die, and in the sections 
 of the spleen and other organs the streptococci are again seen. Baum- 
 garten investigated the same subject, and decided that the streptococcus 
 was identical with Streptococcus pyogenes. 
 
 Small-pox. Hlava has established the presence of Streptococcus 
 pyogenes in the pustules of variola, and Garre found streptococci in the 
 internal organs in a case of variola hsemorrhagica. In a fatal case of 
 variola complicated with pemphigus, Garre found a streptococcus in 
 the pemphigus vesicles. Whether it was identical with Streptococcus 
 erysipelatis Garre left an open question. 
 
 Yellow Fever. Babes observed the presence of streptococci in the 
 vessels of the kidney and liver in yellow fever. Cultivation experiments 
 are wanting. It was probably a case of secondary infection with Strepto- 
 coccus pyogenes. 
 
SUPPURATION, PY^MIA, SEPTICAEMIA, ERYSIPELAS. 183 
 
 />/'//' Babes, in a case of fievre bilieuse typhoide, found 
 
 masses of streptococci filling the vessels of the liver, kidney, and spleen. 
 This was probably another instance of secondary infection with Strepto- 
 coccus pyogenes. 
 
 _l/".f/>7rx. From the blood and inflammatory post-products in measles, 
 Babes isolated a streptococcus, which he describes as closely resembling 
 the Streptococcus pyogenes. 
 
 Ul<-> rtn-> /;//./"/>/ /Y//V/X. Wyssokowitsch found cocci in the internal 
 organs in ulcerative endocarditis, and produced the disease in animals, 
 after injury to the valves, by injection of Streptococcus pyogenes and 
 other organisms. Weichselbaum, by microscopical research and by 
 cultivation experiments, proved the presence of Streptococcus pyogenes 
 in acute verrucous endocarditis. Baumgarten confirmed this. He found 
 
 
 
 
 FIG. 86. Ui.< KKATIN -K ENDOCARDITIS : SECTION OF CARDIAC MUSCLE, x 700 (Kocn). 
 
 Streptococcus pyogenes alone in one case and accompanied by Staphylo- 
 coccus aureus in another. 
 
 Broncho-pneumonia. Thaon found a streptococcus in the lungs of 
 children in fatal cases of broncho-pneumonia, complicating measles, 
 diphtheria, and whooping cough. It was regarded as identical with the 
 streptococcus isolated by Loffler from diphtheria. Frankel discovered 
 a streptococcus in the lungs of a case of true croup complicated with 
 broncho-pneumonia, and by cultivation established its identity with 
 Streptococcus pyogenes. 
 
 Antlinis. Charrin found cocci in rabbits, examined some hours after 
 death from anthrax. These, when isolated, produced death in rabbits 
 from septicaemia, without suppuration. Chains composed of from fifteen 
 to twenty elements were found in all the organs. This was probably 
 another instance of Streptococcus pyogenes. 
 
 >/////'/;*. Kassowitz and Hochsinger found the presence of a strepto- 
 
184 
 
 INFECTIVE DISEASES. 
 
 coccus iu the tissues and internal organs, and especially in the blood- 
 vessels, in fatal cases of congenital syphilis. These observers regarded 
 their discovery as having an important bearing on the etiology of syphilis, 
 but Kolisko pointed out that it was only the result of septic infection 
 with presence of Streptococcus pyogenes, as had already been established 
 in scarlet fever. 
 
 Cerebro-xpinal Mniimjit'i,*. From the meningeal exudation of a case 
 of apparently idiopathic cerebro-meningitis, Banti found Streptococcus 
 pyogenes and Staphylococcus aureus and albus. The cocci probably 
 entered through an abscess of the jejunum. 
 
 FIG. 87. PURE-CULTURES OF STREPTOCOCCUS PYOCJENKS. 
 
 a, On the surface of nutrient gelatine ; b, in the depth of nutrient gelatine ; 
 c, on the surface of nutrient agar. 
 
 BlepJiaradenitis and Dacryocystis. Widmark isolated by cultivations 
 Streptococcus pyogenes and other organisms from cases of blepharadenitis 
 and phlegmonous dacryocystis. In phlegmonous dacryocystis Widmark 
 found Streptococcus pyogenes almost exclusively. 
 
 Leukaemia. Fliigge cultivated a streptococcus from necrotic patches 
 in the spleen of a fatal case of leukaemia. Cultures corresponded very 
 closely with Streptococcus pyogenes. Inoculation in the ears of rabbits 
 produced similar results to Streptococcus pyogenes or erysipelatis. 
 Flitgge calls it Streptococcus pyogenes malignus, but concludes that it 
 is probably dentical with the streptococcus from pus. 
 
MI'ITRATION, PYAEMIA, SKrriC^MIA. ERYSIPELAS. 
 
 185 
 
 BRYHH 
 
 Erysipelas is an acute inflammation of the skin, occurring in 
 connection with wounds, when it is traumatic, and on surfaces 
 apparently sound, when it is idiopathic, as in ery^ip^las of the 
 face. It is highly contagious in surgical wards, and it gives rise 
 to rapidly fatal puerperal fever in lying-in hospitals. In such cases 
 the virus is obviously conveyed from sick to healthy persons by 
 direct contact, or by instruments and sponges, or by the hand 
 of the surgeon, physician, or nurse, and possibly by the air. 
 
 o 
 
 - 
 
 -to. 
 
 ss. .SEX-J -.-;ix IN ERYSIPELAS. 
 
 (lymphatic vessels containing leucocytes and m,m, streptococci ; t, connective 
 tissue ; a, connective tissue and wandering cells, x 600 (CoBNiL and RAXVIEK). 
 
 Streptococcus of Erysipelas. In 1882, Fehleisen isolated a 
 streptococcus in erysipelas, described the appearances on cultivation, 
 and maintained that it could be distinguished from the streptococcus 
 of suppuration. Rosenbach agreed that the two micro-organisms 
 could be distinguished by parallel experiments, and named the one 
 Streptococcus pyogenes and the other Streptococcus erysij^latis. 
 (Fehleisen). Rosenbach asserted that the colonies of the latter 
 were more opaque and whiter than those of Streptococcus py< _ 
 and the growth more marked in the depth of nutrient gelatine, 
 while microscopically the chains were letter marked and larger, and 
 the individual cocci larger than in Streptococcus pyogenes. Others 
 who investigated thi> subject could not distinguish them with 
 certainty, either by their morphological or cultural char 
 effects on inoculation. Passet found that inoculation of Strepto- 
 coccus pyogenes induced a condition very similar to that produced by 
 modulation of Streptoc* Hoffa and Hajek described 
 
 minute differences, but Biondi and Eiselsberg failed to confirm these. 
 
186 INFECTIVE DISEASES. 
 
 Baumgarten failed to prove any essential difference. Mitchell 
 Prudden found that Streptococcus pyogenes injected into the sub- 
 cutaneous tissue of the ears of rabbits, produced in one no effect ; 
 in four, slight transient redness ; in five, local redness followed by 
 abscess; in twelve, well-marked erysipelatous redness, followed by 
 complete resolution in seven, abscess in three, and death in two. 
 Passet, Biondi, Eiselsberg, Baumgarten, and Mitchell Prudden 
 concluded that, in their morphological, biological, and pathogenic 
 characters, so far as animals are concerned, the two organisms are 
 practically identical. 
 
 The author investigated the morphology and cultural characters 
 of the Streptococcus erysipelatis, which he had isolated from a 
 typical case. This result cleared up the conflicting statements 
 which had been made by different observers. By carrying out 
 absolutely parallel experiments, the Streptococcus pyogenes and 
 Streptococcus erysipelatis were unquestionably distinguishable, as 
 Fehleisen and Rosenbach had asserted. In both cases, however, 
 inoculation of a trace of a culture from a solid medium produced 
 only transient redness. Injection hypodermically of a broth-culture 
 produced in both cases a spreading erysipelatous redness, followed 
 by suppuration. It was found that primary cultures of the two 
 micro-organisms, cultivated under precisely the same conditions, 
 differed in the size and character of their chains, in the size of the 
 individual elements, in the greater opacity of the colonies of Strepto- 
 coccus erysipelatis, in a greater tendency to confluence, and in 
 a more rapid growth. The author found that the difference was 
 most marked in broth -cultures. Abundant flocculi were formed by 
 Streptococcus pyogenes ; a powdery deposit with special tendency to 
 form a granular adhesive film at the bottom of the culture flask, 
 in the case of the streptococcus of erysipelas. Lastly, they differed 
 in their power of resisting germicides. 
 
 Fehleisen inoculated patients in hospital suffering with malignant 
 growths, and produced a typical erysipelas with sub-cultures after 
 an incubation of from sixteen to twenty hours. The disease was 
 marked by rigors, fever, and general disturbance. Patients who had 
 recently suffered from erysipelas had an immunity. 
 
 Emmerich succeeded in proving the presence of streptococci in 
 the air of a hospital where erysipelas had broken out. These cocci 
 in their form, their characters on cultivation, and their inoculation 
 results, were identified with the Streptococcus erysipelatis. It is not 
 therefore exclusively parasitic. 
 
 Streptococci identical or agreeing very closely in their description 
 
MT1THATION, I'Y^MIA, SEPTICAEMIA, ERYSIPELAS. 187 
 
 with Streptococcus pyogenes. ha\e heen found in cattle plague, font 
 ami mouth di>rase. wrangles, contagious inaininitis in cows, and 
 progressive tissue necrosis in mice, and they will be referred to fully 
 in subsequent chapters. 
 
 EXAMINATION- AND CULTIVATION OF STREPTOCOCCI. 
 < '<>ver-glass preparations can be stained with the watery solutions 
 of the aniline dyes. In some cases very beautiful preparations can 
 he obtained by using Neelsen's solution, and removing excess of 
 
 FIG. 89. STREPTOCOCCUS PYOGENES HOMINIS. 
 
 gelatine. 
 
 Pure-cultures on nutrient 
 
 a, Sub-culture from agar. 
 r, Sub-culture from milk. 
 
 6, Sub-culture from broth. 
 d, Sub-culture from milk. 
 
 >tain by rinsing in alcohol. To examine pus, milk, or broth, take 
 an ordinary platinum needle bent at the extremity into a booklet. 
 Dip it into the liquid to be examined, and spread it on a cover - 
 irl;i into as thin a film a> po>>ihle ; the preparation is treated 
 in the ordinary way, that is to say. the film is allowed to dry. 
 and the cover is taken up with forcep>. and paed three time> 
 through the flame with its prepared side upperino-t. 
 
 ti i'n in'. < M<>ili<J n-'ith Eosin. -In thi> way th- streptococci are 
 >tained blue, and stand out in mark' d contra>t to the rest of the 
 preparation. 1 >e fre-ldy prepared solution. Float the cover-glasses 
 
188 
 
 INFECTIVE DISEASES. 
 
 011 the solution for ten minutes to half an hour, then transfer them 
 to iodine-potassic-iodide solution, until they assume the colour of a 
 tea leaf ; then immerse them in alcohol until they are decolorised ; 
 dip them in an alcoholic solution of eosin for a few moments, and 
 then transfer them to clove oil to clarify the film ; to remove the 
 clove oil gently press the cover between two layers of clean filter 
 paper, then mount in xylol balsam. 
 
 A good method for cultivating streptococci is to employ a steril- 
 ised looped platinum wire, and to spread a droplet, for example, of 
 pus or blood, over the surface of nutrient agar-agar solidified obliquely. 
 
 a. b. c. 
 
 FIG. 90. STREPTOCOCCUS PYOGENES Bovis. Pure-cultures on nutrient 
 
 gelatine. 
 
 a, Sub-culture from agar. b, Sub-culture from broth. 
 
 c, Sub-culture from milk. d, Sub-culture from milk. 
 
 The tubes are then placed in the incubator at 37 C. ; the strepto- 
 cocci will appear in the course of two or three days in the form 
 of minute dotted colonies. If present alone, arid in considerable 
 quantities, the inoculated surface will exhibit a pure cultivation 
 consisting of a number of such colonies, whilst a flocculent mass is 
 observed in the liquid which collects at the bottom of agar-agar 
 tubes ; this flocculent mass will be found to be composed of chains. 
 From such a tube inoculate a number of the small flasks employed 
 in Pasteur's laboratory for cultivations in liquids. In this way 
 a number of pure- cultivations in milk and broth are established, 
 which can be readily examined from time to time. From a pure- 
 
M PPURATION, PYAEMIA, SEPTICAEMIA, ERYSIPELAS. 189 
 
 cultivation in broth or agar-agar tubes of nutrient gelatine can 
 be inoculated. Cover-glass-preparationfl from the growths on solid 
 media can be made in the usual way, and stained with either a 
 watery solution of t'uchsme or gentian violet : but to stain prepa- 
 rations made from milk or broth, or from the liquid in agar-agar 
 tubes, use the method of Gram ; the stain will then be removed, 
 except from the streptococci, and very beautiful preparations 
 result. 
 
 GONORRHOZA. 
 
 Gonorrhoea is the result of a catarrhal inflammation of the 
 mucous membrane of the urethra, vagina, or conjunctiva caused by 
 a characteristic pyogenic organism discovered by Neisser in 1879. 
 
 Gonococcus of Neisser. Cocci, usually in pairs 1-6 //. in 
 length, '8 p. in width, and tetrads, with those surfaces of the com- 
 ponent elements which are in contact, flattened. The elements 
 are more or less kidney -shaped, and are separated by a clear 
 unstained interval. They are found free in the pus and also in 
 the interior of the pus cells. They stain with the aniline dyes, 
 but art- decolorized by (.Train's solution. They do not grow on 
 the ordinary media, such as gelatine, agar, and potato, in marked 
 contrast to the common pyogenic cocci ; but Bumm succeeded in 
 obtaining a cultivation by using human blood serum, which was 
 procured for the purpose from the placenta. They give rise t< 
 a very delicate growth in the form of an almost invisible film, 
 with a moist appearance, which attains its full development in 
 a few da vs. Steinschneider used human blood serum and agar 
 incubated at 35 C. 
 
 Krall recommended either agar with grape-sugar and blood serum, 
 or the same inixrmv with the addition of 5 per cent, glycerine. 
 Others have employed nutrient agar with the surface moistened with 
 sterilised human blood. More recently Keifer ha> l>een successful 
 with a medium which is prepared in the following way: aseitie 
 fluid is filtered and sterilised by Tyndall's process, to this is added 
 an equal quantity of the following mixture, agar 3'5, peptone 5, 
 glycerine '_'. salt \> (per cent.). The ascitic agar is solidified in a 
 Petri'> dish, and the culture incubated at 36 C. 
 
 They have also been cultivated in albumin from plovers' eggs, 
 and in the fluid obtained from a case of synovitis of the knee joint. 
 
 Inoculation of rabbit-, dogs, horses, and monkeys, has been 
 invariably unsuccessful, but .sub-culture.-, produce the disease in 
 the healthy urethra. 
 
190 INFECTIVE DISEASES. 
 
 The cocci are found in pus from the urethra and other mucous 
 membranes affected by the disease. They have also been found in 
 urethral and inguinal abscesses in association with Staphylococcus 
 pyogenes aureus. 
 
 METHOD OF STAINING. 
 
 Cover-glass preparations are made in the usual way, and 
 double stained with Lomer's methylene blue, and eosin. 
 
 Schiitz recommends floating the cover-glasses for five or ten 
 minutes in a saturated solution of methylene blue in 5 per cent, 
 solution of carbolic acid. They are washed in water, rinsed in very 
 weak acetic acid, and again washed in water. Safranin may be 
 used as a contrast stain. 
 
 & < 
 
 FIG. 91. GONOCOCCUS x 800 (BUMM). 
 a, free cocci ; 6, cocci in pus cells ; c, epithelial cell containing cocci. 
 
 EGYPTIAN OPHTHALMIA. 
 
 There are two forms of ophthalmia in Egypt, one associated 
 with Gonococcus and the other with a bacillus closely resembling 
 the bacillus of mouse-septicaemia, but there are minute differences. 
 
 Bacillus of Ophthalmia (Koch and Kartulis). Minute rods 
 which do not grow on gelatine but readily on blood serum and 
 nutrient agar, forming a plainly visible, whitish-grey shining growth. 
 Animals are insusceptible, but cultures produced the disease in the 
 human conjunctiva in two out of six cases. 
 
CHAPTER XIV. 
 
 ANTHRAX. 
 
 ANTHRAX is a very fatal malady, and most irregular in its be- 
 haviour. At one time it attacks only one or two animals, and 
 at another time it will destroy nearly all the stock on a farm. 
 Farmers formerly regarded the disease as non-communicable, and 
 possibly the result of excessive or improper feeding, or faulty sani- 
 tation, or of climatic conditions over which no control could be 
 exercised. It is obvious that so long as the disease was regarded as 
 the result of unknown conditions, no explanation could be given of 
 its recurrence from time to time, or of certain animals contracting 
 the disease and others not, and no measures of any use could be 
 suggested to cope with an outbreak. 
 
 Anthrax has always been more prevalent on the Continent than 
 in England, and this to some extent accounts for the fact that it has 
 received greater attention abroad. In France, Germany, Hungary, 
 Russia, and in India and Persia, anthrax at times produces wide- 
 spread losses. In Siberia it is still known on this account as the 
 Siberian Plague. 
 
 On the Continent there are certain localities known as anthrax 
 'Usti'icts on account of their reputation for anthrax for example, in 
 the Upper Bavarian Alps in Germany and in Auvergne in France. 
 
 In 1849, Pollender happened to examine the blood of a cow 
 after death from anthrax, and discovered peculiar rod-like bodies 
 among the blood cells. The same observation was made independ- 
 -ntly by Brauell and Davaine about the same time, but the greatest 
 importance must be attached to the publication of Davaine's further 
 researches in iSU.'j. .Many ridiculed the discovery of bacilli, and 
 stoutly maintained that they were only blood crystals or accidental 
 -nurtures of no importance. 
 
 For many years very little progress was made, and the statements 
 of other observers who were able to verify and add to Pollender 's 
 and Davaine's di>coverie-. \\ere still received with scepticism. 
 
 191 
 
192 INFECTIVE DISEASES. 
 
 Within the last few years a great change of opinion has taken 
 place. Bacteriologists have investigated the whole subject, so that 
 at the present day we know exactly the cause of anthrax. 
 
 Bacillus anthracis (Bacteridie du charbon, Bacillus of splenic 
 fever, Wool-sorters' disease, or malignant pustule). Rods 5 to 20 /A 
 long and 1 to 1 '2 5 /A broad, and .threads ; spore-formation present. 
 As a thorough knowledge of the life-history of this bacillus is of the 
 greatest importance, the various steps to be followed in a practical 
 study of it will be successively treated in detail. Its morphological 
 
 FIG. 92. BACILLUS ANTHRACIS, x 1200. Blood corpuscles and bacilli 
 unstained ; from an inoculated mouse (FRANKEL and PFEIFFER). 
 
 and biological characteristics have been very completely worked out, 
 and it" serves as an excellent subject for gaining an acquaintance 
 with most of the methods employed in studying micro-organisms. 
 
 A mouse inoculated with the bacillus or its spores will die in 
 from twenty-four to forty-eight hours, or more rarely in from 
 forty-eight to about sixty hours. 
 
 Examination after Death. The spleen is found to be considerably 
 enlarged, and may be removed, and examined by making cover-glass 
 preparations, inoculations in nutrient media, and subsequently 
 sections. 
 
 Cover -glass Preparations. In cover-glass preparations of the 
 blood of the spleen the bacilli are found in enormous numbers. 
 Preparations should also be made with blood from the heart and 
 with the exudation from the lungs and other organs ; it will be 
 
DESCRIPTION OF PLATE V. 
 Bacillus Anthracis. 
 
 FIG. 1. From a cover-glass preparation of blood from the spleen of a guinea- 
 pig inoculated with blood from a sow. x 1200. Powell and Lealand's 
 apochromatic T V Horn. imm. E. P. 10. 
 
 FIG. 2. From a section of a kidney of a mouse. Under a low power the 
 preparation has exactly the appearance of an injected specimen. Under 
 higher amplification the bacilli are seen to have threaded their way along 
 the capillaries between the tubules, and to have collected in masses in 
 the glomeruli. Stained with Gram's method (gentian-violet), and eosin. 
 x 500. 
 
 FIG. 3. Bacillus antkracis and Microooccus tetragenus. From a section from 
 the lungs of a mouse which had been inoculated with anthrax three days 
 after inoculation with Mlcrococcus tetragenus. A double or mixed infection 
 resulted. Anthrax- bacilli occurred in vast numbers, completely filling the 
 small vessels and capillaries, and in addition there were great numbers 
 of tetrads. Stained by Gram's method (gentian-violet), and with eosin. 
 x 500. 
 
Plate V. 
 
 \- 
 
 Fig 
 
 2. 
 
 Fig 3 
 
 BACILLUS ANTHRACIS 
 
ANTHRAX. 193 
 
 noted that in these the bacilli are present in very small numbers, or 
 altogether absent. The bacilli should be examined both unstained 
 and stained. The rods are straight or sometimes curved ; rigid and 
 motionless. They can be stained with a watery solution of any of 
 the aniline dyes, and are then seen to be composed of segments with 
 their extremities truncated at right angles; between the segments 
 a clear linear space exists, which gives them a characteristic appear- 
 ance (Plate V., Fig. 1). By double staining, with Gram's method and 
 eosin, the rods are seen to consist of a membrane or hyaline sheath 
 with protoplasmic contents. 
 
 Drop-cultures. A little of the blood from the spleen or heart 
 may be employed to inoculate sterilised broth or blood serum. 
 Several of these cultures should be prepared, and some of them 
 placed in the incubator, and examined at intervals of a few hours. 
 It will be observed that the rods grow into long homogeneous fila- 
 ments, which are twisted up in strands, and partly untwisted in long 
 and graceful curves. The filaments begin to swell, 
 become faintly granular, and bright, oval spores 
 develop (Plate 1). The cultures in the incubator 
 develop rapidly. A temperature of 30 to 37 C. 
 is the most favourable for spore-formation. The 
 spores are eventually set free, and by making 
 a fresh cultivation, or by injecting them into a 
 mouse or guinea-pig, they germinate again into 
 the characteristic bacilli, which in their turn 
 grow into filaments and spores. When the spore 
 germinates it swells, the envelope becomes jelly- 
 like, and gives way at one or other pole, and the 
 contents escape and grow into a rod. 
 
 Test-tube Cultivations in Xntrient Gelatine. 
 Typically characteristic appearances are obtained 
 by inoculating a 5 to 8 per cent, nutrient gelatine. 
 A whitish line develops in the track of the inocu- 
 lating needle, and from it fine filaments spread out 
 in the surrounding medium (Fig. 93). The fila- FIG. 93. PURE CUL- 
 ments are more easily <>lrrveil with a magnifying TIVATIOX OF BA- 
 
 ! TJ , . T - ,1 CII.M'S ANTHRACIS 
 
 glass. In a more solid nutrient gelatine the growth ix N ITHIFN 
 appeal's only ;is u thick white thread. As lique- LATINK. 
 faction of the gelatine progresses, these appearances 
 gradually alter, and the growth subsides to the bottom of the 
 tube as a white flocculent mass. In exhausted culture-media, and 
 sometimes in the blood, filaments are seen in a state of degeneration. 
 
 13 
 
194 
 
 INFECTIVE DISEASES. 
 
 This has also been observed in sections of the internal organs of a 
 rabbit which had been inoculated with the anthrax bacillus and had 
 died of septicaemia the following morning. 
 
 Test-tube Cultivations in Nutrient Agar-agar. Cultivated upon a 
 
 FIG. 94. COLONIES OF BACILLUS ANTHRACIS, x 80 (FLUGGE). 
 a, after 24 hours ; I, after 48 hours. 
 
 sloping surface of nutrient agar-agar a viscous snow-white layer is 
 developed, but without access of air no cultivation can be obtained, 
 the bacilli being aerobic. This can be demonstrated by completely 
 
 embedding a piece of lung or spleen 
 pulp containing bacilli, in nutrient 
 agar-agar (p. 22). 
 
 Potato - cultivations. In about 
 thirty-six to forty-eight hours a creamy- 
 white or very faintly yellowish layer 
 forms over the inoculated surface, 
 usually with a translucent edge, and 
 sometimes a strong, penetrating odour 
 of sour milk. 
 
 Plate-cultivations. From the spleen 
 or blood of the heart cultivations may 
 be made in nutrient gelatine on plates. 
 The colonies develop in about two days, according to the temperature 
 of the room. They appear to the naked eye as little white spots 
 
 FIG. 95. IMPRESSION-PREPARA- 
 TION OF A COLONY, X 70. 
 
ANTHRAX. 
 
 195 
 
 or specks, which, on examination with a low power of the microscope 
 and small diaphragm, exhibit two distinct forms. One form, on 
 careful focussing, has the appearance of a little compact ball of 
 
 FIG. 96. MARGIN OF A COLONY, x 250 
 
 twisted threads; in the other, liquefaction of the gelatine has 
 commenced, and the threads spread out like locks or plaits of 
 haii' in the neighbouring gelatine. These appearances are perfectly 
 characteristic (Figs. 94, 96). 
 
 Cover-glass Impressions.- The plate- cultivations should be also 
 examined as soon as the colonies 
 appear, by making cover-glass im- 
 pressions (Fig. 95). The filaments, 
 examined with a high power, will be 
 seen to consist of a number of rods 
 or segments which are perfectly 
 regular in form. On the other 
 hand, filaments from a tube-cultiva- 
 tion in a solid medium will often be 
 found to be composed, not only of 
 rods, but here and there of the so- 
 
 called involution-forms (Fig. 97). FIG. 97. FII.AMKNTS WITH OVAL AMI 
 Prom cultures in gelatine ;.,', 
 
 .irlvcerine agar, very striking preparations are ><>met iim->; obtained, 
 with numerous large spherical and lemon-shaped elements. In' a 
 
196 INFECTIVE DISEASES. 
 
 cover-glass preparation from a potato- culture the individual segments 
 will be found to have a great tendency to be isolated one from the 
 other, and there is copious spore-formation. 
 
 Preservation of Spores. Spores may be preserved simply by allow- 
 ing anthrax blood to dry and then sealing it in a tube. The spores 
 from a potato-cultivation are treated as follows : The inoculated 
 surface bearing the creamy cultivation is sliced off in a thin 
 layer, and is mashed up with distilled water in a glass capsule. 
 Sterilised silk-thread is cut up into lengths of about a quarter of 
 an inch, and allowed to soak in the paste for some hours, under a 
 bell-glass. The threads are then picked out with a pair of forceps, 
 and laid upon a sterilised glass plate, covered with a bell-glass, 
 and allowed to dry. From the plate, when perfectly dry, they 
 are transferred to a small test-tube, which can be plugged with 
 cotton-wool, or sealed in the Bunsen burner. 
 
 Examination of the Tissues. The organs should be hardened 
 in absolute alcohol, and sections prepared and stained by the 
 ordinary methods. The method of Gram is the most instructive, 
 and eosin a very satisfactory contrast stain. The capillaries in 
 the lungs, liver, kidney, spleen, skin, mucous membrane, etc., will 
 be found to contain bacilli. In some cases the bacilli are so 
 numerous that a section under a low power has the appearance 
 of an injected specimen. 
 
 Inoculation of Animals. A thread containing spores, a drop of 
 blood from an infected animal, or a minute portion of a cultivation, 
 introduced under the skin of a mouse or guinea-pig, causes a fatal 
 result, as a rule, in from twenty-four to forty-eight hours. Sheep 
 fed upon potatoes which have been the medium for cultivating the 
 bacillus, die in a few days. Goats, hedgehogs, sparrows, cows, horses, 
 swine, and dogs are all susceptible. Rats are infected with diffi- 
 culty. Frogs and fish have been rendered susceptible by raising the 
 temperature of the water in which they lived. Oats, white rats, 
 and Algerian sheep have an immunity from the disease. 
 
 Attenuation of the Virus.- Toussaint attenuated cultures by 
 exposing them for ten minutes to 55 C. Pasteur obtained a similar 
 result by resorting to lower degrees of temperature ; and Koch, 
 Gaffky, and Loffler concluded from their experiments, that from 
 42 to 43 C. the bacillus was most easily deprived of its poisonous 
 properties. By cultivating the bacillus in neutralised broth at 
 42 to 43 C. for about twenty days, the infecting power is weakened, 
 and animals inoculated with it (premier vaccin) are protected against 
 the disease. To obtain a still more perfect immunity, they are 
 
ANTHRAX. 197 
 
 inoculated a second time with material (deuxieme vaccin) which lias 
 been less weakened. The animals are then protected against the 
 most virulent anthrax, but only for a time. From a weakened 
 culture, according to Klein, new cultures of virulent bacilli can be 
 started, and a culture that can be used as a vaccine for sheep kills a 
 guinea-pig, and then yields bacilli that are fatal to sheep. 
 
 The virulence of the bacillus is also altered by passing the 
 bacillus through different species of animals. The bacillus of sheep 
 or cattle is fatal when re-inoculated into sheep or cattle; but if 
 inoculated in mice, the bacilli then obtained lose their virulence 
 for sheep or cattle, only a transitory illness results, and the animals 
 are protected for a time against virulent anthrax. 
 
 Exposure to a temperature of 55 C., or treatment with '5 to 
 1 per cent, carbolic acid, deprives the bacilli of their virulence. 
 
 Chauveau obtained a similar result by cultivating the bacillus at 
 38 or 39 C. under a pressure of eight atmospheres. The possibility 
 of mitigating the virus depends upon the species of animal ; rodents 
 cannot be rendered immune by any known anthrax vaccine. The 
 nature of the toxic products has been described in a previous chapter 
 (p. 42). 
 
 METHODS OF STAINING THE BACILLUS ANTHRACIS. 
 
 Cover-glass preparations of. blood, etc., can be stained with a 
 watery solution of any of the aniline dyes, or with ISTeelsen's solution 
 and subsequent treatment with alcohol (p. 87 \ The preparations 
 may be dried and mounted permanently in Canada balsam, but the 
 typical appearances are best observed in freshly stained specimens 
 examined in water. 
 
 The sheath and protoplasmic contents can be demonstrated in 
 cover -glass preparations from the 
 blood or spleen which have been 
 stained with eosiii after the method 
 of Gram. 
 
 Spores must be stained by the ^ 
 
 special methods already described. /^^ ''-"'^ 
 
 The most satisfactory preparations 
 a iv obtained by double-staining with KM;. <)8. SPORES OF P>A< -n.i.i s 
 Ziehl-Neelsen solution and methy- ANTHRACIS STAINED WITH 
 
 , . . GENTIAN VIOLET, x 1500. 
 
 lene blue (Fig. / ). 
 
 sections are >>est stained by the method of Grain, and 
 with eosin, picrocarminate of ammonia, or picro-lithinm- 
 cannine. 
 
198 INFECTIVE DISEASES. 
 
 ORIGIN AND MODE OF SPREAD. 
 
 As every outbreak of anthrax is the result of the introduction 
 into the system of the bacilli, the question naturally arises, how 
 are they introduced on the farm ? Where do they come from ? and 
 what are the channels of infection ? 
 
 The spores of the bacilli may get into the soil, and may remain 
 there in a dormant state for many years. The spores were believed 
 by Pasteur to be taken up by earth-worms, carried to the surface 
 and deposited in their castings. Animals grazing are thus liable 
 to be infected ; but Koch's experiments tended to disprove this 
 theory. Anthrax has been known to break out among cattle 
 grazing on a field where several years previously some Russian 
 hides from infected animals had been buried. By some means or 
 other the spores may contaminate the grass, and hay imported 
 from an anthrax district may start the disease on a farm on which 
 it had never been known to occur. The spores may in a similar 
 way be introduced with blood manure and bone manure, and with 
 refuse used as manure. The skin, hair, wool, hoofs, and horns of 
 infected animals, if soiled with blood, are contaminated by the 
 bacillus. 
 
 Another way in which the disease can be communicated may 
 be illustrated by the transmission of the disease to man. Those 
 who handle carcasses, wool or hides of infected animals are liable to 
 contract the disease. Slight scratches, cuts, bites, and pimples, may 
 readily be inoculated with the bacilli or their spores. Veterinary 
 surgeons, butchers, herdsmen, cattle drovers in fact, all those whose 
 occupation leads them to cut open or skin cattle, sheep, or horses, or 
 to handle hides and wool are liable to fall victims to this disease. 
 
 In one case which was brought to the author's notice, a veteri- 
 nary surgeon had been called to see a bullock which had died 
 suddenly in a meadow. A post-mortem examination was made, and 
 the veterinary surgeon wiped his hands, which were soiled with 
 blood, on some rough grass, and then washed them in a stream. 
 The sedgy grass made some small cuts on his fingers, and the 
 result was that he was simultaneously inoculated with the blood 
 of the bullock. Local anthrax followed, two of his fingers were 
 amputated, and he fortunately recovered. In another case a butcher 
 dressed the carcass of a beast which had died suddenly, and while 
 doing so scratched a pimple on his neck. An anthrax pustule 
 developed, and after a very serious illness he also recovered; but 
 in many cases the attack is fatal. " Wool -sorters' disease " is 
 
ANTHRAX. 199 
 
 anthrax of the lungs. Bales of foreign wool contain not only wool 
 from living sheep, but wool which has been clipped from skins of 
 (lra<l sheep. If any of the sheep died from anthrax the wool is 
 sure to be contaminated with blood containing the bacilli, and then 
 wool sorters engaged in picking the wool readily inoculate them- 
 selves through a scratch or pimple, or by inhaling the spores. In 
 many ca>-> Wool-sorters' disease is fatal. 
 
 A farm may become extensively infected by the living animal. 
 Blood containing the bacilli may be discharged from the mouth and 
 nostrils, or be passed with the contents of the intestinal canal 
 and bladder. The droppings contaminate the pasture or byre, and 
 spore formation, especially in warm weather, quickly takes place. 
 From this cause the disease may not only be conveyed to healthy 
 cattle grazing \vith infected animals, but fresh cases may occur, year 
 after year, on the same farm, and if hay is cut and sold off the farm, 
 other cattle at a distance are similarly infected. If the flooring of 
 cattle sheds is once soiled by infected animals it is easy to account 
 for those otherwise mysterious outbreaks which occur when the 
 cattle are taken in for the winter. 
 
 Another source of danger arises when blood from a diseased 
 animal is washed into brooks or streams, for thus the disease may 
 be carried to farms in which it was previously unknown. 
 
 PREVENTIVE MEASURES. 
 
 Early recognition and prompt action are essential to prevent the 
 spread of any communicable dist 
 
 Unfortunately in the case of anthrax only too often the very 
 first indication of the existence of the disease is the sudden death 
 in the pasture or byre of an apparently healthy beast, or possibly 
 of one or more sheep. Nevertheless, the importance of being able 
 to recognisf any early indications is very great, because an im- 
 mediate and careful examination should at once be made of the 
 stock on the farm, and suspicious cases isolated from the rest. The 
 stock- man may notice that one or two animals tend to keep away 
 from the others. They look dull and cease feeding, and possibly 
 shivering may be observed. In horses swelling of the throat may 
 occur, and in some places there is discharge of blood from the 
 orifices. Death follows the appearance of these symptoms in a 
 few hours, and often with startling suddenness. Cattle die rapidly, 
 but sheep, though rapidly contracting the disease, do not as a rule 
 die so suddenly. 
 
200 INFECTIVE DISEASES. 
 
 The characteristic sign after death is enlargement of the spleen 
 to three or four times its natural size. It is not only enlarged, 
 but extremely soft and dark in colour. Blood spots are visible on 
 the internal organs generally, and the intestine often contains a 
 quantity of blood. The examination of a drop of blood will show 
 under the microscope the characteristic bacilli. It is, however, 
 quite unnecessary to make an elaborate post-mortem examination 
 in order to satisfy oneself whether the disease is really anthrax or 
 not. If an animal has died suddenly and has created a suspicion 
 of anthrax, all that it is necessary to do is to cut cff an ear or a 
 foot in the case of a sheep and make a cover- glass preparation at 
 the first opportunity. 
 
 A farmer with a case of anthrax must be made to realise the 
 fact that an enormous quantity of poisonous material has to be 
 dealt with. In fact, an infected animal is more dangerous when 
 dead than alive. The owner or person in charge must immediately 
 notify to a police constable the existence, or even a suspicion of 
 the existence, of the disease. Prompt measures must be taken 
 to destroy the carcass and all traces of the blood, and thus to 
 reduce to a minimum the chance of the disease spreading to the 
 rest of the stock, and of creating fresh outbreaks in the future. 
 Every possible precaution must be taken to prevent the blood of 
 the dead animal from contaminating the pasture, byre, or water 
 supply. The rest of the stock should be removed from the pasture 
 or cowshed where the disease has broken out. It is desirable to 
 give a complete change of food and water, and the w T hole of the 
 stock should be examined every day for a week, and any animals 
 showing a rise of temperature should at once be isolated from the 
 rest. Preventive inoculation has been recommended to protect 
 the rest of the stock, but there is not sufficient evidence of the 
 safety of the process to lead to the adoption of this treatment. 
 Animals ready for the butcher may be removed from the risk of 
 infection by immediate slaughter. To disinfect the pasture the 
 best plan is a heavy top-dressing of lime, and after six weeks 
 stock may be readmitted, though not without some risk. If 
 year after year cases of anthrax occur on a particular pasture, 
 the most obvious precaution is to keep stock from it altogether 
 and convert it into arable land. As roots grown on anthrax- 
 infected soil have been known to convey the disease, the wisest 
 course if we have to deal with a small field or comparatively 
 small tract of land is to throw it out of cultivation or to plant 
 it with trees. 
 
ANTHRAX. 201 
 
 DISPOSAL OF THE CARCASS. 
 
 The surest method to render harmless all the bacilli which 
 exist in the carcass is burning, but cremation offers practical 
 difficulties, especially if several carcasses have to be destroyed. In 
 the case of an animal dying in a town, the local conditions may 
 render it best to adopt destruction by burning or by means 
 of chemicals. In such a case the carcass should be covered 
 with quicklime, and then taken, in charge of an officer of the 
 Local Authority, to a horse -slaughterer's or knacker's-yard, and 
 destroyed by exposure to a high temperature, or by chemical agents 
 especially in the vicinity of chemical works. Under the usual 
 circumstances of death occurring on a farm, fortunately the simple 
 plan of burial, with the addition of lime or other chemical agents, 
 is perfectly efficacious, and even without the use of chemicals if the 
 carcass has been left unopened, as the bacilli die rapidly if air is 
 excluded. 
 
 Some experiments carried out by M'Fadyean clearly indicate the 
 importance of leaving the carcass unopened. 
 
 On July 16th a sheep was infected with anthrax by feeding it with a 
 virulent culture. Five days later it died, and a microscopic examina- 
 tion of blood from the ear, immediately after death, showed very many 
 anthrax bacilli. The carcass was left unskinned and unopened until 
 July 27th, when the various organs were cut out of the chest and 
 abdomen and placed in a tin box. The box was then buried at a depth 
 of about two feet in garden earth, and left there undisturbed until 
 February loth, when it was exhumed. The organs had become con- 
 verted into adipocere, and this was thoroughly mixed up with water and 
 administered to a sheep. The sheep remained perfectly healthy. In 
 another experiment a rabbit was inoculated with anthrax on June 1st. 
 It died on June 3rd, and blood from the ear contained the bacilli. The 
 rabbit was left unopened for three days, and then placed in a flower pot 
 and buried in garden earth at a depth of two feet. It was exhumed on 
 February 15th. The tissues were all destroyed by putrefaction, and the 
 earth in contact with the bones was administered to a sheep without 
 conveying the disease or producing any ill effects. 
 
 Thus, in the first experiment, the lungs and the intestines, in 
 which spore formation was mast likely to occur, were used as a 
 test, and in the second experiment the entire carcass. In both cases 
 there was destruction or disappearance of the bacilli, and these tests, 
 therefore, confirm in a very marked way the opinion that prompt 
 burial of the unopened carcass is a perfectly safe plan to adopt. 
 
202 INFECTIVE DISEASES. 
 
 If an animal has died in a meadow, a pit six feet deep should 
 be dug close to the carcass, and if quicklime can be procured with- 
 out delay the carcass should be buried with a layer about a foot in 
 depth beneath it and with about the same quantity to cover it, and 
 the pit filled up with the excavated soil. 
 
 If there are any traces of blood where the animal lay, the 
 contaminated ground should be covered with quicklime or drenched 
 with strong carbolic acid, and the whole of the site of burial fenced 
 off for six months. If an animal dies near a brook or stream then 
 the carcass must be removed for burial to a sufficient distance to 
 prevent any reasonable probability of contamination of the water. 
 
 If death has occurred in the byre, the carcass must be removed 
 to the nearest and most convenient spot for burial, any fodder or 
 litter which may have been in contact with the deceased animal must 
 be destroyed, and the shed arid cart and any utensils, hurdles, etc., 
 disinfected. For the latter purpose thorough scouring with water 
 and then washing with limewash is recommended. The limewash 
 should be prepared immediately before use, and four ounces of 
 chloride of lime, or half a pint of commercial carbolic acid, be added 
 to each gallon of limewash. 
 
 The following is an illustration of the value of preventive 
 measures based upon a knowledge of the exact nature of the disease. 
 A farm on the banks of the Yeo was repeatedly attacked by 
 anthrax. One morning two sheep died, and other cases followed. 
 The farmer learnt that his predecessor had buried cattle which had 
 died of anthrax on the very spot where the sheep were folded. He 
 removed his flock, and had no further losses among the sheep, but 
 he continued to lose cattle grazing in the pastures by the river. 
 These pastures were occasionally flooded by the Yeo. Another 
 farmer in the same locality heavily manured a field, and shortly 
 afterwards anthrax broke out in a most deadly form on his farm. 
 
 What was the cause of these mysterious outbreaks ? The 
 explanation was forthcoming, and prevention an easy matter. The 
 river Yeo received the washings from the wool factories at Yeovil, 
 and the pastures were contaminated by anthrax spores in the 
 deposit which was left behind when the flood subsided. In the 
 second instance, it was found that the manure used for dressing 
 the pasture consisted of a quantity of refuse from the wool factories. 
 
 Infected wool from foreign countries is one of the principal 
 sources of the disease in this country, and the remedy is to insist 
 upon the factories destroying their refuse instead of its being 
 allowed to contaminate the rivers or to be sold as manure. 
 
ANTHRAX. *>03 
 
 So long us this source of the disease was unknown anthrax 
 continued to be spread through the agency of the wool factories. 
 
 Anthrax spores may also be introduced with foreign oats, hay, 
 and manure, so that it is almost impossible absolutely to prevent 
 the importation of the disease; but the danger of its unlimited 
 extension and disastrous losses can be minimised, and the com- 
 munication of the disease to man and to swine entirely avoided by 
 simple precautions. 
 
 ANTHRAX IN SWINE. 
 
 The occurrence of anthrax in swine is a subject upon which 
 there has long been considerable diversity of opinion. Some of the 
 
 FIG. 99. ANTHRAX IN SWINE. From a photograph taken during life, showing a 
 swollen condition of the neck and tin-oat six days after ingestion of part of the 
 viscera of a bullock which had died from anthrax. 
 
 earliest writers on the diseases of animals speak of outbreaks of 
 anthrax among swine, but whether any or all of these outbreaks 
 were examples of true anthrax has long been a matter of un- 
 certainty; for it is well known that diseases quite distinct were 
 included under the name anthrax. 
 
 Menschel states that in an outbreak in which twenty-four 
 persons were attacked with malignant pustule, many of them from 
 eating the flesh of beasts suffering from anthrax, pigs which were 
 t'rd OH the same flesh also became affected, and a woman who ate 
 some of the diseased pork was subsequently ill. 
 
 Roche-Lubin, while apparently accepting the occurrence of 
 anthrax in swine, taught that the pig resisted inoculation with the 
 blood of a different species. 
 
204 INFECTIVE DISEASES. 
 
 In this country accounts have been published from time to time 
 of a fatal disease in pigs induced by eating the flesh of animals 
 which had died of what was described as " blood-poisoning." 
 
 Some very striking cases occurred in the practice of Mr. Wilson, 
 of Berkhampstead, and were reported in the Veterinarian. A 
 farmer consulted Mr. Wilson respecting an illness with which his 
 pigs were affected, stating that two or three were dead and many 
 others seriously ill. They were strong hogs, ranging from six to 
 nine months old. On inquiry it was ascertained that the farmer 
 had lost a beast suddenly about a week previously, that the carcass 
 had been opened in the yard, and the viscera thrown to the pigs. 
 Mr. Wilson expressed the belief that the disease was anthrax, and 
 stated that he found the pigs exhibiting many of the symptoms 
 observable in cattle, with the additional one of enlargement round 
 the throat from infiltration of a yellow fluid causing discoloration 
 of the skin. 
 
 Also, in the reports of the Agricultural Department of the Privy 
 Council thirteen pigs were reported as suffering from anthrax in 
 1886, and one hundred and fifty-nine in 1887. 
 
 But the question arose whether the disease in the pigs was 
 genuine anthrax or septic poisoning. 
 
 Williams says : " The flesh of animals which have died or have 
 been killed whilst suffering from the disease [anthrax] should not 
 be used as food either for men, pigs, or dogs, as it is apt to cause 
 death by blood poisoning"; and Steel writes: "Pigs, dogs, and 
 poultry should not be allowed to feed on blood, flesh, and ejecta 
 of anthrax victims," but no statement is made as to the nature of 
 the illness produced. No doubt these writers have been greatly 
 influenced by the opinion of many bacteriologists, for Toussaint 
 maintained that pigs could not be infected with anthrax, and a 
 .similar view was at one time upheld in this country by Klein, 
 who stated that pigs were very insusceptible. In Germany also, 
 pigs have been credited with an immunity from this disease. 
 
 In the face of these conflicting statements the author carried 
 out a series of experiments in order to ascertain the nature of the 
 disease in swine resulting from the ingestion of the offal of animals 
 which had died of anthrax ; and the result of inoculation with blood 
 of animals which had died of anthrax, and with pure cultivations 
 of the Bacillus arithracis. 
 
 As a result of thess experiments genuine anthrax was produced 
 in swine (a) by feeding them with anthrax offal ; (b) by injection 
 of blood of a bullock which had died of anthrax ; (c) by passing 
 
ANTHRAX. 
 
 205 
 
 bacilli through the guinea-pig, and transmitting them to swine by 
 injection of blood from the spleen ; (d) by injecting a pure cultiva- 
 tion of the anthrax bacillus ; (e) and lastly, the anthrax bacillus 
 was isolated from swine in which the disease was accidentally induced 
 on a farm, and the disease reproduced by inoculation of guinea-pigs 
 and mice with blood from the spleen. 
 
 The Author's Conclusions. Swine of all ages can be affected with 
 anthrax. If the disease is induced by ingestion of anthrax offal, 
 the tonsils are ulcerated, and constitute the point of access of the 
 bacilli to the blood. In such cases the characteristic symptom is 
 
 FIG. 100. ANTHRAX IN SWINK. From a photograph taken ]w$t-nt<-t< m. Death 
 (K-curred four days after the ingestion of offal from a bullock which had died of 
 anthrax, and there was well-marked oedema of the throat, cheeks, and eyelids. 
 
 enormous swelling around the throat. If the disease is induced by 
 hypodermic injection, the same oedematous infiltration of the tissues 
 occurs at the place selected for inoculation. Death may occur 
 in twenty-four hours, or not until after five or six days. There 
 is a rapid rise of temperature, usually a rash-like discoloration 
 of the skin, sometimes loss of power over the limbs, and general 
 weakness and disinclination to move; the animal may lie helplo- ly 
 on its belly, and utter plaintive cries when disturbed. At the post- 
 mortem the most characteristic feature is the gelatinous oedema 
 which, in the case of ingestion of offal, is found around the throat. 
 There is usually congestion of all the organs and engorgement of 
 
206 INFECTIVE DISEASES. 
 
 the heart and large vessels, fluid in the cavities of the chest and 
 abdomen, and enlargement and haemorrhage into the lymphatic 
 glands. There is in some cases inflammation of the intestines with 
 submucous and subserous haemorrhages. The spleen may be normal 
 in size, pale and flabby, and the liver only slightty congested and 
 friable; in other cases the condition is characteristic, the spleen 
 is the seat of haemorrhage, causing more or less local enlargement, 
 which is superficially of a deep purple colour ; the liver may also 
 be greatly congested, very friable, and marked with purple patches. 
 The examination of the blood of the heart and spleen for anthrax 
 bacilli must be carried out with great perseverance and discrimi- 
 nation, as they are present only in small numbers, arid in some 
 cases have given place entirely to septic organisms. Inoculation 
 with the blood will produce either typical anthrax, or malignant 
 oedema or some other form of septicaemia. Possibly in the cases 
 arising from ingestion of offal the ulcerated condition of the throat 
 affords a nidus and a means of access for septic organisms. It 
 is also well known that blood in a state of putrefaction may 
 contain the bacillus of malignant oedema. In the presence of 
 putrefactive organisms the anthrax bacillus rapidly disappears. If, 
 therefore, inoculation of guinea-pigs or mice is used as a test for 
 ascertaining the nature of an outbreak in swine, it must not be 
 concluded, if Pasteur's or some other form of septicaemia result, 
 that the disease was not anthrax, while, on the other hand, the 
 discovery of the anthrax bacillus in the blood of the pig, or the 
 production of anthrax in guinea-pigs or mice, is positive evidence 
 as to the nature of the original disease. 
 
 Peuch, in France, had obtained similar results by injecting pigs 
 with anthrax blood and anthrax cultures. He also carried out 
 some interesting experiments bearing on public health. The leg of 
 a pig which had died of anthrax was covered with pounded sea-salt. 
 Previously to the curing, a slice of the flesh was squeezed in a meat- 
 press, and the liquid thus obtained was employed for inoculation. 
 The animals inoculated died of typical anthrax. In six weeks the 
 curing was considered to be completed, and a slice was cut from the 
 ham and soaked in filtered water. The juice was extracted in the 
 meat-press, and employed for the inoculation of four guinea-pigs 
 and three rabbits. Slight swelling and a certain amount of redness 
 at the seat of inoculation were the only results. A few drops of 
 the muscle- juice were added to sterilised broth, and produced a 
 mixed cultivation of micrococci and motile bacilli. A rabbit and two 
 guinea-pigs inoculated with the cultivation remained quite healthy. 
 
ANTHRAX. 207 
 
 These experiments demonstrated that salting destroys the viru- 
 lence of the flesh of pigs which have died of anthrax, but in order 
 to obtain this result the salting must be thoroughly carried out. If 
 the process be incomplete the flesh is still virulent. Thus the leg 
 of a pig salted for only fourteen days furnished a juice which 
 possessed a certain amount of virulence. Out of three inoculated 
 rabbits, one died in ninety-seven hours of anthrax, and the others 
 recovered. Three guinea-pigs all succumbed, and a fourth guinea- 
 pig inoculated with a cultivation from the muscle- juice also died. 
 Peuch considers that there is danger in consuming flesh which has 
 not been thoroughly cured. 
 
 As it has been clearly shown that pigs may become infected with 
 anthrax, these animals come under the Anthrax Order of 1886. 
 This provides for the disposal of the carcass; and although Peuch 
 has shown that salting destroys the virulence of the flesh of pigs 
 which have died of anthrax, there can be no doubt that it is quite 
 right that such animals should be condemned as unfit for food. 
 
 Further, the recognition of the occurrence of true anthrax in 
 swine is an additional reason for condemning the Continental practice 
 of eating hams, sausages, etc., in the raw state. Indeed, the viru- 
 lence of anthrax flesh suggests one possible explanation of some 
 of those obscure cases of meat poisoning which have occurred in 
 this country. It is possible that the flesh of animals which had 
 died of anthrax was used in the preparation of sausages, pork-pies, 
 etc., and that the cooking was not sufficient to deprive the meat 
 of its poisonous properties. 
 
 EQUINE ANTHRAX. 
 
 Veterinary authorities have described " Anthrax in the Horse," 
 but it remains to be seen whether there are not two or more affec- 
 tions included under this heading. Fleming says : " The most 
 acute form of anthrax, the apoplectic, is somewhat rare in the 
 horse, and has perhaps been most frequently observed on the 
 Continent. Though cases are recorded, but through an error in 
 diagnosis, under other names in the veterinary literature of this 
 country, I have only witnessed two cases in England ; though during 
 the intense summer heat in the north of China I had several." 
 
 The question to which the author is in a position to give a 
 definite answer is, whether the disease produced by the Bacillus 
 anthracis ever occurs in the horse. Whether that has been pre- 
 viously determined, at any rate in this country, it is difficult to say. 
 
208 INFECTIVE DISEASES. 
 
 Fleming in describing the pathological anatomy of anthrax in the 
 horse, says : " The spleen is double and treble its ordinary volume ; 
 its surface is sometimes bosselated by tumours; its texture is 
 softened and transformed into a viscid reddish-brown or violet mass, 
 and the mesenteric glands are infiltrated. The blood in it has 
 been found to contain bacteridia when examined soon after death." 
 Williams, who says that " anthrax in the horse rarely occurs 
 in this country," adds, that it is prevalent in India, and is 
 there termed " Loodiaiia disease," and in Africa " Horse-sick- 
 ness." But " Horse-sickness," from recent researches, is certainly 
 not anthrax. Williams described a case which occurred in 1879 as 
 one of anthrax. A carriage-horse died suddenly while in harness ; 
 " a large black tumour was found in the lungs, and the pulmonary 
 arteries were engorged with black tarry blood, which, when micro- 
 scopically examined, was found to contain the bacilli in a most 
 perfect form, and very numerous indeed." In 1884, an outbreak 
 of charbonous fever occurred in Liverpool. Williams proceeded 
 to investigate the outbreak, and found two horses dead on his 
 arrival, one having died only a few hours previously. The bacilli 
 from the blood in this case are figured, and the following statement 
 made : " These bacilli seem to differ from those of splenic fever, being 
 rather smaller in diameter, and so far as my observations go, 
 multiply by fission only, not developing spores." 
 
 On the other hand, the author investigated the blood of a mare 
 which was supposed to have died of anthrax, and on examining 
 cover-glass preparations of the blood, it was found to contain large 
 numbers of bacilli with the characteristic microscopical appearances 
 of anthrax bacilli. To place the question beyond any possible doubt 
 a number of tubes of agar-agar were inoculated. These, after three 
 days in the incubator, produced typical cultivations, and on examina- 
 tion by the ordinary methods and by double-staining, yielded very 
 beautiful preparations of filaments and spores. 
 
 At the same time that the cultivations were prepared, two mice 
 were inoculated at the root of the tail with a trace of the blood. 
 Two days afterwards they w^ere both found dead, and with the 
 characteristic post-mortem appearances, spleen much enlarged, and 
 anthrax bacilli in enormous numbers. 
 
 There can be no doubt that true anthrax occurs in the horse ; 
 and the author, in 1887, recommended that it should be scheduled 
 under the Contagious Diseases (Animals) Act, and equine anthrax 
 has been included in the Anthrax Order of 1895. 
 
 More recently Pemberthy has described cases of equine anthrax 
 
ANTHRAX. 
 
 209 
 
 which he talieves to have been the result of infection from feeding 
 on foreign oats or imported hay. 
 
 Preventive Inoculation. The prevention of anthrax by 
 i n< ans of protective inoculation or vaccination has been attempted 
 on a very large scale in France, and it is claimed that the results 
 have been very beneficial to agriculture in that country : 
 
 
 
 
 Animals 
 
 Mortality. 
 
 
 
 
 
 Total 
 
 No. of 
 
 Vacci- 
 
 
 
 Total 
 
 Average 
 
 
 No. of 
 
 Veteri- 
 
 nated 
 
 
 
 
 
 loss 
 
 loaT 
 
 Year. 
 
 Animals 
 Vacci- 
 nated. 
 
 nary 
 Re- 
 ports. 
 
 after 
 Receipt 
 of 
 Reports. 
 
 After 
 1st 
 Vacci- 
 nation. 
 
 After 
 2nd 
 Vacci- 
 nation. 
 
 During 
 rest of 
 Year. 
 
 Total. 
 
 per 
 cent. 
 
 before 
 Vacci- 
 nation. 
 
 
 /1882 
 
 270..040 
 
 112 
 
 243,199 
 
 756 
 
 847 
 
 1,037 
 
 2,640 
 
 1-08 
 
 10% 
 
 
 1883 
 
 268,505 
 
 103 
 
 193,119 
 
 436 
 
 272 
 
 T84 
 
 1,492 
 
 0-77 
 
 
 
 1884 
 
 316,553 
 
 109 
 
 231,693 
 
 770 
 
 444 
 
 1,033 
 
 2,247 
 
 0-97 
 
 
 
 1885 
 
 342,040 
 
 144 
 
 280,107 
 
 884 735 
 
 990 
 
 2,609 
 
 0-03 
 
 
 
 1886 
 
 313,288 
 
 88 
 
 202,064 
 
 652 
 
 303 
 
 514 
 
 1,469 
 
 0-72 
 
 
 
 1887 
 
 293,572 
 
 107 
 
 187,811 
 
 718 
 
 737 
 
 968 
 
 2,423 
 
 1-29 
 
 
 Sheeprf 
 
 
 
 
 
 
 
 
 
 
 
 
 1888 
 
 269,574 
 
 50 
 
 101,834 
 
 149 
 
 181 
 
 300 
 
 630 
 
 0-62 
 
 
 
 1889 
 
 230,074 
 
 48 
 
 88,483 
 
 238 
 
 285 
 
 501 
 
 1,024 
 
 1-16 
 
 
 
 1890 
 
 223,611 
 
 69 
 
 69,865 
 
 331 
 
 261 
 
 244 
 
 836 
 
 1-20 
 
 
 
 1891 
 
 218,629 
 
 65 
 
 53,640 
 
 181 
 
 102 
 
 77 
 
 360 
 
 0-67 
 
 
 
 1802 
 
 269,996 
 
 70 
 
 63,125 
 
 319 
 
 183 
 
 126 
 
 628 
 
 0-99 
 
 
 
 U893 
 
 281,333 
 
 30 
 
 73,936 
 
 234 
 
 56 
 
 224 
 
 514 
 
 0-69 
 
 
 Total . 
 
 3,296,815 
 
 990 
 
 1,788,077 
 
 5,668 
 (0-32%) 
 
 4,406 
 (0-24%) 
 
 6,798 
 (0-38%) 
 
 16,872 
 
 0-94 
 
 
 
 /1882 
 
 35,654 
 
 127 
 
 22,916 
 
 22 
 
 12 
 
 48 
 
 82 
 
 0-35 
 
 ** 
 
 
 1883 
 
 26,453 
 
 130 
 
 20,501 
 
 17 
 
 1 
 
 46 
 
 64 
 
 0-31 
 
 
 
 1884 
 
 33,900 
 
 139 
 
 22,616 
 
 20 
 
 13 
 
 52 
 
 85 
 
 0-37 
 
 
 
 1885 
 
 34,000 
 
 192 
 
 21,073 
 
 32 
 
 8 
 
 67 
 
 107 
 
 0-50 
 
 
 
 1886 
 
 30,154 
 
 135 
 
 22,113 
 
 18 
 
 7 
 
 39 
 
 64 
 
 0-29 
 
 
 Oxen 
 
 
 
 
 
 
 
 
 
 
 
 
 1887 
 
 48,484 
 
 148 
 
 28,083 
 
 23 
 
 18 
 
 68 
 
 109 
 
 0-39 
 
 
 or 
 
 
 
 
 
 
 
 
 
 
 
 
 1888 
 
 34,464 
 
 61 
 
 10,920 
 
 8 
 
 4 
 
 35 
 
 47 
 
 0-43 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 1889 
 
 32,251 
 
 68 
 
 11,610 
 
 14 
 
 7 
 
 31 
 
 52 
 
 0-45 
 
 
 
 ISM 
 
 33,965 
 
 71 
 
 11 057 
 
 5 
 
 4 
 
 14 
 
 23 
 
 0-21 
 
 
 
 1891 
 
 40,736 
 
 68 
 
 10,476 
 
 6 
 
 4 
 
 4 
 
 14 
 
 0-13 
 
 
 
 1802 
 
 41,609 
 
 71 
 
 9,757 
 
 8 
 
 3 
 
 15 
 
 26 
 
 0-26 
 
 
 
 U893 
 
 38,154 
 
 45 
 
 9,840 
 
 4 
 
 1 
 
 13 
 
 18 
 
 0-18 
 
 
 
 438,824 
 
 
 200,962 
 
 177 
 
 82 
 
 432 
 
 691 
 
 0-34 
 
 
 
 
 
 (0-09%) 
 
 (0-04%) 
 
 (0-21%) 
 
 
 
 
 i 
 
 
 
 
 
 
 
 
 I 
 
 14 
 
210 
 
 INFECTIVE DISEASES. 
 
 The vaccine is supplied, by a company in Paris, in two strengths. 
 Reports are supplied by veterinary surgeons, and the results have 
 been tabulated by Chamberland and published, and commented upon 
 by Cope in a report to the Board of Agriculture (1894). The column 
 of deaths, in the above table, includes the animals which died from 
 the vaccination, and those which died from natural infection. 
 
 It is claimed that the percentage of losses has been reduced from 
 10 per cent, to *94 per cent, in sheep, and from 5 per cent, to '34 per 
 cent, in cattle. Cope, in the report just referred to, regards these 
 conclusions as somewhat fallacious, because in order to prove that 
 the animals inoculated received immunity, it should be shown that 
 they were subsequently exposed to the risks of natural infection. 
 This was not the case. But a report obtained from the Bureau in 
 Paris gives the actual number of animals on each of the infected 
 farms, and the number which have died of the disease ; and when 
 compared with Chamberland's statistics it is evident that nine-tenths 
 were not on farms where the disease appeared at least, during 
 1889-92 and that the deaths from anthrax on those farms where 
 it was reported to exist were, if anything, higher than they were 
 supposed to be prior to the introduction of the system of vaccination ; 
 and in spite of the immense number of animals vaccinated the 
 official returns obtained from Paris, by Cope, indicate that the 
 mortality from anthrax, calculated in the ordinary way, remains as 
 high as ever. 
 
 Anthrax in France. 
 
 Year. 
 
 No. of 
 Outbreaks re- 
 ported. 
 
 No. of Animals 
 in Premises. 
 
 No. of which 
 Died. 
 
 Percentage of 
 Loss. 
 
 1889 
 
 618 
 
 22,599 
 
 1,458 
 
 6-5 
 
 1890 
 
 536 
 
 24,073 
 
 1,123 
 
 4-7 
 
 1891 
 
 570 
 
 21,356 
 
 1,444 
 
 6-8 
 
 1892 
 
 607 
 
 28,199 
 
 1,581 
 
 5-6 
 
 CATTLE. 
 
 1889 
 
 
 
 6,059 
 
 700 11-6 
 
 1890 
 
 
 
 5,365 
 
 771 14-4 
 
 1891 
 
 
 
 7,299 
 
 849 11-6 
 
 1892 
 
 
 
 5,058 
 
 804 15-9 
 
 SHEEP. 
 
 1889 i 16,540 
 
 755 4-6 
 
 1890 
 
 18,708 
 
 352 1-9 
 
 1891 
 
 14,057 
 
 545 4-2 
 
 1892 
 
 23,141 
 
 777 
 
 3-4 
 
ANTHRAX. 
 
 In Germany, veterinary and agricultural authorities agree that 
 the results have not met with the success which has been claimed 
 for vaccination in France. Experiments were undertaken for the 
 German Government, and in one set of experiments twenty-five sheep 
 \vriv vaccinated with the first vaccine without an accident, but three 
 died five days after the second vaccine. In another experiment two 
 hundred and fifty-one sheep were vaccinated with only one death, 
 and subsequent inoculation with virulent anthrax proved that they 
 had immunity. 
 
 Six head of cattle were vaccinated .without any loss, and six more 
 were used for a control experiment. Inoculation with virulent virus 
 proved fatal to the control animals, but the vaccinated were pro- 
 tected. These, with other animals similarly vaccinated, amounting 
 in all to two hundred and sixty-six sheep and eighty-three head of 
 cattle, were then turned out to graze on infected pastures with two 
 hundred and sixteen unvaccinated sheep as a control experiment. 
 Within five months four of the vaccinated and eight of the un- 
 vaccinated sheep died of anthrax, and one of the vaccinated and one 
 of the unvaccinated cattle. 
 
 The result of these experiments led to the following conclusions : 
 
 (1) That the first vaccine is mild and harmless. 
 
 (2) That the second vaccine, even in the hands of experts, is 
 dangerous and often fatal. 
 
 (3) That sheep are more affected than cattle by the injections, 
 exhibiting fever and other indications of illness. 
 
 (4) That cattle and sheep which recover from the vaccination 
 have an immunity against anthrax when tested by experimental 
 inoculation. 
 
 (5) That vaccinated cattle and sheep tested by exposure to 
 natural infection by grazing on infected pastures contract the 
 ilist-ase in the ordinary way. 
 
 (6) That the time for which immunity is conferred has not been 
 determined. 
 
 In England, Klein tested the vaccine, with the result that animals 
 either succumbed to the vaccine, or to virulent anthrax after recovery 
 from the vaccine. Protective inoculation has also been employed in 
 a few instances by leading agriculturists, but with very unsatis- 
 factory results. 
 
 Stamping-out System: In Germany the conclusion is that 
 the safest measures are destruction of carcasses and disinfection, and 
 that inoculation will have no effect in lessening the loss caused by 
 this disease. 
 
212 INFECTIVE DISEASES. 
 
 In England the stamping- out system has been advocated for 
 many years, and is still regarded as the only reliable means for 
 suppressing the disease ; and the possible introduction of the disease 
 among healthy stock by vaccination, and especially in localities in 
 which anthrax is unknown, would be contrary to the principles upon 
 which the system is based. These principles are illustrated by the 
 following extracts from the Anthrax Order of, 1895 : 
 
 NOTIFICATION. 
 
 2. (1) Every person having or having had in his possession or under 
 his charge, an animal affected with or suspected of anthrax, shall, with all 
 practicable speed, give notice of the fact of the animal being so affected or 
 suspected, to a constable of the police force for 'the police area wherein 
 the animal so affected or suspected is or was. 
 
 (2) The constable shall forthwith give information of the receipt by 
 him of the notice to an Inspector of the Local Authority, who shall forth- 
 with report the same to the Local Authority. 
 
 (3) The Inspector of the Local Authority shall forthwith give 
 information of the receipt by him of the notice to the Medical Officer 
 of Health of the Sanitary District in which the affected or suspected 
 animal is or was. 
 
 Duty of Inspector to act immediately. 
 
 3. An Inspector of a Local Authority on receiving in any manner 
 whatsoever information of the supposed existence of anthrax, or having 
 reasonable ground to suspect the existence of anthrax, shall proceed with 
 all practicable speed to the place where such disease, according to the 
 information received by him, exists, or is suspected to exist, and shall 
 there and elsewhere put in force and discharge the powers and duties 
 conferred and imposed on him as Inspector, by or under the Act of 1894 
 and this Order. 
 
 Public Warning as to Existence of Disease. 
 
 4. (1) The Local Authority may, if they think fit, give public 
 warning by placards, advertisement, or otherwise, of the existence of 
 anthrax in any shed, stable, building, field,' or other place, with or without 
 any particular description thereof, as they think fit, and may continue to 
 do so during the existence of the disease, and, in case of a shed, stable, 
 building, or other like place, until the same has been cleansed and dis- 
 infected in accordance with this Order. 
 
 (2) It shall not be lawful for any person (without authority or 
 excuse) to remove or deface any such placard. 
 
 Milk of Diseased or Suspected Cow not to be Removed. 
 
 5. Where anthrax exists or has existed in any shed, stable, building, 
 or other place, it shall not be lawful to remove from such shed, stable, 
 
ANTHRAX. 213 
 
 building, or other place the milk of any cow which is affected with or 
 suspected of anthrax. 
 
 Removal of Dung or (] / Things. 
 
 G. It shall not be lawful for any person to send or carry, or cause to 
 be sent or carried, on a railway, canal, river, or inland navigation, or in 
 a coasting vessel, or on a highway or thoroughfare, any dung, fodder, or 
 litter that has been in any place in contact with or used about a diseased 
 or suspected animal, except with a Licence of the Local Authority for 
 the District in which such place is situate, on a certificate of an Inspector 
 of the Local Authority certifying that the thing moved has been, so far 
 as practicable, disinfected. 
 
 Dupotttl of Carcasses. 
 
 7. (1) The carcass of an animal which at the time of its death was 
 affected with or suspected of anthrax shall be disposed of by the Local 
 Authority as follows : 
 
 (i.) Either the Local Authority shall cause the carcass to be buried 
 as soon as possible in its skin in some convenient or suitable place 
 removed from any dwelling house and at such a distance from any 
 well or watercourse as will preclude any risk of the contamination 
 of the water therein, and at a depth of not less than six feet 
 below the surface of the earth, having a layer of lime not less 
 than one foot deep beneath, and a similar layer of lime above, 
 the carca-< : 
 
 (ii.) Or the Local Authority may, if authorised by Licence of the 
 Board, cause the carcass to be destroyed, under the inspection of 
 the Local Authority, in the mode following: The carcass shall 
 be disinfected, and shall then be taken, in charge of an officer of 
 the Local Authority, to a horse-slaughterer's or knacker's-yard 
 approved for the purpose by the Board, or other place so approved, 
 and shall be there destroyed by exposure to a high temperature, 
 or by chemical agents. 
 
 (2) With the view to the execution of the foregoing provisions of this 
 Article the Local Authority may make such Regulations as they think fit 
 for prohibiting or regulating the removal of carcasses, or for securing the 
 burial or destruction of the same. 
 
 (3) Before a carcass is removed for burial or destruction under this 
 Article it shall be covered with quicklime. In no case shall the skin of 
 the carcass be cut, nor shall anything be done to cause the effusion of 
 blood. 
 
 (4) A Local Authority may cause or allow a carcass to be taken into 
 the District of another Local Authority to be buried or destroyed, with 
 the previous consent of that Local Authority, but not otherwise. 
 
 *. It shall not be lawful for any person, except with the Licence of 
 the Board or permission in writing of an Inspector of the Board, to dig 
 
214 INFECTIVE DISEASES. 
 
 up. or cause to be dug up, the carcass of any animal that has been 
 buried. 
 
 Disinfection in Case of Anthrax. 
 
 9 (i) The Local Authority shall at their own expense cause to be 
 cleansed and disinfected in the mode provided by this Article 
 
 () All those parts of any shed, stable, building, or other place in 
 
 which a diseased or suspected animal has been kept or has died or 
 
 been slaughtered ; 
 (&) Every utensil, pen, hurdle, or other thing used for or about any 
 
 diseased or suspected animal ; 
 (c) Every van, cart, or other vehicle used for carrying any diseased 
 
 or suspected animal on land otherwise than on a railway. 
 
 (2) The mode of the cleansing and disinfection of such shed, stable, 
 building, or other place, or the part thereof, shall be as follows : 
 
 (i.) All those parts aforesaid of the shed, stable, building, or other 
 place shall be swept out, and all litter, dung, or other thing that 
 has been in contact with, or used about, any diseased or suspected 
 animal shall be effectually removed therefrom ; then 
 
 (ii.) The floor and all other parts of the shed, stable, building, or other 
 place with which the diseased or suspected animal or its droppings 
 or any discharge from the mouth or nostrils of the animal has come 
 in contact, shall be, so far as practicable, thoroughly washed or 
 scrubbed or scoured with water ; then 
 
 (hi.) The same parts of the shed, stable, building, or other place shall 
 be washed over with limewash made of freshly burnt lime and 
 water, and containing in each gallon of limewash four ounces of 
 chloride of lime or half a pint of commercial carbolic acid, the 
 limewash being prepared immediately before use ; 
 
 (iv.) Except that where any place as aforesaid is not capable of being 
 so cleansed and disinfected, it shall be sufficient if such place be 
 cleansed and disinfected so far as practicable. 
 
 (3) The mode of the cleansing and disinfection of such utensil, pen, 
 hurdle, or other thing, and such van, cart, or other vehicle aforesaid, shall 
 be as follows : 
 
 (i.) Each utensil, pen, hurdle, or other thing, van, cart, or other 
 vehicle, shall be thoroughly scraped, and all litter, dung, sawdust, 
 or other thing shall be effectually removed therefrom ; then 
 
 (ii.) It shall be thoroughly washed or scrubbed or scoured with water ; 
 then 
 
 (iii.) It shall be washed over with limewash made of freshly burnt 
 lime and water, and containing in each gallon of limewash four 
 ounces of chloride of lime or half a pint of commercial carbolic 
 acid, the limewash being prepared immediately before use. 
 
 (4) All litter, dung, or other thing that has been removed from any 
 such shed, stable, building, place, van, cart, or vehicle as aforesaid, shall 
 be forthwith burnt or otherwise destroyed or disinfected to the satisfac- 
 tion of an Inspector of the Local Authority. 
 
ANTHRAX. 215 
 
 (5) The Local Authority may make such Regulations as they think 
 fit for the purpose of carrying out the provisions of this Article. 
 
 Occupiers to give Facilities for Cleansing. 
 
 10. (1) Where the power of causing any place, thing, or vehicle to 
 be cleansed and disinfected under this Order is exercised by a Local 
 Authority, the owner and occupier and person in charge of the place, 
 thing, or vehicle shall give all reasonable facilities for that purpose. 
 
 (2) Any person failing to comply with the provisions of this Article 
 shall be deemed guilty of an offence against the Acttof 1894. 
 
 Regulations of Local Authority as to Movement of Animals, 
 Fodder, etc. 
 
 11. A Local Authority may make such Regulations as they think fit 
 for the following purposes, or any of them : 
 
 (a) For prohibiting or regulating the movement of any diseased or 
 
 suspected animal into or out of any shed, stable, building, field, or 
 
 other place, or any part thereof ; 
 
 (6) For prohibiting or regulating the movement of any animal into 
 or out of any shed, stable, building, field, or other place, or any 
 part thereof, in which there is or has been any diseased or 
 suspected animal ; and 
 
 (c) For regulating the removal out of any shed, stable, building, 
 field, or other place of any fodder, litter, or other thing that has 
 been in contact with or used for or about any diseased or suspected 
 animal ; 
 
 but nothing in any such Regulation shall authorise movement in 
 contravention of any provision of any Order of the Board for the time 
 being in force : and a Regulation under paragraph (6) of this Article 
 shall operate so long only as any animal which in the judgment of the 
 Local Authority is diseased or suspected remains in the shed, stable, 
 building, field, or other place to which the Regulation refers, and, in c:ise 
 of a shed, stable, building, or other like place, until the same has been 
 cleansed and disinfected in accordance with this Order. 
 
 Slaughter in Anthrax and Compen*ati<>n. 
 
 12. (1) A Local Authority may if they think fit cause to be 
 slaughtered 
 
 (/{) Any animal affected with anthrax or suspected of being so 
 affected ; and 
 
 (6) Any animal being or having been in the same field, shed, or other 
 place, or in the same herd or flock or otherwise in contact with 
 animals affected with anthrax, or being or having been in the 
 opinion of the Local Authority in any way exposed to the infection 
 of anthrax. 
 
 (2) The slaughter of animals under this Article shall be conducted 
 in such mode as will so far as possible prevent effusion of blood. 
 
216 INFECTIVE DISEASES. 
 
 (3) The Local Authority shall out of the local rate pay compensation 
 as follows for animals slaughtered under this Article : 
 
 (a) Where the animal slaughtered was affected with anthrax the 
 compensation shall be one-half of the value of the animal 
 immediately before it became so affected ; and 
 
 (&) In every other case the compensation shall be the value of the 
 animal immediately before it was slaughtered. 
 
 (4) Provided, that if the owner of the animal gives notice in writing 
 to the Local Authority, or their Inspector or other officer, that he objects 
 to the animal being slaughtered, it shall not be lawful for the Local 
 Authority to cause that animal to be slaughtered except with the further 
 special authority of the Board first obtained. 
 
 Keeping of Swine in Slaughter-houses. 
 
 16. It shall not be lawful for any person, in any case in which the 
 slaughter of any animal is authorised or required by this Order, to use 
 for such slaughter any slaughter-house in which swine are kept. 
 
 Whether an anthrax virus can be obtained which is absolutely 
 incapable of creating centres of infection, and can therefore be 
 recommended with safety for vaccination as an auxiliary and 
 voluntary measure, is a matter for further investigation. 
 
CHAPTER XV. 
 
 QUARTER-EVIL. MALIGNANT (EDEMA. RAG-PICKERS' SEPTICAEMIA. 
 SEPTIC /EMIA OF GUINEA-PIGS. SEPTICLEMIA OF MICE. 
 
 QUARTER-EVIL in cattle, malignant oedema, and rag-pickers' septicaemia 
 in man, septicaemia in guinea-pigs, and septicaemia in mice, are all 
 varieties of septicaemia produced by bacilli. 
 
 An account of quarter-evil, malignant oedema, and rag-pickers' 
 septicaemia may appropriately follow the chapter on anthrax, as 
 they have certain similarities to that disease. They are, however, 
 not only distinct from anthrax, but must be carefully distinguished 
 from each other. In connection with these forms of bacillary 
 septicaemia in man and cattle we may study bacillary septicaemia 
 in small animals. 
 
 QUARTER-EVIL. 
 
 The disease known in this country as quarter-evil or black-leg 
 is identical with the French Charbon 8y//>fiiif if/tie and the 
 German Rauschbrand. Symptomatic anthrax in a very slight degree 
 resembles anthrax. The disease occurs usually in young cattle from 
 a few weeks to about twelve months old, and attacks sheep and 
 horses, but not swine or poultry. It is characterised by the develop- 
 ment of an emphysematous swelling of the subcutaneous tissue and 
 muscles, generally over the hind quarter. Infected animals cease 
 feeding, the temperature rises, lameness supervenes, and death 
 occurs in about forty-ri^ht hmir>. The tumour on incision is found 
 to contain a quantity of dark sanguineous fluid, with characteristic 
 bacilli; 
 
 Bacillus of Quarter-evil (Jiacille du charbwi syinptomatique, 
 li'i,i*,-1,l,fi,,nl ftocitfu*). Motile rods with rounded ends, 3 to 5 /x in 
 lenirth, -5 to '6 /* in breadth. Spore-formation present. The 
 spores are oval, generally >ituate<l near the extremity of the nU. 
 and when fully developed considerably exceed the rods in diameter. 
 
 217 
 
218 
 
 INFECTIVE DISEASES. 
 
 Involution forms are freely developed in old cultures, and in 
 cultures made in unsuitable media. The bacilli possess numerous 
 
 flagella, and their 
 
 power of movement at 
 
 ^p once disting uishes them 
 
 . * from anthrax bacilli. 
 
 J 4? They can be cultivated 
 
 \ , in the ordinary media 
 
 o _ O o in the absence of oxy- 
 
 gen, but more readily 
 j with the addition of 
 grape-sugar or glyce- 
 rine. Radiating fila- 
 
 01 ments grow out from 
 
 . ^ the more or less spher- 
 ^ ical colonies directly 
 
 FIG. 101. BACILLI OP QUARTER-EVIL x 1000. From 
 an agar culture (FRANKEL and PFEIFFER). 
 
 liquefaction commences. In the depth of 
 nutrient gelatine the growth occurs in two 
 or three days at 20 to 25 0. towards the 
 lower part of the track of the inoculating 
 needle. The gelatine slowly liquefies, and 
 there is considerable formation of gas with 
 the development of a peculiar odour. Spore- 
 formation occurs freely in cultures, but not 
 in the blood of infected animals until after 
 death. 
 
 Guinea-pigs inoculated with a pure- 
 culture, or with spore-bearing threads, die 
 in twenty-four to thirty-six hours. An em- 
 physematous infiltration with sanguineous 
 serum is produced at the seat of inoculation, 
 and the surrounding muscles are of a dark 
 colour. The internal organs are more or less FlG 102 P UBE _CULTUHK OF 
 congested. The bacilli are found in the BACILLI OF QUARTER-EVIL 
 
 local exudation and in the surrounding IN GI^PE-SUGAR GELA- 
 TINE (FRANKEL and 
 tissue, and some hours after death in PFEIFFER). 
 
QUARTER-EVIL. 219 
 
 increasing numbers in the blood of the heart and in the internal 
 organs. 
 
 Quarter-evil and malignant oedema, though possessing points of 
 resemblance, are distinct diseases. Not only do the bacilli in the 
 two cases differ in minute morphological and biological details, but 
 Kitasato showed that guinea-pigs rendered immune against virulent 
 quarter-evil had no immunity against malignant oedema. 
 
 Protective Inoculation. Arloing, Cornevin and Thomas have 
 produced immunity by inoculating healthy cattle with a small 
 quantity of the fluid from the tumour of an infected animal. 
 Recovery takes place, and subsequent inoculation with a strong dose 
 is without effect. Similar results may be obtained by intravenous 
 injection of a few drops of the exudation. For general application 
 of the system of protective inoculation, the virulent liquid and 
 affected muscles are dried at 32 to 35 C., and the dried mass 
 triturated with water and heated to 100C. This is used as the 
 first vaccine. 
 
 An infusion similarly prepared, but only heated to 80 C., forms 
 the second vaccine. The dry powder is a convenient form for 
 general distribution, and y 1 ^ of a gramme is triturated with 5 cc. 
 of water, and | cc. is injected into each animal. In about ten days 
 the second vaccine is employed, and cattle so treated are said to 
 have a complete immunity from fatal doses. 
 
 The place chosen for the injection is the under surface of the 
 tail, a short distance from the extremity. The hair is clipped at 
 this spot, and the point of a syringe is pushed in between the skin 
 and the bone, and the vaccine slowly injected. 
 
 Roux and Chamberland produced immunity by inoculation of 
 filtered cultures. Cultures in broth were deprived of bacilli by 
 heating to 115 C., or by filtration through porcelain. Guinea-pigs 
 were inoculated with three doses of 30 cc. at intervals of two days, 
 and subsequently injected with a solution of virulent black-leg 
 powder and lactic acid, which killed control animals in twenty-four 
 hours. Kitt employed cultures on agar a fortnight old, or fresh 
 cultures sterilised by steam for thirty minutes. It was found 
 possible to confer immunity in oxen, sheep, and guinea-pigs against 
 the most virulent extract. Kitt's method has the advantage over 
 others of only necessitating one single injection. Whether these 
 experiments are of scientific interest rather than of practical value 
 may be regarded as an open question. 
 
 On the Continent, and especially in France, vaccination against 
 quarter-evil has been carried out extensively ; and by comparing the 
 
220 INFECTIVE DISEASES. 
 
 mortality among the vaccinated and unvaccinated in localities where 
 the disease commonly occurs, it has been said that the results are 
 extremely favourable. The matter was investigated in this country 
 by a committee of the Midland Veterinary Medical Association, 
 and in the course of the experiments some surprising results were 
 obtained. Six calves and four sheep were vaccinated, and five 
 calves and two sheep were left unvaccinated as a control experiment. 
 The seventeen animals were subsequently inoculated with virulent 
 virus in the form of dried and pow^dered muscle. In forty-eight 
 hours all the sheep died, and all the calves exhibited a swelling at 
 the seat of inoculation. In another set of experiments, healthy 
 calves inoculated with fresh juice from the tumour in a case of 
 quarter-evil were not materially affected. The possibility of those 
 calves which possess a natural immunity being classed as protected 
 by the inoculation must be admitted, and the efficacy and safety 
 of the process is by no means established. 
 
 MALIGNANT (EDEMA. 
 
 The disease known by surgeons as progressive gangrene, gan- 
 grenous emphysema, or surgical gangrene, has been shown by the 
 researches of Chauveau, Arloing, Rosenbach and Babes, to be 
 due to a bacillus identical with the microbe septique of Pasteur and 
 the bacillus of malignant oedema of Koch. The bacillus or its spores 
 may be spread by the neglect of antiseptics. The disease occurs 
 especially after compound fractures and gun-shot wounds. 
 
 If a guinea-pig is subcutaneously inoculated with earth, putrid 
 fluid, or hay dust, death frequently occurs in from twenty-four to 
 forty-eight hours. At the autopsy the most characteristic symptom 
 is a widespread subcutaneous oedema accompanied by air-bubbles. 
 This originates from the point of inoculation, and contains a 
 clear reddish liquid full of motile and n on- motile bacilli. The 
 internal organs are little changed, the spleen is enlarged and of a 
 dark colour, and the lungs are hypersemic, and have hsemorrhagic 
 spots. Examined immediately after death, few or no bacilli are 
 detected in the blood of the heart, but in that of the spleen, liver, 
 lungs, and other organs, in the peritoneal exudation, and in and 
 upon the serous coating of the abdominal organs, they are present in 
 large numbers. If, on the other hand, the animal is not examined 
 until some time after death, the bacilli are found in the blood of 
 the heart, and distributed all over the body. 
 
 Bacillus CEdematis Maligni, Koch (Pasteur's Septicaemia). 
 
MALK.NANT (EDEMA. 
 
 221 
 
 Rods from 3 to 3'5 /x long and 1 to 1*1 p wide ; they mostly li.- 
 in pairs, and then appear to be double this length. The rods are 
 rotinde I at their ends, and form threads which are sometimes straight, 
 but more commonly curved. In stained preparations they have a 
 somewhat granular appearance. They are motile, p>eing flagella, 
 and form spores. The bacilli are distinguished from anthrax bacilli 
 by their being somewhat thinner, by their rounded ends, and by their 
 motility. Moreover, anthrax bacilli never appear as threads in fresh 
 blood, and are differently distributed throughout the body. They 
 are anaerobic, and can be cultivated on blood serum and on neutral 
 >o!ution of Liebig's meat extract in an atmosphere of carbonic acid. 
 By embedding material containing bacilli in nutrient agar-agar 
 
 \ 
 
 FIG. 103. BACILLI OF MALIGNANT (EDEMA x 950. From the subcutaneous tissue 
 of a guinea-pig. (BAUMGARTEN.) 
 
 and nutrient gelatine, characteristic cultivations are obtained. The 
 following process may be adopted to obtain a pure cultivation. A 
 mouse inoculated sulx-utaneously with dust, as a rule, dies in one 
 to two days. It is then pinned out, back uppermost, on a slab of 
 wood, and the hair sinired with a Paquelin's cautery from one hind 
 leg up to the neck, across the latter, and down again to the opposite 
 hind leg. Following the cauterised line, the skin is cut through with 
 steri!: HOTS, and the tlap turned back and pinned out of the 
 
 way. With curved scissors little pieces of the subcutaneous 
 cwlematous tissue, in the neighbourhood of the inoculated spot, .u 
 cut out, and sunk with a platinum needle in a 1 per cent, nutrient 
 airar-asrar, or 5 per cent, nutrient gelatine. Fragments of tissue may 
 also be embedded by the method already described for anaerobic- 
 bacteria. 
 
222 
 
 INFECTIVE DISEASES. 
 
 The inoculated tubes are placed in the incubator. In a few hours 
 a whitish turbidity spreads out from the piece of tissue, and upwards 
 in the needle track. Examined microscopically, the turbidity is found 
 to be due solely to the development of the bacilli of oedema. The 
 surface exposed to the air exhibits no trace of the bacilli. To 
 investigate the tubes microscopically, a sterilised glass tube with a 
 capillary end may be used, with its neck 
 plugged with sterilised cotton- wool, and 
 provided at the mouth w r ith a suction ball. 
 The capillary end is thrust into the cultiva- 
 tion, and a small fragment removed by 
 aspiration. 
 
 In the course of the first day the bacilli 
 spread throughout a great part of the 
 agar-agar in such a way that a more or less 
 equally diffused cloudiness of the medium en- 
 sues, with subsequent appearance of strongly 
 marked clouds or lines of turbidity. At the 
 same time gas- bubbles develop along the 
 needle track, and a collection of liquid takes 
 place, while spore-formation also commences. 
 The following day these appearances are more 
 marked, the opacity is more pronounced, the 
 development of gas increases, and the liquid 
 contains more spore-forming bacilli and nu- 
 merous free-spores. 
 
 The nutrient-gelatine cultures during the 
 first day show no macroscopic change, but 
 
 FIG. 104. PURE-CULTURE after a few days the piece of tissue is sur- 
 OF BACILLUS OF MALIG- rounded with a white halo. This gradually 
 NANT (EDEMA IN GRAPE- spreads in all directions, and is apparently 
 SUGAR GELATINE (FRAN- \ 
 
 KEL and PFEIFFER). beset with hairs. I he gelatine liquefies, and 
 
 the fragment of tissue, degenerated bacilli, 
 
 and spores, sink to the bottom. The cultivation is also very 
 characteristic in a | per cent, nutrient agar-agar. If placed in 
 the incubator, in a few hours a cloudiness forms around the piece of 
 embedded tissue, which is caused by bacilli gradually spreading in all 
 directions in the nutrient medium. Mice inoculated from these 
 cultivations die more quickly than from the original infection from 
 dust. On potatoes they are cultivated by introducing a piece of liver 
 or other tissue containing the bacilli, into the interior of a sterilised 
 potato, and incubated at 38 C. The bacillus is not deprived of its 
 virulence by cultivation. 
 
\ 
 
 S 
 / ' f 
 
 MALIGNANT (EDEMA. 223 
 
 The spores of the oedema -bacilli appear to be very widely dis- 
 tributed. They are found in the upper cultivated layers of the soil, 
 in hay dust, in decomposing liquids, and especially in the bodies of 
 suffocated animals, which are left to decompose at a high temperature. 
 From any of these sources animals can be successfully inoculated. 
 The bacillus is not only pathogenic in guinea-pigs, rabbits, and 
 mice, but also in man and in farm animals, including calves but not 
 cattle. Pure-cultures inoculated in animals produce oedema at the 
 seat of inoculation 
 without appreciable 
 gas formation and 
 without any putrefac- 
 tive odour. The odour ^ 
 and frothy effusion f 
 resulting from the in- / ^ ^ 
 oculation of earth are 
 
 ) M 
 
 due to other bacteria, \i 
 
 which are introduced 
 simultaneously with *t~ 
 
 the bacilli of malig- ^ 
 
 * ^ ^ .w 
 
 nant oedema. The ^ ^^^ 
 
 spleen is sometimes 
 slightly enlarged. By x 
 
 touching with a cover- 
 glass the capsule of FlG- 105 BACILLI OF MALIGNANT (EDEMA x 10dO. 
 the spleen, or by ex- From an agar culture (FBANKEL and PFEIFFER). 
 amining the serous 
 
 effusion, the bacilli are found in abundance ; but if a preparation 
 is made from the interior of the spleen or from the blood of 
 the heart, no bacilli will be found until several hours after 
 death. In this respect there is a marked difference from 
 anthrax. Another difference is shown in spore-formation, which 
 occurs in the living body in malignant oedema, but never in 
 anthrax. Animals which recover from the disease are said to 
 be protected. 
 
 Protective Inoculation Roux and Chamber-land produced 
 
 immunity by injecting the chemical products in the filtrate obtained 
 from cultures in broth. The serum from fatal cases will, it is said, 
 confer immunity on other animals. 
 
 There is a variety of this bacillus in soil according to Fliigge, 
 agreeing in morphological and cultural but not in pathogenic 
 characters. 
 
224: INFECTIVE DISEASES. 
 
 RAG-PICKERS' SEPTICAEMIA. 
 
 Rag-pickers' disease has a resemblance to anthrax or wool-sorters' 
 disease. After death the spleen is found to be enlarged, the in- 
 ternal organs are congested, and there are haemorrhages on the 
 serous membranes. Bordoiii -Uffreduzzi isolated bacilli which are 
 quite easily distinguished from anthrax bacilli. They were found 
 in the blood and in sections of the internal organs. 
 
 Proteus hominis capsulatus. Rods with rounded ends, singly, 
 in pairs, and in filaments, somewhat smaller than anthrax bacilli, 
 and often irregular in form. Spore -formation not described ; they 
 have a well- marked capsule. Colonies are circular, appearing at 
 first granular, and later possessing a filamentous structure. In 
 the depth of gelatine they grow in the shape of a round-headed 
 nail, like a culture of Friedlander's pneumococcus. On the surface of 
 gelatine they form a shining white layer. On agar the growth is 
 somewhat transparent. On potato a moist, glistening film gradually 
 spreads over the surface. They do not liquefy blood serum, and 
 the growth is similar to that obtained on agar. They prove fatal 
 to mice and dogs, but rabbits and guinea-pigs are not very sus- 
 ceptible. Dogs die usually on the second day after intravenous 
 injection, and after death there is congestion of the internal organs 
 and of the intestinal mucous membrane. (Edema is produced at 
 the seat of inoculation in mice. There are haemorrhages in the 
 lymphatic glands, and congestion of the liver and kidneys. Similar 
 organisms have been described by Kolb and by Babes in purpura 
 hsemorrhagica. 
 
 SEPTICAEMIA OF GUINEA-PIGS. 
 
 Guinea-pigs and mice sometimes die of septicaemia, characterised 
 by congestion of the lungs, liver, and kidneys, inflamed peritoneum, 
 pleural and pericardial exudation, congested spleen, and congestion 
 of the mucous and serous coats of the intestine. Klein isolated 
 a bacillus from the blood and the internal organs in these cases. 
 
 Bacillus of Septicaemia in Guinea-pigs. Rods with rounded 
 ends, motile, with pleomorphic forms, cocci, short rods and filaments. 
 Colonies appear as small, circular, white dots, which enlarge and 
 become irregular in outline. In the depth of gelatine a white 
 filament develops, and on the surface the growth rapidly spreads 
 with a crenated outline. Broth becomes turbid, and after the second 
 day a copious white sediment is deposited. Spore -formation not 
 observed. 
 
DESCRIPTION OF PLATE VI. 
 Bacillus Murisepticus. 
 
 FlG. 1. From a section of a kidney of a mouse which had died after inocula- 
 tion with a pure-cultivation of the bacillus. With moderate amplification, 
 the white blood-corpuscles have a granular appearance, and irregular 
 granular masses are scattered between the kidney tubules. Stained by 
 Gram's method with eosin. x 200. 
 
 FlG. 2. Part of the same preparation with high amplification. The granular 
 appearances are found to be due to the presence of great numbers of 
 extremely minute bacilli, x 1500. 
 

 4 
 
 * - 
 
 * 
 
 Figl 
 
 
 
 
 m 
 
 fc>\i'' 
 
 ^ 
 
 <rvr 
 
 
 .iff 
 
 Fig 2. 
 
 **" 
 
 BACILLUS MURISEPTICUS 
 
 -,iD^- 4r<.ix.^A. 
 
SEPTICAEMIA OF MICE. 225 
 
 According to Wooldridge, the chemical products of this bacillus, 
 separated by filtration, produce on inoculation immunity against 
 virulent bacilli. 
 
 SEPTICJEMIA OF MICE. 
 
 Mice inoculated with a minimum quantity of putrid fluid often 
 die of septicaemia. They rapidly sicken, their 
 eyes inflame, their eyelids stick together, they 
 become soporific, and death occurs in forty to 
 sixty hours. There is slight oedema at the seat 
 of inoculation, and enlargement of the spleen ; 
 the bacilli are found free and in the interior 
 of white corpuscles, both in the oedematous 
 tissue and in the blood capillaries. 
 
 Bacillus of Septicaemia of Mice (Koch). 
 Extremely minute bacilli, -8 to 1 JJL long, and 
 1 to *2 n broad, and filaments. In cultivations 
 in gelatine they do not appear to make threads, 
 but the bacilli lie together in masses. Spores 
 have been observed. The bacilli are probably 
 non-motile. They are most commonly in the 
 interior of white blood corpuscles. In these 
 they increase, and in many cases a white blood 
 cell is represented only by a mass of bacilli. 
 
 A minimal quantity of blood containing the 
 bacilli produces the disease if inoculated in 
 house-mice or sparrows. Field-mice have an 
 immunity. Rabbits and guinea-pigs inoculated 
 
 in the ear suffer only from a local erythema, , 
 
 J J ' FIG. 106. PURE CUL- 
 
 wmch disappears after five or six days, and TIVATIOX OK THK 
 
 renders them for a time immune. Rabbits BACILLUS OF SEPTI- 
 
 inoculated in the cornea suffer from an intense CXUIA OP MICK IN 
 . a ,. j, mi i -IT f NITKIEXT GELATINE. 
 
 inflammation of the eyes. The bacilli form 111 
 
 After two days, 
 plate -cultivations scarcely perceptible cloud-like 
 
 specks, and in a test-tube of nutrient gelatine they form a delicately 
 clouded cultivation along the needle track. 
 
 An identical bacillus has been isolated in swine measles. 
 
 15 
 
jCHAPTEK XVT. 
 
 SEPTICEMIA OF BUFFALOES. SEPTIC PLEURO-PNEUMONIA OF 
 
 CALVES. SWINE FEVER. SEPTICAEMIA OF DEER. SEPTICAEMIA 
 OF RABBITS. FOWL CHOLERA. FOWL ENTERITIS. DUCK 
 CHOLERA. GROUSE DISEASE. 
 
 THERE are several varieties of septicaemia occurring naturally in 
 buffaloes, deer, calves, and birds, and artificially induced by inocula- 
 tion of rabbits with septic material. They are associated with 
 bacteria which agree in their morphological and cultural characters, 
 though in some cases differing in their pathogenic properties. As 
 the differences between the bacteria cultivated from these different 
 sources is not greater than the differences which exist between 
 the morphological, biological, and pathogenic effects of varieties of 
 the tubercle bacillus, it will be convenient and fully justifiable to 
 follow Hueppe and Baumgarten, and regard them as varieties of 
 the bacillus of hcemwrhagic septiccemia. 
 
 EPIDEMIC DISEASE OF BUFFALOES. 
 
 Oreste and Armanni investigated an epidemic among herds of 
 young buffaloes in Italy (Biiffel-seuche). The disease was extremely 
 acute, death occurring in from twelve to twenty -four hours. It was 
 probably identical with an epidemic disease described by Bellinger in 
 deer. The symptoms were fever, rapid pulse, discharge of mucus 
 from the nose and mouth, and a local swelling of the head and face 
 leading to suffocation. The only marked feature after death was 
 hsemorrhagic inflammation of the small intestine. 
 
 The bacilli were identical with those found by Schiitz in swine 
 fever. Cultures inoculated in young buffaloes produced the disease. 
 The bacilli were pathogenic to mice, guinea-pigs, rabbits, pigeons, 
 and fowls, death taking place in from one to three days. 
 
 226 
 
EPIDEMIC DISEASE OF DEER AND BOARS. 227 
 
 SEPTIC PLEURO-PNEUMONIA IN CALVES. 
 
 Septic pleuro-pneumonia is a disease which attacks young calves 
 within the first two months after their birth. Percussion and 
 auscultation reveal lung mischief. The disease is very rapid and 
 fatal, death occurring on the second or third day. In the less 
 acute cases one or more lobes of the lungs are found after death 
 in a state of lobular and inter-lobular pneumonia. The inter-lobular 
 connective tissue is distended with exudation, giving rise to white 
 or yellowish bands between the inflamed lobules, which produce a 
 marbled appearance, recalling the condition of the lungs in infectious 
 pleuro-pneumonia. The internal organs are congested, and there 
 are very often hsemorrhagic spots on the mucous and serous coats 
 of the small intestine. All the organs contain rods identical with 
 those of septicaemia of rabbits. Rabbits, guinea-pigs, and mice were 
 infected. A calf was injected in the pleural cavity with a broth - 
 culture, and died in twenty hours. 
 
 SWINE FEVER. 
 
 This disease will be described in a separate chapter. Several 
 bacteria have been isolated by different investigators. In swine 
 fever in Germany (Schwein-seuche) Lb'mer and Schiitz isolated a 
 bacillus which has been identified with the bacillus isolated by 
 Salmon and Smith from hog-cholera in America, and with the 
 bacillus of rabbit septicaemia and of fowl cholera. 
 
 EPIDEMIC DISEASE OF DEER AND BOARS. 
 
 A very fatal epizootic (Wildseuche) occurred in the royal game 
 preserves near Munich, destroying one hundred and fifty-three deer 
 and two hundred and thirty-four boars (Bellinger). The disease 
 lasted from twelve hours to six days. In the less acute cases pneu- 
 monia and pericarditis supervened. In cattle there was also severe 
 hsemorrhagic inflammation of the small intestine. In another form 
 it produced swelling of the head, face, neck, and tongue. The 
 virus proved fatal to rabbits in six to eight hours, and to sheep and 
 goats in about thirty hours. A pig inoculated with a few drops of 
 blood died in twenty-two hours. Kitt also investigated this malady. 
 The bacteria were found to be identical, in their appearance and 
 pathogenic properties, with extremely virulent bacteria from swine 
 fever. Schiitz distinguished them from the bacteria obtained from 
 swine fever by their pathogenic effect on pigeons, but cultures 
 obtained from swine fever do not act uniformly in this respect. 
 
228 INFECTIVE DISEASES. 
 
 SEPTICAEMIA IN RABBITS. 
 
 Koch minutely investigated a disease of rabbits produced 
 by inoculation with impure river water and with putrid meat 
 infusion. Bacteria are found in the blood in abundance, and may 
 be readily cultivated. 
 
 The smallest quantity inoculated subcutaneously or in the cornea 
 of a rabbit produces a rise of tempera- 
 ture and laboured breathing after ten 
 to twelve hours, and death in sixteen 
 to twenty hours. The spleen and 
 lymphatic glands are found to be 
 
 FIG. 107. -BACTERIUM OF RABBIT en i ar g ed , an d the lungs congested, 
 SEPTICAEMIA; BLOOD OF SPAR- ' 
 
 ROW x 700 (Kocn). but there are no extravasations, and no 
 
 peritonitis. Mice and birds are very 
 susceptible ; guinea-pigs and white rats have an immunity. 
 
 DAVAINE'S SEPTICAEMIA. 
 
 A disease was produced by Davaiiie by injecting rabbits with 
 putrid blood. Rabbits, mice, fowls, pigeons, and sparrows are sus- 
 ceptible, and guinea-pigs and rats are insusceptible to the bacteria 
 found in this disease. Rabbits inoculated with a trace of blood con- 
 taining the bacteria, or with a culture, died in from twenty-four to 
 thirty-six hours. The spleen, liver, lungs, and intestines are highly 
 congested, and sometimes extravasations and peritonitis are found. 
 
 FOWL CHOLERA. 
 
 Fowl cholera is an epidemic disease of the poultry-yard much 
 dreaded in France, and well known through the researches of 
 Perroncito, Toussaint, Pasteur, and Kitt. 
 
 FIG. 109. BACTERIUM OF FOWL 
 
 I'JS 8< If Tr. F F WL CHOLERA ' CHOLERA, x 2500. Muscle juice 
 
 x 1200. From blood of inoculated Fowl. of -p owl 
 
 Fowls suffering from the disease usually die in from twenty-four 
 to forty-eight hours. The disease shows itself by the fowls becoming 
 
FOWL CHOLERA. 229 
 
 somnolent. They suffer from weakness of the legs, and their wings 
 trail. There is frequently diarrhoea, with slimy greenish evacuations, 
 and death usually ensues after a slight convulsive attack. On 
 making a post-mortem examination the viscera will be found to be 
 congested, and there is intense inflammation of the mucous membrane 
 of the intestine, with haemorrhages. 
 
 The blood from the heart, and the intestinal contents, contain 
 the bacilli which were at one time believed to be peculiar to 
 this disease. Inoculation subcutaneously, or administration with 
 food, of a small quantity of a broth cultivation will produce death 
 in twenty-four to thirty-six hours. Pigeons, pheasants, sparrows, 
 
 -' ' > 
 
 -^: "} ^ \ 
 
 ' teMk 
 
 
 FIG. 110. BACTERIUM OF FOWL CHOLERA. Section from liver of Fowl x 700 
 
 (FLUGGE). 
 
 rabbits, and mice are susceptible. In guinea-pigs, sheep, and horses, 
 an abscess develops at the seat of inoculation. Rabbits are readily 
 infected by sprinkling a broth-cultivation on cabbage leaves or any 
 suitable food. It was with this microbe that Pasteur proposed to 
 eradicate the plague of rabbits in Australia. 
 
 Fowl cholera has an additional interest, as it was with this 
 disease that Pasteur first investigated the attenuation of virus. 
 Broth-cultures which were several months old were found, when 
 injected, to produce apparently only a local effect. Tin's weakening 
 of the virus was attributed by Pasteur to exposure to oxygen. 
 After recovery the fowls were protected against the action of 
 virulent cultures, while fowls not immunised died the following day. 
 Kitt, by working with pure-cultures on solid media, showed that the 
 
230 INFECTIVE DISEASES. 
 
 weakening was not due to prolonged exposure to oxygen, but that 
 old contaminated broth- cultures after a time completely lost their 
 power, owing to the antagonism of the bacteria accidentally present. 
 Filtered broth- cultures contain the toxic products of the bacillus, 
 and produce slight illness and subsequent immunity. 
 
 FOWL ENTERITIS. 
 
 Fowl enteritis is an acute infectious disease of fowls, the course 
 and symptoms of which are regarded by Klein as distinct from fowl 
 cholera. The fowls suffer from diarrhoea, with liquid greenish 
 evacuations, but are never somnolent, and death occurs in one or 
 two days. After death the mucous membrane of the intestine is 
 found to be congested, and coated with grey or yellowish mucus; 
 the liver is congested, spleen enlarged, and lungs normal. There are 
 a few bacilli in the blood of the heart, very many in the spleen and 
 liver, and they are in the form of a pure-culture in the mucus of 
 the intestine. Klein says that the bacilli are a little longer and 
 thicker than those found in fowl cholera, which they only slightly 
 resemble, and that the course of the disease, the symptoms and 
 pathological appearances, definitely distinguish it from fowl cholera, 
 but that nevertheless it belongs to the same family of bacilli 
 Pigeons are said to be insusceptible, rabbits only slightly susceptible. 
 By feeding and by subcutaneous inoculation the disease can be 
 communicated to healthy fowls, but there is no sign of illness until 
 the fourth day. As regards attenuation, the bacilli behave like 
 those from cases of fowl cholera. 
 
 DUCK CHOLERA. 
 
 Duck cholera is an epidemic disease of ducks which was investi- 
 gated by Cornil. The symptoms are similar to those of fowl cholera. 
 They suffer from diarrhoea and weakness, followed by death in two 
 or three days. 
 
 The bacillus cultivated from the blood of ducks is pathogenic in 
 ducks but not in fowls or pigeons, and large doses are required to 
 kill rabbits. 
 
 GROUSE DISEASE. 
 
 Grouse disease is an acute infectious disease of red grouse. 
 According to Klein the chief pathological feature is severe pneu- 
 monia ; there is also patchy redness of the serous and mucous linings 
 
GROUSE DISEASE. 
 
 231 
 
 
 I ' 
 
 FIG. 111. BACILLUS OF H^MOBBHAGIC 
 SEPTICAEMIA. Blood of a Rabbit after 
 death from Septicaemia x 950 (BAUM- 
 GARTEN). 
 
 of the intestine, and the liver is congested and dark, but the spleen 
 is not enlarged. The bacilli are found in the heart, lungs, and liver, 
 and in the extravasated blood. 
 Cultures inoculated in mice and 
 guinea-pigs produce pneumonia 
 and death. Sparrows are sus- 
 ceptible, and other small birds. 
 Fowls, pigeons, and rabbits are 
 insusceptible. 
 
 Bacillus of Haemor- 
 rhagic Septicaemia. Very 
 short rods, with rounded ends, 
 "6 to '7 jj. in width and 1 '4 /u, in 
 
 length. In stained preparations the rods 
 are observed to be deeply stained at the 
 ends and to have a clear interval in the 
 middle; they were on this account mis- 
 taken by earlier observers for dumb-bell 
 micrococci or diplococci. They are non- 
 motile, and spore-formation is unknown. 
 They grow readily in the ordinary media. 
 The colonies in nutrient gelatine appear 
 about the third day. They are circular 
 in form, with a sharp dark outline, and 
 of a yellow colour, lighter at the peri- 
 phery. Later, the central zone is finely 
 granular, and of a dark yellowish -brown 
 colour, with the lighter peripheral zone 
 more clearly denned. In the depth of 
 gelatine a delicate filament develops in 
 the track of the needle, composed of 
 minute spherical colonies, somewhat trans- 
 parent, and yellowish-white in colour. 
 At the point of puncture there may be no 
 growth visible, or a flat and very limited 
 growth. Inoculated on the surface of 
 nutrient media a thin layer develops, 
 with an irregular serrated and thickened 
 border. On potato different results have 
 been obtained by different observers. Some maintain that a 
 greyish-white or yellowish film will develop at the temperature of 
 the" blood ; but according to Caneva, the bacilli, whatever their source, 
 
 FIG. 112. BACILLUS <>F 
 H.EMORRHAGIC SEPTK'.K.MIA 
 (Rabbit Septicaemia). Pure- 
 culture in Gelatine after 
 four days (BAUMGARTEN.) 
 
232 INFECTIVE DISEASES. 
 
 will not grow on potato, while Bunzl-Federn maintains that the 
 bacilli from fowl cholera and rabbit septicaemia do grow upon potato, 
 but those from septicaemia in deer, buffaloes, and swine do not. 
 Opinions differ with regard to their action on milk. The reaction 
 for phenol and indol is given in all cases, except with cultures 
 obtained from septicaemia of buffaloes. The virulence of the bacilli 
 may be diminished and attenuated, but it may subsequently be 
 restored by successive inoculation in animals. The pathological 
 lesions vary in different animals. The most common result is con- 
 gestion of the internal organs and haemorrhage. The bacilli culti- 
 vated from cattle or deer produce fata,l results when inoculated 
 in swine. The bacilli from any of these sources inoculated in 
 pigeons will produce fowl cholera, but the bacilli isolated by Schiitz 
 from swine, and those from deer, are not fatal to fowls. Further, 
 the bacilli cultivated from swine fever are fatal to guinea-pigs, 
 while the bacilli from rabbit septicaemia have very little effect upon 
 them. The bacilli have been found in association with diseases of 
 cattle, swine, deer, birds, rabbits, and mice, and have been cultivated 
 from healthy mucous membrane. Yeranus Moore found the bacilli 
 in the mucus from the upper air passages, of 71 per cent, of cattle, 
 85 per cent, of cats, and 33 per cent, of dogs. From these sources 
 inoculations were made in rabbits, and rapidly fatal septicaemia 
 was produced, associated in less acute cases with peritonitis, pleurisy, 
 and pericarditis. 
 
CHAPTER XVII. 
 
 PNEUMONIA. INFECTIOUS PLEURO-PNEUMONIA OF CATTLE. 
 INFLUENZA. 
 
 ACUTE CROUPOUS PNEUMONIA. 
 
 PNEUMONIA is an acute inflammation of the hiDgs with fibrinous 
 infiltration of the air vesicles and interstitial tissue. There are 
 varieties of pneumonia, and one form is commonly believed to be 
 infectious. 
 
 The lung passes through three stages engorgement, red hepati- 
 sation, and grey hepatisation. In the first stage the lung is of 
 a deep red colour, but still vesicular ; in the second stage the 
 affected part is more or less solid, and has the consistency of liver, 
 owing to the fibrinous lymph which is poured out into the alveolar 
 cavities. In the grey hepatisation, the exudation contains more 
 leucocytes and less fibrin, and this is followed by the stage of 
 suppurative softening and final absorption. The sputum at the 
 commencement of the disease is rusty, from the presence of blood, 
 and later on has the appearance of prune juice. Examination of 
 the sputum by Gram's method will reveal numerous micro-organisms, 
 and two of these are deserving of special study the pneumococcus 
 of Friedlander, which is present in a considerable proportion of 
 cases, and Sternberg's micrococcus, which was found in sputum by 
 Talamon. 
 
 In 1888, there was considerable prevalence of pneumonia in 
 Mi<Mlesbrough, with strong tendency to occur in groups of c; 
 but there was admittedly room for doubt whether the clinical 
 and post-mortem appearances were not identical with ordinary 
 pneumonia. Dr. Ballard maintained that there were facts and 
 considerations which appeared to show that the disease was com- 
 municable from the sick to the healthy, and that it was a specific 
 febrile disease, and Klein isolated and described the micrococcus 
 present in these cases. 
 
 Bacterium Pneumonias Crouposae (Pneumococcus, Fried- 
 
 233 
 
234 
 
 INFECTIVE DISEASES. 
 
 FIG. 113. BACTERIUM PNEUMONLE 
 CROUPOS.E, FROM PLEURAL CAVITY 
 OF A MOUSE, x 1500. A, B. 
 Thread-forms. C, D, E. Short 
 rod-forms. G. Diplococci. H. 
 Cocci. I. Streptococci. (Zopf.) 
 
 lander). Cocci ellipsoidal and round, singly, or in pairs (diplococci), 
 
 rods and thread-forms. The cell- 
 membrane thickens, and develops 
 into a gelatinous capsule, which is 
 round if the coccus is single, and 
 ellipsoidal if the cocci occur in pairs 
 or in rod-forms. Cultivated in a 
 test-tube of nutrient gelatine they 
 grow in the form of a round- 
 headed nail, without liquefaction 
 of the gelatine (Fig. 114). The 
 cocci when artificially cultivated 
 have no capsule, but it again 
 appears after their injection into 
 animals, 
 The cocci 
 can also be 
 cultivated 
 
 on blood serum and on boiled potatoes. 
 They occur in pneumonic exudation. In- 
 oculation of dogs with a cultivation of the 
 cocci occasionally gave positive results; but 
 in rabbits no results followed. Guinea- 
 pigs proved to be susceptible in some cases ; 
 but thirty-two mice, after injection of a 
 cultivation diffused in sterilised water, into 
 the lungs, died without exception. The 
 lungs were red and solid, and contained the 
 cocci, which were also present in the blood, 
 and in enormous numbers in the pleural 
 exudation. Inhalation experiments by spray- 
 ing the cocci diffused in water into mouse 
 cages produced pneumonia and pleurisy in 
 three out of ten mice. 
 
 The nail-shaped cultivation is not always 
 
 produced, nor are these conclusions accepted 
 
 ,, . ' FIG. 114. FRIEULANDER'S. 
 
 by all investigators. PNEUMOCOCCUS. Pure- 
 
 culture in nutrient- 
 gelatine four days old 
 (BAUMGARTEN). 
 
 METHODS OF STAINING FRIEDLANDER'S 
 PNEUMOCOCCUS. 
 
 Cover-glass preparations of pneumonic sputum or exudation may be 
 treated as follows : 
 
PNEUMONIA. 235 
 
 () Stain by the method of Gram, and after-stain with eosin. 
 
 (b) Treat with acetic acid, then stain with gentian-violet or Bismarck- 
 brown. Examine in distilled water, or dry and preserve in Canada 
 balsam. 
 
 (c) Float them on weak solutions of the aniline dyes twenty-four 
 hours ; differentiation between coccus and capsule is thus obtained. 
 
 (r/) Stain with osmic acid ; the contour of the capsules is brought 
 out. 
 
 v 
 
 FIG, 115. CAPSULE-COCCI FROM PNEUMONIA, x 1500 (BAUMGARTEN). 
 
 Sections of pneumonic lung should be stained by 
 
 (a) Method of Gram. 
 
 (6) Method of Friedlander. This method is employed to demonstrate 
 the capsules in tissue sections. It consists in placing the sections twenty- 
 four hours in the following solution : 
 
 Fuchsine 1 
 
 Distilled water 100 
 
 Alcohol ........ 5 
 
 Glacial acetic acid 2 
 
 They are then rinsed with alcohol, transferred for a couple of minutes 
 to a 2 per cent, solution of acetic acid, and treated with alcohol and oil 
 of cloves in the usual way, and preserved in Canada balsam. 
 
 Sternberg's micrococcus was first found in the blood of rabbits 
 inoculated with saliva. Three months afterwards, Pasteur encoun- 
 tered the same organism in rabbits inoculated with the blood of a 
 child suffering from rabies. The same organism in 1883 was found 
 by Talamon in pneumonic sputum. It was identified by Sternberg, 
 Two years afterwards further observations were made by Frankel, 
 Gamaleia, and others. It has also been found in purulent meningitis 
 by Netter, and by Monti in cerebro-spinal meningitis, by Weichsel- 
 baum in ulcerative endocarditis, and by others in acute abscess of 
 the middle ear, and in purulent inflammation of the joints following 
 pneumonia. 
 
236 INFECTIVE DISEASES. 
 
 Sternberg's Micrococcus. (Microbe de salive, Pasteur ; Micro- 
 coccus Pasteuri, Sternberg ; Lancet-shaped micrococcus, Talainon ; 
 Streptococcus lanceolatus Pasteuri, Gamaleia ; Diplococcus pneu- 
 rnonice, Weichselbaum ; Bacillus septicus sputigenus, Fliigge ; 
 Micrococcus of sputum septiccemia, Frankel.) Spherical or oval 
 cocci, singly, in pairs or in chains, often lanceolate and capsuled. 
 Stain readily with the aniline colours and by Gram's method ; 
 non-motile. They flourish in alkaline media in the incubator. In 
 broth they produce in twelve hours a cloudiness due to the develop- 
 
 * 
 
 FIG. 116. MICROCOCCUS OF SPUTUM SEPTICAEMIA. From the blood of a 
 Kabbit. x 1000 (FRANKEL AND PFEIFFER). 
 
 ment of cocci and short chains. After a time these subside to 
 the bottom of the tube, and the liquid above becomes clear. In 
 plate -cultivations the colonies are small, circular, white, and granular. 
 In the depth of gelatine, minute white colonies develop along the 
 track of the needle without liquefaction of the gelatine ; and on 
 the sloping surface of nutrient agar or blood serum minute trans- 
 parent drops appear along the line of inoculation. They grow in 
 milk, coagulating casein; but they do not grow on potato. Sub- 
 cultures quickly lose their virulence, but regain it by inoculation. 
 
1'NKUMONIA. 237 
 
 The injection of a minute quantity ('2 cc.) of a virulent culture 
 Mibrutaneotisly proves fatal to mice and rabbits in from twenty- 
 four to forty-eight hours. Immediately afterwards there is a rise of 
 temperature of 2 or 3 C., later it falls, and just before death 
 it is several degrees below normal. After death, the post-mortem 
 appearances of septicaemia are observed, in addition to diffuse 
 inflammatory oedema extending in all directions from the point of 
 injection. The subcutaneous connective tissue contains sanguineous 
 serum and micrococci in abundance. The liver and spleen are some- 
 
 FIG. 117. COLONIES OF STERNBERG'S MICROCOCCUS. Agar plate-cultivation, 
 after 24 hours, x 100 (FRANKEL AND PFEIFFER). 
 
 times dark and engorged, and blood from the heart and internal 
 organs teems with micrococci. 
 
 There is no indication of pneumonia after subcutaneous inocula- 
 tion, but intra-pulmonary injections produce fibrinous pneumonia^ 
 often fatal (Talamon, Gamaleia). The result is usually fatal in 
 rabbits and sheep, but dogs, as a rule, recover. Injection of cultures 
 into the trachea of rabbits is said to induce typical pneumonia 
 (Monti). 
 
 Sternberg concludes that this micrococcus is the cause of acute 
 infectious pneumonia, but the micrococcus is undoubtedly associated 
 with widely different pathological processes, and the possibility of its 
 being a saprophyte, which finds in pneumonia a suitable soil for its- 
 development, must not be overlooked. 
 
238 INFECTIVE DISEASES. 
 
 Klein's Micrococcus. Klein found in pneumonic sputum a 
 diplococcus which does not appear to differ from Sternberg's micro- 
 coccus. In cover-glass preparations the bacilli are surrounded with 
 a halo, but no definite capsule, as in Friedlander's coccus. They 
 appear as short rods constricted in the centre, or dumb-bell forms, 
 and forms intermediate between cocci and bacilli. In gelatine, after 
 two or three days, greyish -white spots appear, which enlarge in the 
 next two or three days into flat, translucent, greyish-white plaques, 
 with irregular serrated outline. Colonies beneath the surface are 
 spherical, and of a brownish -yellow colour. In test-tubes in the 
 depth of the gelatine a whitish-brown filament develops on incuba- 
 tion, composed of minute spherical colonies, and on the surface the 
 growth spreads out into a greyish-white film with serrated margin. 
 On the surface of obliquely solidified gelatine the growth forms a 
 thin whitish film, which enlarges in breadth with irregular outline, 
 reaching its maximum in about a fortnight. The growth on agar 
 is very similar. Broth becomes uniformly turbid in twenty-four 
 hours, then a powdery precipitate makes its appearance. On potato 
 there is a thin, moist, faintly yellowish-brown film. Cultures examined 
 in the fresh state show many rods in a resting stage, and others 
 actively motile. In addition to the dumb-bell forms there are others 
 of greater length, and in old cultures involuted and degenerated 
 forms. Spore-formation has not been observed. A broth-culture 
 inoculated into two rabbits produced a local tumour which subsided 
 in a week. Death ensued in one case in eight days, and in the 
 other in three weeks. There was purulent matter at the seat of 
 inoculation in one ; in the other, pericardial exudation and hypersemia 
 of the lungs. Broth- cultures inoculated intravenously produced no 
 effect. In guinea-pigs there was swelling at the seat of inoculation, 
 or slight indication of disease and recovery. Cultures inoculated 
 in mice produced rapid breathing, drowsiness, and death in from 
 twenty-four to ninety- six hours. The internal organs were con- 
 gested, the lungs inflamed, and the blood and organs in the inoculated 
 animals contained the diplococci in considerable numbers. 
 
 Foa isolated a coccus which he named the Micrococcus lanceolatus 
 capsulatus. It produced in small animals either rapid septicaemia 
 and death, or local oedema and death at a later period. 
 
 Protective Inoculation. Immunity has been produced in 
 rabbits by the intravenous injection of the virus in a diluted form. 
 Blood obtained from immunised rabbits was kept at 10 C. for twelve 
 hours, and then filtered, and animals injected with it acquired 
 immunity against virulent cultures (Emmerich). 
 
INFECTIOUS PLEURO-PNEUMOXIA. 
 
 239 
 
 Filtered cultures are said to confer immunity for six months, and 
 raising the temperature of filtered cultures increases the strength of 
 the substance which gives immunity (Klemperer). The blood serum 
 of immune animals can confer immunity on other animals, and, it is 
 said, will arrest the progress of the disease produced by injection 
 of healthy animals with virulent cultures. The cultures contain a 
 proteid body, for which the name pneumo-toxin has been suggested, 
 and anti-pneumo-toxin has been isolated from immunised blood 
 serum. 
 
 INFECTIOUS PLEURO- PNEUMONIA. 
 
 infectious pleuro-pneumonia is a highly infectious disease 
 peculiar to cattle ; it is characterised by rise of temperature and 
 exudation into the lungs. It is often fatal, and sometimes exists in 
 an extremely chronic form. It is believed to have been unknown in 
 England previously to 1840, and is supposed to have been introduced 
 from Holland, where in one year it destroyed seven thousand cattle. 
 
 Fn;. 118. ACUTE CATARRHAL PNKIMONIA (Ox). 
 
 ", Coagulated mucus with catarrhal cells (c) embedded in it ; b, catarrhal cells 
 sprouting from alveolar wall, x 480. (Hamilton.) 
 
 The disease cannot be conveyed artificially. A living, di- 
 animal must be the medium of infection. The disease is apparently 
 only communicated by cohabitation. Brown injected large quant i- 
 ti.-> of lymph from diseased lungs into the jugular vein, into the 
 
240 
 
 INFECTIVE DISEASES. 
 
 tissue of the lungs, and into the trachea, without any result except 
 a small abscess at the seat of puncture. Administration of the 
 virus by the mouth gave equally negative results. The lungs from 
 a recently killed animal infected with pleuro-pneumonia were placed 
 in a shed occupied by healthy heifers, and left there for several days. 
 Fodder, litter, and manure were taken from places in which there 
 were diseased cattle, and placed in contact with healthy cattle, and sub- 
 sequently all the animals used in these experiments were slaughtered 
 and carefully examined, and the results were absolutely negative. 
 Similarly negative results followed experiments made by Saiider- 
 
 FIG. 119. INFECTIOUS PLEURO-PNEUMONIA OF CATTLE, x 480. 
 
 a,a,a, Exudation in air-vesicles, composed of a network of fibrinous lymph with 
 entangled leucocytes; b,b, the same caseating; c, the air- vesicle filled with 
 leucocytes only. In the centre is a blood-vessel filled with a fibrinous plug. 
 (Hamilton. ) 
 
 son and Duguid, and thus confirmed the conclusion arrived at by 
 Brown, that the disease could only be communicated by actual contact 
 of a living, diseased animal with a healthy one. 
 
 The symptoms of the disease in cattle are a rise of temperature 
 to 105 or 107, and a peculiar dry cough, and later the usual 
 indications of pneumonia, difficulty in breathing, and dulness on 
 percussion. As a rule, death follows from exhaustion ; but the disease 
 may also assume a chronic form, if the animal escapes slaughter, 
 and the lung may become gangrenous or tubercular. The period of 
 incubation is about thirty days, but it is uncertain. The lesions are 
 
fc v; 
 
 U^^Jv^> - .-^ .'- - - ; \- 1 fjr^. ;;? f ; > ^jft^ 
 
 "S?k: 
 
 Fi;. 120. IXFKC TIOUS I'LHUKO-I-NKUMONIA OF CATTLE, x :>". 
 ,,, Spaces in deep layer of pleura and interlobular septa filled with fibrin<>u> 
 lymph; b, deep luyt-r of pleura running down to an interlobular septum ; c,c, 
 air-vehicles filled with fibriuous lymph; <t, bLnxl-vesscls of alvexjlar walls, 
 much congested; e, large congested Uood-v els ; /,/, interlobular Upfel 
 infiltrated with fibrinous lymph ;i .'/, blo<xl- vessel in interlobular septum 
 (Logwood, Eosin and Farrant's solution). HAMILTON. 
 
 16 
 
242 INFECTIVE DISEASES. 
 
 limited almost entirely to the lungs ; congestion is quickly followed 
 by inflammation and effusion into the air vesicles and the intra- 
 lobular fibrous tissue which is so well marked in the lungs of cattle. 
 Leucocytes are entangled in the fibrinous lymph, and the intra- 
 lobular septa are enormously enlarged, so that the red lobules are 
 mapped out by the paler septa, and produce on section of the diseased 
 parts a very striking marbled appearance. A somewhat similar 
 appearance is sometimes observed in septic pleuro-pneumonia in 
 calves. The effusion occurs also in the air vesicles. The stages 
 of grey hepatisation and suppurative softening have not, as a rule, 
 time to develop. Hsemorrhagic infarctions are sometimes produced, 
 which in turn become gangrenous or cheesy, and a capsule may form 
 round the diseased part. Roy found micro-organisms in the lymph, 
 but attached no importance to them. Bruylants and Yerriet also 
 described a micro-organism in the lymph. Later, Poels and Nolen 
 isolated a micrococcus resembling JFriedlander's pneumococcus. 
 Inoculation in the lungs produced a condition in cattle which they 
 considered indicative of pleuro-pneumonia. 
 
 Lustig was unable to confirm these observations, but succeeded 
 in isolating from lymph a bacillus and three species of micrococci. 
 One of the micrococci formed an orange growth when cultivated, 
 and was regarded as the specific micro-organism, as it caused sub- 
 cutaneous tumefaction, and, it is said, some degree of immunity. 
 
 Brown cultivated a number of organisms which on inoculation 
 only produced local irritation. Intravenous injection produced death 
 from septicaemia in one case in thirty-six hours. 
 
 Arloiiig isolated four different organisms, including a bacillus 
 which was named Pneumo-bacillus liquefaciens bovis. Later, he 
 prepared a fluid from broth- cultures, pneumo-bacillin, which pro- 
 duced a more marked rise in temperature in animals suffering from 
 pleuro-pneumonia than in healthy animals, and its use was 
 suggested as an aid in diagnosis. Arloing named the micro- 
 organisms provisionally Pneumo-bacillus liquefaciens bovis, Pneumo- 
 coccus gutta cerei, Pneumococcus lichenoides, and Piieumococcus 
 flavescens. 
 
 Pneumo-bacillus liquefaciens bovis. Short rods, non- 
 motile ; spore-formation not observed. They rapidly liquefy gelatine, 
 and form on potato a white layer, which becomes brownish and 
 sometimes greenish. According to Arloing pure-cultures produce 
 in the ox, when injected subcutaneously or in the lung, the same 
 lesions which are produced by virulent lymph. Guinea-pigs and 
 rabbits are slightly susceptible, dogs are immune. 
 
INFECTIOUS PLEURO-PNEUMONIA. 243 
 
 Nocard does not accept Arloing's conclusions, and expresses the 
 opinion that the virus is participate, but is not due to any micro- 
 organism which cun be detected or cultivated by the methods 
 at present adopted. In the opinion of the author, who has 
 also examined the micro-organisms in pleuro- pneumonia, it is 
 fully justifiable to regard the nature of the contagium as 
 unknown. 
 
 Preventive Inoculation. In 1852 Willems introduced inocula- 
 tion. The liquid from the lungs of an animal with pleuro-pneumonia, 
 which had recently died, was inoculated in the extremity of the tail 
 by a puncture with a lancet. Swelling occurred at the seat of inocu- 
 latiou, and on recovery the animals were believed to be protected. 
 A Dutch Commission reported that the inoculation gave a temporary 
 protection. A Belgian Commission in the following year reported 
 that the phenomena of inoculation could be produced several times 
 in succession in the same animal, and that it was not a certain 
 preventive. A French Commission in 1854 concluded that a power 
 of resisting infection was given, but the period was undetermined. 
 Protective inoculation continued to be employed, and various modi- 
 fications of the method were introduced. Threads soaked in lymph 
 were inoculated, or the lymph subcutaneously or intravenously 
 injected. 
 
 The usual result of the inoculation is swelling and, in about ten 
 or fourteen days, effusion of straw-coloured fluid, which is occasion- 
 ally blood-stained. Gangrene may follow, involving amputation of 
 the tail. Germont and Loire in Queensland adopted the method 
 which was suggested by Pasteur of inoculating calves in the loose 
 cellular tissue behind the shoulder. This produces intense oedema 
 and a quantity of lymph. There has been much controversy with 
 regard to the value of protective inoculation. 
 
 Stamping-out System. Brown maintains that pleuro- 
 pneumonia can be exterminated only by slaughter of the diseased 
 animals, and quotes the results experienced in the Netherlands in 
 support of his views. 
 
 In 1871 slaughter for pleuro-pneumonia was commenced in the 
 Netherlands. There were 6,000 cattle attacked by the disease. In 
 1872 owners were compelled to slaughter not only diseased cattle, 
 but those which hud been in contact with them, unless inoculated, 
 ami the attacks were, in consequence, reduced to 4,000. In 1873 it 
 was forbidden to move cattle out of infected districts, and the attacks 
 were reduced to 2,479. In 1876 slaughter of the whole herd was 
 decreed, and during the first year of this heroic system the cases fell 
 
244 
 
 INFECTIVE DISEASES. 
 
 from 2,227 in 1875, to 1,723 in 1876, to 951 in 1877, to 698 in 
 1878, to 157 in 1879, and to 48 in 1880. 
 
 In England the Pleuro-pneumonia Act came into force on 
 September 1st, 1890. Notification was to be given by the owner 
 to a police constable of the district, who was required to transmit 
 the information to the Local Authority and also to the Board of 
 Agriculture. An inspector, with the aid of the veterinary surgeon, 
 arranged for the slaughter of the suspected animal, and, if the 
 disease proved to be pleuro-pneumonia, of the rest of the herd. The 
 results are shown in the following table : 
 
 
 
 
 
 Diseased Cattle. 
 
 
 Cattle 
 
 YEARS. 
 
 Number 
 of 
 Infected 
 Counties. 
 
 Number 
 of 
 Fresh 
 Outbreaks. 
 
 Number 
 of 
 Cattle 
 Attacked. 
 
 
 Healthy 
 Cattle in 
 contact 
 slaughtered. 
 
 slaughtered 
 as suspected, 
 but found 
 free from 
 Pleuro- 
 Pneumonia. 
 
 Killed. 
 
 Died. 
 
 1890 
 
 36 
 
 465 
 
 2,057 
 
 2,022 
 
 37 
 
 11,301 
 
 
 1891 
 
 27 
 
 192 
 
 778 
 
 778 
 
 
 
 9,491 
 
 232 
 
 1892 
 
 10 
 
 35 
 
 134 
 
 134 
 
 
 
 3,477 
 
 188 
 
 1893 
 
 4 
 
 9 
 
 30 
 
 30 
 
 
 
 1,157 
 
 86 
 
 1894 
 
 2 
 
 2 
 
 15 
 
 15 
 
 
 
 391 
 
 41 
 
 Thus the number of cases was reduced from 2,057 in 1890 to 
 15 in 1894. 
 
 A departmental committee appointed in 1892 to inquire into 
 pleuro-pneumonia and tuberculosis, came to the following conclusions 
 with regard to pleuro-pneumonia : 
 
 (1) That the system of compulsory slaughter be applied not only to 
 all diseased cattle, but also to all cattle which have been in association 
 with them, or otherwise in any manner exposed to the infection of the 
 disease. 
 
 (2) Compulsory slaughter should be accompanied by supplementary 
 measures, such as restrictions on the movement and sale of cattle within r 
 or coming from, infected districts. 
 
 (3) Any exception to, or modification of, the system of compulsory 
 slaughter, as provided in the Slaughter Order, 1888, should only be 
 applicable to cattle in the dairy yards, byres, and cowsheds of large 
 towns, the owners or occupiers of which may claim in writing the privi- 
 lege of exemption for their cattle from immediate slaughter, on the 
 following conditions : 
 
INFECTIOUS PLEURO-PNEUMONIA. 245 
 
 (a) Xo head of cattle that has been brought into such dairy premises 
 shall be removed therefrom, except for the purpose of immediate 
 slaughter. 
 
 (6) In the event of an outbreak of pleuro-pneumonia, all the diseased 
 cattle shall be slaughtered. 
 
 (c) All the remaining cattle on such premises where an outbreak has 
 occurred shall be branded, and regularly subjected to the ther- 
 mometer test ; and whenever a continuous increase of temperature, 
 rising above 104, is shown, they shall be slaughtered. 
 
 (r/) No fresh cattle shall be admitted into such premises while any 
 of the cattle thus branded remain alive. 
 
 (4) Inoculation cannot be recommended as a means of eradicating 
 pleuro-pneumonia, nor as practicable under existing conditions. Although 
 it is open to owners to inoculate their cattle, it should be distinctly 
 understood that that operation shall not give them any immunity from 
 the regulations above suggested. 
 
 The order at present in force is the Pleuro- Pneumonia Order 
 of 1895. In addition to regulations for the movement of cattle, 
 for disposal of carcasses, for markets, and for compensation for 
 slaughter, the Order contains the following provisions : 
 
 NOTIFICATION. 
 Notice of Disease. 
 
 (1) Every person having or having had in his possession or under his 
 charge a head of cattle affected with or suspected of pleuro-pneumonia 
 shall with all practicable speed give notice of the fact of the head of cattle 
 being so affected or suspected to a constable of the police force for the 
 police area wherein the head of cattle so affected or suspected is or was. 
 
 (2) The constable receiving such notice shall immediately transmit 
 the information by telegraph to the Board of Agriculture. 
 
 (3) The constable shall also forthwith give information of the receipt 
 by him of the notice to an Inspector of the Local Authority, who shall 
 forthwith report the same to the Local Authority. 
 
 nf Inxjtector to act immediately. 
 
 (1) An Inspector of a Local Authority on receiving in any manner 
 whatsoever information of the supposed existence of pleuro-pneumonia, 
 or having reasonable ground to suspect the existence of pleuro-pneumonia, 
 shall proceed with all practicable speed to the place where such disease, 
 according to the information received by him, exists, or is suspected to 
 exist, and shall there and elsewhere put in force and discharge the powers 
 and duties conferred and imposed on him as Inspector by or under the 
 Act of 1894 and this Order. 
 
 (2) The Inspector shall forthwith report to the Board of Agriculture. 
 
246 INFECTIVE DISEASES. 
 
 ^o Movement into or out of Pleuro-pneumonia Infected Place without 
 
 Licence. 
 
 Cattle shall not be moved into or out of an Infected Place except with 
 a Movement Licence of an Inspector or officer of the Board, and such 
 cattle shall not be moved except in accordance with the conditions 
 contained in such Licence. 
 
 Pleuro-pneumonia found in a Market, Railway Station, Grazing Park, 
 or other like Place, or during Transit. 
 
 The Inspector of the Local Authority shall cause to be seized all the 
 cattle affected with pleuro- pneumonia, and also all cattle being in or on 
 the market, fair, sale-yard, place of exhibition, lair, landing-place, wharf, 
 railway station, common, uninclosed land, farm, field, yard, shed, park, or 
 other such place as aforesaid, and shall forthwith transmit the information 
 by telegraph to the Board of Agriculture. 
 
 The Inspector of the Local Authority shall cause all such cattle so 
 seized to be detained at the place where they are seized, or to be moved 
 to some convenient and isolated place and there detained. 
 
 Removal of Dung or other Things. 
 
 It shall not be lawful for any person to send or carry, or cause to be 
 sent or carried, on a railway, canal, river, or inland navigation, or in a 
 coasting vessel, or on a highway or thoroughfare, any dung, fodder, or 
 litter that has been in an Infected Place, or that has been in any place in 
 contact with or used about a diseased or suspected head of cattle, except 
 with a Licence of an Inspector or officer of the Board or of an Inspector 
 of the Local Authority. 
 
 Report to Board of Cattle that have been in Contact with Qittle afected 
 with Pleuro-pneumonia. 
 
 Where it appears to a Local Authority that there is within their District 
 any head of cattle which has been in the same field, shed, or other place r 
 or in the same herd, or otherwise in contact, with any head of cattle 
 affected with pleuro- pneumonia, or otherwise exposed to the infection 
 thereof, the Local Authority shall forthwith report the facts of the case 
 to the Board of Agriculture. 
 
 Disinfection. 
 
 An Inspector or officer of the Board may cause or require any shed or 
 other place which has been used for a head of cattle while affected with 
 or suspected of pleuro-pneumonia, and any utensil, pen, hurdle, or other 
 thing used for or about such head of cattle, to be cleansed and disinfected 
 to his satisfaction. 
 
 Occupiers to give Facilities for Cleansing. 
 
 (1) The owner and occupier and person in charge of any shed or other 
 place which has been used for any head of cattle while affected with or 
 suspected of pleuro-pneumonia shall give all reasonable facilities to an 
 
INFLUENZA. 247 
 
 Inspector or officer of the Board for the cleansing and disinfection of such 
 place, and of any utensils, pens, hurdles, or other things used for or about 
 such cattle. 
 
 (2) Any person failing to comply with the provisions of this Article 
 shall be deemed guilty of an offence against the Act of 1894. 
 
 Prohibition to Expose or Move Diseased or Suspected C<ittl< . 
 (1) It shall not be lawful for any person 
 
 (a) To expose a diseased or suspected head of cattle in a market or 
 fair, or in a sale-yard, or other public or private place where cattle 
 are commonly exposed for sale ; or 
 
 (b) To place a diseased or suspected head of cattle in a lair or other 
 place adjacent to or connected with a market or a fair, or where 
 cattle are commonly placed before exposure for sale ; or 
 
 (c) To send or carry, or cause to be sent or carried, a diseased or 
 suspected head of cattle on a railway, canal, river, or inland 
 navigation, or in a coasting vessel ; or 
 
 (//) To carry, lead, or drive, or cause to be carried, led, or driven, 
 a diseased or suspected head of cattle on a highway or thorough- 
 fare : or 
 
 (e) To place or keep a diseased or suspected head of cattle on common 
 or uninclosed land, or in a field or place insufficiently fenced, or 
 in a field adjoining a highway unless that field is so fenced or 
 situate that cattle therein cannot in any manner come in contact 
 with cattle passing along that highway or grazing on the sides 
 thereof ; or 
 
 (/) To graze a diseased or suspected head of cattle on pasture being 
 on the sides of a highway ; or 
 
 (y) To allow a diseased or suspected head of cattle to stray on a 
 highway or thoroughfare or on the sides thereof or on common 
 or uninclosed laud, or in a field or place insufficiently fenced. 
 
 INFLUENZA. 
 
 Influenza is an infectious disease characterised by a catarrh of 
 the respiratory or the gastric mucous membrane, accompanied by 
 great prostration and mental depression, and frequently ending 
 fatally by pneumonic complication. One attack is not protective. 
 The disease has occurred in the form of great epidemics, like the 
 pandemic of 1890, which is said to have started from Bokhara, and 
 travelled to St. Petersburg, Berlin, Paris, and London, whence it 
 >[.iv:ul ;ill over this country. The incubation period is extremely 
 short, only a few hours, so that numbers are attacked almost 
 simultaneously. The occurrence of ca>es iu succession in a family, 
 the importation of the <li--a>e by an infected person, and the escape 
 of persons in completely isolated localities, point to the existence 
 of a living contagium. Pfeiffer claims to have identified it with a 
 
248 INFECTIVE DISEASES. 
 
 bacillus which was found by him in the purulent bronchial secretion, 
 and, by Canon, in the blood. 
 
 Bacillus of Influenza. Very small rods, singly or in leptothrix 
 filaments. They stain with the aniline dyes, but not by Gram's 
 method. They are non-motile and aerobic; they do not grow in 
 gelatine at the temperature of the room. On glycerine-agar very 
 small transparent drop-like colonies develop in about twenty- four 
 hours. In broth there is only a very scanty growth of whitish 
 particles on the surface, which subside and form a woolly deposit. 
 They are found especially in the bronchial secretion, and only in 
 cases of influenza. Canon obtained them by puncturing the finger, 
 
 FIG. 121. BACILLUS OF INFLUENZA. 
 From a culture on gelatine, x 1000. (!TZEROTT AND NIEMANN.) 
 
 and allowing a few drops of the blood to fall upon the surface of 
 glycerine-agar in a Petri's dish. The organism will retain its 
 vitality for fourteen days in sputum, but is quickly detroyed by 
 drying. It is said that by applying the bacillus to the nasal 
 mucous membrane in monkeys, symptoms similar to influenza were 
 produced. 
 
 METHOD OF STAINING. 
 
 To stain the bacilli use Neelseri's solution or Loffler's methylene- 
 blue ; or the following method : 
 
 Canoris method. Spread the blood on cover- glasses, allow them 
 to dry, immerse for five minutes in absolute alcohol, and stain in the 
 following solution : 
 
INFLUENZA. 249 
 
 Aqueous solution of methylene-blue, strong, 40 parts ; | per 
 cent, solution of eosin in 70 per cent, alcohol, 20 parts ; distilled 
 water, 40 parts. 
 
 Float the cover-glasses from three to six hours in a capsule 
 placed in the incubator at 37 0., wash with water, and dry and 
 mount in balsam. The red corpuscles will be stained pink, and 
 the leucocytes, with the bacilli in them, blue. 
 
 FIG. 122. BACILLUS OF INFLUENZA. 
 
 From a cultivation showing filaments composed of long and short rods, 
 cocci-forms and irregular elements, x 1200. 
 
 EQUINE INFLUENZA. 
 
 Equine influenza, or " pink-eye," has been noticed to be prevalent 
 at the same time as epidemics of influenza in man, but there does 
 not appear to be any evidence of intercommunicability or of any 
 relation between the two diseases. 
 
CHAPTER XVIII. 
 
 ORIENTAL PLAGUE. RELAPSING FEVER. TYPHUS FEVER. YELLOW 
 
 FEVER. 
 
 THE PLAGUE. 
 
 THE plague is a highly infectious disease, having its origin in 
 putrefaction and filth, in tropical climates. The virus in its effects 
 resembles that of typhus. The period of incubation varies from a 
 few hours to a week. The disease produces high temperature and 
 decomposition of the blood, and dark hseinorrhagic patches appear 
 on the skin, but there is no eruption. Lymphatic inflammation and 
 buboes almost invariably occur. The virus is intensified by warmth 
 and overcrowding in houses, and dissipated by exposure to fresh air. 
 
 When the plague occurred in this country it was recognised as a 
 foreign pestilence from the East, and once imported it was fostered 
 and intensified in virulence wherever there was filth, putrefaction, 
 and overcrowding. The disease, like the small-pox, was communicated 
 from one person to another. If a case occurred in a house other 
 inmates were liable to suffer from the disease, while visitors to the 
 house ran a similar but less risk. There was a good deal of variation 
 both in the infectivity of the virus and in the susceptibility of 
 individuals, so that one contemporary writer remarked that " no 
 one can account for how it comes to pass that some persons shall 
 receive the infection and others not.' ; 
 
 Medical men were credited with enjoying an extraordinary degree 
 of immunity, though there were members of the medical profession 
 who undoubtedly died of the plague. This tradition has been 
 supported, to a certain extent, by the experience of the plague 
 in modern times. In the epidemic in Egypt, in 1835, of the ten 
 French physicians engaged there, only one died ; and while those 
 who buried the victims of the plague were liable to suffer from 
 it, and many did so, yet the medical men made more than one 
 hundred post-mortem examinations without any death resulting. 
 
 250 
 
THE PLAGUE. 251 
 
 The clothes and coverings of the infected often spread the disease, 
 and yet there are numerous examples of persons who without having 
 adopted any method of protection occupied the beds of plague patients 
 without contracting the malady. 
 
 The plague is transmissible from one country to another 
 by sea. An infected ship becomes an infective centre as readily 
 .is an infected house. Once imported, whether by land or sea, the 
 virus from infected persons or merchandise spreads wherever the 
 environment is favourable for its development and extension. 
 
 Old London afforded in every way a suitable environment for 
 the plague. The situation of the city was unhealthy, and the old 
 town ditch was a receptacle for all kinds of filth. The houses 
 projected over the roadway, and the streets were saturated with 
 constant contributions of slops and of excrement from animals and 
 human beings. The houses were often filthy and unventilated, and 
 the floors strewn with rushes, which were seldom changed. Erasmus 
 goes so far as to say that the rushes were piled the new upon the 
 old for twenty years, and were fouled with spillings of beer, 
 fragments of fish, expectoration, vomit, excrement, and urine. 
 Another very striking insanitary feature of Old London was the 
 overcrowded state of the graveyards, which was well calculated to 
 predispose to pestilence, if not actually to produce it. The burials 
 were so frequent in St. Paul's Churchyard that a new grave could 
 scarcely be dug without bodies being exposed in all stages of 
 putrefaction. 
 
 In 1894 the plague broke out in China, with all the symptoms 
 of the fatal bubonic pest of Old London. The disease was confined 
 to the poorest classes and the most overcrowded and most filthy 
 localities. In Canton the deaths exceeded one hundred thousand, 
 and in Hong-Kong numbered about ten thousand. The disease 
 was contagious, and mainly diffused by personal contact. Death 
 occurred, as a rule, in from twenty-four hours to five days. The 
 English and European community escaped, with the exception of a 
 very few out of a large number, mostly soldiers, employed in cleansing 
 the houses. The disease was a specific fever, intensely fatal, accom- 
 panied by high temperature, cerebral congestion, delirium, and the 
 formation of buboes. The buboes consisted of exquisitely painful and 
 swollen lymphatic glands. All the glands, in some cases, were affected. 
 
 According to Cantlie the glandular swelling when first recognised 
 was almond-shaped in the inguinal region, and globular in other 
 regions, with peri-glandular redema. The swelling rapidly increased 
 in size, becoming softer, less definite in outline, and less tender, until 
 
252 INFECTIVE DISEASES. 
 
 by the end of five or six days it consisted of an elevated mass, doughy 
 to the touch, almost circular, with a diameter of six inches. The 
 skin over the swelling was livid and dimpled. The swelling was in 
 some cases due to purulent effusion, but more frequently on incision 
 there was only an escape of sero-sanguineous fluid. The cervical 
 glands in very severe cases sometimes attained an enormous size. 
 
 Three out of seven Japanese medical men were attacked and one 
 died, but none out of eleven English doctors, though they were equally 
 exposed to infection. Of eight Englishmen attacked seven were among 
 the soldiers employed, and only two died. No nurses or attendants 
 on the sick were attacked. The virus appeared to be intimately 
 connected with filth in the soil. According to the Chinese, rats, 
 poultry, goats, sheep, cows, and buffaloes are susceptible. In the 
 houses and hotels dead rats were found in great numbers : it was said 
 that they emerged from their haunts in sewers and drains, appeared 
 to be dazed, and limped about, owing to the formation of buboes in 
 their hind legs. Rats, mice, and guinea-pigs inoculated with virus 
 from a human lymphatic gland died with development of buboes. 
 It appears to be clearly proved that rats suffer from the plague in 
 common with man, and it has also been suggested that they may 
 serve to spread the disease. Bacilli were found in human blood and 
 in the swollen lymphatic glands by Kitasato, and independently by 
 Yersin. 
 
 Bacillus of Plague. Short rods with rounded ends. They 
 stain with aniline dyes, but not by Gram's method. The stain 
 collects at the ends of the rods, leaving a clear space in the middle. 
 Sometimes the rods are surrounded by a capsule. They are found 
 in abundance in the buboes, and in small numbers in the blood in 
 very serious and rapidly fatal cases. 
 
 Material from the buboes inoculated on agar gives rise to white 
 transparent colonies, which have an iridescent edge when examined 
 by reflected light. 
 
 The bacilli grow more readily on glycerine-agar and on solidified 
 serum. In broth cultures the liquid remains clear, and a flocculent 
 deposit forms on the sides and at the bottom of the vessel. 
 
 An alkaline solution of peptone 2 per cent., with from 1 to 2 per 
 cent, of gelatine, is the best nutrient medium. In cultures the 
 bacilli develop chains of short rods and well-marked involution 
 forms. Swollen and degenerated forms are found most abundantly 
 in old cultures, and stain with difficulty. 
 
 Mice, rats, and guinea-pigs, inoculated with bubonic tissue, die in 
 a few days, numerous bacilli being found in the lymphatic glands. 
 
THE PLAGUE. 253 
 
 spit -t'li. and blood. Guinea-pigs die in from two to five days, and 
 mice in one to three days. 
 
 In guinea-pigs after some hours there is oedema at the seat of 
 inoculation, and the lymphatic glands are swollen. After twenty- 
 four hours the animal refuses to eat, has a staring coat, and after 
 a time suddenly falls on its side, and is attacked by convulsions, 
 which become more and more frequent until death occur.-. 
 
 After death the seat of inoculation is found to be extensi vely 
 u-.U-inatous, and the neighbouring lymphatic glands enlarged and 
 tilled with bacilli. The intestine is often congested, and the liver i< 
 
 FIG. 123. BACILLI OF PLAGUE AXD PHAGOCYTE*, x 800.' 
 From human lymphatic gland. (AOTAMA.) 
 
 r'ii.L f '->tfd and enlarged. In less acute cases an abscess of the 
 abdominal wall occasionally results. 
 
 The bacilli are sometimes found in the pleural and peritoneal 
 exudation. The liver and spleen also contain many bacilli. Those 
 in the blood are a little longer than those in the lymphatic glands. 
 
 Inoculations can readily be made from guinea-pig to guinea - 
 pig by using the pulp of the spleen, or the blood. Cultures lose their 
 virulence gradually, but the virus can IK- intensified by Boom 
 inoculations in animals. The disease is infectious to ink-- 
 well as inoculable. Pigeons are insusceptible. Rats and flies may 
 convey the bacilli. 
 
 >rdiiig to Aoyama, the bacilli found in the blood of plague 
 
254 INFECTIVE DISEASES. 
 
 patients and in the buboes are not identical. The bacilli in the 
 buboes are different in form, and they stain by Gram's method. 
 
 There is no doubt that the micro-organism which was found in 
 blood is very similar to the bacillus of fowl cholera, and it is quite 
 possible that the so-called plague bacillus is really identical with the 
 bacillus of hsemorrhagic septicaemia, and that the real nature of the 
 contagium in bubonic plague is unknown. 
 
 Protective Inoculation. Yersin, Calmette, and Borrell claim 
 not only to have produced immunity, but to have cured animals 
 after infection. Cultures on agar heated to 58 C. for an hour 
 were attenuated, and rabbits after intravenous or subcutaneous 
 inoculation were protected against virulent cultures. The serum 
 of immunised rabbits was capable of protecting from subsequent 
 virulent cultures, and neutralising the effect of a previous inoculation 
 of a virulent culture. A horse was inoculated with cultures which 
 killed mice in two days, and after six weeks a serum was obtained 
 which produced immunity in mice and guinea-pigs. 
 
 Stamping -out System. It is not until the sixteenth century 
 that we hear of preventive measures being attempted in England, 
 and then they appear to have been adopted only when an outbreak 
 threatened to be very serious. 
 
 Early in the sixteenth century all those who had the plague in their 
 houses were ordered to put up wisps, and to carry white rods in their 
 hands. 
 
 In 1543 the Plague Order of Henry VIII. was issued. In place of 
 wisps the sign of the cross was to be made on every infected house, and 
 to remain there for forty days. Persons afflicted with the disease were 
 to refrain, if possible, from going out of doors, or for forty days to carry 
 a white ro'd in the hand. All straw from the infected houses was to be 
 carried into the fields and burnt. Churchwardens were directed to keep 
 beggars out of churches on holy days, and all streets and lanes were to 
 be cleansed. 
 
 In 1547 the means of notification was a blue cross with the addition 
 of the inscription Lord have mercy upon us. Later on the colour of the 
 cross was changed to red. 
 
 With the outburst of the plague in 1563, came an attempt to enforce 
 a terrible system of compulsorily shutting up infected families. The 
 doors and windows in such houses were to be closed, and no inmates were 
 to leave the premises and no visitors to be allowed for forty days. No 
 better incubator on a large scale could possibly have been devised for 
 both breeding and intensifying the virulence of the plague bacillus, or 
 whatever may be the contagium vivum of this disease. 
 
 This compulsory shutting up of the sick with the healthy amounted 
 to a compulsory infection of many of the unfortunate inmates who might 
 
THE PLAGUE. 255 
 
 otherwise have escaped, and very naturally the order was frequently 
 infringed. 
 
 In 1568 the Lord Mayor of London drew up instructions for the 
 Aldermen for dealing with the plague. It was enacted that constables 
 and officers should search out infected houses and report to the authorities. 
 In other words, that there should be notification by the police. All infected 
 houses were to be shut up, and no person to be allowed to come out for 
 twenty days. All bedding and clothes used by the victims were to be 
 destroyed. 
 
 At Westminster these instructions were to be enforced under a penalty 
 of seven days in the stocks, with imprisonment to follow, making in all 
 a punishment of forty days. 
 
 In 1581 the Lord Mayor transferred notification from the constables 
 to searchers. Two honest and discreet matrons in every parish were to 
 search the body of every such person that happened to die in the parish. 
 They were ordered to make a true report to the clerk of the parish, and 
 the said clerk had to report to the wardens of the parish. For failing 
 to notify, the penalty was an exemplary term of imprisonment. The 
 searchers were of course likely to be offered heavy bribes by the people 
 to suppress reports, owing to their anxiety to avoid the shutting up of 
 infected houses. 
 
 The continued prevalence of the plague led to the publication, in 
 1593, of a book by Simon Kellwaye. One chapter " teacheth what 
 orders magistrates and rulers of citties and towns should cause to be 
 observed," which included among other regulations that no dunghills 
 were to be allowed near the city, and the streets were to be watered 
 and cleansed. 
 
 Xo surgeons or barbers who let blood were to cast the same into the 
 streets. All those visiting and attending the sick to carry something in 
 their hand to be known from other people ; and if the infection were in 
 few places, all the people were to be kept in their houses during the time 
 of their visitation, and when this was over, all clothes, bedding, and 
 other such things used upon the sick, were to be burnt. 
 
 In 1603 Thomas Lodge recommended that discreet and skilful men 
 should be appointed in every parish to notify sickness to the authorities, 
 and so cause them to be visited by expert physicians, and that such as 
 were sick should be separated from the whole, and that isolation hospitals 
 should be built outside the City in separate and unfrequented places. 
 
 In 1665 the Great Plague of London occurred, and was attributed by 
 some to the importation of an infected bale of silks from the Levant. 
 
 According to Hodges the disease stayed among the common people, and 
 hence was called The Poor's Plague. He criticised the system of shutting 
 up infected houses, and strongly recommended that those who were 
 untouched in infected houses should receive " accommodation outside the 
 city.'' The sick were to be removed to convenient apartments provided 
 on purpose for them. To quote his own words, " Timely separation of 
 the infected from the well is absolutely necessary to be done." 
 
 For the purification of houses his directions were to place " a chafing 
 
256 INFECTIVE DISEASES. 
 
 dish in the middle of the room, where proper things were burnt and 
 exhaled all around." The use of sulphur and quicklime was mentioned. * 
 
 Preventive measures were drawn up and published by the Lord Mayor 
 and Aldermen. Examiners in Health, watchmen, and searchers were 
 appointed. Surgeons were selected to assist the searchers in making their 
 reports, and a fee of twelve pence was allowed for every case. The disease 
 was immediately notified to the Examiner of Health. Rules for disinfec- 
 tion were made, and every infected house was shut up, and 110 one removed 
 except to a pest-house or tent. Orders were issued for cleaning and sweep- 
 ing the streets. Hackney coaches were not to be used after conveying 
 patients to the pest-house until they had been well aired. Regulations 
 were also made dealing with loose persons, assemblies, and drinking taverns. 
 
 The plague was scarcely over before the whole city was in flames. A 
 new city speedily rose upon the ashes of Old London. A few sporadic 
 cases of plague are given in the London Bills of Mortality down to 1679, 
 when they finally ceased. London was sterilised by the great fire. " Great 
 as this calamity was," wrote Thomas Pennant, "yet it proved the provi- 
 dential cause of putting a stop to one of far more tremendous nature. 
 The plague, which, for a series of ages, had, with very short intervals, 
 visited our capital in its most dreadful forms, never appeared there again 
 after the rebuilding of the city in a more open and airy manner ; which 
 removed several nuisances, which if not the origin of a plague, was 
 assuredly one great pabulum, when it had seized our streets." 
 
 In the years 1720-22 there was a terrible outburst of plague in France. 
 It was attributed at Marseilles to importation by a ship from Syria. This 
 caused a panic in England, and the Lords Justices considered it necessary 
 for the public safety that measures should be taken to defend the country 
 from a fresh invasion of this disease. Dr. Richard Mead was entrusted 
 with drawing up the required recommendations. Mead laid it down as 
 an essential doctrine that the plague was not native to this country, and 
 therefore the first thing was to prevent importation, and if such a misfor- 
 tune occurred, it was to be prevented from spreading. How was this to 
 be accomplished ? Briefly stated, his system was as follows : Lazarettoes 
 were to be provided for the reception of infected men and merchandise. 
 The healthy were to change their clothes and to be kept in quarantine, and 
 the sick were to be kept remote from the healthy and their clothes 
 destroyed. 
 
 If, through a miscarriage in the public care, by the neglect of officers 
 or otherwise, the disease was imported, then " the civil magistrates were 
 to make it as much for the interest of the afflicted families to discover 
 
 * During outbreaks of the plague amulets were extremely popular. Walnuts 
 filled with mercury, pieces of cloth coated with arsenic, and arsenical cakes, 
 were very generally worn. The College of Physicians recommended issues on 
 the arms and legs. Dr. Hodges wrote, that the more of the ulcers that were 
 made the better, although their largeness answered as well as more in number. 
 IE two issues were preferred, it was recommended to make one on the left arm 
 and the other on the opposite leg. A somewhat similar plan was adopted in 
 Circassia by small-pox inoculators. 
 
KKLAl'SING FEVER. 257 
 
 their misfortune, as it was when a house was on fire, to call in the 
 assistance of the neighbourhood." The shutting up of infected houses 
 was condemned in the strongest terms, and a system of notification and 
 isolation was proposed on the lines originally suggested by Dr. Hodges. 
 
 1. A Gmnril of Heoltli was to be established, and entrusted with such 
 powers as might enable them to see all their orders executed with im- 
 partial justice. 
 
 2. \',t>ji<;it'nn. The ignorant old women employed as searchers were 
 to be replaced by understanding and diligent men, who were to report 
 cases immediately to the Council of Health. 
 
 3. /.sn/W/on. Physicians were at once to be despatched to visit the 
 suspected cases, and when the suspicion of plague was confirmed, all the 
 families in which the sickness occurred were to be isolated. The sick 
 were to be separated from the sound, and isolation houses to be provided 
 three or four miles out of the town. 
 
 The removal of the sick was to be made at night, so as to avoid the 
 danger of spreading infection, and all possible care was to be taken to 
 provide such means of conveyance for the sick that they might receive no 
 injury. The poor were to be isolated in houses provided for the purpose, 
 but the rich were to be allowed to be in their own homes provided that 
 care was taken to separate the healthy from the sick, and no pains were 
 to be spared to provide clean and airy apartments. All expenses were to 
 be paid by the public, and a reward was to be given to the person who 
 made the first discovery of infection in any place. 
 
 Mead further pointed out that general sanitation must be carefully 
 attended to. Officers were to see that the streets were washed and kept 
 clean from filth, carrion, and all manner of nuisances. Beggars and idle 
 persons were to be taken up, and such miserable objects as were fit 
 neither for the hospitals nor for the workhouses, were to be provided for 
 in an establishment for incurables. Houses also were to be kept clean, 
 and sulphur was to be used as a disinfectant. 
 
 After centuries of experience we have learnt that the necessary 
 conditions for avoiding the plague are more accurate knowledge on 
 the part of the profession and the public of the way in which the 
 disease spreads, and the adoption of sanitary precautions, which jnu>t 
 include personal cleanliness, sa nitary dwellings, absence of overcrowd- 
 ing, immediate notification, prompt separation of the sick from the 
 healthy, disinfection of infected dwellings, destruction of infected 
 clothing, and extra-mural burial or, better still, cremation. It \va> 
 because the very reverse of these sanitary conditions exi>te.l that 
 the virus of the plague found a suitable environment in Old London 
 and in recent times in Hong-Kong. 
 
 RELAPSING FEVER. 
 
 Relapsing or famine fever is a contagious disease producing a 
 Mate of high fever lasting about seven clays, followed by apparent 
 
 17 
 
258 
 
 INFECTIVE DISEASES. 
 
 recovery, and in about fourteen days by another attack of fever, 
 which may be repeated after another week. 
 
 Starvation, in association with overcrowding and filth, is intimately 
 connected with the causation of the disease. The subjects of the 
 disease contaminate the air around them, and the virus is principally 
 conveyed by tramps and dirty people. 
 
 Obermeier discovered spirilla in the blood during the paroxysms 
 of fever. The constant occurrence of the spirillum in relapsing 
 fever, and the fact of its not being found in any other conditions, 
 render it very probable that it is the cause of the disease. 
 
 FIG. 124. SPIRILLUM OBERMEIERI IN BLOOD OF MONKEY INOCULATED WITH 
 SPIRILLA AFTER REMOVAL OF THE SPLEEN (SOUDAKEWITCH). 
 
 Spirillum Obermeieri (Spirochceta Obermeieri, Cohn). 
 Threads similar to the Spirillum plica tile. In length they are mostly 
 16 to 40 /A, with regular screw-curves. They move very rapidly, 
 and exhibit peculiar wave-like undulations. They are absent from 
 the blood during the non-febrile intervals, but are found in the 
 interior of leucocytes in the spleen. In blood serum and 50 per 
 cent, salt solution, they preserve their movements. In cover-glass 
 preparations they are readily stained by any of the aniline dyes, 
 and in sections, by preference, with Bismarck brown. They are not 
 
TYPHUS FEVER. YELLOW FEVEK. 259 
 
 t'ouml in the urine. > \\-eat. or saliva. They have not been cultivate! 
 artificially on any nutrient media. Monkeys have been successfully 
 inoculated with blood contaiiiing the spirilla by Koch, Carter and 
 Soudakewitch ; and Koch found the spirilla in the vessels of the 
 brain, liver, and kidneys, after death. According to Soudakewitch 
 a fatal result is produced in monkeys if the spleen is removed, 
 and the spirilla are found in great numbers in the blood ; but if 
 the spleen is not excised the spirilla rapidly disappear, and recovery 
 follows. Munch and Motschutkowsky transferred blood containing 
 the spirilla to healthy persons, and produced typical relapsing fever. 
 
 TYPHUS FEVER. 
 
 Typhus fever is a highly contagious disease, which lasts for two 
 or three weeks, and produces a measly eruption. Like the plague, 
 it is intimately associated with overcrowding and filth, and is liable to 
 occur where the^e conditions exist in cities, in armies, and in prisons. 
 The virus produces profound changes in the blood, and after death 
 the internal organs are found to be congested, especially the lungs, 
 which are very friable. The spleen is softened and often enlarged, 
 and the blood is dark and imperfectly coagulated. 
 
 The virus is dissipated by fresh air. It is given off by the 
 breath of patients, and possibly from the skin. It clings to the 
 clothes of patients, and the disease may be conveyed by their agency. 
 One attack, as a rule, confers immunity. Some persons are naturally 
 insusceptible, failing to contract the disease though daily exposed to 
 it. Hlava has described a bacterium which he believes to be the 
 specific micro-organism. Thoinot and Calmette found the same 
 bacterium with others, but there was no particular micro-organism 
 constantly present. There can be little doubt that the nature of 
 the contagium is unknown. 
 
 Stamping-out System. Sanitary precautions, and especially 
 the operation of the Public Health Acts in relation to lodging- 
 houses, prisons, and the better housing of the working classes, have 
 been instrumental in almost completely stamping out the disease in 
 this country. 
 
 YELLOW FEVER. 
 
 Y'ellnw fever is a disease of tropical climates, dial-art -.-riMMl l.y 
 abdominal tenderness, hsemorrhagic vomiting (black-vomit), and 
 jaundice. The disease may end fatally, or recovery occur in about 
 t\v<> or three weeks. It is especially prevalent in the West In 
 and in parts of North and South America. 
 
260 INFECTIVE DISEASES. 
 
 The virus may be conveyed by infected ships, and has in this 
 way made its appearance at British and French seaport towns. 
 The disease is generally believed to be contagious, but the source 
 of the virus is not known. According to Sternberg the virus is not 
 conveyed by water, but spreads where there is overcrowding and 
 filth. 
 
 Bacteria in Yellow Fever. Freire asserts that there is a 
 specific micrococcus in yellow fever which can be grown on all 
 ordinary nutrient media, and that cultures can be used for protective 
 inoculation with satisfactory results. Carmonay Yalle also claims to 
 have discovered the contagium ; but Sternberg, who has carried on 
 investigations extending over several years, maintains that there is no 
 characteristic micro-organism present in the blood or in the tissues 
 after death. Aerobic and anaerobic cultures were made from the 
 blood, liver, kidney, urine, stomach, and intestines. The liver was 
 found to contain after death a number of bacilli, most frequently 
 Bacillus coli communis and Bacillus cadaveris. Blood or fresh 
 liver does not produce any disease in rabbits or guinea-pigs, but 
 liver tissue kept for forty-eight hours and inoculated subcutaneously 
 in guinea-pigs is extremely pathogenic. Similar results occur after 
 inoculation of healthy liver which has been kept in the same way. 
 We may conclude from these experiments that the nature of the 
 contagium is unknown. 
 
 Stamping-out System. Sternberg states that there are many 
 facts relating to the origin and extension of yellow fever epidemics 
 which support the theory that the virus is present in the evacua- 
 tions, and that accumulations of faecal matter and of organic 
 material of animal origin furnish in certain climates a suitable soil 
 for the development of the contagium. According to this view the 
 evacuations should be thoroughly disinfected, and with other sanitary 
 precautions and efficient quarantine at seaports, the disease may 
 be stamped out, and the danger of importation from the natural 
 home of the disease reduce;! to a minimum. 
 
CHAPTEK XIX. 
 
 SCARLET FEVER. MEASLES. 
 
 SCARLET FEVER. 
 
 >. \KLET FEVER is a highly contagious disease peculiar to man. It 
 produces inflammation of the tonsils and adjoining parts, fever, and 
 a general punctiform eruption. The period of incubation is about 
 a week, and the rash usually appears on the second day. In some 
 cases the disease manifests itself in an extremely inild form, known 
 as /"tent xrtirli-f firer, in which there is only a slight febrile attack, 
 or a mild sore throat, with very little or no rash. Many c 
 would not be recognisable as such if they were not capable of 
 conveying scarlet fever, or unless other cases followed or occurred 
 simultaneously which were undoubtedly typical eases of the disease. 
 The occurrence of such cases in the early history of an epidemic 
 often causes the greatest diflieulty in tracing the origin of the 
 outbreak, and indeed in some cases renders it quite impossible to 
 
 The virus is Driven off by the skin, in desquarnation, and possibly 
 by the urine. It maintains its vitality in clothing for months, and 
 sometimes longer. It may also be conveyed by the hands of the 
 physician to w r oineii during parturition. The disease may be 
 transferred by subcutaneously inoculating persons, who have not 
 previously contracted scarlet fever, with virus obtained by puncturing 
 the eruption on the skin. 
 
 After death the internal organs appear to the naked eye more or 
 less lie .ilthy. The liver is soft, the kidneys are congested, the ileum 
 i- inflamed, and Fryer's patches enlarged and congested ; but these 
 conditions are also produced by other causes. There are inflam- 
 matory changes in the lymphatic follicles of the tonsils, and the 
 larynx and trachea. Other morbid Je-ioiis. (-specially in the kidi 
 are associated with the seijuehe and complications, and though 
 commonly occurring in scarlet fever are al.-o found in other diseases. 
 
 261 
 
262 INFECTIVE DISEASES. 
 
 These changes appear to be due to the poison which is in the blood, 
 and is excreted -by the kidneys. The epithelium is in a state of 
 cloudy swelling, a condition found in other febrile diseases and in 
 septic poisoning. 
 
 Bacteria in Scarlet Fever. The occurrence of micro-organ- 
 isms in cases of scarlet fever has been observed by several investi- 
 gators Coze and Feltz, Crooke, Lb'ffler, Babes, Heubner and Bahrdt, 
 and notably by Frankel and Freudenberg, and more recently bv 
 Klein, the author, Raskin, and others. 
 
 Coze and Feltz found cocci in the blood, and Crooke, in cases of 
 scarlet fever with severely affected throat, found bacilli, cocci, and 
 streptococci in the organs of the throat, and cocci in the internal 
 organs. Crooke left it an open question whether these cocci were 
 the specific organisms of scarlet fever, or were to be regarded as 
 diphtheritic or septic associates. He inclined, for clinical reasons, 
 to the. latter view. 
 
 Loffler,. in cases of scarlatinal diphtheria, found the same chain - 
 forming micrococcus which he had found in typical diphtheria. 
 
 Babes was able constantly to prove the presence of a strepto- 
 coccus in inflammatory products secondary to scarlatina. 
 
 Heubner and Bahrdt, in a fatal case of scarlet fever in a boy, 
 complicated with suppuration of the finger and knee-joints, and 
 with pericarditis, found a streptococcus identical in form with Strepto- 
 coccus pyogenes, but cultivations were not made. The secondary 
 infection started from diphtheritically affected tonsils, which were 
 followed by retro-pharyngeal abscesses. 
 
 Frankel and Freudenberg examined, for micro-organisms, three 
 cases of scarlatina with well-marked affection of the throat. In all 
 three cases they obtained cultivations of cocci from the submaxillary 
 lymphatic glands, spleen, liver, and kidney. These cocci could not 
 be distinguished from Streptococcus pyogenes derived from pus, nor 
 from the undoubtedly identical streptococcus which one of them 
 (A. Frankel) had repeatedly cultivated in large numbers from 
 puerperal affections. In two of the cases Streptococcus pyogenes 
 was the only organism present, and in all three cases it was far in 
 excess of other colonies which developed. The organisms were also 
 found in sections of the organs by microscopical examination. 
 Frankel and Freudenberg could in 110 way distinguish the strepto- 
 coccus in scarlatina from the streptococcus in pyaemia and septi- 
 caemia. The identity of this streptococcus with Streptococcus pyogenes 
 and Streptococcus puerperalis was established by comparison of their 
 macroscopical and microscopical appearances in cultivations on 
 
a AHLET FEVER. 263 
 
 nutrient agar-agar, nutrient gelatine, and in broth, both at the 
 ordinary and at higher temperatures, and also by experiments on 
 animals. They concluded that it could be stated with certainty 
 that the organisms in question did not stand in causal relation to 
 scarlet fever. They considered that special methods of microscopical 
 and biological research were apparently needed for demonstrating 
 the true scarlet fever contagiuui, which probably was especially 
 present in the skin. They considered that the presence of the 
 
 a. b. c. 
 
 FIG. 125. PURE-CULTIVATIONS OF STREPTOCOCCUS PYOGENKS. 
 
 <n) On the surface of nutrient gelatine; (b) In the depth of nutrient gelatine : 
 (c) On the surface of nutrient agar. 
 
 streptococcus was due to a secondary infection, to which the door 
 was opened by the lesions of the throat a view which was sup 
 ported by the fact that the organisms were found iu subinaxillary 
 lymphatic glands. They preferred to use the term "secondary 
 to " complicated " or "combined" infection, because this <-\].t 
 the fact that by the effect of the scarlatinal virus the soil is 
 rendered suitable for this ubiquitous microbe when it has once 
 gained an entrance. 
 
 This streptococcus was found by Klein in five out of eleven 
 
264 
 
 INFECTIVE DISEASES. 
 
 of scarlet fever in man, twice in association with certain other 
 micro- organisms, and three times alone. The micro-organisms were 
 isolated by inoculating tubes of nutrient gelatine, solidified obliquely, 
 by streaking the surface with blood taken from the finger, the arm, 
 or the heart after death. Those cases from which the organism was 
 obtained were all cases with ulcerated throat, and the culture 
 experiments, from the living patient, were made on or about the day 
 at which the temperature was at its maximum. 
 
 
 Name of Patient. 
 
 Condition of 
 Tonsils. 
 
 Source Micro-Organisms 
 Blood. Isolated. 
 
 Death or 
 Recovery. 
 
 1 
 
 T T? 
 
 Severely 
 
 Finger 
 
 Streptococcus 
 
 Ultimately 
 
 lj .T . 
 
 
 aged 5 
 
 ulcerated 
 
 
 
 recovered. 
 
 
 
 
 ! ( Staphylococcus 
 
 
 2 
 
 K F , 
 
 aged 2 
 
 Much 
 ulcerated 
 
 
 pyogenes aureus 
 \ Liquefying micro- 
 
 Died of 
 
 
 
 
 
 coccus 
 
 pyaemia. 
 
 
 
 
 
 ^Streptococcus 
 
 . 
 
 3 
 
 4 
 
 TT T 
 
 Ulcerated 
 Much 
 
 Arm 
 
 f Staphylococcus 
 \ Streptococcus 
 None 
 
 j- Recovered. 
 [Not stated.] 
 
 aged 8 
 (a woman), 
 
 
 aged 40 
 
 ulcerated 
 
 
 
 
 5 
 
 (a girl), 
 
 
 
 
 5> 
 
 
 aged 19 
 
 
 
 
 
 6 
 
 B M , 
 
 Ulcerated 
 
 Finger 
 
 Streptococcus 
 
 Recovered. 
 
 
 aged 15 
 
 
 
 
 7 
 
 E W . 
 
 Much 
 
 ., None 
 
 [Not stated.] 
 
 
 aged 22 
 
 ulcerated 
 
 
 
 8 
 
 R H , 
 
 Ulcerated 
 
 
 ?j 
 
 
 aged 8 
 
 
 
 
 9 
 
 F G . 
 
 n 
 
 Heart Streptococcus 
 
 Died. 
 
 
 aged 2* 
 
 
 
 
 10 
 
 
 
 None 
 
 
 
 aged 3 
 
 
 
 
 11 
 
 R_ B , 
 
 Advanced 
 
 95 
 
 n 
 
 
 
 aged 20 months 
 
 ulceration 
 
 
 
 
 Klein regards this streptococcus as the actual cause of scarlet 
 fever in man. 
 
 The author, Raskin, Holmes, and others who have investigated this 
 subject agree with the conclusions of Fraiikel and Freudenberg. The 
 author is convinced that the streptococci in suppuration, puerperal 
 septicaemia, pysemia, and septicaemia, and in certain cases of measles, 
 scarlatina, and diphtheria, are identical ; and from overwhelming 
 evidence we are justified in concluding that (1) The nature of 
 the contagium of scarlet fever is unknown. (2) The streptococcus 
 regarded by Klein as the contagium is the Streptococcus pyogenes. 
 
MILK-SCARLATINA. 265 
 
 (o) This streptococcus is found, sometimes in company with Staphylo- 
 coccus pyogenes aureus, as a secondary result in scarlet fever and 
 many other disea-e>. and its exact relation to scarlet fever and its 
 identity with the streptococcus from pus and puerperal fever, were 
 definitely established in 1885 by Frankel and Freudenberg. 
 
 MlLK-SCARLATINA. 
 
 It would not be iiec->.Niry to say any thing further on the etiology 
 of scarlet fever if the generally accepted belief, that scarlet fever is 
 a disease peculiar to man, were accepted by the Medical Department 
 of the Local Government Board ; but the theory is officially held 
 that scarlet fever is in its origin a disease of cows. Bovine scarlatina 
 is supposed to be an eruptive disease of the teats, and it is maintained 
 that the virus, by contaminating the milk, produces scarlet fever in 
 the human subject. As this theory is very naturally accepted by 
 many medical officers of health, and is mentioned in English medical 
 text-books, it will be necessary to discuss this question in considerable 
 detail, and especially as these opinions were promulgated in this 
 country with official support, and have since been proved to be 
 erroneous. 
 
 The theory of the origin of the exanthemata in diseases of the 
 lower animals is a very old one. The Arabians imagined that small- 
 pox arose from the camel. Jenner adopted a similar theory, and 
 expressed his belief that small-pox originated in the horse, being 
 generated )>\' horses suffering with "greasy" hocks. Thus Jenner 
 wrote : " May not accidental circumstances have again and again 
 arisen, still working new changes upon it, until it has acquired 
 the contagious and malignant form under which w r e now com- 
 monly sea it, making its devastations among us? and from a 
 consideration of the change which the infectious matter undergoes 
 from producing a disease in the cow, may we not conceive that 
 many contagious diseases now prevalent amongst us may owe their 
 present appearance, not to a simple, but a compound origin ? For 
 example, is it difficult to imagine that measles, scarlet fever, ulcerated 
 sore throats, and spotted skin, all spring from the same source, 
 a .-sinning some variety in their forms according to the nature of 
 their new combinations^" Uaron informs us that this idea was 
 prevalent in Jenner's mind as e:irlv as 1787. It is related that in 
 that year he accompanied his n-'plu-w. George Jenner, into a stable 
 to look at a hors. with diseased heels, and, pointing to them, he 
 remarked: "Then- is the xmrce of small-pox. I have much to >ay 
 
266 INFECTIVE DISEASES. 
 
 on that subject which I hope in due time to give to the world." 
 And again in 1794, when writing in connection with this subject, 
 he adds : " Domestication of animals has certainly provided a prolific 
 source of diseases among man." 
 
 Jenner's views were found to be incorrect, and it was shown by 
 Lov and others that the grease bears no relation to cow-pox, and 
 it is now known that Jenner mistook horse-pox for the disease 
 known as the grease. No one at the present day supports Jenner's 
 theory of small-pox in man arising from any disease of the horse. 
 Indeed, the origin of small-pox from a disease of the horse was not 
 upheld even by Jenner's pupil and nephew, Henry Jenner. The 
 latter promulgated the idea that small- pox originated from the cow. 
 He believed that small-pox, in fact, was cow-pox intensified in its 
 virulence by being passed through man. He thus expressed himself 
 " Nor may it, perhaps, be too hypothetical to suppose that the 
 cow-pox may possibly be the small-pox in its original unadulterated 
 state, before it became contaminated by passing through the impure 
 and scrofulous habits of human constitutions." The theory of the 
 origin, in animals, of human febrile diseases was, later, advocated by 
 Copland, who stated, firstly, that scarlet fever in man was originally 
 a disease of the horse, and that it formerly occurred, and had recently 
 occurred, epidemically as an epizootic among horses; secondly, that 
 it was communicated in comparatively modern times from horses to 
 man ; thirdly, that it might be, and had been, communicated to the 
 dog. But this opinion has not been accepted, for the disease called 
 scarlatina in the horse is a non-infectious disease, generally attack- 
 ing but one or two horses in a large stud. It neither spreads by 
 contagion nor infection ; and Williams states that it is impossible to 
 transmit it from the horse to any other animal, and that many cases 
 of the so-called scarlatina of the horse are in reality identical with 
 purpura. 
 
 The theory was again revived, but in another form, and has 
 been adopted by the Medical Department of the Local Government 
 Board. Owing to failure in tracing, in some cases of milk scarla- 
 tina, the contamination of the milk from a human source, the 
 theory was started that in such cases the disease is derived from 
 the cow that, in other words, there is a disease, scarlet fever 
 in the cow, which is responsible for outbreaks of scarlet fever in 
 man. 
 
 In 1882 an epidemic of scarlatina in St. Giles and St. Paiicras 
 was investigated by Mr. W. H. Power for the Board. The disease 
 \vasdistributedwith a milk supply from a Surrey farm. In this case 
 
MILK-SCARLATINA. 267 
 
 two facts were MOertained : the one, that a cow recently come into 
 milk had been suffering from some ailment from the time of her 
 parturition, of which loss of hair in patches was the most con- 
 spicuous manifestation ; the other, that there existed no discoverable 
 means by which the milk could have received infective quality from 
 the human subject. 
 
 In 1885 an outbreak of scarlet fever occurred in Marylebone in 
 connection with milk from a farm at Hendon, and again Power 
 failed to establish infection from any human source in any commonly 
 accepted way such, for example, as handling of milk, or milk utensils, 
 by persons carrying scarlatina infection. But on examining the 
 cm\- with a view to ascertain any new condition pertaining to 
 them, it came to light during the inquiry that some of them, 
 which had recently been introduced from Derbyshire, were suffering 
 from a vesicular disease of the teats. 
 
 At this stage Klein became associated with Power in the 
 inquiry; and their belief in the existence of a disease among the 
 cows on the farm capable of producing scarlatina among human 
 consumers of the cow's milk, became unreserved. Klein took away 
 with him samples of milk, contents of vesicles, and discharges 
 from ulcers, and afterwards two of the cows were purchased and 
 kept under observation. 
 
 Dr. Cameron of Hendon has given a detailed description of the 
 clinical history of this disease. He expressed his belief that it was 
 a specific disease capable of being communicated to healthy cows 
 by direct inoculation of the teats with virus conveyed by the milker 
 from a diseased animal. 
 
 The condition of the teats is described as follows : The teats 
 became enlarged, swollen to nearly twice their natural si/.e, and 
 oedematous. On handling them there was no feeling of induration. 
 Vesicles appeared on the swollen teats and upon the udder between 
 or near the teats. These varied in number from two to four on a 
 teat, and in si/e from a pea to a horsebean. The vesicle contained 
 a clear fluid. The vesicles were rubbed and broken in milking, siiid 
 left raw sores, sometimes red, in other cases pale in colour, with 
 rai>ed, ulcerated edges. Sometimes a few accessory vesicles formed 
 around the margin of these ulcerated sores. After the rupture of 
 the vesicle a brown scab formed, which might remain attached for 
 five or six weeks, or fall off in ten days or a fortnight, a >mal lei- 
 one forming afterwanl>. A thin, watery fluid exuded from under 
 the scab, and the sore ultimately healed. 
 
 Cameron examined the teats of several cows five or six weeks 
 
268 INFECTIVE DISEASES. 
 
 after they were attacked. The scabs then varied in size from a 
 shilling to a florin ; they were about one-eighth of an inch thick in 
 the centre, thinning off towards the edges. 
 
 Some of the cows were also suffering from an eruption on the 
 rump and hind quarter, consisting of patches of eczematous crusts. 
 When a crust was picked off, the hair came off with it, exposing a 
 raw, moist sore, the crusts and sores looking exactly like eczematous 
 scabs and sores ; but this condition corresponds in description with 
 eczema, the result of ringworm which is very common in young 
 stock. 
 
 In addition to his own observations, Cameron obtained infor- 
 mation from the farmers, and others familiar with cows, who 
 thought they recognised in the disease at the farm one stage of a 
 disease which they were able to describe. Cameron thus gives an 
 account of what he arid his informants together would regard as 
 a connected clinical history of the disease. 
 
 He did not see the earlier symptoms, and hence these were 
 of necessity learnt from other persons. The account, therefore, 
 of these symptoms was to be held liable to future correction or 
 modification. 
 
 Cameron stated that he learnt this disease was capable of 
 being communicated to milkers by inoculation with virus from the 
 vesicles on the teats, though the milkers on the Hendon farm 
 escaped. " A trusty informant received the virus into a recent 
 scratch 011 the forefinger while milking a diseased cow. General 
 weakness, malaise, and loss of appetite resulted, and after about 
 four or five days a vesicle or small blister appeared on the finger. 
 This broke, and several others formed on the back of the hand. 
 The whole hand and fingers became swollen and inflamed, the 
 inflammation extending in broad lines as far as the elbow. The 
 general disturbance lasted a fortnight." 
 
 In the course of the inquiry, Cameron adds that it was 
 strongly asserted by several people, who examined the cows, that 
 they were suffering from cow-pox. He, however, dismissed the 
 diagnosis of cow-pox on the ground that no papule had been 
 observed or subsequent formation of pustule, areola, or pitting, and 
 because the vesicles were not umbilicated. These reasons given 
 for dismissing the diagnosis of cow-pox at Hendon were totally 
 inadequate ; a comparison having been made between the characters 
 of the eruption of vaccinia as it appears on an infant's arm, instead 
 of the eruption of the natural or so-called spontaneous disease on 
 the teats of the cow. 
 
MILK-SCARLATINA. 
 
 Klein stated tint on the teats and udders of two cow> which 
 he investigated there were several flat irregular ulcers, varying in 
 diameter from one-quarter to three-quarters of an inch. Some 
 ulcers were more or less circular, others extended in a longitudinal 
 direction on the teat. The ulcers were covered with a brownish or 
 reddish-brown scab. The animals looked thin, but not strikingly so. 
 In feeding capacity, milking power, and body temperature there 
 was nothing abnormal. 
 
 Four calves were inoculated in the coriuin of the groin and the 
 inside of the ear, with scrapings from the ulcers after removal of 
 the crusts. In one, which may be taken as an example of the result 
 obtained, there was vesicuiation at the margin of the spot inoculated, 
 and in the centre the commencement of the formation of a crust. 
 On the seventh day each sore on the ear had enlarged to about 
 half an inch in breadth, and was covered in its whole extent by 
 a brownish crust. On the eighteenth day they had all healed up 
 and become converted into flat scars. 
 
 To search for micro-organisms, Klein removed the crust 
 from an ulcer on the teat, scraped off the most superficial layer, 
 squeezed the ulcer, and made cover-glass preparations. Tubes of 
 nutrient gelatine and nutrient agar-agar were also inoculated, and 
 a streptococcus was isolated which in morphological and cultural 
 characters agreed with those of Streptococcus pyogene-. 
 
 Two calves were inoculated in the groin with the cultivated 
 micro-organism. One calf died in twenty-seven days. At the 
 necropsy there were found peritonitis, and haemorrhage spot- on 
 the omentum ; the liver, kidneys, and lungs were congested, and 
 there were petechiae under the pleura, and pericarditis. The second 
 calf was killed, and at the necropsy the lungs and kidneys were 
 congested, and there were haemorrhagic patches on the spleen. 
 
 In these cases, the post-mortem appearances and anatomical 
 features recalled to Klein the lesions of >carlatina. In the 
 kidney, for example, the cortex was congested, and there were 
 haemorrhages, glomerulo- nephritis, and granular or opaque > \velling 
 of the epithelial cells and infiltration with round cells. From the 
 blood of the heart the streptococcus, which had been used in the 
 inoculation, was recovered. In view of this evidence it" wa- con- 
 cluded that the streptococcus was the virus of the cow disease, and 
 that it produced in calves a disease very closely resembling that of 
 >carlatina in man. 
 
 Two of the cows selected from the Ilendon farm were killed, 
 and it was observed in one that the lungs were counted, and 
 
270 INFECTIVE DISEASES. 
 
 that there were numerous adhesions by recent soft lymph between 
 the .lower lobes of the lung and the costal pleura. In the 
 liver there were several reddish streaks and patches. The spleen 
 and kidneys, with the exception of slight congestion, appeared 
 normal. 
 
 Sections of the kidney showed well-marked glomerulo-nephritis 
 and infiltration of the sheath of the cortical arterioles with numerous 
 round cells. The epithelium of the convoluted tubules was swollen, 
 opaque, and in many places disintegrating. 
 
 In the other cow there was great congestion of the lungs and 
 pleural adhesions ; the cortex of the kidney was congested, but its 
 medulla was pale. 
 
 On microscopic examination there was a good deal of round- 
 celled infiltration in the walls of the inf undibula and bronchi in the 
 lung, and round the arterioles in the kidney. 
 
 In sections of the ulcers on the teats, the corium was found to be 
 infiltrated throughout the whole extent of the ulcer with round 
 cells. In the superficial layers of the stratum Malpighi, close to 
 the stratum lucidum, as also in the stratum lucidum itself, there 
 were numerous cavities of different sizes. These cavities lay side by 
 side, the most superficial ones being covered by the stratum lucidum, 
 or extending between the layers of this stratum. 
 
 At the marginal parts the cavities, although placed side by side, 
 were well separated from one another by thicker or thinner 
 trabeculse, composed of epithelium, while at or near the centre of 
 the ulcer these trabeculse were destroyed, the cavities had become 
 confluent, and the covering layers of the cuticle having here also 
 given way. their contents extended on to the free surface of the 
 ulcer. In short, Klein states that all the anatomical details of 
 the distribution and arrangement of these cavities recalled vividly 
 the conditions observed in the vesicles of cow-pox. Yet as a result 
 of this investigation he concluded that the cow disease at Hendoii 
 was bovine scarlatina, and that towards its study and supervision 
 every effort ought to be directed in order to check the spread of 
 scarlet fever in man. 
 
 As a result of this conclusion, the Board of Agriculture resolved 
 to have the whole subject fully investigated, and the author was 
 directed to study the bacteriology and micro-pathology of this disease 
 and to report thereon. Professor Axe investigated the origin of 
 the outbreak of the disease in the cows, and Professor M'Fadyean 
 carried out an investigation into the possibility of inoculating cows 
 with the virus from cases of scarlet fever in man. 
 
MILK-SCARLATINA. 271 
 
 THE AUTHOR'S INVESTIGATION . 
 
 An outbreak of an eruptive disease of the teats, alleged to be 
 identical with the so-called Hendon cow disease, was raging in some 
 farms in Wiltshire. In this case every facility was given by the 
 owner of the estate for a thorough investigation into the disease. 
 Not only were animals sent from the farm to London, but the 
 author was allowed to visit the farms, to inspect all the infected 
 animals, and to make every investigation, with the hearty co- 
 operation of the bailiff of the farms, and the voluntary assistance 
 of the head cowmen and those under them. Some of these cowmen 
 were unusually intelligent, while two had had experience of cows 
 for more than half a century. Thus, there was not only every 
 opportunity for studying the disease on the lines indicated by 
 Klein, but it was possible by repeated visits to the farms to enter 
 into the clinical history of the disease in the cow, to study very fully 
 the nature of the disease on the hands of the milkers, and to trace 
 the probable mode of its introduction on the estate, and the way in 
 which it spread from one part of the herd to another. 
 
 Two cows were sent to London with disease of the teats and of 
 the udder between the teats. On the right teats of one there were 
 numerous sores, covered with crusts varying in size and in thickness, 
 and generally fissured. In some they were flat, in others conical ; 
 some were with difficulty removed with forceps, others were readily 
 detached. The crusts varied in colour from reddish-brown to very 
 dark brown or almost black. On detaching or scraping a crust 
 there was a granulating and somewhat indurated base. On the 
 right anterior teat there were several ulcers, from which appa- 
 rently the thick crusts had been detached, and new scabs were 
 forming. On the left posterior teat there were unusually large, 
 dark brown, or blackish crusts, covering a very extensive area 
 of ulceration, extending over the whole of the lower third of the 
 teat. 
 
 In the other cow from Wiltshire there was the same disease on 
 the teats, but not in such a severe form. The sores were covered 
 with thick crusts, but though varying in size they were more 
 regular in form, and more circumscribed. 
 
 Having entirely removed the crusts from some of the ulcer>. a 
 number of inoculations in nutrient gelatine and nutrient agar-agjn- 
 were made from the discharge, and cover-glass preparations were 
 made and stained in the ordinary way. Cultures were obtained of 
 the organisms commonly found in pus. 
 
272 INFECTIVE DISEASES. 
 
 With the discharge and with scrapings from the ulcers two 
 calves were inoculated. 
 
 Of the two calves, one was inoculated by scarification in both 
 ears ; the other, a small calf, was scarified in the left ear. Scrapings 
 from the ulcers were rubbed into the places thus prepared. In 
 addition, in the small calf an incision was made through the corium 
 in the left groin, scrapings from different ulcers on the teats were 
 well rubbed in with the blade of the scalpel, and a portion of crust 
 inserted into a small pocket in the subcutaneous tissue. In the ears 
 and the groin there were positive results. In the large brown calf 
 one of two places inoculated in the right ear passed through the 
 following changes : On the third day there was apparent vesiculation 
 and commencing formation of crust. From day to day the crust 
 thickened, and on the eighth day the crust was at its height . and 
 detached at its edges. By removing the scab an ulcer was exposed ; 
 there was slight inflammatory thickening. About the thirteenth 
 day the ulcer had quite healed. 
 
 Very similar appearances resulted in the ear of the smaller calf. 
 The result of inoculation in the groin was of a very much severer 
 character. In the course of two or three days the incision had 
 apparently commenced to heal by scabbing, but there was a surround- 
 ing area which was inflamed, and painful on manipulation. The 
 inflammatory thickening which resulted continued to increase around 
 the seat of inoculation, and the thickening could be felt to extend 
 deeply into the groin. Suppuration followed, and on firm pressure 
 pus welled up through the wound. The wound then showed very 
 little disposition to heal, and the calf began to exhibit marked 
 constitutional symptoms. During the second week after inoculation 
 the animal became very dull, and was reported by the attendant as 
 refusing to feed. Diarrhoea supervened, and lasted for several days, 
 and bloody urine was passed. The calf was also noticed to cough, 
 and the cough gradually increased in severity. Thirty-six days after 
 the date of inoculation it was decided to kill the calf and examine 
 the condition of the viscera. The appearances which were found 
 at the post-mortem examination were as follows : 
 
 The upper and middle lobes of each lung were adherent to the walls 
 of the chest ; there was congestion, especially of the middle lobe, and 
 patches of recent adherent lymph. Posterior parts of the upper lobes of 
 both lungs were completely consolidated, and on section varied in colour 
 from brick-red to greyish-white. The interlobular tissue was infiltrated 
 with inflammatory products, which mapped out the tissue of the lung in 
 small indurated areas, in which the tissue was granular-looking and friable. 
 
- THE AUTHORS INVESTIGATION. 
 
 These appearances in the upper lobes were due to septic pleuro- pneumonia. 
 They closely resembled, and were supposed to be due to, infectious 
 plenro-pnenmonuL They were, however, found identical with the con- 
 dition observed in septic pleuro-pneumonia in calves, and the disease was 
 not conveyed by infection to other animals in the same stall. Scattered 
 through the other lobes of both lungs were white, mostly firm, nodules 
 raised above the level of the surface of the lung. They were surrounded 
 by a zone of congestion, and in some cases sections were composed of 
 indurated, in others of friable, lung tissue. In the posterior part of the 
 right upper lobe there was a recent infarct. The bronchial glands at the 
 roots of each lung were enlarged to two or three times their natural size, 
 and were firm and hard on section. The parietal surface of the pericardium 
 was covered with recent adherent lymph. The visceral surface of the 
 pericardium was normal. Along the external surface of the aorta were 
 chains of enlarged lymphatic glands connected by dilated lymphatic 
 vessels. These glands were dark red or purplish in colour, from haemor- 
 rhage into their substance. The heart was normal, and the endocardium 
 not stained. There were chains of red glands on the oesophagus similar 
 to those along the aorta. The appearance of the mesenteric glands was 
 very striking. The mesentery, along the lymphatic vessels, was dotted 
 with glands, varying in size from a large shot to a pea, which were deep 
 red or prune-coloured. In addition, there were here and there enlarged 
 glands without haemorrhage into their substance, and greyish in colour. 
 There were scattered petechia3 on the spleen. The kidneys were firm 
 on section, and there was marked congestion in both, while it was more 
 pronounced in one kidney than the other. The liver was congested, the 
 congestion being more marked in patches. 
 
 Sections from the consolidated upper lobes showed under the micro- 
 scope thickening of the pleura and infiltration with round cells. The 
 exudation filled the alveoli, and was breaking down in some cases in the 
 centre. The vessels were injected, and there were hemorrhages into the 
 alveoli. The periphery of the lobules was infiltrated with round cells. 
 In sections of the kidney there was slight infiltration around glomeruli 
 and arterioles with round cells ; the epithelium in the convoluted tubules 
 was granular and disintegrating ; there was haemorrhage in the straight 
 tubules, and engorgement of vessels. In sections of liver the inter- and 
 intra-lobular vessels were engorged ; there were interlobular collections 
 of round cells displacing the liver cells, and the interlobular connective 
 tissue was infiltrated with round cells ; the liver cells were granular and 
 cloudy. 
 
 There can be no doubt from the symptoms and post-mortein 
 jipjKMiMiicvs that this calf had been suffering from septicaemia MS 
 tin* ivsult of introducing the septic virus and crust subcutaneously 
 in the groin. 
 
 The two Wiltshire cows were killed, and there was nothing of 
 importance to note in one, but in the other an incision into the 
 udder revealed an enormous abscess. 
 
 18 
 
274 INFECTIVE DISEASES. 
 
 Though the naked- eye appearances of the kidney in this case 
 were practically healthy, the results of examining sections of the 
 kidney under the microscope were extremely instructive and interest- 
 ing, as they showed that marked changes had taken place which 
 were indicative of septic complication. 
 
 The sections showed glomerulo- nephritis ; there was infiltration 
 of the capsule of Bowman with round cells ; there was infiltration 
 also of the sheaths of the vessels with round cells, especially in 
 the cortex. The blood-vessels in the boundary zone of the medulla 
 were engorged, the arterioles of the glomeruli were also engorged, 
 and there were slight haemorrhages into the capsule. The epithelium 
 of the convoluted tubules was granular, opaque, and in some parts 
 breaking down. 
 
 Sections of the ulcers of the teats of these cows were also 
 carefully examined, and the appearances corresponded exactly with 
 the description given by Klein. 
 
 On visiting the farms it was found that there were altogether 
 about a hundred and sixty cows. Only a few had proved refractory, 
 and had not taken the disease at all. The rest had contracted the 
 disease in varying degrees of severity. About fifty at a time were 
 dry, and they escaped until they were in milk again. The milk 
 was drunk on the farms and in the village, and a quantity was 
 supplied to a large town. Most careful inquiries were instituted to 
 ascertain the existence of scarlatina among consumers of the milk. 
 So far the research was completely analogous to the Hendon 
 investigation ; but, in spite of the contamination of the milk, no 
 cases of scarlatina were found either on the farms or in the village, 
 and there was no epidemic in the town in which the milk was 
 distributed. 
 
 The disease, in fact, was cow-pox, and in no way connected with 
 scarlet fever ; and to assist others who may undertake a similar 
 inquiry the details will now be given of the author's investigation 
 into the nature of the outbreak in Wiltshire. 
 
 THE DISEASE PROVED TO BE COW-POX. 
 
 Locality of the Wiltshire Outbreak. There is considerable interest 
 attached to the fact that the farms were situated a few miles from 
 Cricklade. They are close to the borders of Gloucestershire, and about 
 twenty-five miles from Berkeley. They are, therefore, within that 
 district in which in Jenner's time cow-pox was particularly prevalent. 
 
 Time of Year. The outbreak commenced about the end of September 
 1886, and lasted until about the middle of December. In an outbreak 
 
AN OUTBREAK OF COW-POX. 275 
 
 in 1885, a few miles from these farms, but on a separate estate, the 
 disease appeared in June and July. 
 
 Origin of the Outbreak. The author made careful inquiries as to the 
 origin of the outbreak, but beyond ascertaining with certainty that the 
 disease appeared first at one farm, and was conveyed from this to the 
 other farms, all evidence was negative. The milkers were unable to 
 say whether it commenced in one particular cow or whether it broke 
 out in several simultaneously. 
 
 The only information which could be obtained, which was very 
 suggestive, was that the milkers were in the habit of receiving their 
 friends from neighbouring farms on Sundays. The friends would assist 
 in the milking, to get the work done as quickly as possible on these 
 occasions. As it was reported that the same disease had occurred 
 that summer on a neighbouring farm, it is quite possible that it was 
 introduced by one of the milkers' friends. 
 
 Mode of Dissemination* When the disease made its first appearance, 
 the bailiff, attributing it to the farm being, for some reason, unhealthy, 
 decided to remove the cows to other farms. The herd was therefore 
 divided and sent to two other farms. From these cows the disease was 
 communicated to healthy cows, and, as this interchange was repeated, 
 not only of the cows, but of the milkers, the disease was communicated 
 to four separate farms. 
 
 In all cases the disease was limited to the teats, and was conveyed 
 from the teats of a diseased cow to the teats of a healthy cow by the 
 hand of the milker. In no case was there any evidence of the disease 
 being produced in healthy cows by other means than contact. 
 
 Bulls and dry cows remained free from the disease, while the cows 
 in milk, numbering about a hundred and twenty, were all attacked, with 
 the exception of about a dozen, which proved to be entirely refractory. 
 
 These facts explain how it is that the disease has been known from 
 time immemorial as the "cow-pox." We never hear of cattle-pox or 
 bull-pox. We have not, in other words, to deal with an infectious 
 disease like cattle-plague or pleuro-pneumonia, but with a disease which 
 is communicated solely by contact. 
 
 The disease was only observed in the cows in milk, and was limited 
 to the parts which come in contact with the hand of the milker. The 
 virus was mechanically transferred from diseased to healthy cows, being 
 communicated to all, or nearly all, the animals in the same shed, 
 whether the milker had vesicles on his hand or not. 
 
 Chin-iH'tiT of tin 1'lrujttinn <>,/ //,< Cow. In a recent case which was 
 carefully examined the teats were visibly inflamed, partly red and partly 
 livid in colour. On each teat there were vesicles, some broken, and 
 others which appeared to be just forming. In other cases there was 
 nothing more than the remains of broken and dried vesicles, and more 
 or less characteristic crusts on the teats. 
 
 On visiting a byre at the time that the cows were brought in to 
 be milked, it was a striking sight to look along the line and see one 
 animal after another affected with the eruption ; and thus one character 
 
276 INFECTIVE DISEASES. 
 
 of the disease was clearly shown the tendency to spread through a 
 whole herd. 
 
 On examining the eruption carefully, the degree of severity was 
 found to differ very much in different animals. In a few cases the 
 condition was most distressing, both to the cow and to the observer. In 
 such cases the teats were encrusted with huge, dark brown or black 
 crusts, which, when handled in milking, were broken and detached, 
 exposing a bleeding, suppurating, ulcerated base. Such ulcers varied 
 in size from a shilling to a florin, and in form were circular, ovoid, or 
 irregular. Weeks afterwards, when the animals had recovered, the 
 site of these ulcers was marked by irregular scars. 
 
 All the milkers agreed as to the general characters of the malady, 
 laying particular stress on the teats being red, swollen, and painful when 
 handled. Yesicles would then appear on the teats two, three, four, or 
 more on each teat. They were soon broken in milking, and irritated into 
 sores, which became covered with thick crusts. From four to six weeks 
 elapsed before they had entirely healed. Other more observant milkers 
 insisted that before the teats were red and swollen, spots or pimples first 
 appeared which came to a head. This head increased if it was not broken, 
 which might be the case if it was situated between the bases of the teats, 
 until it formed a greyish vesicle of the size of a threepenny-piece or 
 even larger. 
 
 General Symptoms in the Cow. As to the general condition of the cows 
 nothing abnormal was observed. They appeared in the best of health, 
 and in only one particular was any difference from their condition in 
 health stated to exist. This was, that in the majority of the cases there 
 could be no doubt that the milk had diminished. This might escape 
 notice by inexperienced milkers in any particular animal, but the total 
 amount of milk supplied by the herd was considerably below the average. 
 
 History of the Eruption communicated to the Milkers. The most striking 
 characteristic of this outbreak was the communicability of the disease 
 to the milkers. A milker, with vesicles which presented typically the 
 characters of casual cow-pox, was taken to London and kept under 
 observation. The various cases will be described in the order in which 
 they first presented themselves, their history being given as much as 
 possible in their own words. 
 
 CASE I. J. R., milker, informed the author that he was the first to catch 
 the eruption from the cows. He stated that it came as a hard, painful spot, 
 which formed "matter " and then a " big scab." He had been inoculated 
 about seven weeks previously. He pointed to the scar which remained 
 on his right hand. This scar presented the characters of an irregular 
 cicatrix, indicating considerable loss of substance. He stated that he had 
 also two places on his back, where he supposes he had inoculated himself 
 by scratching. He had continued milking ever since, but had had no 
 fresh places. 
 
 CASE II. W. H., milker. He stated that he was inoculated from the 
 cows about the same time as J. R. They were the two milkers of 
 the herd in which the cow-pox first made its appearance. The eruption 
 
AN OUTBREAK OF GOW-POX. 277 
 
 appeared in one place on each hand. He pointed to two irregular scars 
 as the remains of the eruption. 
 
 r\-i: III. J. L., milker, stated that he also caught the disease from 
 the cows. On his right hand a spot appeared which formed a blister, 
 then discharged matter and produced a bad sore. Lumps formed at the 
 bend of his elbow and in his armpit. He lost his appetite, felt very 
 poorly, and was obliged to leave off work for two or three days. 
 
 CASE IV. W. K., a labourer on the farm, was put on as a milker to 
 take the place of one of the others with bad hands. After his fifth or 
 sixth milking that is to say, about three days after first milking the cows, 
 pimples appeared on his hands, which became blistered and then ran on 
 to bad sores. He pointed to three irregular scars on the first and third 
 fingers and palm of the right hand. Lumps appeared in his elbow and in 
 his armpit, but he did not feel very poorly in consequence. 
 
 CASE V. J. F., milker, stated that about a month ago he noticed 
 spots which appeared on both hands. His fingers swelled and were pain- 
 ful. He said it came first like a pimple, and felt bard. Then it " weeped 
 out " water in four or five days. There were red marks creeping up to 
 his arm. There was a sort of throbbing pain, and he could not sleep at 
 night. On the right hand there was a scar, but on the left hand there 
 was an ulcer about the size of a shilling covered with a thick black crust. 
 The crust was partially detached, and exposed a granulating ulcer. It 
 was in this stage the exact counterpart of the ulcers on the cow's teats. 
 
 CASE VI. W. H., junior, milker, stated that he had both hands bad 
 about a month previously : first on the index finger of the left hand, 
 and then on the right hand on his knuckle and between the first and 
 second fingers. He said that it came up like a hard pimple, and the 
 finger became swollen and red. After a few days it " weeped out " 
 water, and then matter came away. Both his arms were swollen, but 
 his left arm was the worst. About a fortnight after, he noticed kernels 
 in his armpits, which were painful and kept him awake at night. His 
 arms became worse, he could not raise them, and he had to give up 
 milking. He also had had a " bad place " on the lower lip. On examina- 
 tion, I found that the axillary glands were still enlarged and tender. 
 He volunteered the statement that the places were just like the sore 
 teats. 
 
 CASE VII. J. H., the bailiff's son, also milked the cows. He had 
 a sore on the upper lid of his right eye and on his left hand. In both 
 cases he had been previously scratched by a cat, and the scratches were 
 inoculated from the cow's teats. The right hand also had been inoculated. 
 The eruption broke out a fortnight previously. His hands were swollen, 
 red, and hot. He felt very poorly and went to bed. Little spots like 
 white blisters appeared on the back of his right hand. His mother 
 remarked that they " rose up exactly as in vaccination." Thick dark 
 brown scabs formed. He was very ill for two or three days, but did not 
 send for a doctor. He had painful lumps at the bend of his arm and in 
 the armpit. He gave up milking, and had not taken to it since. 
 
 On examining him, the thick crusts on his right hand were identical 
 
DESCRIPTION OF PLATE VII. 
 Casual Cow-pox. 
 
 FIG. 1. Case of W. P , a milker, infected from the teats of a cow with 
 
 natural cow-pox. There was a large depressed vesicle with a small 
 central crust and a tumid margin, the whole being surrounded by a 
 well-marked areola and considerable surrounding induration. 
 
 FIG. 2. The same case a week later, showing a reddish-brown crust on a 
 reddened elevated and indurated base. 
 
Plate VII. 
 
 CASUAL COW-POX. 
 
 Hncenl Avtfelto *5oa,lii. 
 
AN OUTBREAK OF COW-POX. 279 
 
 quite so severe a character as in some of the other milkers. Possibly 
 this may be accounted for to some extent by the fact that the pock was 
 covered with a simple dressing instead of being subjected to the irritation 
 and injury incidental to working on the farm. 
 
 /{> i-'ii-clmdion of the Milker*. There were in all eight milkers, varying 
 in age from seventeen to fifty-five, who had vesicles on their hands 
 from milking the cows. Seven had been vaccinated in infancy, but not 
 since : one had been revaccinated on entering the navy at fifteen. They 
 were all revaccinated by a public vaccinator after complete recovery 
 from the casual cow-pox (that is to say, from three to four months 
 afterwards), and were all completely protected. On the other hand, 
 two of the three milkers who had escaped infection from the casual 
 cow-pox were also vaccinated, with the result in one of typical 
 revaccination, in the other of very considerable local irritation. 
 
 Ri trn-rucriniitinii <>f C'llce*. The result of retro-vaccinating calves with 
 the humanised lymph was strictly in accordance with the experience of 
 Ceely, who has pointed out that in retro- vaccination from the milker's 
 hands the results are doubtful, and depend greatly on the animals selected. 
 "Those of a light colour and with thin skins were generally preferred, 
 but often without avail, scarcely one-half of the operations succeeding." 
 "Vaccine lymph, in passing from the cow to man, undergoes a change 
 which renders it less acceptable and less energetic on being returned to 
 many individuals of the class producing it ; some refuse it altogether." 
 Two cases out of four succeeded, and an eruption was produced with 
 all the typical characters of vaccinia, but running rather a rapid course, 
 and the protection passing off after a few weeks, while the result 
 obtained in calves inoculated with pus or scrapers from ulcers was in 
 accordance with what is well known to occur if pus instead of lymph 
 is taken for carrying on calf to calf vaccination. 
 
 That the cow-pox in Wiltshire was identical with the so-called 
 Heiidon cow-disease there can be little room for doubt, for in both 
 cases we find that 
 
 1. The disease spread through a whole herd of milch cows. 
 
 2. The disease was characterised by the appearance of vesicles, 
 which were broken by the hand of the milker, and irritated into 
 Wp ulceratious. 
 
 3. The disease was conveyed from one cow to another by the 
 hand of the milker. 
 
 4. The vesicular eruption was communicable to the hand of 
 the milker. 
 
 5. The di>.-a>e was not fatal, nml in cows which were killed ami 
 examined the post-mortem appearances could not be distinguished 
 from accidental complicate 
 
 G. The naked-eye appearances and the duration of the ulcers of 
 the teats were the same. 
 
280 INFECTIVE DISEASES. 
 
 7. Sections of the ulcers showed under the microscope identical 
 appearances of a cellular character, and the purulent discharge of 
 the ulcers contained pyogenic cocci. 
 
 8. The results produced by inoculation of calves with the septic 
 virus were identical. 
 
 If we examine the chain of argument which has been brought 
 forward to maintain the existence of cow -scarlatina at Hendon, we 
 find that it was urged : 
 
 1. That the Hendon cow disease was a disease in which the 
 post-mortem appearances resembled scarlatina. 
 
 2. That this disease was associated with a streptococcus, which 
 produced, by inoculation in calves, a disease with post-mortem 
 appearances similar to those of the Hendon cows. 
 
 3. That a streptococcus regarded as identical with the one 
 above mentioned was found in certain cases of scarlatina in man, 
 which when inoculated in calves produced post-mortem appearances 
 similar to the post-mortem appearances in the original Hendon cows 
 and in certain cases of scarlatina in man. 
 
 But the microscopical appearances of the kidney of a Wiltshire 
 cow were identical with those which were regarded as indicating 
 scarlatina in a Hendon cow ; and, indeed, the statements as to the 
 post-mortem appearances in the Hendon cows, when studied, not 
 only do not necessarily indicate scarlatina, but they cannot even be 
 considered of primary importance, or as throwing much light on the 
 question of scarlatina at all. The description of the naked-eye 
 appearances in both cows only suggests coincident pleurisy or 
 pleurisy with pneumonia. The microscopical appearances in both 
 were suggestive of septic complication. 
 
 A careful examination of the post-mortem appearances of calves 
 inoculated with scraping of an ulcer of a Hendon cow, or with 
 cultivations of the streptococcus from certain cases of scarlatina, 
 brings to light much more striking changes. These appearances, 
 however, cannot be regarded as indicative of scarlatina. They are 
 in reality the post-mortem appearances of septic poisoning, and 
 occur commonly in many diseases. This is clearly shown by com- 
 paring the post-mortem appearances in the calf which was killed 
 while suffering from septicaemia as the result of inoculation from 
 the ulcers of a Wiltshire cow. These visceral changes are not to 
 be distinguished from the post-mortem appearances described in 
 the calves inoculated by Klein, Consequently, that the strepto- 
 
AN OUTBREAK OF COW-POX. 281 
 
 coccus found in certain cases of scarlet fever should produce on 
 inoculation in calves certain post-mortem appearances which are 
 found in many diseases, and should fail to produce fever, ulceration 
 of the tonsils, or scarlatinal rash, or any condition in the least 
 resembling, clinically, the disease in man, and yet that the result 
 should be regarded as scarlatina in the calf, is a conclusion quite 
 untenable. 
 
 It is true that visceral lesions similar in character were produced 
 in calves whether inoculated with scrapings or with streptococci from 
 ulcers of the Hendon cows or with streptococci from certain cases of 
 scarlet fever. In both cases the streptococcus is pathogenic, and 
 inoculation of Streptococcus pyogenes or the inoculation of septic 
 virus, is liable to produce septicaemia. These facts constitute a mass 
 of evidence which justifies the conviction that the pathological data 
 which appeared to support the theory that the vesicular disease of 
 the t?ats of cows at Hendon was scarlatina in the cow, admit of an 
 entirely different interpretation, and there can be no longer any 
 doubt that the milk was not infected by the cows but with the virus 
 of scarlet fever from some human source which Mr. Power failed to 
 discover. 
 
 All the other evidence reported to the Board of Agriculture pointed 
 to the same conclusion. The disease at Hendon w r as admittedly 
 introduced from Derbyshire; and from Professor Axe's report it 
 appears that only a part of the herd was sold to the farmer at 
 Hendon; other cows with the same eruption were transferred to 
 other dairy farms, and the disease communicated to healthy cows as 
 at Hendon, but in no instance did scarlet fever occur among the 
 consumers of the milk. At the farm of the brother of the dealer the 
 disease was communicated to three of the milkers, and the eruption 
 diagnosed by Dr. Bates as vaccinia. 
 
 All this evidence must be regarded as conclusive. The con- 
 tamination of the milk at Hendon with scarlet fever must neces- 
 sarily have been a mere coincidence; and the conclusion that 
 the milk could not possibly have become infected from any 
 human source is untenable. Professor Axe even ax-ert.-iineil 
 that scarlet fever existed at Hendon during several months of 
 1880, and that the dwellings where cases occurred stood within six 
 hundred yards of the cowsheds which contained the incriminated 
 cows, and that out of fourteen men on the farm six lived in 
 a di>trirt where cases occurred. Professor Axe has also stated 
 that the father and brother of a girl with scarlet fever, visited the 
 dairy during her illness. Whether any of those engaged on the 
 
282 INFECTIVE DISEASES. 
 
 farm suffered from latent scarlet fever does not appear to have 
 been ascertained. 
 
 There is, it is true, no evidence to show that any one daily carried 
 infection to the milk, but the exact path of infection is not always 
 easy to trace ; and because it was not actually traced it was hardly 
 reasonable to assume that the possibility of contamination from a 
 human source could be altogether eliminated. 
 
 In attempting to communicate scarlet fever to cows Professor 
 M'Fadyean confirmed the negative results which had been experi- 
 enced in some earlier experiments by Klein. In 1882 Klein 
 inoculated and fed cows and yearling heifers with diseased products 
 from human patients, using desquamated cuticle and the discharges 
 from the throat ; but the experiments all failed. M'Fadyean's 
 failures were still more marked. Cows and calves were inoculated 
 with blood from scarlet fever patients, and they were made to drink 
 water thickened with desquamated cuticle, but all the experiments 
 proved unsuccessful. 
 
 The author believes that the outbreak at Hendoii was one 
 of cow-pox, which was prevalent in this country in 1886. The 
 outbreak in Wiltshire could not be distinguished bacteriologically 
 or clinically or in its micropathology, from the disease at Hendon, 
 and the Wiltshire outbreak proved on investigation to be true cow- 
 pox. This conclusion was questioned at the time, as cow-pox was 
 generally believed to be extinct in England ; but that view is 
 entirely fallacious, and the author's conclusions have since been fully 
 confirmed by independent observers, whose work will be referred to 
 in another chapter (p. 321). 
 
 Stamping-out System. The Notification Act of 1890 may be 
 voluntarily adopted in sanitary districts, but it would be a great 
 advantage if notification were carried out uniformly all over the 
 country. Prompt information may lead to detecting the origin of 
 cases of scarlet fever, and isolation and disinfection will assist in pre- 
 venting its spread. Epidemics have occurred on a large scale owing 
 to scarlet fever existing among those engaged in dairy work, and 
 the precaution not being taken of stopping the milk supplied to the 
 consumers. Scarlet fever cannot be so readily controlled as small- 
 pox, for it may be spread by mild cases before the nature of the 
 disease is suspected, and small-pox cannot be conveyed in milk. 
 
MEASLES. 283 
 
 MEASLES. 
 
 M-ji>lfs is a contagious disease peculiar to man. It lasts for 
 one or two weeks, and produces fever, catarrh of the respiratory 
 mucous membrane, and a characteristic rash. It is highly contagious, 
 especially before the nature of the disease is revealed; there is 
 consequently great difficulty in preventing its spread in schools 
 and households. The contagium appears to be given off from the 
 body, principally if not entirely, by the breath. One attack is pro- 
 tective against future attacks. The whole population of a country 
 may acquire a certain degree of immunity. Measles introduced into 
 countries where it was previously unknown assumes a most malig- 
 nant form. There are no characteristic post-mortem appearances. 
 
 Bacteria in Measles. Micrococci have been found in the 
 blood, catarrhal exudation, and skin, by Keating, Babes, and others, 
 but they are accidental epiphytes of no importance, or associated 
 with secondary complications, as in scarlet fever. 
 
 Canon and Pielicke have found in the blood small bacilli varying 
 in form. They do not grow on nutrient agar or blood serum, but 
 cultures were obtained by pricking the finger of a patient suffering 
 from measles, and allowing the blood to drop into sterilised broth. 
 After a few days the broth became cloudy, and later, a flocculent 
 deposit formed. The bacilli were also obtained from the nasal and 
 conjunctiva! secretions. The nature of the contagium of m 
 is unknown. 
 
 Stamping-out System. Measles is not easily controlled by the 
 stamping-out system ; it is, in fact, extremely difficult, almost impos- 
 sible, to prevent its spread, as it is especially infectious during the 
 period of incubation. Notification, isolation, and disinfection assist in 
 controlling an epidemic, but the value of the system does not apply 
 to the same extent in measles as in other infectious diseases. 
 
CHAPTER XX. 
 
 SMALL-POX. CATTLE PLAGUE. 
 
 SMALL-POX. 
 
 SMALL-POX is an infectious and inoculable disease of man, charac- 
 terised by sudden and severe fever, followed in forty-eight hours by a 
 characteristic papular eruption which gradually becomes vesicular 
 and then pustular. The virus is contained in the vesicles, and in a 
 concentrated form in mature pustules. It also passes into the air 
 from the breath and skin. Infection may occur from the dead body, 
 and clothes and bedding may retain the contagium for months. 
 One attack, as a rule, gives immunity against future attacks. 
 
 Small-pox is undoubtedly a disease foreign to this country. Its 
 home is in the East. Some of the old writers held that it spread to 
 Europe from Alexandria about the year 640 A.D., following in the 
 wake of the Arab conquests in Egypt, Palestine, Persia, along the 
 Asiatic coast, throvigh Lycia, Gallicia, along the coast of Africa, 
 and across the Mediterranean to Spain ; others maintained that it 
 was not introduced until the end of the eleventh or beginning of the 
 twelfth century, by the returning Crusaders. At any rate, small- 
 pox was imported from the East, and probably from Egypt. Hero- 
 dotus, who visited Egypt, leads us to infer that epidemics were 
 unknown there during the rule of the Pharaohs ; but Egypt 
 undoubtedly became a hotbed of pestilence during the Mohammedan 
 occupation. Prosper Alpinus imagined that both the plague and the 
 small-pox were concocted in the putrid waters of the Nile, but he 
 would probably have been more correct if he had suggested that 
 they arose from the insanitary condition of the Arab conquerors 
 and their filthy camp followers, who did their best to destroy all 
 that remained of that magnificent civilisation which had existed in 
 the days of the ancient Egyptians. 
 
 We do not know the exact period at which small-pox was first 
 imported into England, and the records of the disease are very 
 meagre until the sixteenth century. 
 
 284 
 
SMALL-POX. 285 
 
 In 1593 Simon Kelhvaye appended to his work on the Plague 
 .1 .-hort treatise on the small-pox. u Oftentimes," he wrote, "those 
 that are infected with the plague are in the end of the disease 
 sometimes troubled with the small pocks or measels, as also by good 
 observation it hath been seen that they are fore-runners or warnings of 
 the plague to come." According to Kellwaye the disease arose from 
 the " excrements of all the foul humours in our bodies, which striving 
 with the purest doth cause a supernatural heat and ebullition of our 
 blood, always beginning with a feaver in the most part." 
 
 Small-pox steadily increased in the seventeenth century until it was 
 a formidable scourge, for no advantage was taken of all the experi- 
 ence which had been gained in dealing with the plague. No public 
 measures were adopted to cope with the disease, and the people came 
 to regard the new pestilence as a visitation which was unavoidable. 
 Early in the eighteenth century, small-pox inoculation was introduced, 
 and this was superseded in the nineteenth century by vaccination. 
 
 Examination of small-pox cases after death does not reveal any 
 characteristic lesions in the internal organs, but sections of small- 
 pox vesicles show an important structure. A vesicle is formed by 
 the exudation raising up the outer layer of epidermis, and the chief 
 feature is the formation of a vacuolated structure in which, especially 
 in the later stages, bacteria are found in abundance. 
 
 Bacteria in Small-pox. Cohn and Weigert found cocci in 
 variolous lymph. Hlava found Streptococcus pyogenes in the pustules, 
 and Garre streptococci in the internal organs in a case of variola 
 hsemorrhagica. In a fatal case of variola complicated with pemphigus 
 Garre found a streptococcus in the pemphigus vesicles. Klein 
 and Copeman have found a small bacillus which they regard a> 
 characteristic, but its biological characters are unknown, as it will 
 not grow on any nutrient media. The bacteria commonly found in 
 variolous pus are the usual pyogenic organisms. The nature of 
 the contagium of small-pox is unknown. 
 
 Protective Inoculation. Experience had taught that a person 
 was not. as a rule, attacked with small-pox a second time; but when 
 and how the method of artificially inducing a mild form of the 
 li>ea>e was discovered, or when this preventive treatment wa> lirM 
 employed, is unknown. Aviccnna of Bokhara was credited with the 
 discovery, and it was supposed that the practice was carried by 
 Tartar and Chinese traders' to Surat, Bengal, and China, and by 
 the Mahommedan pilgrims to Mecca. In Constantinople it was 
 supposed by some to have been introduced from the Morea by 
 an old woman, and by others by the women of Circassia. The 
 
286 INFECTIVE DISEASES. 
 
 Circassian women fastened three needles together, and pricked the 
 skin over the pit of the stomach and heart, the navel, the right 
 wrist, and the left ankle. The variolous matter was applied to the 
 bleeding points, and the eruption came out in five or six days. In 
 Constantinople scarifications were made on the forehead, wrists, 
 and legs, and carefully selected virus applied to the incisions. The 
 needle used was a three-edged surgeon's needle, or the operation was 
 performed with a lancet. The virus was obtained by pricking the 
 vesicles, and pressing out the matter into a clean glass vessel. The 
 Armenians preferred to be inoculated in both thighs. In Barbary 
 a slight wound was made between the thumb and forefinger, and 
 the virus obtained from a mild form of small-pox. In Hindustan 
 the operation was performed at certain seasons of the year, and a 
 preparatory regimen enforced. The inoculators were very careful 
 in the selection of the virus, as they had learnt its varying intensity, 
 and they were credited with being able to control the amount of 
 the eruption. They preferred to inoculate the outside of the arm, 
 midway between the wrist and the elbow in males, and between the 
 elbow and the shoulder in females. The skin over the part to be 
 inoculated was first well rubbed with a piece of cloth ; then, with 
 slight touches of a small instrument, little wounds were made over 
 an area which might be covered by a small coin, and sufficient to 
 cause just an appearance of blood. A pledget of cotton -wool 
 charged with the variolous matter, and moistened with water, was 
 applied to the wound. This virus was obtained from inoculated 
 pustules of the preceding year. 
 
 In China the contents of the variolous pustules were dried and 
 kept for several years. If the virus was to be used from fresh 
 pustules the " acrimony " of the matter was corrected by steaming. 
 The dried powder was made into a paste, which was wrapped up in 
 cotton-wool and introduced into the nostril. 
 
 The Greeks were more cautious in their procedure, and were 
 said to inoculate tens of thousands without an accident. They 
 operated only upon those in perfect health, punctures were made 
 with needles, and the virus was used in the crude state, freshly 
 obtained from the " kindly " pustules of a young child. They were 
 particularly careful in the choice of the " ferment." 
 
 Dr. Perrot Williams, in 1722, wrote that the practice of com- 
 municating small-pox had long been employed in South Wales. The 
 oldest inhabitants said that it had been a common practice with 
 them " time out .of mind," but Lady Mary Wortley Montagu was 
 responsible for the general adoption of small-pox inoculation in 
 
SMALL-POX. 287 
 
 England by persuading physicians in London to employ it. Ladv 
 M:iry had her child inoculated in Turkey. An old Greek woman inocu- 
 lated one arm, and Mr. Maitland, surgeon to the Embassy, the other. 
 The disease endued in due course with an eruption of a hundred 
 pustules. This was the first time that the Byzantine method of 
 inoculation was performed upon an English subject. In 1721 Dr. 
 Harris delivered a lecture before the College of Physicians, and 
 described the successful inoculation of four children of the French 
 consul at Aleppo, by means of a thread imbued with variolous pus. 
 A daughter of Lady Mary was inoculated in England by Maitland in 
 1721, and subsequently a number of criminals were inoculated by 
 him. Incisions were made through the cutis, and pledgets which had 
 l>e -n steeped in variolous pus from ripe pustules, were applied to the 
 wound. This was known as Maitland's or the reformed operation, 
 but it was soon modified, as troublesome ulcers resulted. Shortly 
 afterwards Maitland encountered another obstacle. The child of a 
 Mr. Batt was inoculated, had plenty of pustules, and soon recovered, 
 but six of Mr. Batt's domestic servants, " who all in turn were 
 wont to hug this child while under this operation, and whilst the 
 pustules were out, never suspecting them to be infectious, were all 
 seized at once with the right natural small-pox of several and very 
 different kinds." 
 
 Dr. Jurin in 1729 reverted to the Eastern method, and recom- 
 mended virus from a mild case of small-pox, but the virus was still 
 taken from perfectly maturated pustules, and the operation continued 
 to be followed by bad results. In order to diminish the risks, Burgess 
 in 1766 advocated certain improvements. An incision about an inch 
 long was made on each arm through the cuticle, but not so deep as 
 to wound the cellular tissue. A variolous thread was laid along the 
 whole length of the wound and fixed with plaster. Ulcerations and 
 other accidents continued to take place, arid a new epoch in the 
 history of inoculation was the introduction of the Suttonian method, 
 in 1764-6. 
 
 It was said that Mr. Sutton, with his assistants, inoculated 
 one hundred thousand persons. The method was kept secret at first, 
 but the essential points were all discovered and published by Dr. 
 Dimsdale. Dimsdale recommended a very slight puncture with a 
 lancet wet with variolous matter. Subsequently, Sutton published 
 an account of his method, and the result of his operation may be 
 given in \i\^ own words. 
 
 'The lancet being charged with the anwllftt perceivable r////////V// 
 (and the smaller the better) of um-ijn . /-/-///A . or n-'ifn-y mtttti-r, immediately 
 
288 INFECTIVE DISEASES. 
 
 introduce it by puncture, obliquely, between the scarf and true skin, 
 barely sufficient to draw blood, and not deeper than the sixteenth part of 
 an inch. Neither patting, nor daubing of the matter, in or over the 
 punctured part, is at all necessary to its efficacy. This practice, indeed, 
 is rather prejudicial than otherwise, as it may affect the form of the 
 incision, and thus be apt to confound our judgment upon it. 
 
 " Indications of the Incision. In the incipient state of variolous 
 increase in the incision, a small florid spot appears on the part of access, 
 resembling a flea-bite in size ; and on passing the finger lightly over it a 
 hardness is felt not larger than a small pin's head. This florid appearance 
 and hardness denote that the variolous principle is effectually imbibed, 
 and their indications point no farther, unless the progress to vesication 
 be very slow, in which case an uncomfortable number of pustules may be 
 expected to follow. The florid spot in most instances of inoculation is 
 somewhat larger, or more extended on the second, than on the third day 
 after the insertion. , 
 
 "About the fourth day from inoculation, should the incision begin 
 to vesicate, an itching sensation will be complained of on the place of 
 insertion the occurrence of which symptom is the first indication of a 
 favourable event, yet not of sufficient importance to justify any present 
 relaxation in the preparatory proceedings. 
 
 u The vesication of the incision in most instances will begin to be visible 
 on the fourth or fifth day after the insertion of the matter ; the sooner 
 it becomes so, the more favourable may be expected to be the event. The 
 extent or diameter of the vesication at this stage does not usually exceed 
 that of a large pin's head, and it has invariably a dint or small depression." 
 
 Adams obtained still more striking results by inoculating with 
 variolous lymph from pearl-pox, a mild variety of small-pox. Starting 
 with lymph obtained from this benign form of small-pox, and 
 selecting the cases, and carrying on arm to arm variolation, the 
 results obtained were practically identical with the phenomena 
 obtained by inoculation of the arm with cow-pox lymph. Similar 
 results were obtained by Guillou, but more rapidly. In 1827 there 
 was an epidemic of variola, and Guillou, having no vaccine virus, 
 took variolous lymph from a girl fifteen years of age on the fifth day 
 of the eruption. The case was one of varioloid or mild sin all-pox, attri- 
 buted to previous vaccination. The variolous lymph was inserted in 
 ten places on the arm of a healthy infant still at the breast. This 
 inoculation produced ten beautiful " vaccine " vesicles, from which, on 
 the ninth day, forty -two infants were inoculated under the eyes of 
 two of the local authorities. These furnished virus for the inoculation 
 of one hundred, who were inoculated in the presence of magistrates 
 and many medical men. This experiment was repeated with success. 
 Variolous lymph was taken from two lads at school, and in ten 
 
SMALL-POX. 289 
 
 - produced appearances with a perfect similarity to ordinary 
 vaccination. 
 
 Tliiele produced a benign vesicle in the following manner. 
 Yarii>lou> lymph was diluted with warm cow's milk, and inoculated 
 like ordinary vaccine lymph. Large vesicles resulted. There were 
 t'<-l>rile symptoms from the third to the fourth day, and a secondary 
 onset of fever much more pronounced between the eleventh and 
 fourteenth days. The areola was strongly marked, and not con- 
 fined to the inoculated place, which was occasionally surrounded 
 by minute secondary vesicles. After watching through ten removes, 
 the vesicles finally assumed the characters of an ordinary vaccination 
 with cow-pox lymph. As soon as the secondary fever ceased to 
 occur inoculation was practised from arm to arm without diluting 
 the lymph with cow's milk. The lymph was designated lacto-varioline, 
 and the result was variolation in its mildest form. The result of 
 variolating the cow will be discussed in another chapter. 
 
 Small-pox inoculation, or variolation, protected the individual 
 when genuine small-pox was produced, and .endangered the com- 
 munity. Persons inoculated became centres of infection, and con- 
 veyed the disease to others. Haygarth, although in favour of 
 inoculation, strongly condemned its use without precautions to 
 prevent the spread of the disease. " The most serious and solid 
 objection," he wrote, " that has been advanced against inoculation 
 is deduced from a comparison of the Bills of Mortality for a series 
 of years in various places. They show that a larger proportion 
 of inhabitants have died of the small-pox in towns where it is prac- 
 tised than in the same before it was known, or in others where it is 
 prohibited." 
 
 Even Dr. Dimsdale, an ardent inoculator, admitted that more 
 lives were lost in London than before inoculation commenced, and the 
 practice was more detrimental than beneficial to society ; and he 
 added : " The disease by general inoculation throughout London 
 spreads by visitors, strangers, servants, washerwomen, doctors, and 
 inoculators, by means of hackney coaches in which the sick are sent 
 out to take the air, or by sound persons approaching them in the 
 streets. The poor in London are miserably lodged ; their habitations 
 are in close alleys, courts, lanes, and old dirty houses ; they are often 
 in want of necessaries, even of bedding. The fathers and mothers 
 are employed constantly in laborious occupation abroad, and cannot 
 attend the inoculated sick." In 1798 Jenner, who had practised 
 small-pox inoculation, proposed the use of a benign non-infectious 
 lymph obtained from a disease of the cow or horse as a substitute 
 
 19 
 
290 INFECTIVE DISEASES. 
 
 for variolous lymph, and in 1840 small-pox inoculation was prohibited 
 by Act of Parliament. 
 
 Stamping-out System. The disappointing and dangerous 
 results of small-pox inoculation led to a widespread demand for 
 some new method for dealing with small-pox. This induced Haygarth 
 to turn his attention to the subject, arid towards the end of the 
 eighteenth century to bring before the medical profession and the 
 public a plan for stamping out the disease. Haygarth, who was a 
 close observer and an able physician, studied the question of the 
 communicability of the disease from one person to another, and its 
 conveyance by infected clothing and other means, and ultimately 
 drew up rules and regulations for its prevention, the importance of 
 which we are only now beginning to fully acknowledge. Haygarth's 
 essential doctrine was " that mankind was not necessarily subject to 
 the small-pox, and that it was always caught by infection from a 
 patient or the poisonous matter," and might be avoided by observing 
 his Rules of Prevention. 
 
 These rules comprised a regular system of notification and isolation. 
 Inspectors were to be provided to report cases of small-pox, and people 
 were to be rewarded for carrying out the instructions. Several examples 
 were given of the results at Chester, where the plan was adopted. 
 
 Haygarth met with considerable encouragement from some of the 
 leaders of the profession. Dr. Fothergill wrote to him in 1778, saying, 
 " I have mentioned the intention of freeing this country from the 
 small-pox to divers of the faculty, and shall continue to do so as it falls 
 in my way. The proposal is variously received, but in exact proportion 
 to their humanity." 
 
 In 1793 Haygarth made considerable addition to his rules, and urged 
 that legislation should follow to make them compulsory. Provision was 
 to be made to reward the poor for observing the rules, and public thanks 
 to the wealthy for their support were to be published in the parish church 
 and newspapers. Transgression of the rules was to be punished by a fine of 
 from 10 to 50, one half to go to the informer and the other half to the 
 fund which supplied the expense of rewards to the poor, and all details were 
 to be supplied to the press. It was further suggested that Great Britain 
 should be divided into districts, including a certain number of parishes 
 or townships, and that to each of them a surgeon or apothecary should 
 be appointed as inspector to see that the regulations were exactly 
 observed. In addition, there were to be directors of inspectors, superin- 
 tended by a commission of Physicians in London and in Edinburgh. All 
 salaries were to be paid by the county rates, and the rewards for observing 
 the rules of prevention were to be guaranteed out of the parish funds. 
 On the requisition of the director and inspector of a circuit, power 
 was to be given to two or more justices of the peace to appoint a separate 
 house for the reception of patients with the small-pox. In conclusion, 
 
SMALL- POX. :><J1 
 
 Haygarth maintained that the plague had been completely exterminated 
 from this country, for above a century, by civil regulations, and that 
 there seemed to be little doubt that the small- pox was propagated on 
 principles similar to the plague, and that it also might certainly be 
 exterminated from this island. 
 
 Hay garth's teachings had a profound influence upon both the profession 
 and the educated public, but his system of compulsory notification was 
 never carried out, for no legislation followed to enforce his recommenda- 
 tions. This is a matter deeply to be regretted, for towards the end of the 
 eighteenth century small-pox was declining in London, general sanitation 
 was making rapid advances, small-pox inoculation, which created fresh 
 centres of infection, was falling into disfavour, small-pox hospitals were 
 built, which served to limit centres of infection, and the profession and 
 the public were influenced by the teaching of Haygarth with regard to 
 the various ways of avoiding the spread of the disease. 
 
 It only required the compulsory adoption of Haygarth's system 
 uniformly all over the country to have kept the disease in control, 
 if not to have entirely extirpated it from Great Britain. That a 
 similar conviction existed at the time is evidenced by an article which 
 appeared in 1779 in the Medical and CMruryioal Review, in which 
 the following statements were made : 
 
 ' Plans for the extirpation of the small-pox have been suggested. . . . 
 To do this, however, the exertions of the physician are incompetent unless 
 they be aided by the powerful hand of Governments, but this has hitherto 
 been withheld. The grand means, however, of extirpating this destructive 
 malady is an early and strict separation of the infected from those that 
 are sound." 
 
 Small-pox in the present century has been largely controlled by 
 legislation, especially in recent years, by the Public Health Acts for 
 England and Wales, for Scotland, and for Ireland ; the Epidemic and 
 other Diseases Prevention Act ; the Public Health Amendment Act ; 
 the Labouring Classes' Dwellings Acts ; the Housing of the Working 
 < 'lasses Act ; the Public Health (Ships) Act; the Local Government 
 Board Act and various orders and memoranda of the Local Govern- 
 ment Board ; the Infectious Diseases Notification Act ; the Infectious 
 IM-eases Prevention Act ; and the Public Health (London) Act. 
 
 By the Public Health Act of 1875 England was divided into 
 Vrlian and Rural Sanitary Districts, and powers were given to 
 Mifnrce regulations of the Local Government Board for guarding 
 jtirainst thf spreading of infectious diseases; to provide medical aid 
 and accommodation for infected persons, to promote cleansing, 
 ventilation, and disinfection, to provide hospitals, to provide for 
 
292 INFECTIVE DISEASES. 
 
 destruction or disinfection of infected bedding, clothing, and other 
 articles, and to appoint Medical Officers of Health. 
 
 As to the value of notification and isolation in cities such as 
 London we have the evidence of the Metropolitan Asylums Board. 
 In their Report for 1889 we read in reference to the diminution of 
 small-pox : " These very satisfactory results confirm the view taken 
 by the Committee two years ago to the effect that the rapid and 
 systematic removal from crowded districts of infected persons, each 
 of whom might have become a centre of contagion, is an important 
 factor in stamping out small-pox from the metropolitan population. 
 The notification of cases will also greatly facilitate the action of the 
 managers in this direction." 
 
 More recently there has been a most striking confirmation of 
 these statements. An outbreak of small-pox occurred in Maryle- 
 bone, and by the energy of the officials of the Board this outbreak 
 was suppressed in a few days by means of notification and 
 immediate isolation. 
 
 The Isolation Hospitals Act of 1893 gives power to County 
 Councils to provide, or cause to be provided, an isolation hospital 
 in any district within their county. An application to a County 
 Council for the establishment of an isolation hospital may be made 
 by any one or more of the authorities defined as local authorities 
 having jurisdiction in the county or any part of the county. 
 Further, the County Council may direct an inquiry to be made by 
 two medical officers of health in the county as to the necessity of an 
 isolation hospital being established for the use of the inhabitants 
 of any particular district in the county, and in the event of such 
 medical officers reporting that such a hospital ought to be 
 established for the use of the inhabitants of a district, may take the 
 same proceedings in all respects for the establishment of such 
 hospital, as if a petition had been presented by a local authority for 
 the establishment of an isolation hospital for the district named in 
 the report of such medical officers of health. 
 
 Lastly, the Local Government Act of 1894 provides for the 
 formation of District Councils ; and the powers, duties, and 
 liabilities are principally those which were conferred by the Public- 
 Health Act of 1875. 
 
 In the opinion of the author the Government of this country should 
 enter into friendly negotiations with the Governments of other countries, 
 so that there might be concerted action to prevent an avoidable 
 disease like small-pox. Much good might result from the formation of 
 a permanent International Board of Health. If civilisation is not yet 
 
CATTLE PLAGUE. 293 
 
 sufficiently advanced to admit of a system of international notification, 
 our Consular authorities should be instructed to give immediate notifica 
 tion of the existence of small-pox in other countries, and every measure 
 should be enforced to diminish the possibilities of importation. The 
 duties of a Central Health Office, presided over by a Minister of Health, 
 should include the collection of information as to the existence of small- 
 pox in other countries, and details should be published in the Annual 
 Reports of the Department. Regulations, for example, for dealing with 
 the importation of rags from small-pox stricken places should be enforced, 
 as in the case of cholera : and if, in spite of these precautions, isolated 
 cases occurred in this country, they should be dealt with promptly. 
 
 Notification should be enforced uniformly all over the country, and 
 there is not the slightest reason why the authorities and the public should 
 not immediately receive information of the existence of small-pox, whilst 
 to procure immediate isolation we have only to imitate the excellent 
 ambulance system of the Metropolitan Asylums Board. To procure 
 prompt notification there must be no loophole for evading the Act, and 
 there should be a heavy penalty for failure to notify not only small-pox, 
 but any ra*e tchich //<"/// rf<i*n<il)l ,/ lj s>ij>j><>x<>d to be <nic of *i nail-pox. 
 
 The police should be required to report any case of small-pox in 
 common lodging-houses or shelters : they should have power to require 
 any tramp suffering from small-pox, or from any disease which may 
 reasonably be supposed to be small-pox, to be examined by the medical 
 officer of the Union, and kept under observation, or transferred at once 
 to the isolation hospital : and inmates of the workhouse should be daily 
 inspected, and no case allowed to leave when there is the least suspicion 
 of small-pox infection. 
 
 Objections no doubt will be raised to this proposal, but the frequency 
 with which small-pox is spread by tramps fully justifies these measures. 
 All these measures should be carried out as a matter of routine, and 
 without the semblance of panic. 
 
 Isolation should be uniformly enforced all over the country, and 
 vaccination should be relegated to the position of a voluntary auxiliary 
 measure, which should never be allowed to take the place of sanitary 
 regulations to stamp out the disease. 
 
 CATTLE PLAC;I K. 
 
 < ';t trie plague is a highly contagious disease of bo vines producing 
 high fever, and characterised by an eruption with a resemblance to 
 human small-pox. The disease is transmissible to other ruminant-, 
 and is inoculable in man. One attack gives immunity against 
 future attacks. Cattle plague and small -pox are not intercom - 
 mimicahle, and are specifically distinct diseases, but the resemblance 
 1 et ween them was recognised from early times. Ramaz/ini published 
 an account of the cattle pest in Italy in 1711, and described the 
 pustules which broke out over the body as >inniar to those of variola in 
 
294 INFECTIVE DISEASES. 
 
 kind and appearance. Dr. Layard, in 1780, described this disease of 
 horned cattle as an eruptive fever of the variolous kind, with the 
 appearance and stages of small-pox. This resemblance was endorsed 
 by Murchison, one of the Commissioners appointed in 1866 to 
 inquire into the origin and nature of cattle plague. 
 
 Murchison pointed out that in both diseases the eruption con- 
 sisted of pustules and scabs, and that in both it extended from 
 the skin to the interior of the mouth and nostrils ; in both, the 
 pustules and scabs were preceded or accompanied by patches of 
 roseola ; in both, they were occasionally interspersed with petechise ; 
 and in both, they sometimes left behind pitted scars and discolora- 
 tions on the cutis. The other prominent symptoms of rinderpest 
 were also those of small-pox viz., pyrexia, lumbar pain, salivation, 
 and running from the nostrils, alvine flux, albuminuria, hsematuria, 
 and " the typhoid state." The anatomical lesions of the internal 
 organs in rinderpest and unmodified small-pox were identical viz., 
 congestion or inflammation of the mucous membranes of the air 
 passages and digestive canal, patches of ecchymosis and even 
 gangrene of the stomach and other mucous surfaces, and darkly 
 coloured blood. In both rinderpest and srnall-pox the duration 
 of the pyrexial stage was on an average about eight days. In 
 both diseases a peculiarly offensive odour was exhaled from the 
 body before and after death. The two diseases resembled one 
 another in their extreme contagiousness, and in the facility with 
 which the poison was transmitted by fomites. Both diseases were 
 easily propagated by inoculation, and in both cases the inoculated 
 disease was milder and less fatal than that resulting from infection. 
 In both diseases there was a period of incubation, which is shorter 
 when the poison has been introduced by inoculation than when it 
 has been received by infection. 
 
 Ceely described the result of an accidental inoculation of cattle- 
 plague virus in the human subject. A vesicle was produced which 
 so closely corresponded with the result of inoculated cow-pox that 
 Ceely inclined to the belief that cattle plague was a malignant form 
 of cow-pox. The following is the account of this case as reported 
 by Ceely. Mr. Hancock, a veterinary inspector at Uxbridge, was 
 engaged in superintending the autopsy of a bullock recently dead of 
 cattle plague. His assistant, who was performing the operation, 
 while occupied in removing the skin from the scrotum, accidentally 
 punctured the back of Mr. Hancock's hand with the point of the 
 knife. The puncture being slight was disregarded at the time, but 
 was washed as soon as practicable, and thought of no more. Five 
 
CATTLE PLAGUE. 
 
 days afterwards, a small, slightly elevated, hard pimple was felt 
 and >een on the site of tin' puncture. This gradually advanced 
 till the ninth day of the puncture, the fourth from papula tion, 
 when the enlargement became distinctly vesicular. At that time 
 there were hut slight constitutional symptoms. On the next day, 
 tin- tenth from the receipt of the puncture, the fifth from papulation, 
 and the x'cnnd from vesiculation, Mr. Hancock consulted Mr. Rayner, 
 of Uxl -ridge, who, on seeing the hand, inquired if the patient had 
 heeu handling the udder of a cow, as he thought he could recognise 
 a cow-pox vesicle of the ninth day. The vesicle was distended 
 with thin lymph, its margin elevated and slightly brown, its centre 
 depressed and brownish, and the whole surrounded with a large 
 bright red areola. There was then considerable tumefaction extend- 
 ing from the knuckles to above the wrist. The absorbent ve>-< -'t- 
 were considerably inflamed, and, like the axillary glands, were tender 
 and painful; the pulse, naturally slow, was accelerated; there was 
 much pain in the back and limbs, severe distracting headache, etc. ; 
 all of which symptoms continued to increase during the two following 
 <lays. At the end of that time the diffused areola had extended as 
 far as the elbow. Fifteen days after the puncture, and ten days after 
 papulatioii, the local inflammation and constitutional symptoms had 
 partially subsided. The vesicle contained a rather turbid brownish 
 fluid, and there were present all the indications of a declining vaccine 
 vesicle. 
 
 Murchisnii also saw and described the case, and gave practically 
 the same account of it. He pointed out that the appearances and 
 the entire history were \-ry different from the results of a poisoned 
 wound, but coincided with the appearances seen after vaccination. 
 
 In 1832 Macpherson, in Bengal, inoculated eleven native children 
 with cattle-plague cru>t>. There was no result in six, others 
 suffered from local inflammation, and in one a v->icl- formed. 
 \Vith lymph from this vesicle other children were inoculated. The 
 results in all were similar in appearance to those of vaccination. 
 Two children were subsequently inoculated with human variola, and 
 were said to be protected. 
 
 In 1834 Macpherson's example was followed by Mr. Fun ell 
 in ASM m. Furnell inoculated four children with cattle-plague 
 crusts without result, but hi> awtttnl sucn-eded with crusts taken 
 from the back and abdomen of the di>ens-d cattle, and carried on 
 the lymph from child to child. In one case there was a general 
 eruption. Furnell inoculated his own child from one of the native 
 children: a copious eruption followed, and the child died. Furnell 
 
296 INFECTIVE DISEASES. 
 
 after this misfortune issued a strong warning against taking the 
 virus from the cow. The experiments were made in the belief that 
 cattle plague was really small-pox in cattle, and that the virus 
 would protect against human variola. 
 
 Similar results were obtained by Mr. Wood at Gowalpara 
 in 1838. 
 
 Bacteria in Cattle Plague. Semner cultivated streptococci 
 from the blood and lymphatic glands of a sheep suffering from 
 cattle plague. A calf inoculated with a cultivation died in seven 
 days. The cocci were stated to lose their virulence by cultivation, 
 and the weakened cultivation to protect against the virulent disease. 
 
 The micro-organism was very probably Streptococcus pyogene-i, 
 and the calf may have died of septic infection. Theie can be no 
 doubt that the nature of the contagium of cattle plague is unknown. 
 
 Protective Inoculation. In the great epidemic of cattle 
 plague in England in 1866, owing to a belief that the analogy 
 between cattle plague and small-pox was closer than it really is, 
 vaccination with cow-pox was attempted as a preventive measure, 
 but was proved to be absolutely useless. 
 
 Stamping-out System. When cattle plague was imported in 
 1865 into London, dairymen and stock-Owners made no attempts 
 to prevent the extension of the disease, so that it spread rapidly 
 all over the country through disposal of infected cattle. The losses 
 were enormous, and ari Order in Council was passed in July 1865, 
 directing dairymen and others to notify outbreaks of any contagious 
 or infectious disease among the animals under their charge. A 
 Veterinary Department of the State was formed, and inspectors 
 appointed in various parts of the country. A short Act was passed 
 in February 1866. A stamping-out system, consisting of compulsory 
 notification and the slaughter of diseased animals, was soon brought 
 to the notice of the public. There was violent opposition, but 
 nevertheless, after some delay, the system was carried out. The 
 number of cases of cattle plague had reached 18,000 weekly, and 
 on the introduction of the stamping-out system the disease rapidly 
 declined. The disease was again imported into Great Britain in 
 1872, and there were outbreaks in 1877. In each instance the 
 disease was promptly stamped out, and ever since that year the 
 disease has been kept out of this country. 
 
CHAPTER XXI. 
 
 SHEEP-POX. FOOT-AND-MOUTH DISEASE. 
 
 SHEEP-POX. 
 
 SHEEP-POX, or variola ovina, is an acute febrile disease accompanied 
 by a general vesiculo-pustular eruption, highly infectious, and 
 capable of being propagated by inoculation or clfivelisation. It is 
 a common disease in some parts of Europe. In France the disease 
 is called la clavelee, and in Italy vaccuolo. It has been introduced 
 on several occasions into this country, but has been effectually 
 stamped out. As in human small-pox, there are varieties the 
 benign and the malignant, the discrete and the confluent ; and one 
 attack is protective against the disease in future. 
 
 It is very closely analogous to human small-pox. Vaccination 
 with cow-pox lymph has been employed to protect sheep from sheep- 
 pox, but unsuccessfully, and lymph for vaccination has been rai>rd 
 from sheep-pox to protect human beings from small-pox. These 
 experiments were first performed in Italy. 
 
 Marchelli, in 1802, took lymph from the vesicles of sheep-pox, a ml 
 inoculated children. Sacco repeated these experiments, and found 
 there was no appreciable difference from the results obtained with 
 cow-pox lymph. Dr. Legni carried on the inoculations with ovine 
 virus from arm to arm for several years, and when small-pox 
 <>c< -in-red in Pesaro, it was said that all those who were inoculated 
 with the sheep virus were protected. 
 
 Inoculation of children with ovine virus, direct from sheep, was 
 r !; i ted by Sacco and Magnani in 1806. 
 
 M arson in England succeeded in producing on the human 
 subject a vesicle with the physical characters of the vaccine vesicle. 
 The vesicle had a bluer tinge, and subsequent inoculation of the 
 patient with human variola was int-ilVctual. Other experiment <-r> 
 \\PIV unsuccessful, but their failures, as in tin- ca<c of variolation of 
 the cow, do not invalidate the results of those who were successful. 
 
 297 
 
298 INFECTIVE DISEASES. 
 
 Sheep-pox and cow-pox are quite distinct diseases. Sheep-pox 
 is highly infectious, whereas cow-pox is only conveyed by direct 
 inoculation, and is never infectious, and further, cow-pox inoculated 
 in sheep does not produce sheep -pox. 
 
 Bacteria in Sheep-pox. Hallier and Zurn, Klein, and others, 
 have found micrococci and bacteria in the lymph of the vesicles of 
 sheep-pox, but they are only accidental epiphytes. The nature of 
 the contagium is unknown. 
 
 Protective Inoculation. Extensive experiments were carried 
 out in England to test the protective power of vaccination against 
 sheep-pox. According to Marson and Simmonds, it was very difficult 
 to get cow-pox to take on sheep, and when an effect was produced, 
 the resulting affection, even when developed to its fullest extent, was 
 very unlike the same disease in the human subject. In the sheep 
 it seldom produced anything more than a small papule, which occa- 
 sionally resulted in the formation of a minute vesicle, or more 
 commonly, a pustule, which was sometimes, although very rarely, 
 surrounded by a slight areola. Generally, however, neither vesica- 
 tion nor pustulation followed, but a small scab was produced, which 
 soon fell from the site of the puncture, leaving no trace behind. The 
 disease passed quickly and irregularly through its several stages, 
 and terminated by the eighth or ninth day, and not unfrequently 
 even before that time. Lymph was but rarely obtainable, and then 
 only in the smallest quantity, and this on the fifth or sixth day suc- 
 ceeding the vaccination. The effects were only local, and the animal's 
 health was not impaired. 
 
 Sheep were found to be just as susceptible of the cow-pox virus 
 on subsequent repetition of the inoculation as they were in the 
 first instance, and hence the conclusion that cow-pox was utterly 
 worthless as a protective against sheep-pox. According to Depaul, 
 however, cow-pox takes characteristically on sheep, and sheep- pox 
 lymph inoculated on cows produces a result indistinguishable from 
 the appearances obtained with the inoculation of cow-pox lymph. 
 It is impossible to say whether these conflicting results depended 
 upon the employment in the experiments of different breeds of sheep 
 or different stocks of vaccine lymph. 
 
 The objection to clavelisation or ovination is that the disease 
 may be introduced in localities where it was previously unknown. 
 By ovination, as in the analogous case of variolation, fresh centres 
 of infection are created, whereas every precaution should be taken 
 to prevent the introduction of the disease. 
 
 Stamping-out System. Sheep-pox has been imported into this 
 

 FOOT-AND-MOUTH DISEASE. 299 
 
 country on several occasions. It was introduced in 1847, and again 
 in 1862; in 1865 it was introduced again, and active measures of 
 repression \\cre at once taken. The diseased flocks were carefully 
 isolated, and day by day as fresh cases occurred the diseased animals 
 were killed and buried. Owing to the adoption of these precaution- 
 ary measures, the affection did not extend beyond the flock among 
 which it first appeared. It was introduced again in 1866 at Long 
 Buckby, in Northamptonshire. In this case the disease was exter- 
 minated by the slaughter and burial of the whole flock, and imme- 
 diate application of disinfectants to the hurdles and other things with 
 which the sheep had been in contact. Then it was introduced 
 a.irain in Cheshire, and strict isolation being enforced the infection 
 died out. Since 1866 we have had no outbreak of sheep-pox in 
 this kingdom, but foreign sheep have been landed with sheep-pox 
 in 1868, 1869, 1870, 1871, 1875, 1876, 1878, and 1880, but the 
 disea.se lias been prevented from spreading. 
 
 The Sheep-pox Order of 1895 provides for the notification of 
 the disease, for disinfection and for compulsory slaughter of infected 
 sheep, and prohibits the movement of diseased or suspected sheep, 
 and the local authority may, if they think fit, order the slaughter 
 of suspected sheep and of sheep which have been in contact with 
 diseased sheep. 
 
 FOOT-AND-MOUTH DISEASE. 
 
 Foot-and-mouth disease is a highly contagious arid infectious 
 febrile disease, characterised by a vesicular eruption affecting the 
 lips, tongue, roof of the mouth, and feet of sheep, cattle, and pigs. 
 and according to some observers it also attacks horses, poultry, 
 hares, and rabbits. Sometimes the mouth only is affected, in other 
 cases the principal seat of the eruption is in the feet. The vesit 1. - 
 soon break and give rise to ulcers. When these occur in the mouth 
 they cause pain and difficulty in taking food. Extensive ulceration 
 may occur on the feet, causing great pain and lameness. In milch 
 cows it sometimas happens that the eruption occurs on the udder and 
 . and it is this manifestation of the disease which has received 
 so much attention from Rayer. The milk is contaminated by the 
 discharge of the vesicles, and is unfit for use, either as food for the 
 human t>eing or for the lower animals. It induces a vesicular 
 eruption in the mouth, larynx, pharynx, and intestinal canal. It 
 ftcta most vigorously when administered warm to young animal>, 
 and calves occasionally die quite suddenly after sucking cows 
 
300 INFECTIVE DISEASES. 
 
 affected with the eruption on the teats. Fatal effects also result 
 when the milk is administered to young pigs. 
 
 It has been stated that no injurious consequences arise from the 
 consumption of the milk by human beings, but there is abundant 
 evidence to the contrary, and the conflicting opinions probably arise 
 from the fact that milk is seldom drunk direct from the cow, and 
 rarely in an undiluted form. Hertwig experimented upon himself 
 with milk freshly drawn from a cow with the eruption. He drank 
 a pint, and two days afterwards experienced slight fever, restless- 
 ness, and headache. The mouth was dry and hot, and there was 
 tingling in the skin of the hands and fingers. These symptoms 
 continued for seven days after taking the milk. On the ninth day 
 vesicles had formed on the tongue, principally on the edges, and on 
 the mucous membrane of the cheeks and lips (the largest being 
 about the size of a lentil). They were yellowish-white in colour, and 
 contained a whitish turbid liquid, which flowed when the vesicles 
 were pricked with a needle. At the same time a number of vesicles 
 developed on the hands and fingers ; and most of them at the 
 time of their first appearance were the size of a millet seed. They 
 were firm to the touch, yellowish -white, and occasioned a slight 
 tingling. The vesicles of the mouth increased in size and eventually 
 broke, and the epithelium detached itself completely from the affected 
 parts, leaving dark red spots, which disappeared gradually. The 
 slight fever present during the first days ceased after the appearance 
 of the eruption ; but from this time, until the disappearance of 
 the red spots, Hertwig felt a continual burning pain in the mouth, 
 and speaking and deglutition caused considerable uneasiness. On 
 the lips the vesicles dried up, and were covered with thin brownish 
 crusts, which fell off ten days after the appearance of the first 
 vesicles. The vesicles which developed on the hands ran a slower 
 course. From the tenth to the thirteenth day they filled with a 
 liquid, like turbid lymph. They were large and confluent, and 
 finally broke and dried up. 
 
 Bacteria in Foot-and-mouth Disease. Klein in 1885 isolated 
 from the vesicles a streptococcus which in its microscopical and its 
 cultural characters on gelatine, agar and blood serum resembled 
 Streptococcus pyogenes. Minute differences in the size of the 
 colonies and in their rate of growth, and in the character of the 
 chains, were observed 011 making comparative cultures with 
 Streptococcus pyogenes from a human source, but no comparison 
 was made with Streptococcus pyogenes from acute suppuration in 
 cattle. Baumgarten regarded this micro-organism as Streptococcus 
 
FOOT-AND-MOUTH DISEASE. 301 
 
 pyogMie>. and not as the eontagium of the disease. The author has 
 pointed out the variation which exists in the size of the chains and 
 of the colonies, and the difference which is found in tin- rate of 
 growth of cultures of Streptococcus pyogenes. mid these variations are 
 especially marked in Streptococcus pyogenes ho\i>. Klein believe-* 
 that tlie administrations of broth cultures produced the disease in 
 sheep, hut the results were very probably due to accidental infection. 
 It is well known how very readily foot-and-mouth disease is spread. 
 The appearance of a ca>e in a Hock of sheep or a herd of cattle will 
 !> almost certain to be followed by all or nearly all of the other 
 animals being infected with great rapidity. The virus chugs to the 
 clothe-: of shepherds and others who have been in contact with 
 infected sheep, and may be readily conveyed to healthy animals bv 
 tli<e who have l>eeii visiting infected premi>e>. 
 
 Schottelius described chain- composed of rounded element.-, xmie 
 of which resembled an amoeba or plasmodium. The chains were said 
 to be motile, and delicate growths were obtained in blood serum and 
 a gai-. and in broth and on potato. Inoculation in sheep and pigs 
 and numerous small animals gave negative results. These organisms 
 were described as >treptocytes. tn distinguish them from bacteria. 
 
 Piani and Fiorentini investigated the contents of the veside>. and 
 also described corpuscular elements exhibiting auia-boid movements. 
 They regarded these bodies as protoxoa, and concluded that foot-and- 
 mouth disease is due to their presence. 
 
 I'ntil a micro-organism is cultivated which will produce .sheep- 
 pox in sheep on a farm or on premises where the disease does not 
 exist, and where there can be no possibility of accidental infectio 
 we are fully justified in concluding that the nature of the contagiti 
 of this disease is unknown. 
 
 Stamping-OUt System. Foot-and-mouth disease was impcr 
 into this country in 1839. It has been successfully dealt with by the 
 stamping-out system, which in this case is very difficult to apply 
 because of the very short period of incubation, and the value of 
 the stamping-out method very greatly depends upon the length 
 of the incubation periml. Foot-and-mouth disease very often, 
 from infection to recovery, does not exceed ten days; yet according 
 to the reports of the Board of Agriculture, when foot-and-mouth 
 di-ase exi>ts in a manageable state, perfect isolation and effectual 
 disinfection have proved equal to the complete control of the 
 spreading of the infection, and the final extinction of the dis. 
 Nothing more is nec.-ss.-n-y in any case than to clo>e up all the 
 channels through which infected matter can be conveyed; but in 
 
302 INFECTIVE DISEASES. 
 
 order that this may be done close supervision by conscientious and 
 responsible officers is required; without it the case is hopeless. 
 
 The Foot-and-mouth Disease Order of 1895 enforces notification, 
 isolation, and disinfection, and the question of slaughter is left to 
 the local authority. 
 
 (1) A local authority may, if they think fit, cause to be slaughtered 
 () Any cattle, sheep, or swine affected with foot-and-mouth disease 
 
 or suspected of being so affected ; and 
 
 (6) Any cattle, sheep, or swine being or having been in the same field, 
 shed, or other place, or in the same herd or flock or otherwise in 
 contact with animals affected with foot-and-mouth disease, or being 
 or having been, in the opinion of the local authority, in any way 
 exposed to the infection of foot-and-mouth disease. 
 
CHAPTER XXII. 
 
 HORSE-POX. COW-POX. 
 
 CONSTITUTIONAL GREASE OR HORSE-POX. 
 
 HORSE- POX is a vesicular disease of the horse communicable from 
 animal to animal by inoculation, but never infectious. It is 
 communicable by inoculation to man, and the attenuated virus 
 pro luces phenomena indistinguishable from the results of vaccina- 
 tion with cow-pox lymph. 
 
 The existence of this disease of the horse had long been known 
 to farmers and farriers, but Jenner was the first to draw attention 
 to it in writing. " There is a disease to which the horse from his 
 state of domestication is frequently subject. The farriers have 
 termed it the grease ; it is an inflammation and swelling in the heel 
 accompanied at its commencement with small cracks and fissures, 
 from which issues matter possessing properties of a very peculiar 
 kind." Jenner gave several instances in which this disease \\a> 
 communicated to man and to cows. 
 
 Thus, a man named Merret attended to some horses with sore 
 heels and also milked the cows. The cows were infected, and the 
 man had several sores upon his hands. 
 
 William Smith, on another farm, attended to horses with sore 
 heels and milked the cows also. The cows were infected, and on one 
 of Smith's hands there were several ulcerated sores. 
 
 Simon Nicholls applied dressings to the sore heels of one of his 
 uiastt-r'- liur>'s and at the same time milked the cows, and the o>u- 
 were infected in consequence. 
 
 A mare, the property of a dairy farmer, had sore heels, and 
 was attended to by the men of the farm, Thomas Virtue, William 
 Wiu-nvt. and William Haynes. They contracted "aores on their 
 hands, followed by inflamed lymphatic glands in the arms and 
 axilla-, -liivcrin^s Micr.-cdrd by heat, lassitude, and gi-iH-ral pains in 
 the limbs," and the disease wa> al>< communicated to the co\\>. 
 
 908 
 
304 INFECTIVE DISEASES. 
 
 But Jeimer's experience of this disease was not limited to cases 
 in which the eruption occurred in the heel. 
 
 He mentions a case in which 
 
 " An extensive inflammation of the erysipelatous kind appeared 
 without any cause upon the upper part of the thigh of a sucking 
 colt. The inflammation continued several weeks, and at length 
 terminated in the formation of three or four small abscesses." Those 
 who dressed the colt also milked the cows on the farm, and communi- 
 cated the disease to them. 
 
 Subsequently, Jenner gave a more comprehensive description of 
 this disease. 
 
 11 The skin of the horse is subject to an eruptive disease of a 
 vesicular character, which vesicle contains a limpid fluid, showing 
 itself more commonly in the heels. The legs first become cedematous r 
 and then fissures are observed. The skin contiguous to these fissures r 
 when accurately examined, is seen studded with small vesicles sur- 
 rounded by an areola. These vesicles contain the specific fluid. It 
 is the ill-management of the horse in the stable that occasions the 
 malady to appear more frequently in the heel than in other parts. 
 I have detected it connected with a sore on the neck of the horse, 
 and on the thigh of a colt." 
 
 Mr. Moore, of Chalford Hill, described a case in 1797, and re- 
 garded the disease as virulent grease. His horse was attacked with 
 what was supposed to be ordinary "grease." A cow was subse- 
 quently infected, and the disease communicated to the servant, who 
 had " eruptions on his hands, face, and many other parts of the 
 body, the pustules appearing large, and not much unlike the small- 
 pox, for which he had been inoculated a year and a half before, and 
 had then a very heavy burden." 
 
 In 1798, Mr. Fewster, of Thornbury, met with a case of this 
 equine malady, and wrote a very full account to Jenner of its 
 transmission to the human subject. 
 
 " William Morris, aged thirty-two, servant to Mr. Cox of 
 Almonsbury in this county, applied to me the 2nd of April, 1798. 
 He told me that four days before he found a stiffness and swelling 
 in both his hands, which were so painful it was with difficulty he 
 continued his work ; that he had been seized with pain in his head, 
 small of the back, and limbs, and with frequent chilly fits succeeded 
 by fever. On examination I found him still affected with these 
 symptoms, and there was great prostration of strength. Many 
 parts of his hands on the inside were chapped, and on the middle 
 joint of the thumb of the right hand there was a small phagedsenic 
 
CONSTITUTIONAL GREASE OR HORSE-POX. 305 
 
 ulcer, about the size of a large pea, discharging an ichorous fluid. 
 On the middle finger of the same hand there was another ulcer of a 
 similar kind. These sores were of a circular form, and he described 
 their first appearance as being somewhat like blisters arising from a 
 burn. He complained of excessive pain, which extended up his arm 
 into the axilla. On the 5th of April I again saw him, and found 
 him still complaining of pain in both his hands, nor were his febrile 
 symptoms at all relieved. The ulcers had now spread to the size of 
 a seven-shilling gold coin, and another ulcer, which I had not noticed 
 before, appeared on the first joint of the forefinger of the left hand, 
 equally painful with that on the right. I ordered him to bathe his 
 hands in warm bran and water, apply escharotics to the ulcers, and 
 wrapped his hands up in a soft cataplasm. The next day he was- 
 much relieved, and in something more than a fortnight got welL 
 He lost his nails from the thumb and fingers that were ulcerated." 
 
 Mr. Tanner, a veterinary surgeon, was the first to succeed in 
 experimentally transmitting horse-pox to the teats of a cow by 
 inoculating some of the liquid matter from the heel of a horse. From 
 handling the cow's teats he became infected himself, and had two 
 pustules on his hand, which brought on inflammation, and made 
 him unwell for several days. The matter from the cow and from his 
 own hand proved efficacious in infecting both human subjects and 
 cattle. 
 
 In 1801 Dr. Loy published his experiments. A butcher had 
 painful sores from dressing a horse suffering from * grease,' and Dr. 
 Loy succeeded in transmitting the disease to the udder of a cow. 
 Matter was taken from the cow and inserted into the arm of a child. 
 Dr. Loy also inoculated a child direct from a horse suffering from 
 * grease,' and subsequently five other children from this child. 
 
 From his experiments and observations Dr. Loy was led to 
 differentiate constitutional grease from the merely local affection 
 commonly known as the grease, and thus he explained the failure 
 on the part of many experimenters to transmit this disease to 
 the cmv. 
 
 " This fact induces me "he says " to suspect that two kinds 
 of grease <-xi>t. differing from each other in the power of giving 
 di>t-iM' to tin- human or brute animal ; and there is another circum- 
 stance which renders this supp<itk>n prohal>l. The horses that 
 communicated the infection to their dressers were affected with a 
 ral a> \\vll as a topical dis-a>e. The animals at the commence- 
 ment of their disease were evidently in ;i tc\cri>h state, from which 
 they were relieved as soon as the complaint appeared at their heels, 
 
 20 
 
306 INFECTIVE DISEASES. 
 
 and an eruption upon the skin. The horse, too, from which the 
 infectious matter was procured for inoculation, had a considerable 
 indisposition, previous to the disease at his heels, which was attended, 
 as in the others, with an eruption over the greatest part of his 
 body; but those that did not communicate the disease at all, had 
 a local affection only. From this perhaps may be explained the 
 want of success attending the experiments of the gentlemen I have 
 mentioned." 
 
 Experiments with horse-pox were also made about this time on 
 the Continent. Sacco made some observations upon this disease at 
 Milan. Several horses were suffering from what was called giardoni, 
 and Sacco's servant was attacked on both arms, from dressing one 
 of his horses troubled with this disease. Several children and cows 
 were inoculated from the horses, but without success. In another 
 instance, a coachman went to the hospital with the eruption on his 
 hands, and the disease was successfully communicated to three out 
 of nine children. 
 
 In 1803 Dr. Marcet described some experiments which had been 
 made at Salonica by M. La Font. The disease was known to the 
 farriers in Macedonia as javart. In one case, a horse was attacked 
 with feverish symptoms that ceased as soon as the eruption appeared. 
 The fore legs were much swelled and several ulcers formed. M. La 
 Font took some of the discharge from an ulcer and inoculated a cow 
 and three children, and succeeded in transmitting the disease to two 
 of the latter. 
 
 Vaccinogenic grease was observed in Paris in 1812, and Baron 
 cites the case of a coachman who, after dressing a horse with the 
 " grease," had a crop of pustules on his hands, from which the disease 
 was experimentally transmitted by inoculation to two children. A 
 series of inoculations was started from an infant who was infected 
 from one of the scabs taken from the pustules on the hand of the 
 coachman. 
 
 In 1813 Mr. Melon, a surgeon at Lichfield, met with vaccinogenic 
 grease in the horse, and some of the virus was sent to Jenner, who 
 carried on a series of arm to arm equiriations for some months. And 
 again in 1817, vaccinogenic grease broke out in a farm at Wansell. 
 The farm-servants and the cows were infected, and Jenner employed 
 this equine matter for a series of inoculations for eight months. 
 
 In 1817 Baron described a case of a young man who had not less 
 than fifty pustules on his hands and wrists from dressing a horse with 
 this disease, and in the following year Baron obtained some fresh 
 equine virus from the hands of a boy who had been infected directly 
 
CONSTITUTIONAL GREASE OR HORSE-POX. 307 
 
 from a horse. The disease assumed a pustular form, and extended 
 over both arms. 
 
 In 1818 Kahlert met with this equine disease in Bohemia, and 
 confirmed the experiments made by Loy and Sacco. Kahlert noticed 
 that the joint of the foot was swollen, and moisture exuded from it, 
 and tliat the posterior part of the pastern was slightly red, swollen 
 and hotter than the neighbouring parts, and a clear yellowish fluid 
 with a peculiar odour escaped. At the slightest touch the animal 
 showed signs of pain ; the hair was stuck together. The disease was 
 successfully transmitted to cows and from cows to children. 
 
 In 1860 the horses at Rieumes, near Toulouse, were attacked by 
 an epizootic malady ; in less than three weeks there were more than 
 one hundred cases. According to the veterinary surgeon, M. Sarnins, 
 the animals suffered from slight fever, rapidly followed by local 
 symptoms, the most marked of which were swelling of the hocks, and 
 an eruption of small pustules on the surface of the swollen parts, 
 which were, at the same time, hot and painful. After three to five 
 days there was a discharge from the pastern which continued for eight 
 to ten days, during which the inflammation gradually diminished. 
 The pustules dried up, and in about a fortnight the crusts with patches 
 of hair fell off, leaving more or less marked scars. The eruption 
 appeared at the same time on different parts of the body, especially 
 on the nostrils, lips, buttocks, and vulva. Sarrans believed that the 
 mares taken to the breeding establishment at Rieumes had been 
 infected from the ropes which had been used in tying up other affected 
 animals, and had become thereby infected with the virus of this 
 disease. One of the mares was taken by the owner, M. Corail, to 
 the veterinary school to be examined by M. Lafosse. About eight 
 days after this visit significant symptoms appeared : loss of appetite, 
 lameness, stiffness of both pastern joints, and a hot, painful swelling 
 of the left pastern joint. The hair was staring, and there were 
 vesicles on the skin, from which a liquid exuded having an ammoniacal 
 odour but less foetid than the secretion in eaux auxjambes. 
 
 M. Lafosse successfully transmitted the disease to cows, and from 
 cows to children and to a horse. 
 
 In 1863 the subject of vaccinogenic grease or horse-pox a.irain 
 received great attention in France. A student named Amyot wa> 
 engaged in dressing a horse on which an operation had been per- 
 formed. The leg which had been operated on became the seat of a 
 \f-ry confluent eruption of horse-pox, which was followed by such 
 an abundant flow of serosity that at first the nature of the affection 
 wa> mistaken, and it was thought to be a complication of eaux aux 
 
308 INFECTIVE DISEASES. 
 
 jambes. Amyot had a wound on the dorsal aspect of the first inter- 
 phalangeal joint of the little finger of his right hand ; in spite of 
 this, he continued to dress the horse entrusted to his care. The 
 wound on his finger became accidentally inoculated with the virus, 
 which flowed in great abundance from the horse's leg. 
 
 The wound was made on August 3rd, and the next day it was 
 swollen, and rather painful. On the 5th, Amyot suffered from malaise 
 and great weakness; on the 6th, 7th, and 8th, vesicles appeared 
 successively on the fingers of his left hand, and on his forehead 
 between the two eyebrows. On the 9th, these vesicles were fully 
 developed ; those of the fingers consisted of very large epidermic 
 bullse on a bluish-red base. On opening them, a perfectly limpid 
 fluid escaped in such abundance that small test-tubes might have 
 been filled with it. The vesicle on the forehead was surrounded by 
 a bluish-red areola, within which, the epidermis, of a leaden-grey 
 hue, was raised, and had a slight central depression. The liquid 
 which flowed from it when it was opened, and which continued to 
 ooze, was also very abundant and of a deep citrine colour. 
 
 The vesicles which had developed on the dorsal side of Amyot's 
 fingers were extremely painful. The incessant shooting pains, of 
 which they were the seat, prevented him from getting any rest for 
 three days. On the 10th, inflammation of the lymphatics followed ; 
 both arms were swollen and very painful, with red lines indicating 
 the course of the lymphatic vessels. The glands of the axillae were 
 also enlarged. 
 
 The lymphatic glands behind the jaws were also swollen and pain- 
 ful. Amyot's chief sufferings were occasioned by the intense local 
 pain caused by the vesicles on the fingers, and by the inflammation 
 of the lymphatic vessels and glands, which continued in this state up 
 to the 18th of August. It was only at the end of the month that the 
 vesicles were completely cicatrised. 
 
 Bouley felt very great anxiety in the presence of the grave 
 symptoms which accompanied the eruption. The eruption on the 
 forehead was especially a cause of great uneasiness, because glanders 
 manifests itself in a similar way. 
 
 With virus from Amyot's vesicles the disease was transmitted to 
 cows and to children. 
 
 Further, this outbreak enabled exhaustive experiments to be 
 made, by which it was definitely established that horse-pox is never 
 infectious, but, like cow-pox, is transmitted solely by contact. 
 
 In 1880, M. Baillet, Director of the National Veterinary School 
 of Toulouse, was informed that a contagious malady had developed 
 
CONSTITUTIONAL GREASE OR HORSE-POX. 309 
 
 in the mares, which had been served by the stallions at the breeding 
 establishment at Rieumes, belonging to M. Mazeres. M. Peuch 
 was delegated to investigate this outbreak, and he visited for that 
 purpose Berat, Rieumes, and Labastide-Clermont. 
 
 At Berat three mares were examined. In one, there were soar* 
 and crusts, the remains of an eruption on the lips and in the vicinity 
 of the vulva ; in another, there were several reddish circular ulcers 
 in the same region ; and in a third, there were dried pustules with 
 blackish adherent crusts at the circumference of the vulva and 
 extending over the perinseum. On the lower part of the left flank 
 a voicle was discovered surmounted by a crust, and when the latter 
 was detached a sero-sanguinolent liquid oozed from the exposed 
 surface. M. Peuch recognised the true nature of this disease, 
 having several times previously had the opportunity of examining 
 mares with a vesicular eruption round the vulva after coition, 
 which eruption he had studied from its first appearance to complete 
 cicatrisation, and had ascertained to be horse-pox. 
 
 On proceeding to Rieumes, M. Peuch inspected eleven stallions, 
 .six horses, and five asses. In one ass there were several vesicles 
 on the right side of the penis scattered about from the base to the 
 glans. In another ass there was a trace of a vesicle on the penis 
 and a characteristic vesicle on the left nostril. 
 
 In an old bay mare there were the remains of an eruption 
 on the circumference of the vulva, and in an old white mare there 
 were not only vesicles on the vulva, but in addition vesicles on the 
 inner side of the lower lip. M. Peuch drew special attention to 
 these cast's as likely to be confounded with aphthous stomatitis, but 
 the existence of the same eruption on other parts of the body is an 
 important aid in making a diagnosis of horse-pox. 
 
 At Labastide-Clermont one mare was particularly noticed. 
 This mare had been served on the 19th and 21st of April, and on the 
 occasion of the inspection, May 1 1th, there were the remains of an 
 eruption around the vulva,' and lymphangitis existed in the right 
 posterior limb, which was engorged, hot, and painful in its whole 
 extent, so that the animal walked with difficulty. The proprietor 
 had contracted the disease in attending to his mare, and exhibited 
 a vesicle on the thumb of the right hand, excoriated and blackened, 
 but still recognisable. 
 
 Some of the crusts collected from the cases at Berat were used 
 for inoculating a cow. The result was successful, and the disease 
 was transmitted by inoculation to a heifer and several students and 
 children. 
 
310 INFECTIVE DISEASES. 
 
 M. Peuch ascertained that horse-pox caused considerable alarm 
 from the fact that the breeders regard this eruptive affection as 
 syphilitic, and this alarm consequently brings discredit upon the 
 breeding establishment whence the illness has spread. He was also 
 led to appreciate the great necessity for further study of this disease 
 in relation to dourine or maladie du coit. 
 
 In 1882 M. Peuch had the opportunity of investigating a case 
 of horse-pox in Algeria. The disease occurred in a thoroughbred 
 Arab. There was an eruption of vesicles, and there was also an 
 ulcer the size of a five- franc piece in the nostril. In the mouth and 
 on the lips there were a number of small vesicles about the size of 
 a pea. The sublingual glands were engorged, hot, and painful on 
 pressure. The coat, in patches, on the lateral aspect of the neck, 
 on the shoulders, the flanks, and in the hollow of the heel, was 
 staring, giving the appearance of small paint-brushes. On passing 
 the hand over these, vesicles could be detected partly dry and partly 
 secreting. 
 
 The disease was transmitted to cows, and from cows to about 
 one thousand five hundred persons. 
 
 Cases similar to the one just described, in which there is more or 
 less marked ulceration of the nostril or nasal septum, must be care- 
 fully distinguished from glanders. And again, when the sublingual 
 glands are affected the disease may be mistaken for strangles. 
 
 NATURE AND AFFINITIES. 
 
 Horse-pox and human small-pox are quite distinct diseases, 
 and the theory that horse-pox is derived from grooms or other 
 attendants suffering from small-pox may be dismissed without 
 further comment. 
 
 Horee-pox is never infectious, but is communicated solely by 
 contact either by grooms inoculating the virus with their hands, 
 sponges, or brushes, or by horses coming into contact with each other, 
 and in breeding establishments by coition. Auzias Turenne, who 
 wrote exhaustively on this subject, maintained that horse-pox came 
 into the same category of diseases as syphilis in. man. 
 
 "A un point de vue, le grease pustuleux inocule off re la plus 
 parfaite ressemblance avec la verole inoculee, par le produit des 
 accidents secondaires. Des deux cotes nous voyons, absence de 
 contagion par la voie de Fatmosphere, travail local, retentissement 
 lymphatique et ganglionaire, fermentation universelle de Forganisme, 
 eruption generale et immunite acquise contre de nouvelles atteintes. 
 
 " A un autre point de vue, la ressemblance avec la variole est 
 
NATURE AND AFFINITIES. 311 
 
 frappante. Mais il s'en distingue enorniement par Tabsence de la 
 contagiosite atmospherique." 
 
 Human small-pox belongs to a different group of diseases, and 
 has affinities rather with small-pox of sheep and cattle plague, 
 diseases which are not only inoculable, but highly infectious. Human 
 small-pox is an infectious disease characterised by sudden and severe 
 fever, followed after forty-eight hours by a generalised eruption ; horse- 
 pox commences as a local affection, and constitutional symptoms 
 follow. Auzias Turenne, guided by analogy, described the general- 
 ix-il eruptions following "grease" or horse-pox as greasides ("comme 
 on dit syphilides"). 
 
 Horse-pox and human syphilis are absolutely distinct diseases; 
 and there is no more ground for believing that horse-pox originates 
 in human syphilis than there is for accepting the theory that it arises 
 from grooms suffering from small-pox. Syphilis artificially inocu- 
 lated on the human subject only resembles the casual or intentional 
 inoculation of virulent horse-pox. The stages of papulation, vesicu- 
 lation, ulceration, scabbing, and the formation of a permanent scar, 
 occur in inoculated syphilis, and if we examine Ricord's illustrations 
 and study the experiments of Auzias Turenne, \ve cannot fail to 
 be struck with the remarkable similarity to the results obtained and 
 depicted by Jenner. 
 
 But in order to follow the argument of Auzias Turenne we 
 must study the natural and casual horse-pox. And if we are not 
 familial with what has been written on this subject, and if we 
 restrict our knowledge to the artificially cultivated horse-pox, we 
 shall fail to recognise the disease when we meet with it, and we shall 
 be liable to attribute the results of the full effect of the virus to 
 accidental contamination. 
 
 Another question of very great interest is the relation of horse- 
 pox to cow-pox. Jenner first of all propounded the theory that all 
 cu\v-pox arose from horse- pox, or as he termed it " the grease," and 
 thus cow-pox and horse-pox were manifestations of the same disease. 
 But it \va> established that cow-pox also arose quite independently 
 of horse-pox, and Jenner was led to distinguish between cow-pox, 
 a disease peculiar to the cow, and the eruptive affection transmitted 
 to the cow from the horse, which farmers and others, by a strange 
 ])"i-ver>ion of terms, called the cow-pox. Whether the eruption of 
 cow-pox can be distinguished from the eruption of horse-pox com- 
 municated to the cow, and whether cow-pox and horse-pox are 
 identical, or only analogous, are questions which call for further 
 investigation. 
 
312 INFECTIVE DISEASES. 
 
 Bacteria in Horse-pox. Outbreaks of horse-pox have not 
 been investigated from the bacteriological point of view, and the 
 nature of the contagium is unknown. 
 
 COW-POX. 
 
 Cow-pox is a vesicular disease of the teats of cows. It is never 
 infectious, and only attacks cows in milk, the virus being transferred 
 from cow to cow by the hand of the milker. The disease is com- 
 municable to milkers, and the virus artificially inoculated produces 
 what is commonly known as vaccinia. In its clinical history and 
 epidemiology cow-pox is totally distinct from human small-pox, and 
 the hypothetical and entirely erroneous suggestion that the disease 
 arises from milkers suffering from human small-pox is responsible 
 for the belief which prevailed until recently, that cow-pox was an 
 extinct disease in this country ; but, by the author's researches, this 
 has been shown to be a mistake. 
 
 Cow-pox is not a rare disease, and it has never been found to 
 arise from a milker suffering from small-pox. As this is a matter 
 of great importance in discussing the etiology of the disease, the 
 history of outbreaks and the clinical characters of cow-pox will be 
 given in considerable detail. 
 
 According to Jenner, cow-pox had been known among farmers 
 from time immemorial. He refers to cases occurring in 1770, 1780, 
 1782, 1791, 1794, 1796, and 1798. In 1799 cow-pox was raging in 
 the dairies in London, and outbreaks were investigated by Woodville, 
 Pearson, and Bradley. In the same year cow-pox broke out at 
 Norton Nibley, in Gloucestershire. Pearson and Aikiri referred to 
 the prevalence of cow-pox in Wilts, Somerset, Devon, Bucks, Dorset, 
 Norfolk, Suffolk, Leicestershire, and Staffordshire ; and Barry men- 
 tioned its prevalence in Ireland. 
 
 From this time onwards, for a long period, natural cow-pox 
 received little or no attention in this country. Fresh stocks of lymph 
 were raised for the purposes of vaccination, but no further attention 
 was given to studying the disease in the cow. In 1836 Leese 
 described an outbreak of cow-pox, and in 1838 Bstlin discovered an 
 outbreak in Gloucestershire. In 1838-39 cow-pox was met with 
 by Mr. Fox, of Cerne Abbas, and again in 1839, in Dorsetshire, by 
 Mr. Sweeting. Ceely frequently met with cow-pox in the Yale of 
 Aylesbury, and particularly refers to outbreaks in 1838, 1840, 1841, 
 and 1845. But after this, outbreaks of this disease in the cow were 
 not recorded, though several medical practitioners met with the 
 disease and raised fresh stocks of vaccine lymph. Thus, when 
 
COW-POX. 313 
 
 inquiries were made in 1857, it was found that Mr. Donald 
 Dalrymple, of Norwich (on two occasions), Mr. Beresford, of Nar- 
 borough, in Leicestershire, Mr. Gorham, of Aldeburgh, Mr. Alison, 
 of Great Retford, Mr. Coles, of Leckhampton, Mr. Rudge, of 
 Leominster, and one or two others, had met with outbreaks of 
 cow-pox. 
 
 In 1885 cow-pox was discovered by the author in Wiltshire. The 
 publication of the fact led to the recognition of the disease in the 
 same year in many parts of England, and cases were met with in 
 man in 1888 by Mr. Forty in Gloucestershire, and by Mr. Bucknill 
 near London in 1894. 
 
 In Italy, cow-pox was found by Sacco in the plains of Lombardy 
 in 1800, and by other practitioners in 1808-9. In 1812 it was 
 observed at Naples by Miglietta; in 1830 in Piedmont ; and in 1832 
 and 1843 at Rome, by Dr. Maceroni. More recently, several out- 
 breaks of cow-pox have been met with in this country, and the stocks 
 of vaccine lymph renewed. 
 
 In France, in 1810, cow-pox was found in the department of 
 La Meurthe, and in 1822 at Clairvaux ; at Passy, Amiens, ;m<l 
 Rambouillet in 1836 ; at Rouen in 1839 ; at St. Illide, at St. Seine, 
 and at Perylhac, in 1841 ; in 1842 at Pagnac ; in 1843 at Deux 
 Jumeaux, where, during the previous thirty years, several fresh 
 stocks of lymph had been raised and circulated. The disease occurred 
 in a cow belonging to M. Majendie in 1844, and it was found at 
 Wasseloune, in the department of Bas Rhin, in 1845 ; it occurred in 
 three other departments in 1846 ; at Rheims, and in the department 
 of Eure et Loire, in 1852 ; in the arrondissement of Sancerre, and at 
 Beziers in 1854; and at Guyonville in 1863. It broke out on farms 
 in three villages near Nogent in 1864 (the disease was introduced by 
 newly purchased cows ; milkers were infected, and from one of these 
 milkers a lymph stock was established) ; it also occurred in 1864, at 
 Petit Quevilly, near Rouen; and in April 1866 at Beaugency ; in 
 1881 at Eysines, near Bordeaux, and again at the same place in 
 1883; and in 1844 at Cerons. 
 
 In Germany, as soon as attention had been drawn to the dis :!>. 
 cow-pox was frequently discovered. There were as many as thirty- 
 eight outbreaks reported in one year in Wurtemberg. 
 
 It is hardly necessary, after reciting these instances, to insist 
 that cow-pox is far from being a rare disease, as many have sup- 
 posed who are unacquainted with the literature of the subject and 
 unfamiliar with the appearances of the natural disease in the 
 cow. 
 
314 INFECTIVE DISEASES. 
 
 NATURAL AND CASUAL COW-POX. 
 
 To appreciate the characters of the natural disease in the cow r 
 we must dismiss from our minds the artificial disease vaccinia, for 
 the ordinary results of vaccination stand in much the same relation 
 to the natural disease cow-pox as the benign vesicle of variolatiori to 
 natural small-pox. 
 
 The description of cow-pox given by Jenner, in 1798, was the 
 first published account. The disease in the cow was described as 
 consisting of irregular pustules on the teats, of a palish blue colour, 
 surrounded by an erysipelatous inflammation, and characterised by 
 a tendency to degenerate into phagedsenic ulcers. The animals were 
 indisposed and the secretion of milk lessened. 
 
 In referring to an outbreak which occurred epizootically in 
 London in February 1799, Dr. Bradley gave a coloured plate of the 
 disease on the arm and fingers of a milker. The cow-pox, he said, 
 in this instance, " appears to have been very mild, for no loss was 
 experienced by the farmers from the deficiency of milk, as usually 
 happens." 
 
 These early descriptions were supplemented by an account of 
 cow-pox by Mr. Lawrence, author of A Philosophical and Practical 
 Treatise on Hwses, and on the Moral Duties of Man toward the Brute 
 Creation. Lawrence's article on cow-pox not only affords evidence 
 that this disease was known to those who had the care of cattle 
 before Jenner's paper was published, but it shows that it had also 
 been made the subject of practical observation and study by veteri- 
 narians. Lawrence concluded by saying : " Whatever may be the 
 fate of cow-pox inoculation, it has and will give further occasion to 
 a pretty large arid open discussion, which is always beneficial as 
 having a tendency to produce discovery and promote improvement ; 
 and when the public ardour for the present topic shall have become 
 a little cool and satisfied, I hope it will be turned by enlightened 
 men towards another, perhaps of nearly as great consequence 
 namely, the prevention of the original malady in the animals them- 
 selves. Those who have witnessed it and only reflected upon the 
 excessive filth and nastiness which must unavoidably mix with the 
 milk in an infected dairy of cows, and the corrupt and unsalubrious 
 state of their produce in consequence, will surely join me in. that 
 sentiment." 
 
 Lawrence was almost a century before his time. Cow-pox was not 
 again brought forward in this light until 1887-88, when the author 
 reported the contamination of the milk at the Wiltshire farms, and 
 
NATURAL AND CASUAL COW-POX. 315 
 
 advocated the advisability of placing this disease under the Con- 
 tagious Diseases (Animals) Act. 
 
 The numerous pathological details wanting in the early accounts 
 of cow-pox were supplied by the painstaking and laborious re- 
 > -n-ches of Robert Ceely. From his classical papers in the Trans- 
 actions of the Provincial Medical Association, we can obtain a 
 complete picture of the natural disease in the cow. 
 
 In Ceely's experience in the Yale of Aylesbury, outbreaks 
 occurred at irregular intervals, most commonly appearing about 
 the Ijegi lining or end of the spring, rarely during the height of 
 summer. There were outbreaks at all periods from August to May 
 and the beginning of June, cases being met with in autumn and 
 the middle of winter, after a dry summer. The disease was occa- 
 sionally epizootic, or occurring at times at several farms at no great 
 distance from each other, but was more commonly sporadic or 
 nearly solitary. It was to be seen sometimes at several contiguous 
 farms ; at other times at one or two farms. Many years might 
 elapse before it recurred at a given farm, although all the animals 
 might have been changed in the meantime. Cow-pox had broken 
 out twice in five years in a particular vicinity at two contiguous 
 farms, while at an adjoining dairy, in all respects similar in local 
 and other circumstances, it had not been known to exist for forty 
 years. It was sometimes introduced into a dairy by recently 
 purchased cows. Twice it had been known to be so introduced by 
 milch heifers. It was considered that the disease was peculiar to 
 the milch cow ; it came primarily while the animal was in milk, 
 and it was casually propagated to others by the hands of the milkers. 
 Sturks, dry heifers, dry cows, and milch cows milked by other hands, 
 grazing in the same pastures, feeding in the same sheds, and at 
 contiguous stalls, remained exempt from the disease. 
 
 For ^ many years, the "spontaneous" origin of cow-pox had 
 not been doubted in the Yale of Aylesbury. In all the cases that 
 Ceely had noticed he could never discover the probability of any 
 other origin. 
 
 Condition of Animal primarily affected. There was much diffi- 
 culty in determining at all times, with precision, whether this 
 disease arose primarily in one or more individuals in the same dairy. 
 Most commonly, however, it appeared to be solitary. The milkers 
 believed they were able to point out the infecting individual. In 
 two instances, there could be very little doubt on this point. In 
 August 1838, three cows were affected with the disease. The first 
 was attacked two months after calving and seven weeks after 
 
316 INFECTIVE DISEASES. 
 
 weaning. This animal was considered in good health, but it looked 
 out of condition. Heat and tenderness of the teats and udder were 
 the first noticed signs. The other two were affected in about ten 
 days. In December 1838, in a large dairy, a milch cow slipped her 
 calf, had heat and induration of the udder and teats, with cow-pox 
 eruption, and subsequently leucorrhcea and greatly impaired health ; 
 the whole dairy, consisting of forty cows, became subsequently 
 affected, and also some of the milkers. In another dairy, at the 
 .same time, it first appeared in a heifer soon after weaning, and in 
 about ten or twelve days extended to five other heifers and one cow, 
 milked in the same shed, affecting the milkers. And in another 
 dairy thirty cows were severely affected, and also one of the milkers. 
 It appeared to originate in a cow two months after calving. The 
 only symptoms noticed were that the udder and teats were tumid, 
 tender, and hot just before the disease appeared. 
 
 Condition of Animals casually affected. In some animals, it was 
 less severe than in others, depending on the state .and condition of 
 the skin of the parts affected, and the constitution and habit of the 
 animal. It was sometimes observed to diminish the secretion of 
 milk, and in most cases it commonly did actually affect the amount 
 artificially obtained ; with this exception, and the temporary trouble, 
 and accidents to the milk and the milkers, little else was observed ; 
 the animal continued to feed and graze apparently as well as before. 
 The topical effects varied very much in different individuals ; the 
 mildness or severity being greatly influenced by temperament and 
 condition of the animal, and especially by the state of the teats and 
 udder, and the texture and vascularity of the skin of the parts 
 affected. Where the udder was short, compact, and hairy, and the 
 skin of the teats thick, smooth, tense, and entire, or scarcely at all 
 chapped, cracked, or fissured, the animal often escaped with a mild 
 affection, sometimes with only a single vesicle. But where the 
 udder was voluminous, flabby, pendulous, and naked, the teats 
 long and loose, and the skin corrugated, thin, fissured, rough, and 
 unequal, then the animal scarcely ever escaped a copious eruption. 
 Hence, in general, heifers suffered least, and cows most, from the 
 milkers' manipulations. 
 
 Progress of the Disease. Cow-pox once arising or introduced, and 
 the necessary precautions not being adopted in time, appeared in ten 
 or twelve days on many more cows in succession, so that among 
 twenty-five cows perhaps by the third week nearly all would be 
 affected ; but five or six weeks or more were required before the 
 teats were perfectly free from the disease. 
 
NATURAL AND CASUAL COW-POX. 317 
 
 Propagation by the Hand of the Milker. Ceely was able to confirm 
 the way in which the disease was said to spread. In December 
 l v ."> s . ui a large dairy farm, where there were three milking-she U. 
 cou -]>ox broke out in the home or lower shed. The cows in this 
 >he 1 bring troublesome, the milker from the upper shed, after 
 milking his own cows, came to assist in this for several days, morning 
 and evening, when in about a week some of his own cows began to 
 exhibit the disease. It appears that, having chapped hands, he 
 neglect I washing them for three or four days at a time, and thus 
 conveyed the disease from one shed to another. During the progress 
 of the disease through this shed, one of the affected cows, which had 
 been attacked by the others, was removed to the middle shed, where 
 all the animals were perfectly well. This cow, being in an advanced 
 >tage of the disease, and of course difficult to milk and dangerous 
 to the milk-pail, was milked first in order by the juvenile milker for 
 three or four days only, when, becoming unmanageable by him, its 
 former milker was called in to attend exclusively to it. In less than 
 a week, all the animals of this shed showed symptoms of the disease, 
 tlMjugh in a much milder degree than it had appeared in the other 
 >he Is, fewer manipulations having been performed by an infected 
 hand. 
 
 Topical Symptoms of the Natural Disease. For these, Ceely was 
 almost always, in the early stage, compelled to depend on the obser- 
 vations and statements of the milkers. They stated that for three 
 or four days, without any apparent indisposition, they noticed heat 
 and tenderness of the teats and udder, followed by irregularity and 
 pimply hardness of these parts, especially about the bases of the 
 tf its and adjoining the vicinity of the udder; these pimples on skins 
 not very dark are of a red colour, and generally as large as a vetch 
 <>r a pea. ami quite hard, though in three or four days many of 
 tlitM- increase to the size of a horse-beau. Milking is generally 
 very painful to the animal; the tumours rapidly increase in >i/e. 
 vesicate, and are soon broken by the hands of the milker. Milking 
 HOW becomes a troublesome and occasionally a dangerous pr< 
 < V:-ly adds: ''It is very seldom that any person competent to 
 judge of the nature of the ailment has access to the animal before 
 the appearance of the disease on others of the herd, when the cow 
 tir>t atl'erted presents on the teats acuminated, ovoid, or globular 
 ve.xications. some entire, others broken, not infrequently two or three 
 interfluent; those broken have evidently a central d-jin->in with 
 marginal induration; tho>e entire, being punctured, ditl'use a more 
 or ! vi>cid ainl.er-coloured fluid, collapse, and at once indicate the 
 
318 INFECTIVE DISEASES. 
 
 same kind of central and marginal character. They appear of 
 various sizes, from that of a pin's head, evidently of a later date, 
 either acuminated or depressed, to that of an almond or a filbert, or 
 ever larger. Dark brown, or black, solid, uniform crusts, especially 
 on the udder near the base of the teats, are visible; at the same time, 
 some much larger are observed on the teats ; these, however, are 
 less regular in form and less perfect. Some are nearly detached, 
 others quite removed, exhibiting a raw surface with a slight central 
 slough. On the teats, the crusts are circular, oval, oblong, or 
 irregular ; some flat, others elevated, some thin and more trans- 
 lucent, being obviously secondary. The appearance of the disease 
 in different stages, or at least the formation of a few vesicles at 
 different periods, seems very evident. The swollen, raw, and en- 
 crusted teats seem to produce uneasiness to the animal only while 
 subjected to the tractions of the milkers, which it would appear are 
 often nearly as effectual as usual." Referring again to the character 
 of the vesicle, Ceely says, that " those fortunate enough to have an 
 opportunity of watching the disease in its progress may observe 
 that, when closely examined, they present the following characters : 
 In animals of dark skin, at this period, the finger detects the 
 intumescent indurations often better than the eye, but when closely 
 examined the tumours present at their margins and towards their 
 centres a glistening metallic lustre or leaden hue ; but this is not 
 always the case, for occasionally they exhibit a yellowish or yellowish- 
 white appearance." 
 
 In describing the crusts in detail, Ceely says that " large black 
 solid crusts, often more than an inch or two in length, are to be 
 seen in different parts of these organs, some firmly adherent to a 
 raw elevated base, others partially detached from a raw, red, and 
 bleeding surface ; many denuded, florid, red, ulcerated surfaces, with 
 small central sloughs secreting pus and exuding blood, the teats 
 exceedingly tender, hot. and swollen. ... In some animals, under 
 some circumstances, this state continues little altered till the third 
 or fourth week, rendering the process of milking painful to the 
 animal, and difficult and dangerous to the milker." 
 
 " In many, however, little uneasiness seems to exist. The parts 
 gradually heal ; the crusts, although often partially or entirely 
 renewed, ultimately separate, leaving apparently but few deep 
 irregular cicatrices, some communicating with the tubuli lactiferi, 
 the greater part being regular, smoothly depressed, circular, or oval." 
 
 Ceely illustrated his classical memoir with a series of valuable 
 coloured drawings. One plate is a faithful picture of the disease on 
 
NATURAL AND CASUAL COW-POX. 319 
 
 the teats as it is ordinarily met with ; the other is a composite picture, 
 consisting of the disease as ordinarily observed in the cow, to which 
 is superadded a number of depressed vesicles as they occur in inocu- 
 lated cow-pox. It is, however, an improvement on a plate published 
 by Sacco. The latter is an elaborate drawing, representing the udder 
 and teats of a cow, with an eruption purporting to be the natural 
 <-o\\-pox. Jenner had described a bluish tint in the vesicles in 
 natural cow-pox, and Sacco deliberately represents the natural disease 
 by a highly coloured diagrammatic illustration in which he depicts 
 clusters of vesicles of inoculated cow-pox, coloured blue, and with 
 a silvery lustre. 
 
 Hering has given a coloured plate of the natural cow-pox. On 
 the teats are a number of oval and circular bttllous vesicles and 
 crusts. More recently, Layet has pointed out the same characters 
 in the cow-pox discovered near Bordeaux in 1883 and 1884. The 
 classical characters of the inoculated disease were wanting, particu- 
 larly the central depression. In Wiltshire, the author could only 
 distinguish, on the cow's teats, globular and broken vesicles and 
 thick prominent crusts and ulcers, appearances which had very 
 little in common with the ordinary results of vaccination. 
 
 The early accounts of the severe character of the disease will 
 appear by no means exaggerated to those who have had an oppor- 
 tunity of studying the effects on the hands of the milkers, or indeed 
 t<> those who have made themselves familiar with the descriptions 
 given by Jenner, in some of his cases : 
 
 Joseph Merret had several sores on his hands, swelling and stiffness 
 in each axilla, and much indisposition for several days. 
 
 Mrs. H. had sores upon her hands which were communicated to her 
 nose, which became inflamed and very much swollen. 
 
 " Sarah Wynne had cow-pox in such a violent degree that she was 
 confined to her bed, and unable to do any work for ten days. 
 
 William Rodway was so affected by the severity of the disease that 
 he was confined to his bed. 
 
 " William Smith had several ulcerated sores on his hands, and the 
 usual constitutional symptoms, and was affected equally severely a second 
 and a third time. 
 
 William Stinchcomb had his hand very severely affected with several 
 corroding ulcers, and a considerable tumour in the axilla. 
 
 " Sarah Xelmes had a large pustulous sore on the hand, and the usual 
 symptoms. 
 
 "A girl had an ulceration on the lip from frequently holding her 
 finger to her mouth to cool the raging of a cow-pox sore by blowing 
 upon it. 
 
320 INFECTIVE DISEASES. 
 
 " A young woman had cow-pox to a great extent, several sores which 
 maturated having appeared on the hands and wrists. 
 
 " A young woman had several large suppurations from cow-pox on the 
 hands." 
 
 Pearson in his investigations encountered, and was informed of,. 
 similar experiences. 
 
 " Thomas Edinburgh was so lame from the eruption of cow-pox on 
 the palm of the hand as to necessitate his being for some time in hospital. 
 For three days he had suffered from pain in the armpits, which were 
 swollen and sore to the touch. He described the disease as uncommonly 
 painful, and of long continuance. 
 
 " A servant at a farm informed Pearson that in Wiltshire and 
 Gloucestershire the milkers were sometimes so ill as to lie in bed for 
 several days. 
 
 " Mr. Francis said that cow-pox was very apt to produce painful sores 
 on the hands of milkers. 
 
 " A servant of Mr. Francis said that cow-pox affected the hands and 
 arms of the milkers with painful sores as large as a sixpence. 
 
 " Mr. Dolling describes the disease as ' a swelling under the arm, chilly 
 fits, etc., not different from the breeding of the small-pox. After the 
 usual time of sickening, namely, two or three days, there is a large ulcer, 
 not unlike a carbuncle, which discharges matter.' 
 
 " Dr. Pulteney described the disease as causing ' a soreness and swell- 
 ing of the axillary glands, as under inoculation for the small- pox, then 
 chilliness and rigors and fevers, as in the small-pox. Two or three days 
 afterwards abscesses, not unlike carbuncles, appear generally on the hand a 
 and arms, which ulcerate and discharge much matter.' 
 
 " Mr. Bird wrote a short account : ' It appears with red spots on the 
 hands, which enlarge, become roundish, and suppurate, tumours take 
 place in the armpit, the pulse grows quick, the head aches, pains are felt 
 in the back and limbs, with sometimes vomiting arid delirium.' 
 
 " Annie Francis had pustules on her hands from milking cows. These 
 pustules soon became scabs, which, falling off, discovered ulcerating and 
 very painful sores, which were long in healing. Some milk from one of 
 the diseased cows, having spurted on the cheek of her sister and on the 
 breast of her mistress, produced on these parts of both persons pustules 
 and sores similar to her own on her hands." 
 
 In more recent times these descriptions have been confirmed. 
 
 In 1836 cow-pox was discovered at Passy, near Paris. A black 
 cow, in very poor condition, had cow-pox six weeks after calving. 
 Bousquet had no opportunity of seeing the eruption in the early 
 stage, but on examination he found reddish-brown crusts on the 
 teats, which later gave place to puckered scars. The milk-woman, 
 Fleury,' who had had small-pox, nevertheless contracted the disease 
 from the cow. She had several vesico-pustules on the right hand 
 
NATURAL AND CASUAL COW-POX. 321 
 
 ;ml on her lips. A vesieo-pustule, when opened with a lancet, 
 di*rhari:ed like an abscess. 
 
 In a letter to Mr. Badcock, dated April 3rd, 1845, Ceely referred 
 t> .mother now stock of lymph raised from a milker's hand. He 
 added : 
 
 " In the enclosed lymph I see nothing unusually severe, except 
 on very thin skins; although the milker's hand exhibits now rough 
 ulcers, one on the hand deep enough to encase a bean." 
 
 Recent discoveries of coiv-pox in England. After Ceely's cases in 
 1840-41, no cases of casual cow-pox on the hands of milkers were 
 recognised as such mid recorded in this country for nearly fifty years. 
 In the outbreak of cow-pox discovered by the author in December 
 1887, in Wiltshire, the disease was communicated to nearly all the 
 milkers. The reader is referred to the account of this outbreak, 
 which has already been given in the chapter on scarlet fever (p. 274). 
 
 The author's researches were confirmed by Mr. Forty in 1888, 
 and Mr. Bucknill in 1895. 
 
 In June 1888 Mr. Forty, in practice at Wotton-under-Edge, 
 Gloucestershire, reported to the Local Government Board, that at 
 a farm at Alderley, an eruptive disease on the udder and teats was 
 occurring amongst cows, and that the farmer's son, and other persons 
 engaged as milkers, had contracted an eruption like that of the cows. 
 The farmer's son had been under Mr. Forty's care suffering from an 
 eruption, ami circum-aiial piles. Mr. Forty had watched the course of 
 the eruption from papules to vesicles and scabbing, and concluded 
 that the eruption could not be distinguished from vaccinia. Klein 
 visited the farm, and found a number of cows with sores on the teats 
 and udders. The sores were of various si/.e-s and outline. mo>tlv 
 irregular, and covered with brown or black scabs. Those on the 
 teats were larger and more irregular than those on the udder. 
 Klein wa.* shown several milker* who had had soree on one or more 
 tinkers; one had had a had arm with swollen axillary gland-. 
 The farmer had al>o contracted the eruption: but in the-e peiv-on-. 
 only scabs were visible as the remnants of their >oiv>. 
 
 A girl of about twenty had taken the place of an incapacitated 
 milker, and noticed a red pimple form on the dor>:l sm fan- of her 
 right thumb. Eight da\> afterward* tin re was a .*lightly n 
 circular ve-icle. with dark centre and pale periphery : the rent re of the 
 \esicle wa* slightlv depreed. It \va* ju-t under half an inch in 
 diameter; there was peripheral redne. but no marked an-ola. 
 irii-1 had three good vaccination mark*. 
 
 Klein experimented on calve* with lymph from the vehicle and 
 
322 INFECTIVE DISEASES. 
 
 crusts from the cow's teats, with the result that from both sources 
 an eruption was produced, which in appearance and course was like 
 vaccinia. With lymph from one of the calves, a public vaccinator 
 inoculated a number of infants, and fine vesicles developed, indis- 
 tinguishable from vaccinia. 
 
 In 1894 Mr. Bucknill met with a case in a milkman. He had 
 been milking a cow affected with cow-pox, and on the ninth day after 
 exposure to infection, and the seventh day after the eruption of the 
 first papule, there were three pocks on the fore-arm. The pocks 
 were elevated, circular, and umbilicated, with a dull, creamy-white 
 ring at the circumference, and there was well-marked induration 
 and extensive areola. There were four excellent marks of primary 
 vaccination. The vesicles contained clear lymph, and re-inoculation 
 of the arm failed to take. An attempt to re-vaccinate the man 
 with current calf lymph produced only topical irritation. 
 
 INOCULATED COW-POX. 
 
 Natural or Virulent Lymph. Severe symptoms are not limited to 
 milkers casually infected from the cow. Under certain conditions, 
 artificial inoculation of fresh virus from the cow reproduces the 
 disease without any mitigation. Thus, in Jenner's cases : 
 
 " James Phipps. The incisions assumed at their edges rather a 
 darker hue than in variolous inoculation, and the efflorescence around 
 them took on more of an erysipelatous look. They terminated in 
 scabs and subsequent eschars. 
 
 " Susan Phipps was inoculated from the cow by inserting matter 
 into a superficial scratch on December 2nd. The child's arm now 
 showed a disposition to scab, and remained nearly stationary for 
 two or three days, when it began to run into an ulcerous state, 
 and then commenced a febrile indisposition, accompanied with an 
 increase of axillary tumour. The ulcer continued spreading near 
 a week, during which the child continued ill, when it increased to 
 a size nearly as large as a shilling. It began now to discharge pus ; 
 granulations sprung up, and it healed." 
 
 Jenner's lymph was employed by Mr. Cline with similar results. 
 
 " The child sickened on the seventh day, and the fever, which 
 was moderate, subsided on the eleventh. . . . The ulcer was not 
 large enough to contain a pea." 
 
 Precisely similar experiences have since been encountered, in the 
 early removes of fresh stocks of virulent lymph. Bousquet in France, 
 in his first trials with a new lymph, in 1836, made three punctures, 
 but he had soon to abandon this practice, because the intensity 
 
INOCULATED COW-POX. 
 
 of the inflammation was sometimes so great tli.-it it >pread over the 
 entire aim as far as the glands of the axilla. In one case, tin- 
 vehicle* were enormou>. and the inflammation so violent, that bath>. 
 poult ice>. fomentations, and antiphlogistic diet scarcely sufficed 
 to reduce it. The crusts when they fell off left ulcerations which 
 were very >low to undergo cicatrisation. In some cases, the vesicles 
 which resulted hollowed out the skin so deeply that they left regular 
 holes. 
 
 In the following year Estlin, in England, started a stock of fresh 
 \arrine virus from the cow, and found on inoculating children that 
 the new lymph was extremely active. 
 
 In 52 the disease was regular, 
 ., 1 severe erysipelas, 
 
 ,, 4 er ythematous eruptions of a violent character, 
 ,, 2 highly inflamed ulcerated arms, 
 ,, 1 no effect after twice vaccinating, 
 8 result unknown ; supposed to have been favourable. 
 
 68 
 
 Cultivated or Attenuated Lymph. When cow-pox lymph has 
 been mitigated by successive transmission through the human subject, 
 or by cultivation on the belly of the calf, with careful selection 
 of vesicles, it will produce effects which are as follows : About 
 the end of the second day after insertion, or early on the third day, 
 a slight papular elevation is noticeable. By the fifth or sixth day, 
 it has become a distinct vesicle, of a bluish-white colour, with raised 
 margin and central cup- like depression. By the eighth day, the 
 vesicle is perfect. It is circular, pearl -coloured, distended with clear 
 lymph, and the central depression is well marked. On the same day, 
 <>r a little earlier, the areola begins to appear, and gradually extends 
 to a diameter of from one to three inches, accompanied with 
 induration and tumefaction of the subjacent connective tissue. After 
 the tenth day, the areola begins to fade, and the vesicle at the same 
 time begins to dry in the centre; the lymph becomes opaque arid 
 gradually concretes, and by the fourteenth or fifteenth day, a hard 
 mahogany-coloured seal> is formed which contracts, dries, blackens, and 
 fall.- off between the twentieth and twenty-fifth days. A circular, 
 depressed, foveated, and sometimes radiated scar remains behind. 
 By >e]ectin,i,' ch:ir,-icteri>tie ve>icles on the calf or on the human 
 Millet, and by collecting the lymph at an early Maire on the fifth, 
 sixth, or seventh day, this artificial disease, commonly known ifl 
 
324 INFECTIVE DISEASES. 
 
 vaccinia, can be kept up in this comparatively mild form. But 
 under certain conditions, such as a peculiarity in the subject inocu- 
 lated, or if lymph be taken too late, there will be, just as in variolation, 
 tendency to revert to the full intensity of the natural virus. 
 
 Bacteria in Vaccine Lymph. Cohn, Sanderson, and Godlee 
 described micrococci in vaccinal vesicles. Quist and Ferre in 1883 
 investigated the same subject. Yoigt in 1885 distinguished three 
 species of micrococcus a diplococcus, a large coccus, and a third form. 
 Bauer in the same year described the presence of bacilli and sphsero- 
 cocci. Marotta in 1886 regarded a tetracoccus as the specific micro- 
 organism, and Tenhot in 1887 distinguished a dozen micrococci, two 
 bacilli, and two yeasts. In the same year Garre isolated a micrococcus 
 which appeared to him to be the contagium, but inoculated on a child 
 it neither produced local vesicles nor immunity ; while Guttmann 
 pointed out three micro-organisms which appeared to be rather more 
 constantly present than others. Pfeiffer much more fully investigated 
 the bacteriology of vaccine lymph, and found Saccharomyces vaccinse, 
 which was seldom present in human lymph but constantly found in 
 calf lymph ; sarcinse, both in human and calf lymph, including Sarcina 
 lutea, Sarcina tetragonus, Sarcina aurantiaca, Sarcina muscopus ; 
 bacteria and bacilli were found only exceptionally in human lymph, 
 but frequently in calf lymph. These included a bacterium corre- 
 sponding with Proteus vulgaris. 
 
 Three mice were inoculated subcutaneously with a drop of the 
 liquefied gelatine, but the result was negative. The injection of a 
 considerable quantity proved fatal to guinea-pigs and rabbits, a 
 result which was probably due to ptomaine poisoning. 
 
 There were also several bacilli which did not liquefy gelatine ; 
 these were not investigated. 
 
 Staphylococcus cereus albus was found very frequently, and 
 Staphylococcus pyogenes aureus occasionally. Pure- cultivations of 
 these micrococci inoculated on the skin of calves produced a rapid 
 local irritation, followed by vesiculation, but without the classical 
 characters of the vaccine vesicle. The inoculated part was com- 
 pletely healed in three to five days. According to Pfeiffer they 
 explain the so-called false vaccine. 
 
 Micrococcus pyogenes albus was almost constantly present. 
 Numerous other micrococci were found, but not constantly present ; 
 vaccine lymph being a splendid medium for the growth of micrococci. 
 Pfeiffer pointed out that the effects of Staphylococcus pyogenes aureus, 
 albus, and citreus, and of Streptococcus pyogenes on rabbits had an 
 important bearing upon the practice of vaccination, and he recom- 
 
INOCULATED COW-POX. 325 
 
 mended that calf lymph should be tested before use upon children 
 by inoculation of the ear of a rabbit. If after two days no erysipelas 
 occurs in the inoculated rabbit, the absence of streptococci may be 
 considered as almost proved. Two or three rabbits should be inocu- 
 lated at the same time. 
 
 The author's researches into the bacteriology of vaccine lymph 
 extended over some years. They independently confirmed and 
 extended the results obtained by Pfeiffer. Having on several 
 occasions examined vaccine lymph and vaccine pus, and failed to find 
 a specific bacterium, the author proceeded to make a more systematic 
 examination of the different species of bacteria in samples of current 
 vaccine lymph. Pure-cultivations were obtained by plate-cultivation, 
 and inoculation of the surface of nutrient agar, obliquely solidified 
 in test tubes. Various current stocks of lymph were used in the 
 investigation. Among the specimens of calf lymph, No. 1 yielded a 
 torula, Bacillus pyocyaneus and Bacillus subtilis; No. 2, a bacterium, a 
 variety of proteus, Staphylococcus pyogenes aureus, and yellow bacte- 
 rium ; No. 3, a bacterium, micrococcus, yellow bacterium, and torula ; 
 No. 4, yellow micrococcus, white micrococcus, white torula, yellow 
 sarcina, white diplococcus, Staphylococcus cereus albus, and a mould 
 fungus; No. 5, yellow sarcina, Staphyloceccus pyogenes aureus, yellow 
 micrococcus, white bacillus, Staphylococcus pyogenes albus, large 
 white micrococcus, yellow bacterium, and a white micrococcus. Among 
 the specimens of human vaccine lymph, No. 1 contained a white 
 micrococcus, proteus, and Staphylococcus pyogenes aureus ; No. 2, a 
 micrococcus, a tetracoccus, a white liquefying micrococcus, and a 
 yellow bacterium ; No. 3, white micrococcus, yellow micrococcus, 
 Staphylococcus aureus and flavus, a bacterium, a white micrococcus, 
 a bacillus resembling Bacillus subtilis, Staphylococcus pyogenes cereus 
 and a brown tetracoccus. The author is familiar with these different 
 species of bacteria, and not one of them is peculiar to vaccine lymph ; 
 there was no bacterium constantly present in human and calf vaccine, 
 and there was not one which could be regarded as the contagium. 
 To >um up, most of them are well known saprophytic bacteria, and 
 some were identical with bacteria commonly found in suppuration. 
 Vaccine lymph is a most suitable cultivating medium for micro- 
 organism^ and bacteria invariably got access to the contents of the 
 vaccine vesicle. There is no evidence to be obtained by the present 
 methods of research as to the bacterial nature of the contagium of 
 vaccine lymph. Copeman obtained similar iv>ults. and thus con- 
 firmed the author's conclusions. 
 
 Klein and Copeman have also observed minute bacilli in CM If- 
 
326 INFECTIVE DISEASES. 
 
 lymph and in variolous lymph. Numerous attempts to cultivate them 
 in nutrient media and in the living animal failed entirely, and the 
 identity of the bacilli could not be determined. Pfeiffer, Guarnieri, 
 Monti, Buffer, and Plimmer have drawn attention to structures in 
 lymph, which they believe to be of the nature of parasitic protozoa. 
 These bodies have been studied, more especially in the tissues. They 
 are four times the size of ordinary micrococci, and are found in the 
 clear vacuole in the protoplasm of epithelial cells. Whether they 
 ;uv really parasites or altered anatomical elements has not been 
 determined. No other conclusion can be drawn from all these 
 observations, except that the nature of the contagium of cow-pox 
 is unknown. 
 
 ORIGIN OF COW-POX. 
 
 Jenner's original theory was that cow-pox was derived from 
 " grease," but subsequently he distinguished between cow-pox, a 
 disease peculiar to the cow, and "grease," a disease transmitted to 
 the cow from the horse, and the mistake of confounding these two 
 diseases was attributed to farmers and farriers. Thus he wrote : 
 
 " From the similarity of symptoms, both constitutional and local, 
 between the cow-pox and the disease received from morbid matter 
 generated by a horse, the common people in this neighbourhood 
 when infected with this disease, through a strange perversion of 
 terms, frequently called it the cow-pox." 
 
 Jenner's theory of the origin of cow-pox has been discouraged ; so 
 also has the view of its being a " spontaneous " disease in the cow, 
 though Ceely, after many years of research in the Yale of Aylesbury, 
 could never discover the probability of any other origin. Both 
 opinions have given way to the theory that cow-pox is small-pox 
 transmitted to the cow an opinion advocated by Baron, and 
 supported by an erroneous interpretation of Ceely's and Badcock's 
 variolation experiments. Thus the cow-pox and grectse of farmers 
 and farriers no longer attracted attention in this country, and as 
 natural cow-small-pox has never been discovered, cow-pox has been 
 credited with being extinct. 
 
 For a full discussion of this subject the reader is referred to the 
 work by the author on the History and Pathology of Vaccination, 
 but the variolation experiments alluded to will be briefly mentioned. 
 
 In 1801 Gassner inoculated eleven cows with small-pox lymph, 
 and succeeded in one in producing phenomena indistinguishable from 
 the results of ordinary vaccination with cow-pox, and children were 
 inoculated from the cow. 
 
ORIGIN OF CoW-l'oX. i',27 
 
 Iii 1828 Dr. McMichael reported tli.-tt several physicians in 
 had obtained similar results, and children weiv .-successfully 
 vaccinated." 
 
 In 1836 Dr. ]\Iartin. in America, inoculated the cow's udder with 
 variolous Ivmph. and l>y inoculating children produced an epidemic 
 of small-pox with fatal cases. In 1839 Reiter of Munich, after fifty 
 unsueces>ful attempts, succeeded in producing a vesicle, and a child 
 inoculated from the vesicle contracted small-pox. 
 
 In 1839 Dr. Thiele , af ter a num]?er of unsuccessful attempts to 
 inoculate cows with variolous virus, succeeded in producing a vesicle 
 with the physical characters of the vaccine vesicle, and from it a 
 stock of lymph was raise 1 from which over three thousand^ persons 
 were inoculated. Thiele 's method was to inoculate the udder with 
 lymph, and to select for the purpose young cows which had recently 
 calved and had delicate skins. In England Ceely succeeded by 
 inoculating the vulva of a heifer. One of the punctures developed 
 into an enormous vesicle, which was undoubtedly variolous. His 
 a>^ i-tant punctured his hand with the lancet which had been used 
 to open the vesicle, and febrile symptoms appeared, followed by an 
 eruption on the face, neck, trunk, and limbs, at first papular, then 
 vesicular, and finally pustular. The lymph was used in children, 
 and " vaccine " vesicles were produced. One child suffered from 
 vomiting delirium, and extensive roseola, but there was no eruption 
 in any other ca 
 
 In 1840 Badcock of Brighton inoculated a cow successfully, and 
 later succeeded in variolating thirty-seven out of two hundred cows 
 14 ton which he experimented. 
 
 In 1847 va isolation of the cow was successfully performed at 
 Berlin, but the virus produced variola, and one of the children 
 inoculated died of confluent small-pox. 
 
 In 1864 Chauveau inoculated seventeen animals with virulent 
 small-pox lymph. Very small papules resulted, and the virus from 
 the papules produced variola in a child, which was infectious to others. 
 Klein in this country until recently was uniformly unsuccessful. 
 Voiirt. Fischer, King, Eternod, Hacciu>. Hime and Simpson, have 
 all succeeded in inoculating cows arid producing variola-vaccine. 
 
 The results of these experiments have been very generally misin- 
 t.'i-pretrd. and claimed by some as conclusive evidence of the identity 
 of cow-pox and small-pox. Instead of the vesicle beini: regarded as 
 the most attenuated form of variola, the experimenters are said to 
 have succeeded in producing cow-pox. 
 
 It is quite true that they produced phenomena indistinguishable 
 
328 INFECTIVE DISEASES. 
 
 from the phenomena of an ordinary vaccination, but that does not 
 mean that they produced the disease cow-pox. The vesicle which 
 followed the inoculation, whether papular or vesicular, was small-pox. 
 Ceely, Badcock, Voigt, and others, succeeded in ingrafting the cow 
 with small-pox, and when suitable lymph and suitable subjects were 
 employed, the virus was so attenuated that a benign vesicle resulted. 
 Similar results were obtained by Button and Dimsdale, and identical 
 results by Adams, Guillou, and Thiele, by inoculating the human 
 subject with variolous lymph without first ingrafting the disease 011 
 the cow. 
 
 Vaccination with variola -vaccine is simply a modification of the 
 Suttonian system of small-pox inoculation, only in the first remove 
 the cow is substituted for the human subject. All those who were 
 inoculated with Ceely's, Badcock's, or Simpson's variola-vaccine, 
 were not in the usual meaning of the word vaccinated ; they were 
 not inoculated with cow-pox but they were variolated, and in such 
 an extremely attenuated form that the persons so variolated do 
 not convey the infection. By judicious selection it is thus possible 
 to obtain a strain of lymph from variola which, by direct inoculation 
 of the human subject or by first inoculating a cow, is deprived of 
 infectious properties, and produces on the arm the physical characters 
 of an ordinary vaccine vesicle. This has been regarded as a proof 
 of the identity of small-pox and cow-pox, but it is not so. Variola, 
 and cow-pox are not the only diseases caused by a virus which can 
 be attenuated until only a vesicle is produced with the characters of 
 an ordinary vaccine vesicle. The results which have been obtained 
 with the virus of cattle plague and of sheep-pox and horse-pox have 
 been given in previous chapters ; and no one would urge on this 
 account that human small-pox, cattle plague, cow-pox, sheep-pox, 
 and horse-pox are all manifestations of the same disease. Cow-pox 
 has never been converted into human small-pox, and, in their clinical 
 history and epidemiology, natural cow-pox and human small-pox 
 are so different, that the comparative pathologist is no more pre- 
 pared to admit their identity than he is prepared to admit the 
 identity of cow-pox and sheep-pox, or small-pox and cattle plague. 
 
 Protective Inoculation. Whether vaccination of all heifers 
 on a farm would protect them from cow-pox when they came into 
 milk is not known, the duration of the immunity in calves afforded 
 by vaccination having not been determined. Calves undoubtedly 
 have an immunity after vaccination, lasting for some weeks. 
 
 In 1896 Beclere, Chamber), and Meriard experimented upon 
 the immunising power of the serum of vaccinated calves. They 
 
COW-POX AND SMALL-POX. 329 
 
 concluded from experiments on animals and children that the serum 
 of a vaccinated calf from ten to fifty days after vaccination will give 
 immunity against inoculated cow-pox. They further stated that, 
 whereas the immunity given by vaccination in the ordinary wax- 
 is not complete until the eighth day, the immunity obtained by 
 injection of the immunising serum is immediate. The serum has 
 also been credited with therapeutic properties and has, it is said. 
 proved efficacious in cases of small-pox. 
 
 Jenner believed that cow-pox did not protect against itself but 
 protected against small-pox, and for a century this has been a subject 
 of much controversy. The reader is referred to the Reports and 
 conclusions of the Royal Vaccination Commission. 
 
 Stamping-out System. It would undoubtedly be an advan- 
 tage if cow-pox were scheduled under the Contagious Diseases Animals 
 Act. Cow-keepers and dairy-men, being anxious that their trade 
 should not be interfered with, very commonly conceal the existence 
 of the disease, and perhaps nothing is known about it, unless a milker 
 infected from the cows seeks for medical advice. The contamination 
 of the milk with lymph, pus, crusts, and sometimes blood, renders 
 it unwholesome, and therefore precautions ought to be taken to 
 prevent its occurrence. If the infected cows in a herd are the last 
 to be milked, and the milker washes his hands after the milking, the 
 disease will not spread. 
 
CHAPTER XXIII. 
 
 DIPHTHERIA. 
 
 DIPHTHERIA is a specific infectious disease, especially of children, 
 characterised most commonly by inflammation, and infiltration with 
 lymph cells and fibrine, of the mucous membrane of the fauces, 
 pharynx, larynx and trachea, followed by necrosis of the mucous 
 membrane and the formation of a greyish-white false membrane, 
 the diphtheritic membrane. In some cases a diphtheritic membrane 
 forms in the stomach, intestine, the urinary organs and in wounds. 
 After the separation of the membrane an ulcer remains, which may 
 gradually heal. In the superficial part of the diphtheritic membrane 
 there are masses of bacteria including cocci, streptococci, and bacilli. 
 The diphtheria bacilli are not found in the blood or in the 
 internal organs. There is no doubt of the fact that diphtheria 
 is a disease which can be communicated from one person to 
 another ; but the question of its origin is still a vexed one. There 
 is a close association with insanitary conditions and decaying 
 animal and vegetable refuse, and dampness. Localities with damp 
 houses, defective drainage, and a cold exposure, are favourable 
 to the development of diphtheria; but that does not necessarily 
 indicate that these conditions can originate it. On the other 
 hand, assuming the disease to be due to a living contagium, 
 these insanitary conditions would afford a suitable environment 
 predisposing to the development, and facilitating the spread, of the 
 disease. Scarlet fever and measles predispose to diphtheria ; and 
 defective sanitary conditions, causing sore throat, may indirectly 
 act as a predisposing cause. A great many cases have been quoted 
 to illustrate the possibility of the conveyance of diphtheria, by milk, 
 and the theory which best harmonises with all these observations is 
 the existence of a specific bacillus, which may be readily transferred 
 from the throat of the diseased to the healthy ; which finds also in 
 milk a suitable soil for its growth, and by its agency may be trans- 
 mitted to the consumer. Such a bacillus was discovered by Loffler, 
 
 330 
 
DIPHTHERIA. 
 
 331 
 
 and may be easily 
 obtained from the 
 throat of diphtheritic 
 patient- in the fol- 
 lowing manner : 
 ifure Outfit. 
 Steel rods like or- 
 dinary knitting 
 needles, about MX 
 inches in length, are 
 beaten out or rough- 
 ened at one end, and 
 a pledget of wool is 
 twisted round so as 
 to form a swab. 
 These swabs are 
 placed in clean 
 tubes, which are then 
 plugged with cotton- 
 wool. The test-tubes 
 and swabs are steri- 
 lised by heating in 
 the hot air steriliser 
 f or an hour at 1 50 C. 
 The so-called culture 
 outfit conM>t> of 
 a small box con- 
 taining a test -tube of 
 blood serum and a 
 swab. They can be 
 always kept ready for 
 use, and after u>e 
 should be conveyed 
 by hand for further 
 examination. The 
 danger of trans- 
 Ed i 1 1 i n g virulent 
 diphtheritic material 
 by po>t i> obvious. 
 When the examina- 
 tion of the tube has 
 been completed, the 
 
 
 
 
 _- v^ * - : -V ^ *^V / 
 
 W&Z&Mi 
 
 ''* * V 
 
 ,'3>A* \V'.>yf:- 
 
 FIG. 12G. FKKK SUKFACF. OF DIPMTHKKITK LARYNX 
 x 350 (HAMILTON). A, Deposit of diphtheria iKwillu* 
 on surface of false membrane ; B, false memhr;uit : 
 C, mucosa; I, lymph-cells and false membr.ui. 
 surrounded by meshes of fibrine ; e, surface of mucosa 
 deprived of its epithelium ; l,v, lymph-cells containing 
 shed epithelium. 
 
332 INFECTIVE DISEASES. 
 
 culture outfit and its contents should be disinfected or destroyed. 
 To inoculate the tubes the patient, if it is possible, should be turned 
 to the light, the mouth well opened, the tongue depressed, and the 
 swab, without touching the teeth or the tongue, should be passed 
 straight to the tonsils or pharynx, and especially to the membranous 
 exudate. The swab is carefully and quickly withdrawn, and at 
 once very gently rubbed over the surface of the blood serum. The 
 culture outfit is then sent to the laboratory with full particulars, 
 and the tubes are placed in the incubator at 37 C., and can be 
 examined after twelve hours. If the throat has been disinfected 
 
 FIG. 127. BACILLUS OF DIPHTHERIA ; FROM A CULTIVATION ON BLOOD 
 SERUM, x 1000 (FRANKEL and PFEIFFER). 
 
 before examination, this must be taken into account, as the failure 
 to find bacilli would not then necessarily indicate a wrong diagnosis. 
 In all undoubted cases of diphtheria, growths will be obtained either 
 in the form of a pure-culture of the bacillus, or far more commonly 
 there will alsolbe colonies of various bacteria, especially Streptococcus 
 pyogenes. 
 
 Bacillus of Diphtheria Rods, straight or slightly curved, 
 "3 to '8 fj, in breadth, and 1-5 to 6*5 //, in length. They occur 
 singly, in pairs, sometimes in chains, and sometimes as short 
 leptothrix forms. In some cultures very irregular forms are 
 observed, the bacilli being swollen at one or both ends or thicker 
 in the middle portion, or the bacillus may contain oval or spherical 
 
DESCRIPTION OF PLATE VIII. 
 Bacillus diphtherias and Bacillus typhosus. 
 
 FIG. 1. Cover-glass preparation from a pure-cultivation of Bacillus diph- 
 
 therire on blood serum ; obtained from the throat in a typical case of 
 
 diphtheria. Stained with gentian- violet, x 1200. 
 FIG. 2. Cover-glass preparation from a pure-cultivation of Bacillus typhosus 
 
 on nutrient-agar ; from the spleen in a case of typhoid fever Stained 
 
 with gentian-violet, x 1200. 
 
MP'jK/J ^ ^ \- ' v f ^ 
 
 v%^\ .XXVvjVv 
 
 TV.V.v- -- : >UC -^ :*: : 
 :-^&O^ :: '^xft- 
 
 ^t^-i A "^ - ^ i ^ 
 
 ^ 
 
 BACILLUS DIPHTHERIA 
 
 ',-BFt, 
 
 Hg2.BAC ILLUS TYPHOSUS 
 
DIPHTHERIA. 
 
 333 
 
 elements. They differ greatly in si/.e and shape, often in the same 
 cultures, ami still moiv in cultures obtained from different sources. 
 S{x>re formation is unknown. In unstained preparations there are 
 hiirhly refractive elements which correspond with the deeply stained 
 parrs of the bacillus. They stain readily with the ordinary aniline 
 dyes. At certain stages of their growth they stain irregularly, the 
 protoplasm of the rod being broken up into irregular segments. 
 The bacillus is non-motile, and does not liquefy gelatine ; it grows 
 at 20 C., but much more readily at higher temperatures. Colonias 
 in gelatine plate-cultivations are 
 yellowish-brown, and opaque, 
 jH'anular, and circular, but with 
 more or less irregular margin. 
 
 In plate -cultivations on agar 
 and on glycerine agar the same 
 description applies. 
 
 On the surface of gelatine 
 the appearances depend greatly 
 on the method of inoculation. 
 The growth may occur in the 
 form of a whitish film, but if a 
 sub-culture has been prepared 
 from broth the growth is often 
 composed of a number of iso- 
 lated white colonies (Fig. 128, a). 
 
 On blood serum, after twelve 
 hours the colonies appear in the 
 form of little elevated grey is] i- 
 \vhite or pearl-grey dots, which 
 coalesce, forming a film if the 
 serum is moist. On the surface 
 of 1 per cent, alkaline glycerine 
 agar, the appearances are found 
 
 to vary, and this medium is not so suitable for the cultivation of the 
 bacillus. In slight Iv alkaline broth, with or without the addition of 
 1 per cent, grape-sugar, the culture is cloudy, or a fine granular 
 deposit occurs along the sides and bottom of the tube, while the 
 broth remains clear. 
 
 On potato the growth is almost invisible, in the form of a dry. 
 thin glaze. Irregular forms are very numerous on microscopical 
 examination, whilst the rods are thicker than usual (Welch and 
 Abbott). In milk the organisms grow readily. 
 
 FlG. 128. PURK-CULTURES OF BACILLI > 
 DIPHTHERIA ON GELATINE : a, isohttt-d 
 
 colonies ; b, filmy growth. 
 
334 INFECTIVE DISEASES. 
 
 
 Dried diphtheritic membrane and cultures dried 011 silk threads 
 retain their vitality for several months. 
 
 A broth-culture in forty-eight hours may be used for inoculating 
 guinea-pigs. A few drops will cause death in from three to five 
 days ; there is hypersemia and oedema at the seat of inoculation, the 
 lymphatic glands are enlarged, there is fluid in the peritoneal, 
 pleural and pericardia! cavities, and the lungs are congested. The 
 bacillus is found at the seat of inoculation, but not, as a rule, in the 
 blood or internal organs. Inoculation of rabbits produces extensive 
 local oedema, enlargement of the lymphatic glands, and death in 
 from four days to three weeks. Roux and Yersin pointed out that in 
 less acute cases there was paralysis of the hind limbs. Mice and 
 rats have an immunity. Cultures lose their virulence with age, but 
 the nitrate from old cultures contains more toxic substance than 
 that from fresh cultures. The toxin has been described in a 
 previous chapter (p. 46). 
 
 Old cultures sterilised by heating for an hour to 60 C. or 
 70 C. will render guinea-pigs immune in two weeks. The toxic 
 substance is believed to be destroyed by this process, while according 
 to Frankel the immunity-giving substance which is also present in 
 the culture is not affected. 
 
 According to Behring's researches, the blood of immune animals 
 contains diphtheria antitoxin, consequently the blood of an im- 
 mune animal is capable of neutralising the toxic properties in a 
 filtered culture, not only in the living animal but when added to the 
 culture in a test-tube. These researches led to the employment of 
 the serum of an immune animal as a therapeutic agent in the treat- 
 ment of diphtheria in man (p. 58). 
 
 Bacteriological Diagnosis. The diphtheritic bacilli are not only 
 found in the throat while the lesions exist, but they are found after 
 all sign of the disease has disappeared. In some cases they 
 persist for a few days, in others for three or four weeks, and 
 in rarer cases they have been found several months afterwards. 
 They have also been found in the throats of persons in health, 
 especially of those who have been in contact with cases of diph- 
 theria, such as healthy children in infected families and healthy 
 nurses. 
 
 The bacilli which persist in the throat after recovery may be 
 virulent up to the time of their disappearance, or they may gradu- 
 ally become attenuated, and entirely lose their pathogenic properties. 
 The value of a microscopical examination as an aid in the diagnosis 
 of diphtheria has been considerably exaggerated, and unless the 
 
DIPHTHERIA. 335 
 
 bacillus when isolated is tested by inoculation the test mav prove 
 to be entirely fallacious. 
 
 Loffler and Von Hoffman both found bacilli in healthy throats, 
 ami tliu.v created doubt as to the importance of the Loffler bacillus. 
 Hotl'inan found this bacillus in the throats of twentv-six out of 
 forty-five individuals, some of them suffering from scarlet fever, 
 measles or some other non- diphtheritic affections, while the rest were 
 healthy. The bacilli from these sources showed slight differences 
 in morphological and cultural characteristics, and Hoffman was 
 unable to decide whether these bacilli were diphtheria bacilli, which 
 had become harmless, or whether they were accidental epiphytes. 
 belonging to a closely allied but different species. 
 
 Roux and Yersin confirmed these observations. In a hospital 
 for children in Paris without any question of the existence of 
 diphtheria they found the so-called pseudo-diphtheria bacilli in 
 fifteen cases out of forty-five. In a school, in a seaside place 
 entirely free irom diphtheria, the same bacilli were found in 
 twenty- six out of fifty-nine children. They were also found in 
 children with simple sore throats, and in five out of seven cases 
 in measles. Roux and Yersin concluded that these bacilli were 
 not distinct from the Loffler bacillus. There were slight variations, 
 but there was no constant difference except in their pathogenic 
 properties. The appearance of the colonies, the growth in broth, 
 and the peculiar morphological elements showed characters common 
 to both, and there was, in fact, less difference than there is 
 between attenuated anthrax and virulent anthrax in form and in 
 cultures; but inoculations of the bacillus did not cause death, 
 though in some cases in guinea-pigs there was marked oedema at 
 the seat of inoculation. On the other hand, Loffler's bacilli 
 j )()>>-> different degrees of virulence, some cultures producing only 
 temporary redema, while others cause death in twenty-four hours. 
 
 Virulent diphtheria bacilli subjected to a current of air can 
 in two weeks be deprived of their virulence partially, and in four 
 weeks entirely. Weakened bacilli can be raised in virulence by the 
 simultaneous injection of the streptococcus of erysipelas, but bacilli 
 deprived of their virulence and bacilli originally non- virulent cannot 
 be made to assume virulent properties. Escherich maintained 
 that they could be distinguished by comparative cultures; that the 
 pseudo-diphtheria bacilli made broth alkaline, so that in forty-eight 
 hours litmus was turned red by Loffler's bacilli and blue by the 
 false bacilli. The bacilli themselves, according to Hoffman, are, as 
 a rule, shorter, wider and more uniform in size. 
 
336 INFECTIVE DISEASES. 
 
 Parke and Beebe, with a view to clearing up this question, made 
 cultures from three hundred and thirty healthy throats. They 
 found bacilli of three varieties : bacilli characteristic in growth 
 producing acid reaction in broth, but having no virulence ; bacilli 
 not characteristic in growth producing an alkaline reaction in broth, 
 not virulent ; and bacilli producing acid reaction in broth and 
 virulent. The virulent characteristic diphtheria bacilli were found 
 in eight cases, non- virulent diphtheria bacilli in twenty-four, and 
 non-virulent false diphtheria bacilli in twenty-seven. They con- 
 cluded that the eight cases must have been in contact with diphtheria, 
 although the throats were healthy. With regard to the bacillus 
 in the twenty-four cases they regarded it as the true diphtheria 
 bacillus which had lost its virulence, and the bacillus found in the 
 twenty-seven cases showing differences in size and manner of staining 
 and the reaction produced in broth was properly designated pseudo- 
 diphtheria bacillus. 
 
 
 DIPHTHERITIC DISEASES IN ANIMALS. 
 
 There are diptheritic diseases of the lower animals which are in 
 some respects similar to, and, some observers maintain, identical 
 with, human diphtheria. 
 
 In pigeons there is a disease accompanied w r ith the formation of 
 false membranes associated with a bacillus described by Lbffler. 
 
 Bacterium of Diphtheria of Pigeons (Bacillus columbarum, 
 Loffler). Short rods with rounded ends, mostly in irregular masses. 
 In plate-cultivations on nutrient gelatine they formed whitish patches 
 on the surface, and compact, ball-like masses when embedded in the 
 gelatine. They were also cultivated on blood serum and potatoes. 
 Subcutaneous inoculation of a pure- cultivation produced in pigeons 
 local inflammation and necrosis ; inoculation in the mucous mem- 
 brane of the mouth gave the appearances of the original disease. 
 Other animals were only locally affected, except mice, in which 
 characteristic symptoms and death resulted. They were isolated 
 from the diphtheritic exudations in pigeons, and in sections were 
 found in the vessels of the lungs and liver. 
 
 A similar disease is known to attack fowls, and there are also 
 diseases with development of false membranes of the respiratory 
 passages in horses, cats and swine. Outbreaks of these diseases 
 have been said to occur in times of prevalence of diphtheria in 
 man, and their intercommunicability has been suggested. 
 
 Dr. Turner supposes that diphtheria in man originates in diseases 
 
DIPHTHERITIC DISEASES IN ANIMALS. 337 
 
 simulating diphtheria in cats, pigs, and horses; and Klein, who 
 accepts this theory, maintains that cats suffer from genuine 
 diphtheria, and that after death the lungs are found full of grey, 
 consolidated lobular patches, and the kidneys are enlarged and 
 white. 
 
 Human diphtheritic membrane inoculated subcutaneously in cats 
 produces a painful swelling in the groin, and fever, and a fatal 
 termination in a week. The subcutaneous and muscular tissues at 
 the seat of inoculation are haemorrhagic and redematous. The 
 internal organs are congested, and in the kidneys the medulla is 
 congested, while the cortex is fatty. 
 
 A recent culture produces illness in twenty-four hours, a painful 
 tumour forms at the seat of inoculation, and death ensues in from 
 two days to a week. Pneumonia and fatty white kidney are found 
 after death, and the tissues at the seat of inoculation are haemor- 
 rhagic, and in parts almost gangrenous. 
 
 Klein found that diphtheritic membrane or a pure-culture in- 
 oculated into the cornea after removal of the superficial epithelium 
 produced ulceration, and in two cases perforation of the cornea,. 
 and purulent panophthalmitis. Bacilli were again recovered from 
 the ulcer similar in cultural characters, but conspicuously shorter 
 and thinner. 
 
 An epidemic occurred amongst cats at the Brown Institution. 
 Five out of fourteen died. The symptoms were, running from the 
 eyes, sometimes a muco-purulent discharge, sneezing, coughing, and 
 pulmonary trouble, resulting in emaciation and death in from one 
 to three weeks. After death lobular pneumonia and large white 
 kidney were found ; and in one case a diphtheritic condition of the 
 trachea, preparations of which showed diphtheria bacilli in crowds 
 under the microscope. 
 
 Klein regarded this disease as an epidemic of cat-diphtheria, 
 and believed that the disease was possibly induced accidentally by 
 the cats drinking milk, which was infected in the course of some 
 other experiments with diphtheria. He states that on account 
 of the very definite , results obtained by inoculating diphtheritic 
 membrane and cultures of the bacillus, subcutaneously and on 
 the cornea, and of the condition of the lung and kidney in cats 
 naturally or experimentally infected, the disease must be con- 
 si, lered as equivalent to human diphtheria, and the cat capable 
 of communicating the disease to other cats, and also to human 
 beings. These conclusions have not yet met with the acceptance cf 
 veterinary authorities. The results of the experimental inoculations 
 
 22 
 
338 INFECTIVE DISEASES. 
 
 fire certainly by no means conclusive. It does not follow from these 
 experiments that the disease diphtheria naturally occurs in the 
 cat or that under ordinary circumstances cats may contract the 
 disease from the human subject ; but the experiments show that, 
 like guinea-pigs and rabbits, cats are susceptible to the toxic effects 
 of the extremely poisonous principles developed during the growth 
 of Loffler's bacillus. 
 
 MILK DIPHTHERIA. 
 
 It has been shown that milk infected with diphtheria has been 
 the cause of epidemics among the consumers ; there have also been 
 epidemics apparently associated with the milk supply, in which it 
 has not been possible to trace the source from which the milk was 
 infected. A difficulty in tracing the origin in no way excludes the 
 possibility of contamination from a human source. In the light of 
 recent researches we should expect that it would be easy to overlook 
 the source of the virus, if it be true that diphtheria may exist without 
 .any symptoms indicating its presence, and be unrecognised until the 
 throat has been examined for diphtheria bacilli. As this fact was 
 unknown until quite recently, the absence of an acknowledged 
 case of diphtheria was taken as evidence that no diphtheria existed, 
 and consequently that the milk must have been infected by a, 
 diseased condition of the cow. Mr. Pow r er, whose views upon milk 
 scarlatina have already been referred to, endeavoured to trace the 
 origin of a milk epidemic to the very common disease of " garget," 
 or mammary abscess. This idea may be dismissed without further 
 consideration ; but the theory of some disease existing in the cow 
 capable of producing diphtheria in man was resumed by Dr. 
 Cameron, who suggested that there might be an eruptive disease of 
 the teats producing diphtheria, and by Mr. Power, who supported the 
 theory in an investigation of a milk-diphtheria outbreak in 1886 
 At Oamberley. Diphtheria in this case existed in the neighbourhood, 
 but as the source of human infection could not be traced, attention 
 was drawn to two cows in the herd which had recently calved, and 
 especially to one with chapped teats. Following this line of inquiry, 
 Klein investigated the behaviour of milch cows to the diphtheria 
 bacillus. Two cows were injected subcutarieously under the skin of 
 the shoulder with a Pravaz' syringe filled with a sub-culture in broth. 
 There was a rise of temperature, and on the third day a painful 
 tumour, which enlarged to the size of a child's head. In about a 
 fortnight the tumour began to decrease, and ultimately one cow 
 
MILK DIPHTHERIA. 339 
 
 died and the other was killed. Such results might have been 
 anticipated as the result of injecting a large quantity of the toxic 
 products of the bacillus, but certain other phenomena were observed 
 to which importance was attached. On the fourth day, on one of 
 the cows an eruption on the teat was first noticed, consisting of 
 small vesicles passing into pustules and crusted ulcers. Examina- 
 tion of the contents of the vesicle revealed the bacillus. With 
 matter from the vesicles and pustules two calves were inoculated, 
 and a similar vesiculation produced at the seat of inoculation. 
 The milk of the cows was inoculated on nutrient gelatine, and 
 produced a culture of Bacillus diphtheria. The question naturally 
 arose whether this eruption had any connection with the original 
 experimental inoculation. No other cows in the locality from 
 which these cows were obtained had a similar eruption, and it was 
 taken for granted that it was the result of the experimental inocu- 
 lation. By accepting the possibility of this eruption being identical 
 with the chaps on the teats of the Camberley cows, the theory was 
 gradually built up that cows suffer from diphtheria, which manifests 
 itself in the form of an eruptive disease of the teats, and that the 
 disease is conveyed in the milk to the consumers. 
 
 In the original experiment the bacilli were found to have 
 multiplied abundantly in the tumour at the seat of inoculation. The 
 eruption might have been, as admitted by Klein, a symptom of the 
 work of the chemical poison, and the elimination of the bacilli by the 
 milk is also possible ; but that there is in cows a vesicular disease of 
 the teats which is the origin of human diphtheria is not accepted by 
 veterinarians, and there is not sufficient evidence to justify the con- 
 clusion that the infectivity of the milk in epidemics of milk diphtheria, 
 has been proved to be due to a morbid condition of the cow. 
 
CHAPTER XXIV. 
 
 TYPHOID FEVER. 
 
 TYPHOID FEVER is a specific febrile disease peculiar to man, with 
 characteristic pathological lesions in the intestine, mesenteric 
 glands, and spleen. The Peyer's glands pass through three stages. 
 They become swollen from infiltration of round cells in lymph 
 follicles, due, it is supposed, to the presence of the typhoid fever 
 bacillus. The enlargement of the lymph follicles is followed by 
 coagulation necrosis until the entire patch becomes necrosed, and 
 sloughs away, leaving an ulcer. The disintegration of the patch 
 may extend in depth, and result in perforation and peritonitis, or 
 the ulcer may heal, and a pigmented scar take the place of the 
 Peyer's patch. The lymphatic glands are found more or less 
 enlarged, and may be easily felt in the groin, axilla, and neck. In 
 some cases there is a tendency to haemorrhage, followed by infarctions 
 in the spleen and lungs, which may develop into pysemic abscesses 
 In the mesenteric glands similar changes take place, but without 
 ulceration. Pneumonia may occur as a pulmonary complication. 
 The bacteria of pneumonia and Streptococcus pyogenes may be 
 found in association with the bacillus of typhoid fever. It is 
 now generally accepted that the disease is conveyed by water and 
 food which have become contaminated with the virus contained 
 in typhoid evacuations. This has been practically proved by the 
 number of cases which have been shown to have been intimately 
 connected with contamination of drinking water from wells and 
 other sources, by sewers, cesspools and faulty drains, the sewage 
 presumably having been infected with typhoid excreta ; but whether 
 sewage independently of typhoid contamination can originate 
 typhoid is still an open question. Accepting the former theory 
 as a working hypothesis, we must assume that a typhoid fever 
 bacillus exists in the intestinal evacuations, and that it must be able 
 to retain its vitality under very varying conditions until it gains 
 
 310 
 
TYPHOID FEVER. 
 
 341 
 
 access by the mouth to a fresh host, and by its development in the 
 intestine, and by the absorption of its toxic products, produces the 
 phenomena which we recognise as typhoid fever. 
 
 m 
 
 ;Mita 
 
 ' 
 
 FIG. 129.-TYPHOID FEVER. ILEUM OF ADULT, SHOWING SLOUGHY ANE 
 INFILTRATED PATCHES (HAMILTON). 
 
 Typhoid fever is also disseminated by milk ; sewage- contaminated 
 water having been added to the milk, or used for washing the i 
 cans and other vessels. 
 
342 INFECTIVE DISEASES. 
 
 Typhoid fever cannot be communicated to the lower ^ animals. 
 Numerous experiments have been made by feeding and by injecting 
 typhoid stools, but with absolutely negative results. Murchison 
 gave typhoid fever discharges to pigs, Klein experimented with 
 rabbits, monkeys, and other animals. Motschutkowsky injected the 
 blood from cases of typhoid into monkeys, rabbits, and other animals, 
 but with negative results. 
 
 FIG. 130. TYPHOID BACILLI FROM A COLONY ON NUTRIENT GELATINE, x 1000 
 (FRANKEL AND PFEIFPER). 
 
 Various micro-organisms have been described in typhoid, but the 
 one to which most importance is attached is a bacillus which was 
 first discovered by Eberth, but cultivated and fully described by 
 
 Gaffky. Gaff ky cultivated it from typhoid 
 evacuations, from typhoid ulcers, from 
 the mesenteric glands, and from the 
 spleen. It is found in scattered colonies 
 
 in the spleen, and is rarely if ever present 
 FIG. 131. TYPHOID BACILLI, . ,, f. , 
 x 950 (BAUMGARTEN). 
 
 Bacillus of Typhoid Fever Rods 
 
 1 to 3 /M, in length, and -5 to '8 ^ in breadth, and threads (Plate VIII., 
 Fig. 2). Spore-formation has not been observed, but the protoplasm 
 may be broken up, producing appearances which may be mistaken 
 for spores. They are actively motile, and provided some with a single 
 and others with very numerous flagella, which are from three to five 
 times as long as the bacilli. They stain well with aqueous solutions 
 
TYPHOID FEVER. 
 
 34S 
 
 of aniline d\vs. and grow well at the temperature of the room. In 
 plat (--cultivations minute colonies are visible in thirty-six to forty- 
 eight hours ; they are circular or oval, with an irregular margin ; they 
 appear granular by transmitted light, and are yellowish -brown in 
 colour. Cultivated in the depth of gelatine a well-defined shiny film 
 forms at the point of puncture, and a greyish -white filament, com- 
 1 ><>>-. I of closely packed colonies, develops in the track of the needle 
 (Fi<r. 134). On the surface of gelatine a greyish- white translucent 
 film forms, with sharply defined margin (Plate II., Fig. 2). On agar 
 there is a whitish transparent layer. They flourish in milk. On 
 potato at the temperature of the blood there is no culture visible, but 
 
 
 Fiu. 132. FLAGELLA OF TYPHOID BACILLI, x 1000, STAINED BY LOFFLEU'S 
 MKTHOD (FRANKEI, AND PFEIFFER). 
 
 the inoculated area appears moist and shining, and cover-glass pre- 
 parations made from the potato will demonstrate that there is really 
 a copious growth of the bacillus. This almost invisible growth is not 
 peculiar to this micro-organism. 
 
 Whether this bacillus is really peculiar to typhoid is much dis- 
 puted. Bacilli very closely resembling it, if not actually identical, 
 have been found under other conditions. These pseudo-typhoid bacilli 
 are regarded by some bacteriologists as \ari<-tir> resulting from the 
 different environment afforded by a saprophytic existence. Numer- 
 ous experiments have been made on animals with pure-cultures of 
 the bacillus, but in the production of typhoid fever they have 
 been no more successful than the experiments with typhoid stools. 
 
.344 
 
 INFECTIVE DISEASES. 
 
 Frankel and Simmonds inoculated a number of rabbits in the vein of 
 the ear, producing death, in some cases in forty-eight hours. Seitz 
 .administered broth -cultures by Koch's method of introducing them 
 
 into the stomach after 
 the administration of 
 opium in guinea-pigs, 
 and death resulted in 
 several instances. 
 But in all these cases 
 the results depended 
 upon the poisonous 
 products found in the 
 
 FIG. 133. COLONIES OF TYPHOID BACILLUS. 
 Three days old. x 100 (FRANKEL AND PFEIFFER). 
 
 cultivations, a similar result following the 
 injection of sterilised cultures. An account of 
 the products has already been given (p. 41). 
 
 Cassedebat isolated three species of bacilli 
 from water, which could be distinguished 
 with great difficulty, and only after the most 
 careful comparison. The bacillus which most 
 closely resembles it is the Bacillus coli com- 
 munis ; in fact, Roux regards it as a non- 
 pathogenic variety of the typhoid bacillus. 
 Others claim to be able to distinguish it by 
 careful comparison and the application of 
 tests. Special importance is attached to 
 potato cultures, the typhoid bacillus forming 
 an invisible film, and Bacillus coli communis 
 .a well-marked yellowish growth. Terni 
 pointed out that Bacillus typhosus retains its 
 motility in media containing hydrochloric 
 acid, while Bacillus coli communis and other bacilli resembling those 
 of typhoid lost their motility. Schild maintained that Bacillus typhosus 
 
 FIG. 134. PUEE- CULTURE 
 OF TYPHOID BACILLI 
 
 INOCULATED IN THE 
 
 DEPTH OF NUTRIENT 
 GELATINE (BAUMGAR- 
 
 TEN). 
 
TYPHOID FEVER. 345 
 
 was destroyed by exposure to the vapour of formalin, while Bacillus 
 coli communis and similar bacilli isolated from water gave subcul- 
 tures after exposure for two hours. Typhoid bacilli do not give the 
 reaction for indol, and there is no development of gas in cultures in 
 the depth of nutrient agar containing 2 per cent, of grape-sugar. 
 According to Muller, sterilised milk is coagulated in twenty-four 
 hours, at 37 C., by Bacillus coli communis, but not by the Bacillus 
 typhosus until several weeks have elapsed ; and, further, cultures on 
 acid potato give different results. The typhoid bacillus on micro- 
 scopical examination shows marked polar staining, but Bacillus 
 coli communis only shows a slight tendency of the protoplasm to 
 break up. 
 
 
 
 
 FIG. 135. TYPHOID BACILLI IN A SECTION OF SPLEEN, x 800 (FLUGGE). 
 
 Kitasato suggested the negative indol test, and recommended 
 Salkowski's method. Broth-cultivations are treated with a solution of 
 sodium or potassium nitrite : 1 cc. of the nitrite solution ('02 gr. to 
 100 cc. of water) is added to 10 cc. of a broth-culture after twenty- 
 four hours in the incubator, and on adding a few drops of strong 
 sulphuric acid, the typhoid cultures remain colourless, but cultures 
 of bacilli apparently identical give the red colour. On the other 
 hand, Losener maintains that he has cultivated from earth, water 
 and healthy human evacuations bacilli which could not be distin- 
 guished from typhoid bacilli by any of these tests. 
 
 The detection of the typhoid bacillus in water has been 
 
346 
 
 INFECTIVE DISEASES. 
 
 described in another chapter (p. 147) ; but sufficient has been said to 
 show that bacteriological reports in which it is stated that the typhoid 
 fever bacillus has been found in water causing typhoid epidemics 
 must be accepted with great reserve ; and further, no one is justified 
 in stating that the typhoid fever bacillus is undoubtedly the cause 
 of typhoid fever. It is not found in every case of typhoid, it is 
 not found in the blood, but it is found in those tissues which are 
 
 ;($ , $y^v ,T l& ^^ w (^^^ w 
 
 |^|^f^;r:;^,, 
 
 FIG. 136. TYPHOID BACILLI IN A SECTION OF INTESTINE, INVADING THE SUBMUCOUS 
 (T.J.) AND MUSCULAR LAYERS (M.), x 950 (BAUMGARTEN). 
 
 commonly the seat of secondary invasion of epiphytic bacteria, 
 whose normal habitat is the intestinal canal. 
 
 Lastly, as the disease does not exist in the lower animals, the 
 crucial test cannot be applied. The etiology of typhoid fever is 
 still enveloped in doubt, and the nature of the contagium has not 
 yet been determined. 
 
CHAPTER XXV. 
 
 SWINE FEVER. 
 
 PIG TYPHOID, or swine fever, is a highly contagious disease peculiar 
 to swine, causing death in from ten to thirty days, associated with 
 a fibriiious pneumonia, enlargement of, and haemorrhage into, the 
 lymphatic glands, and characteristic ulcers of the mucous membrane 
 of the stomach and intestines. The lesions may assume the form of 
 extensive croupous or diphtheritic deposit, which may fill the intes- 
 tinal tube. But the most characteristic appearance results when 
 the lower part of the ileum and commencement of the colon is 
 dotted all over with elevations of the mucous membrane, resembling 
 leather buttons or mix vomica seeds, and sometimes with concentric 
 rings, so that they have been compared to slices of calumba root. 
 
 Swine fever is difficult to detect in the early stage, and sometimes 
 symptoms are absent altogether in animals suffering from the 
 disease and quite capable of transmitting it ; or nothing may be 
 noted except cough, and possibly enlargement of the inguinal glands. 
 In typical cases the animals are noticed not to feed, to exhibit 
 dull less, and to have occasional rigors. Partial paralysis may 
 follow, producing unsteady gait or loss of power over the hind legs. 
 Diarrhoea sets in, and the evacuations become blood stained. There 
 is occasionally a diffused or patchy reddish or purplish rash on 
 the skin. After death the appearances most commonly found are 
 inflammation of the peritoneum, and redness and enlargement of the 
 n;- ntcric glands and the lymphatic glands in the lungs. There 
 is generally ulceration, especially of the colon and ileo-caecal valve, 
 or a diphtheritic exudation, sometimes pale yellow, more commonly 
 iriwish or Mack. >imilar to the centres of necro>i> within the ul 
 Tin- spleen is enlarged and liver congested, and there are haemorrhages 
 in the kidneys. As the lungs are so commonly affected, Klein 
 proposed the name pneumo-enteritis ; but the pulmonary lesions 
 aiv not constant. Indeed, the caflefl in whifh the intestines and 
 
 347 
 
348 
 
 INFECTIVE DISEASES. 
 
 lungs are simultaneously affected are not numerous, and sometimes 
 the lungs may be found to be perfectly healthy in cases with 
 
 YIG. 137. ULCERATION OF THE INTESTINE IN A TYPICAL CASE OF SWINE-FEVER. 
 
 long-standing lesion of the intestine. Old pigs may linger on for 
 weeks, and ultimately recover, and in the meantime act as centres 
 for the dissemination of the disease. 
 

DESCRIPTION OF PLATES IX. AND X. 
 Swine Fever. 
 
 PLATE IX. Part of intestine from a typical case of swine fever, showing 
 
 scattered ulcers and ulceration of the ileo-csecal valve. 
 PLATE X. From the same case of swine fever. The lungs were extensively 
 
 inflamed and partly consolidated, and the lymphatic glands were enlarged 
 
 and of a deep red or reddish-purple colour. 
 

 
 i A. .*. _. /^^^^^^^^_ 
 
 // : *33Hi^^ 
 
 FT 
 
 '* 3 
 
 
 
 
 SWINE FEVER 
 
 .-..:. v. 
 
SWINE FEVER. 
 
 Vincent Bmokf.Day * Son., J 
 
SWINE FEVER. 
 
 349 
 
 Biuld first pointed out that this disease might be compared 
 to human typhoid, both diseases being attended by a peculiar 
 ulceration of the intestinal follicles ; but the diseases are not to 
 be considered in any sense identical or interchangeable. 
 
 Bacteria in Swine Fever. 
 In 1877 Klein published a 
 ivM-aivh iii a Report to the Local 
 
 ff 
 
 FIG. 138. BACILLUS OF SWINK-FKVKK 
 No. 1. (KLKIX.) 
 
 Government Board, in which 
 
 he claimed to have discovered 
 
 bacilli characteristic of the 
 
 disease. They were described as 
 
 similar to Bacillus subtilis, or 
 
 Bacillus anthracis, but smaller 
 
 in size. These bacilli developed 
 
 into long leptothrix filaments, and formed spores. It was further 
 
 asserted that 011 inoculation, cultures produced lesions indicative of 
 
 swine fever ; the bacilli were also pathogenic in mice and rabbits. 
 
 Later this bacillus was re- 
 nounced in favour of another. 
 In the following year Det- 
 mers described a bacillus, but 
 subsequently renounced it in 
 favour of a micrococcus. 
 
 In 1882 Pasteur maintained 
 that the virus of swine fever in 
 France (rouget) was a dumb-bell 
 micrococcus, which produced the 
 same effect in pigeons as the 
 microbe of fowl-cholera. Though 
 
 rouget or swine measles is probably a different disease, the occurrence 
 
 of this micro-organism is of interest in this connection. 
 
 In 1883 Klein again investigated swine fever, and discovered 
 
 Bacillus No. 2, and maintained 
 
 that these bacilli were found in 
 
 the blood, in the peritoneal and 
 
 bronchial exudations ; and in the 
 
 air vesicles of the lungs, in the 
 
 FIG. 139. BACILLUS No. 2. FROM A 
 
 PRKPARATION OF BRONCHIAL Mrcrs 
 OF A PIG. (KLKIX.) 
 
 Fi;. 140. BACILLUS No. 2. FROM AX 
 ARTIFICIAL CULTURE. (KLKIX.) 
 
 form of leptothrix filaments ten 
 
 or twenty times the length of 
 
 single rods. Cultivations were 
 
 made on solid media. The organisms in these cultures were minute 
 
 rods actively motile, occurring singly or forming chains, two or three 
 
350 
 
 INFECTIVE DISEASES. 
 
 times as long as Bacterium termo ; and in preparations made from 
 diseased organs they were found to possess a very narrow trans- 
 parent halo, a sort of hyaline gelatinous capsule. Inoculation of 
 cultures failed to produce the lesions found in animals naturally 
 infected. Two pigs were inoculated, one with a sub-culture from the 
 swollen bronchial gland of a pig that had died of pig-typhoid, and 
 a second with a culture obtained from the spleen of a mouse that 
 had bean inoculated from another case of swine fever. After two 
 days the inguinal glands near the seat of inoculation became swollen, 
 and the temperature rose slightly. After three or four weeks the 
 animals recovered. 
 
 Mice on the fifth or sixth day after inoculation showed symptoms 
 
 of illness, then respiration became 
 superficial and slow. Death 
 occurred on the sixth or seventh 
 day. 
 
 Rabbits showed a rise of 
 temperature, and death followed 
 between the fifth and eighth days, 
 the temperature falling before 
 death. At the post-mortem ex- 
 amination there was usually 
 peritonitis, with copious exuda- 
 tion. The kidney, spleen, and 
 liver were enlarged and dark in 
 many cases, there was red hepa- 
 tisation of the lobes of the lungs, 
 and generally pericarditis and haemorrhage under the pericardium. 
 
 In 1885 Salmon, in the annual report of the United States 
 Bureau of Animal Industry, published the result of his investigations 
 into American hog cholera, which is identical with English pig typhoid. 
 A motile figure-of-eight bacterium was isolated, each part being 
 about twice as long as broad. The bacterium grew on nutrient 
 gelatine without liquefying it, and on potato produced a brownish 
 growth ; broth tubes became turbid on the following day. Colonies 
 in plate- cultivations were oval or circular, and brownish in colour. 
 Six pigs inoculated subcutaneously were all said to have died of hog 
 cholera, and the bacterium was again obtained from the blood of 
 the heart and spleen. The bacteria proved fatal to mice, rabbits, 
 guinea-pigs, and pigeons. 
 
 In 1893 Welch and Clement described the hog-cholera bacillus 
 as variable in form, and they further stated that a culture obtained 
 
 FIG. 141. BLOOD OF FRESH SPLEEN OF 
 A MOUSE, AFTER INOCULATION WITH 
 BACILLUS No. 2. (KLEIN. ) 
 
MVINE FEVER. 351 
 
 from Klein, while not possessing the characters originally ascribed 
 to it, could not in its form, biological characters, or pathogenic pro- 
 perties, be distinguished from the American hog-cholera bacillus. 
 
 In 1887 pig typhoid was investigated at Marseilles by Rietsch, 
 Jobert, and Martinaud, and a bacillus found. This grew rapidly 
 on all the nutrient media. In gelatine a growth was obtaine 1 i'n 
 twenty-four hours at 18 C. ; on blood serum and agar an opaque 
 growth developed; and on potato the growth was yellowish. It 
 was asserted that a young pig was killed by a culture in twenty- 
 two days, and that the characteristic ulcerations were observed in 
 the intestines. 
 
 In 1887 pig typhoid was prevalent in Sweden, and Bang and 
 Selander experimented with cultures from a rabbit that died after 
 inoculation with a fragment of spleen from a diseased pig. The 
 bacilli were motile, varying from rods to cocci, without spore-formation 
 and pathogenic in mice, guinea-pigs, and rabbits, but not in pigeons. 
 Pigs fed on broth-cultures were said to have succumbed to genuine 
 pig typhoid. In the blood they were generally found in the form 
 of short oval bacteria, but in the blood of the heart longer rods 
 were sometimes found. Metchnikoff described a bacillus isolated by 
 Chantemesse from an outbreak in France, as highly polymorphic. 
 
 Smith identified the hog-cholera bacillus with the bacillus found 
 by Schiitz, and this in turn has been identified with the bacillus 
 of haemorrhagic septicaemia. 
 
 From these researches it would appear to be probable that one of 
 the bacteria isolated by Klein, and those found by Salmon, Smith, 
 Bang, Welch and Schiitz are identical ; and further, that they have 
 been identified with the bacillus of haemorrhagic septicaemia. We 
 may sum up the characters thus : 
 
 Bacillus of Klein, Salmon, Smith and Schtitz. Very small 
 rods, actively motile ; spore-formation not observed ; colonies circular 
 and brown by transmitted light. Inoculated in the depth of gelatine 
 faintly yellowish- white colonies develop along the track of the needle ; 
 on the surface an opalescent film ; on potato they produce a straw- 
 coloured layer, changing to brown. There is absence of indol in 
 cultures containing peptone; the bacilli are fatal to mice, guinea- 
 pigs, rabbits, and pigeons. Swine die after intravenous injection, 
 but not, as a rule, after subcutaneous injection. 
 
 According to Caneva, the bacillus obtained from the Marseilles 
 epidemic would appear to be closely allied, if not identical, with 
 the bacillus of ferret disease (Eberth and Schimmelbusch), and the 
 bacillus of Texas fever (Billings). 
 
352 INFECTIVE DISEASES. 
 
 Billings appears to have isolated two bacilli, one identical with 
 the Marseilles bacillus, and the other with the hog-cholera bacillus. 
 
 Bacillus of Rietsch. and Jobert. Rods about twice as- 
 long as broad, rather shorter than the bacillus of typhoid fever, 
 longer and thicker than the American bacillus. They exhibit 
 end-staining. They possess flagella, and are actively motile. 
 
 They grow rapidly in nutrient media. They are only feebly 
 pathogenic. They are also said to be distinguished from the bacilli 
 of hog-cholera by producing indol in solutions containing peptone, 
 and by causing an acid reaction in milk. 
 
 Bacillus of M'Fadyean. M'Fadyean investigated swine fever 
 in 1895, and found bacilli which he differentiated from hog-cholera 
 bacilli. The method employed was to inoculate the surface of 
 nutrient agar and of potato, with fragments torn out of the centre 
 of a lymphatic gland, with specially constructed forceps. Inocula- 
 tions were also made from the spleen pulp, and blood, in the usual 
 way. They are from 1 to 2 /A in length, and '6 /A in breadth. 
 They never grow into filaments, they do not form spores, and 
 they are actively motile. They are readily stained by the watery 
 solutions of the aniline dyes, and are decolorised by Gram's- 
 method ; with methylene-blue they show end-staining. The bacilli 
 grow on gelatine without liquefaction, forming a thin white line- 
 along the needle track. On agar a thin, transparent pellicle forms, 
 which is not easily visible at first, but gradually acquires a faint 
 greyish tint. More characteristic appearances result in plate- 
 cultivations of gelatine-agar, at 37 C. The colonies are distinctly 
 visible in eighteen hours, appearing when viewed by transmitted 
 light as bluish-white, circular specks ; each colony has a dark centre- 
 arid a granular margin. In broth the bacilli produce turbidity after 
 twenty-four hours. On potato there is no visible growth, even when 
 the surface is inoculated with an abundance of material. On solid 
 blood serum the growth is scanty. They grow in milk without 
 producing coagulation. They are harmless to guinea-pigs and feebly 
 pathogenic to rabbits. 
 
 Several experiments were carried out upon swine. In the first 
 series the most rigid precautions were taken to prevent accidental 
 infection with swine fever. Four young pigs were inoculated, upon 
 a farm where there was no previous history of the disease. These 
 pigs were killed, and the post-mortem examinations were said to 
 show indications of swine fever, principally patches of diphtheritic 
 material in the colon, and healing ulcers. The next series of pigs 
 inoculated at the Royal Veterinary College. Cultures were 
 
KKYKK. ;;;,;; 
 
 admini>tere.l with milk to two pigs. Five days afterward* on,- ^ 
 .lied; and the mesenteric glands were congested, and the mu 
 membrane showed spots of necrosis. The other pig was killed, an. I 
 there were ulcers in the colon. In a third experiment, eight pii:> 
 were fed with milk and broth cultures. These pigs were all killed 
 at different dates, and most of them had ulceration of the colon 
 in control experiments the intestine was normal. 
 
 M'Fadyean compared his bacillus with a culture of the hog- 
 cholera bacillus, and found that the American organism grows at a 
 lower temperature in gelatine, and colonies appear in plates much 
 earlier. They produce a less transparent and thicker growth, and 
 much greater turbidity in broth and a more abundant sediment. 
 On potato they form an abundant growth at 37 C., at first 
 yellow, later brown, with considerable resemblance to a glanders 
 culture. 
 
 Colonies upon gelatine-agar are distinguished by their opacity 
 and sharp outline. Agar, potato, and broth cultures of the American 
 organism consist of short ovoid forms like the bacilli of fowl cholera, 
 while the bacillus isolated by M'Fadyean has a closer resemblance 
 to the bacillus of glanders. M'Fadyean asserts that the American 
 organism is not pathogenic to the pig. Pigs after feeding on broth 
 cultures remained healthy, and showed no trace of swine fever when 
 killed from one to three weeks afterwards. On the other hand, 
 broth cultures of his bacillus produced the characteristic ulceration 
 of the bowel. M'Fadyean claims, therefore, to have discovered the 
 true pathogenic organism of swine fever. He does not appear to 
 have compared this bacillus with that obtained from the epidemic 
 of swine fever at Marseilles. From the description of the mor- 
 phological and other details there seems to be a close resembl 
 between the two. 
 
 Not less than three and possibly four species of bacilli have been 
 cultivated from swine fever, two at different times by Klein, one 
 ly Reitsch Jobert and Martinaud, and one by M'r'adyran : and 
 cultures of all these bacilli have been credited with producing swim- 
 fever in experimental animals, and each one has been pronounced 
 t< be the contagium of the disease. We must conclude either that 
 contaminated cultures were inoculated in some cases, or, uli.-'t M 
 far more probable, the swine fever which resulted in experimental 
 animal- was due to accidental infection; and until a bacillus has 
 been cultivated from swine fever from which another investigator 
 can prepare sub-cultures, and with those sub-cultures produce t lie 
 typical ulcerations of swine fever in pigs on a farm, or on premises 
 
354 INFECTIVE DISEASES. 
 
 in which swine fever is unknown, we are justified in concluding that 
 the contagium has not yet been discovered. 
 
 The mistakes which are likely to occur when the same investi- 
 gator isolates bacilli from cases of swine fever, and subsequently 
 inoculates or feeds healthy swine, cannot be better illustrated than 
 by quoting from a leaflet issued by the Board of Agriculture, 
 pointing out the great precautions necessary to prevent accidental 
 infection. 
 
 "There seems reason to believe that the disease is not infre- 
 quently introduced by means of persons who have been in contact 
 with diseased animals. Pig owners, therefore, are advised to prevent 
 strangers from at any time approaching their pigs, and should the 
 admission to the premises of spayers or castrators be necessary, 
 those persons should be required, before approaching the animals, to 
 thoroughly wash their hands with soap and water, and to wash and 
 disinfect their boots with a solution of carbolic acid and water, or 
 some other suitable disinfectant. Such persons might also with 
 advantage be required to wear, while operating, a waterproof apron, 
 which should be washed and disinfected before the wearer is per- 
 mitted to approach the animals to be operated on." 
 
 Protective Inoculation. The experiments of Salmon and of 
 Schweinitz have been referred to in another chapter (pp. 41, 46). 
 A method of protective inoculation was attempted in America, but 
 the experiments were unsuccessful, and the plan was abandoned. 
 
 Stamping-out System. Notification is compulsory, and the 
 order in force is the Swine Fever Order of 1896, but the stamping- 
 out system has not been applied in a thoroughly satisfactory manner, 
 .and the disease is still very prevalent. 
 
CHAPTER XXVI. 
 
 SWINE MEASLES. DISTEMPER IN DOGS. EPIDEMIC DISEASE OF 
 FERRETS. EPIDEMIC DISEASE OF MICE. 
 
 SWINE MEASLES. 
 
 SUINE MEASLES, or swine erysipelas, is described as an acute, in- 
 fectious disease of swine which is very prevalent in France and 
 Germany, but is included in this country in the term " swine fever." 
 According to some, it is a distinct disease, and distinguished from i>iV 
 typhoid by absence of the ulceration of the intestines which i 
 characteristic of that disease ; while, according to others, ulceration 
 of the intestine and ileo-ccecal valve may be found post-mortem. The 
 onset of the symptoms, as in pig typhoid, is very rapid ; the anim.-iN 
 cease to feed, and show other general signs of illness ; the voice is 
 hoarse, and there is a rapid rise of temperature. On the neck, 
 chest, and abdomen, red patches make their appearance, which 
 extend and coalesce, and change to a dark reddish or brownish colour. 
 These symptoms may be followed by convulsions, and sometimes by 
 paralysis of the hind legs; and death occurs in from one to four 
 days. It is especially a disease of young pigs, and from 50 t< 
 per cent, of infected animals die. 
 
 On post-mortem examination there is haemorrhage and n-di-ma 
 in the patches of the skin, the lymphatic glanl> are >wll*-ii and 
 dark red, the peritoneum is ecchymosed, the intestinal mumus 
 membrane is congested and swollen, and the solitary follicles and 
 Peyer's patches are prominent, and in the neigh bourhood of the il<-.> 
 ccecal valve there are, according to Fliigge, ulcers of con.-idfiahl.- >i/.-. 
 The liver and spleen are congested and enlarged. Pasteur in 
 gated swine measles or rouyet, and described a figure-of-eight micn.- 
 coccus, which he believed to be the contagium of thi> di-e. 
 organism appears to be identical with the hacu-riuni "t hsi-n 
 septicaemia, which is also commonly found in pig-typhoid 
 
 In experimenting with the- virus obtained fnun the sple.-n 
 
 355 
 
356 INFECTIVE DISEASES. 
 
 Pasteur found that, by successive inoculation of rabbits, the virulence 
 was exalted for rabbits, but attenuated for swine, and the virus 
 which had thus been passed through the rabbit was used as a 
 vaccine for swine, to protect them against virulent erysipelas. 
 
 Pasteur found that by passing the virus through pigeons it was 
 made more virulent for swine. 
 
 In the blood, and the juice of the internal organs, and of the 
 lymph glands, Schiitz found a minute bacillus identical with the 
 bacillus of mouse septicaemia. 
 
 Bacillus of Swine Erysipelas (Schiitz). Extremely minute 
 rods -6 to 1*8 /A in length, morphologically and in cultural charac- 
 ters identical with the bacillus of mouse septicaemia. Filaments 
 and involution forms. Spore-formation present. 
 
 House mice if inoculated with a pure culture die in two to four 
 days. Pigeons are also very susceptible. Fowls and guinea-pigs 
 
 FIG. 142. BACILLI OF SWINE FIG. 143. BLOOD OF PIGEON INOCULATED 
 ERYSIPELAS (BAUMGARTEN). WITH BACILLI OF SWINE ERYSIPELAS, x 600 
 
 (SCHUTZ). 
 
 are immune. Rabbits after inoculation of the ear suffer from 
 erysipelatous inflammation, identical with that produced by inocu- 
 lation of the bacillus of mouse septicaemia. The bacilli are also 
 pathogenic in swine and sheep. 
 
 Protective Inoculation. With Pasteur's vaccine immunity is 
 xi id to be produced which lasts about a year. Schiitz and Schottelius 
 found the minute bacilli in Pasteur's vaccine, which they had already 
 found in cases of swine erysipelas in Germany. 
 
 The results of vaccination in France are said to be very satis- 
 factory, but in test experiments in Germany they were riot so 
 favourable. Out of 119 vaccinated swine 5 per cent, died as the 
 result of the inoculation, while the average loss in the ordinary way 
 is 2 per cent. 
 
 Metchnikoff found that the blood of immunised rabbits was 
 antitoxic, and Lorenz maintains that the serum of swine which 
 
SWINE Fl-:\KR. 
 
 357 
 
 have recovered from swine erysipelas is also antitoxic, and will 
 produce immunity in other animals. The treatment introduced 
 by Lorenx is to inject serum in the proportion of 1 cc. to every 
 10 kilogrammes of the weight of the animal's body. Two days 
 afterwards -5 to 1 cc. of virulent culture is injected, and 
 twelve days the dose is doubled. Lorenz inoculated 294 
 12 were suffering from swine erysipelas, and 
 of these 6 recovered and 6 died. 
 
 In the opinion of the author this disease 
 requires re-investigation, for if it be true that 
 rowjet or schweinrothlauf is associated with 
 ulceration of the intestines, the recognition 
 of it as a disease distinct from our English 
 swine fever apparently rests upon the pres- 
 ence of a bacillus, which cannot be distin- 
 guished from the bacillus of mouse septicaemia. 
 The question arises whether this bacillus 
 is really the cause of a distinct disease, swine 
 erysipelas, or, on the other hand, whether 
 the bacillus is really the bacillus of mouse 
 septicaemia which has been isolated from 
 certain cases of swine fever. The bacillus of 
 mouse septicaemia is widely distributed, and 
 it may only be an accidental concomitant in 
 rouyet or scltwe'mroildauf. The presence of 
 the bacterium of haemorrhagic septicaemia in 
 both rouget and pig typhoid would not prove 
 identity, as this micro-organism is un- 
 doubtedly only secondary in both diseases. 
 There is great need, therefore, for further 
 careful investigation. Clinical and patho- 
 logical observations must be made in this 
 country, to determine whether there are 
 really two dix-ases included under the name 
 
 "swine fever." If this prove to be the case, we must ax-ertain th- 
 clinical and pathological differences between rouget and pig typhoid. 
 How can rouget be distinguished from cases of swine fever in which 
 there is a patchy rash, paralysis of hind legs, but no ulceration of the 
 intestine ? Further, how is swine erysipelas with ulceration of the 
 intestine and ileo-cojcal valve to be distinguished from an onlin.ny 
 case of pig typhoid ? 
 
 FIG. 144. PuuK-cri.Ti UK 
 
 IN XUTKIKXT (iKI.A- 
 
 TINE OF BACIM.I 
 
 IN SWIXK Kl<VSIl'KI.A> 
 
 (BAUMGARTEN). 
 
358 INFECTIVE DISEASES. 
 
 DISTEMPER IN DOGS. 
 
 Distemper is an infectious febrile disease of dogs, characterised by 
 bronchial catarrh and discharge from the eyes. Bronchitis and 
 pneumonia may supervene, or there may be intestinal catarrh ter- 
 minating in dysenteric diarrhoea, sometimes complicated by jaundice. 
 The disease may affect the nervous system, and produce convulsive 
 contractions of the muscles of the nose, ears, lips, and limbs. 
 Occasionally there is an eruption, especially in animals which are 
 out of condition. The virus exists in the discharge from the nostrils 
 and eyes, and is given off from the lungs and the skin. 
 
 One attack of the disease does not confer entire immunity ; 
 and some dogs are completely insusceptible. 
 
 Bacteria in Distemper. Millais has isolated a micro-organism 
 resembling the pneumococcus of Friedlander, which he believes to be 
 the cause of the disease. The bacillus occurs with other bacteria and 
 micrococci in the nasal discharge. 
 
 Protective Inoculation. Mixed cultures of these bacteria 
 liquefy the gelatine, and the liquid has been used as a vaccine. 
 When applied to the nose, it is said to produce a mild attack of 
 distemper, which protects as much as an attack of the disease 
 contracted naturally. These results require confirmation. 
 
 Inoculation of the nasal discharge in healthy dogs has been 
 practised, so that they may have the disease under favourable con- 
 ditions ; but the system should not be encouraged, as dogs need not 
 necessarily contract distemper. Vaccination with cow-pox lymph 
 has been advocated, but it is perfectly useless. 
 
 Stamping-out System. Dogs suffering from distemper must be 
 completely isolated. Any straw or litter which has been in contact 
 with a diseased dog should be burnt. Clothing, collars, chains, and 
 the kennel or premises inhabited, must be thoroughly disinfected. 
 The animal after recovery should be washed with carbolic soap. 
 
 EPIDEMIC DISEASE OF FERRETS. 
 
 Eberth and Schimmelbusch investigated an epidemic disease of 
 ferrets (frettchen-seuche), and isolated a bacillus, which in mor- 
 phology and cultivation agrees very closely with the bacillus of 
 htemorrhagic septicaemia. 
 
 EPIDEMIC DISEASE OF MICE. 
 
 Loffler investigated an epidemic disease which occurred in mice 
 kept in confinement, and isolated a bacillus resembling Bacillus 
 typhosns. 
 
EPIDEMIC DISEASE OF Ml< 359 
 
 Bacillus Typhi Murium. Rods varying in length ; and fila- 
 ments; motile; flagellated. The colonies are circular, brownish 
 and granular on the surface of obliquely solidified gelatine. The 
 bacteria inoculated on the surface produce a greyish-white semi- 
 transparent growth, and on agar and potato the appearance of the 
 growth is very similar. They can be cultivated readily in milk and 
 in broth. White and field mice are killed in from one to two 
 weeks, when given bread moistened with a culture. 
 
 Lbffler claims to have used this method with success in Thessaly, 
 where there was a plague of field mice causing great losses to 
 agriculturists. 
 
CHAPTEE XXVII. 
 
 ASIATIC CHOLERA. CHOLERA NOSTRAS. CHOLERAIC DIARRHOEA 
 FROM MEAT-POISONING. DYSENTERY. CHOLERAIC DIARRHOEA 
 IN FOWLS. 
 
 ASIATIC CHOLERA. 
 
 THERE are several diseases in man associated with diarrhoea, which 
 have certain characters in common, but are totally distinct. They 
 include Asiatic cholera, cholera nostras, dysentery, and choleraic 
 diarrhoea. Asiatic cholera is an endemic disease of the Delta of 
 the Ganges, a locality which has become notorious as the home 
 of cholera. Cholera is a filth disease ; and the accumulation .of 
 filth on the banks of the Ganges, with contamination of the 
 water, and the climate, afford most favourable conditions for the 
 development of the cholera virus. 
 
 Four great cholera epidemics have originated in, and spread 
 from, India : in 1817, in 1826, in 1846, and in 1865. Cholera 
 follows the routes of pilgrims and caravans, and now, owing to 
 the rapid means of communication by steamers and railways, it 
 spreads to the most distant parts of the world, covering in a few 
 weeks or days distances which in former times could only be traversed 
 in several months or even years. 
 
 In 1892 the epidemic passed from India, through Afghanistan, 
 to Russia in Asia, and quickly spread westwards along the route 
 of the trans-Caspian railway ; and all this occurred within the space 
 of a few weeks. By Russian emigrants it was carried to Hamburg 
 and Antwerp ; and the virus, finding a suitable environment in the 
 former place, produced a severe epidemic there. Thus, in about three 
 months, it was brought into close proximity with England. Mecca 
 is one of the great infective centres of the world, for there all 
 the conditions are found for the propagation of cholera, including 
 filth, overcrowding, and the water of the famous Holy Well, which 
 is used for ablutions and drinking purposes. The return of 
 
 360 
 
ASIATIC THOLERA. 361 
 
 the pilgrims to Egypt, and the proximity of England to Egypt, 
 necessitate the greatest possible precautions to prevent the intro- 
 duction of the disease into this country. 
 
 In 1884 a German Commission was sent out to India, and Koch 
 discovered a micro-organism which he described as a curveil or 
 comma-shaped bacillus, and pronounced to be the contagium of 
 this dise; or. 
 
 , 
 
 FIG. 145. COVER-GLASS PREPARATION* OF A DROP OP MEAT INFUSION, containing 
 a pure-cultivation of comma-bacilli, with (a) spirilliform threads, x 600. (Kocn.) 
 
 Spirillum cholerse Asiatic (Comma-bacittw, Koch). Curved 
 rods, spirilla, and threads. The curved rods or commas are about 
 half the length of .a tubercle-bacillus. They occur isolated, or 
 attached to each other forming S'skiP 6 ^ organisms or longer 
 screw-forms, the latter resembling the spirilla of relapsing fever. 
 
 FIG. 140. ARTHROSPORES ; () Comma-bacillus breaking up into spheres; (b, r), 
 formation of spheres in spiral forms; (d, e\ groups of spheres; (/) .spirilla 
 with spheres from an old cultivation; (g) germination of UK- spli.-n-. 
 
 (HUEPJ'K.) 
 
 Finally they may develop into spirilliform threads. In old cult 
 tions threads are found with swellings or irregularities (Fig. 1 
 The commas are actively motile, and possess flagella (Fig. 147). 
 Their movements) and development into spirilla may be stu<li<><l in 
 drop-cultivations. Arthrospore formation has been described by 
 Hueppe (Fig. 146). In plate-cultivations, at a temperature of from 
 
362 
 
 INFECTIVE DISEASES. 
 
 16 to 20 C., the colonies develop as little specks, which begin to 
 be visible after about twenty -four hours. Examined with a low 
 power, and a small diaphragm, these colonies have the following 
 characteristics. They appear as little masses, granular, and 
 
 FIG. 147. FLAGELLA OF COMMA-BACILLI ; STAINED BY LOFFLER'S METHOD 
 (FRANKEL AND PFEIFFER). 
 
 yellowish-white in colour, and sometimes very faintly tinged with 
 red, which have liquefied the gelatine, and sunk down to the bottom 
 of the resulting excavations. 
 
 In test-tubes of slightly alkaline nutrient gelatine (10 per cent.), 
 
 (X, 
 
 FIG. 148. INVOLUTION FORMS, x 700 FIG. 149. COLONIES OF COMMA-BACILLI 
 (VAN ERMENGEM). ON NUTRIENT GELATINE, NATURAL 
 
 SIZE (KOCH). 
 
 the appearance of the growth is very striking. In typical cultures 
 it begins to be visible in about twenty-four hours. Liquefaction 
 sets in very slowly, commencing at the top of the needle track 
 
ASIATIC CHOLERA. 
 
 368 
 
 around an enclosed bubble of air, and forming a funnel continuous 
 with the lower part of the growth ; the latter preserves for several 
 lays its resemblance to a white 
 thread (Plate II., Fig. 1). In 
 about eight days, however, lique- 
 faction takes place along the 
 whole of the needle track. 
 
 On the surface of agar-agar 
 the cultivation develops as a 
 white, semi-transparent layer, 
 with well-defined margin. The 
 
 appearance on blood serum is very similar; liquefaction very slowly 
 takes place. In broth they form a wrinkled film on the surta< . 
 there is a rapid and abundant growth at the temperature of tin- 
 
 FIG. 150. COLONIES OK KOCH'S COM M.\ 
 BACILLI, x 60. 
 
 ct 
 ** 
 
 f'tir 
 
 j 
 
 !,\" j > .'/ 
 //VS , " 
 l'^ ^ 
 
 ^f t;' ; "'^ft 
 
 Mftrfr^wfl 
 
 )^>//j( ^ ' 
 
 FIG. 151. COVER-GLASS PREPARATION 
 FROM THE CONTENTS OF A CHOLERA 
 INTESTINE, x 600. (a) Remains of the 
 epithelial cells ; (b) Comma-bacillus ; 
 (c) Group of comma-bacilli (Koch). 
 
 FIG. 152. COVER-GLASS PRKPAK 
 OF CHOLERA DEJECTA ON DAMP I 
 (two days old), x 600. Great prolife- 
 ration of the bacilli with spirilla (a) 
 (Koch). 
 
 blood, and the same applies to sterilised milk ; and they will even 
 multiply in sterilised water. In potato-cultivations tin- micro!*- 
 will only grow at the temperature of the blood (37 C.), forming a 
 slightly brown, transparent layer. Inoculation of a cultivation of 
 the bacillus in the duodenum of guinea-pigs, with and without 
 
364 
 
 INFECTIVE DISEASES. 
 
 ligation of the bile-duct, has given positive results. More recently 
 these results have been confirmed by the following method : Five 
 cc. of a 5 per cent, solution of potash were injected into the 
 stomach of a guinea-pig, and twenty minutes after, 10 cc. of a 
 cultivation of comma -bacilli, diffused in broth, were similarly intro- 
 duced. Simultaneously with the latter, an injection of tincture of 
 opium was made into the abdominal cavity, in the proportion of 
 1 cc. for every 200 grammes weight of the animal. Those who 
 have had success with inoculation experiments maintain that choleraic 
 symptoms were produced without any trace of peritonitis or putrid 
 infection, and that the comma-bacilli of Koch were again found 
 in the intestinal contents, and fresh cultivations established. 
 
 
 FIG. 153. SECTION OF THE Mucous MEMBRANE OF A CHOLERA INTESTINE, x 600. 
 A tubular gland (a) is divided transversely ; in its interior (b) and between 
 the epithelium and the basement membrane (c) are numerous comma-bacilli 
 (Koch). 
 
 On the other hand, these results have been disputed, the fatal 
 effects of the inoculation attributed to septicsemic poisoning, and 
 the proliferation of the bacilli considered to be dependent upon an 
 abnormal condition of the intestines, induced by the injection of 
 tincture of opium. It has, however, been shown that these organisms, 
 like several others which have been isolated from intestinal dis- 
 charges, produce definite poisonous substances. The comma-bacilli 
 were found in the superficial necrosed layer of the intestine, in 
 the mucous flakes and liquid contents of the intestinal canal of 
 cases of Asiatic cholera. It is stated that they were also detected 
 
ASIATIC CHOLERA. 
 
 in a tank which contained the water supply of a neighbourhood 
 where cholera cases occurred; but commaihaped organisms are 
 frequently present in sewage-contaminated water. Korh> comma- 
 bacilli are aerobic, and their development is arrested by deprivation 
 of oxygt^ are destroyed by drying on a cover glass, but 
 
 retain their vitality longer when dried on silk threads. Cultures 
 are sterilised by exposure for fifteen minui c., and l.v 
 
 various antiseptic substances. 
 
 FIG. 154. PCKE-CULMVATIOXS ix NUTRIENT GELATINE. , KOCH'S CHOUEKA 
 BACILLUS, twenty-four hours old. s FIKKLEE'S BACILLUS, twenty-four 
 
 :- : .. 
 
 METHODS .} STAINING THE COMMA-BACILLI OF KOCH. 
 
 In cover-glass preparations they may be well stained in the ordinary 
 way, with an aqueous solution of methyl-riolet or fuchsine, or by the 
 rapid method, without passing through the flame (p. 85, Babes' method). 
 
 A small quantity of the stools, or of the scraping of the intestinal 
 mucous membrane, is spread out on a glass slide and dried, then steeped 
 during some seconds in sublimate solution, or in osmic acid (I to 100). 
 It is then stained by immersion in fnchsine-aniline solutioi. 
 grammes of Bale fuchsine dissolved in a saturated aqueous solution of 
 aniline), washed, dried, and mounted in Canada balsam. 
 
366 INFECTIVE DISEASES. 
 
 In sections of the intestine their presence may be demonstrated by : 
 
 (a) Koch's method. 
 
 Sections of the intestine, which must be well hardened in absolute 
 alcohol, are left for twenty-four hours in a strong, watery solution of 
 methylene-blue, or for a shorter time if the solution is warmed ; then 
 treated in the usual way. 
 
 (&) Babes method. 
 
 Sections, preferably from a recent case of cholera, and made as soon as 
 possible after death, are left for twenty-four hours in an aqueous solution 
 of f uchsine, then washed in distilled water, faintly acidulated with acetic 
 acid, or in sublimate solution 1 in 1000, passed rapidly through alcohol, 
 and finally treated in the usual way. 
 
 Klein investigated cholera in India, and does not accept Koch's 
 conclusions. With regard to the inoculation experiments, Klein 
 believes that the living choleraic comma -bacilli, even if introduced 
 in large numbers into the small intestine, are quite innocuous, 
 but capable of great multiplication if the intestine is previously, 
 from some cause or another, diseased ; the chemical products of the 
 comma- bacilli then act as poisons analogous to the ptomaines 
 obtained from other putrefactive bacteria. The observations made 
 by Roy, Brown, and Sherrington, in Spain, tended to confirm 
 Koch's views. Comma-bacilli were found to be present, in some 
 cases, in enormous numbers, and the frequency of their occurrence 
 led these observers to believe that they must bear some relation to 
 
 the disease. At the same time, 
 as they failed to find them in all 
 cases, they regarded the existence 
 of a causal relation as not proven. 
 They failed to find the Naples 
 bacterium, or a small, straight 
 
 *^K& ^ 
 
 bacillus noted by Klein ; and 
 they drew attention to certain 
 FIG. 155. -COMMA-SHAPED ORGANISMS peculiar mycelium -like threads 
 
 WITH OTHER BACTERIA IN SEWAGE- 
 CONTAMINATED WATER, x 1200. m the mucous membrane of the 
 
 intestines ; but these cannot be 
 
 considered to have any significance. Methylene-blue has been 
 employed by Koch and others, including the author, for staining 
 sections of the intestine from cholera cases, and had they been 
 constantly present, it is hardly possible that such striking objects 
 could have been overlooked. Again, we must bear in mind that 
 hypho-mycetous fungi occasionally have been found to occur sapro- 
 phytically in the intestinal canal, as well as in the lungs, external 
 auditory meat us, and elsewhere. Cunningham, of C Icutta, maintains 
 
ASIATIC CHOLERA. 
 
 
 that Koch's comma -bacilli are not constantly found; and that 
 the comma-bacilli obtained from typical cholera cases show I 
 great variation in cultivation, and cannot be distinguished from 
 comma-bacilli from other sources. 
 Cunningham asserts that 
 comma-bacilli resembling Koch's 
 are found in the intestine in 
 health. Sternberg, on the other 
 hand, made a number of examina- 
 tions of the evacuations of yellow 
 
 fever patients and healthy indi- ^^^^-BAoauoraBMora, 
 
 1 * x 700 (VAN ERMENGEM). 
 
 victuals, and failed to find anv 
 
 micro-organism resembling the cholera spirillum. 
 
 Various comma-bacilli have been isolated from different s. 
 and compared with Koch's comma-bacillus. Comma-bacilli have been 
 
 found in the mouth by Le\\ i> ; 
 
 ( jty, in cholera nostras by Finkler and 
 
 Prior ; in cheese by Deneke ; 
 in hay infusion and sewage hv 
 Weibel ; in the intestines of fowls 
 by Gamaleia, and in water hy 
 Sanarelli. 
 
 Whether the comma -ha (-ill us 
 
 i> the cause of cholera ) 
 
 not, its detection is an aid in 
 
 ~* f/ ^ ^AvN^-t^V-. 
 
 >\ cC^-^-^-^d. 
 
 ,$ %p&&* 
 
 FIG. 157. FINKLER'S COMMA-BACILLI ; 
 FROM CHOLERA NOSTRAS, x 700 
 (FLUGGE). 
 
 diagnosis. If we are dealing 
 with a case alleged to be one of Asiatic cholera, and a micro- 
 organism is found in the intestinal evacuations, which can be 
 differentiated from the comma-bacillus described by Finkler in 
 cholera nostras, and identified 
 
 with the comma -bacillus de- 
 
 ^ 
 
 scribed by Koch, we are justified 
 in regarding the case as one of 
 Asiatic cholera. But we cannot 
 diaL r ii(- Koch's comma -bacill u >, 
 with certainty, unless we know 
 
 the source of the culture. The Fj( . m _ DEXEKE > 8 COMMA-HAHM... 
 
 I--K..M f'HKESE, x 700 
 
 ^ \ 
 
 clinical symptoms of cholera in 
 
 man, and especially the presence 
 
 of rice-water stools, must be taken into account, together with 
 
 the biological, morphological, and chemical cha, 
 
 the bacilli which are found to be present. There are several 
 
368 INFECTIVE DISEASES. 
 
 chemical tests which can be applied to cultures. According to 
 Frankel, the Bujwid- Dunham test can be relied upon to distin- 
 guish Koch's comma-bacillus from the comma-bacillus of Finkler - 
 Prior (cholera nostras), and from those found by Gamaleia. The 
 comma-bacilli are inoculated in broth containing peptone, and, 
 after twelve hours in the incubator, a drop of strong sulphuric 
 acid added to the culture will produce a red colour, owing to the 
 presence of indol. A test which distinguishes Koch's comma - 
 bacillus from Finkler- Prior's and Deneke's was introduced by Cahen. 
 A solution of litmus is added to the broth, and the culture placed 
 in the incubator, until the following day ; in the case of Koch's 
 commas, the colour will have disappeared. 
 
 Koch points out that in the bacteriological diagnosis of cholera 
 the first step is to examine the mucus in the evacuations, or in the 
 intestine if the examination is made after death. Cover-glass 
 preparations should be stained with dilute Ziehl-Neelseii solution. 
 Cultures are next made in peptone, and in eight hours will give the 
 indol reaction. In twenty-four hours the colonies may be examined 
 on plate-cultivations. The peptone cultures are prepared by adding 
 a trace of the choleraic evacuations, or of mucus containing the 
 bacilli, to a sterilised 1 per cent, solution of peptone, with *5 to 1 
 per cent, of common salt. The solution must be alkaline, and the 
 culture is placed in the incubator at 37 C. The pathogenic effects 
 can be ascertained by diffusing the bacilli from an agar- culture in 
 broth, and injecting it into the peritoneal cavity. 
 
 Toxic Products. Brieger isolated several toxic products which he 
 had found in association with putrefaction, such as cadaverin and 
 putrescin ; but there were also present two new toxic substances, one 
 producing cramps and muscular tremors in inoculated animals, and 
 the other lowering the temperature and depressing the action of 
 the heart. Later, Brieger in conjunction with Frankel, succeeded 
 in isolating a tox-albumin from pure cultures. Guinea-pigs were 
 killed in two or three days, but rabbits had an immunity. Pfeiffer 
 found that cultures contained a poisonous principle which proved 
 fatal to guinea-pigs in extremely minute doses. It is broken up 
 by alcohol and by boiling, and secondary products formed, of very 
 much mitigated virulence. Similar toxic products were obtained 
 from cultures of both Finkler- Prior's and Metchnikoff's commas. 
 
 Protective Inoculation. Haffkine has introduced a system of 
 protective inoculation, which is founded on the principle of inducing 
 the formation of antitoxins, or defensive proteids. Comma-bacilli 
 when first cultivated from a cholera patient are not sufficiently 
 
ASIATIC CHOLERA. 3(59 
 
 virulent, and the virulence is increased by cultivation in tin- 
 peritoneal cavities of a succession of guinea-pigs. This succc 
 cultivation is carried on until a virus is obtained which proves fatal 
 in a few hours when inoculated into the peritoneum. A culture 
 from the peritoneum is obtained on an agar plate-cultivation, and 
 a pure sub-culture on agar is thoroughly shaken up with broth. 
 This constitutes the vaccinating fluid. It maybe used as a living 
 vaccine, or the comma-bacilli killed by the addition of carbolic 
 acid. 
 
 Haffkine, having studied the pathological and physiological effects 
 on some sixty persons, mostly scientists interested in the subject, and 
 finding the treatment to be harmless, transferred his operations to 
 localities in India affected by cholera. The inhabitants of the 
 northern part of India were the first. to come forward and submit 
 themselves to the inoculation. In the course of the first vcar 
 2 2, 703 were inoculated in the North- West Provinces and Oudh. 
 and in the Punjab. All classes of the population were included. 
 In the second year operations were carried out in those parts of 
 the country where cholera always prevails, and where, therefore, the 
 method could be more satisfactorily tested. 
 
 From March 1894, to July 1895, 19,473 individuals \\vn- 
 inoculated in some of the most affected localities. 
 
 From observations made at Calcutta by Dr. Simpson, from March 
 1894 to August 1895, cholera occurred in 36 houses containing 
 inoculated people. There were 521 inhabitants in the infected 
 houses, of whom 181 were inoculated from 1 to 45!) days U-fuiv tin- 
 occurrence, while 340 remained timnoculated. The uninoculated had 
 45 cases with 39 deaths from cholera; the inoculated had 4 death-. 
 1 occurring 451 days after the first inoculation, and 3 others from 
 1 to 4 days after the first inoculation. These four cases had not 
 been re-inoculated. If the occurrences in inoculated and non-inocu- 
 lated during the first 10 days were set aside, and those considered 
 that occurred after the 10 days expired, then, according to Dr. 
 Simpson, the proportion of cases was 19'27 and that of deaths 17 _' I 
 times smaller in the inoculated then in the uninoculated. 
 
 Cholera broke out in the Gya gaol, and inoculations were mad*- 
 after 6 cases, with 5 deaths, had occurred. During the stay of the 
 prisoners in the gaol, there were 209 uninoculated, with 7 cases and 
 5 deaths, and 211 inoculated, with 5 cases and 4 deaths. 
 
 In July and August in the same year cholera attacked tlu 
 Lancashire Regiment. Out of 773 men there were 13:5 inocnlat. d 
 and 640 uninoculated, 
 
 34 
 
370 
 
 INFECTIVE | DISEASES . 
 
 The occurrences of cases and deaths were : 
 
 In 640 unmoculated 120 cases (18'75 %), 79 deaths (12-34 %). 
 
 In 133 inoculated 18 cases (13-53 %), 13 deaths (9'77 %). 
 
 These results were, it is said, due to the weakness of the vaccines 
 procurable at that period of the work, and to the small doses used. 
 
 There were a great many records kept of the results of inocula- 
 tion of coolies on tea estates in different localities. After a summary 
 of the results Haffkine concludes, in his Report to the Government 
 of India, that, in his opinion, the experimental stage was not yet 
 in so advanced a condition as to be completely closed; but that 
 the observations made and records collected justified steps being 
 taken to give the inoculations a more 
 extended trial. 
 
 CHOLERA NOSTRAS. 
 
 Cholera nostras, English cholera, or 
 English dysentery, produces an inflamma- 
 tion of the mucous membrane of the 
 bowels with croupous exudation. The 
 large intestine is commonly affected, and 
 the mucous membrane may be covered 
 with small superficial ulcers. The disease 
 is associated with severe diarrhoea. 
 
 Tinkler and Prior obtained a comma- 
 bacillus from the evacuations, which they 
 believed to be identical with the comma- 
 bacillus found by Koch in Asiatic cholera. 
 Koch pointed out that there were marked 
 differences in the biological character of 
 the two micro-organisms. 
 
 Spirillum. Pinkler-Prior (Comma- 
 bacillus in Cholera nostras). Curved rods, 
 thicker than the comma-bacillus of Koch, 
 and spirilla. The colonies on plate- 
 cultivations are very much larger than 
 
 those of the comma- bacillus of Koch of the same age. They have a 
 very faint yellowish-brown tinge, a well-defined border, and a distinctly 
 granular appearance. They liquefy nutrient gelatine very rapidly, 
 so that the first plate of a series is, as a rule, completely liquefied on 
 the day following inoculation, and the second plate in two or three 
 days more. In a test-tube cultivation in nutrient gelatine the 
 appearances are especially characteristic : the gelatine is very rapidly 
 
 FIG. 159. PURE-CULTIVATION 
 OF THE SPIRILLUM FINKLER- 
 PRIOR, IN NUTRIENT GELA- 
 TINE. In thirty -six hours. 
 
CHOLERAIC DIARRHOEA ~ FROM MEAT POISONING. 371 
 
 liquefied along the whole track of the needle, so that the cultivation 
 resembles a conical sack, or the finger of a glove turned inside out. 
 On a sloping surface of nutrient agar-agar a white moist layer forms 
 very quickly. On potato they grow at the ordinary temperature 
 of the air, producing a brownish layer and corrosion of the surface 
 of the potato. They have been shown to be pathogenic. 
 
 CHOLERAIC DIARRHCEA FROM MEAT POISONING. 
 
 There are two varieties of choleraic diarrhoaa from meat poisoning, 
 and both are associated with vomiting, diarrhoea, pain in the abdo- 
 men, in severe cases followed by suppression of urine, collapse, and 
 death. These conditions are brought about by poisonous foods, and 
 include those cases of poisoning by tinned meats, pork pies, hams, 
 cheese, sardines, and other articles of food improperly prepared. In 
 most cases putrefaction has taken place, owing to the action of 
 various bacteria. Associated with their growth we find highly 
 poisonous substances, but no bacteria are found in the body in these 
 cases. They are all due to chemical poisoning ; but Klein has also 
 described cases of poisoning due to the growth of bacteria without 
 the presence of putrefaction. The latter were of the nature of an 
 infectious disease. In the Welbeck poisoning cases, described by 
 Ballard, the poisonous hams contained a short bacillus, which was 
 also found in the kidney and spleen in the fatal cases in man. In 
 the Carlisle epidemic, which was due to poisonous pork pies, the pork 
 and gravy stock proved fatal to mice, and from the infected mice 
 a bacillus was cultivated, which, administered to mice by feeding or 
 subcutaneous inoculation, produced enteritis, diarrhoea, and congestion 
 of the lungs. 
 
 Gartner cultivated Bacillus enteritidis from the spleen in a 
 fatal case of meat poisoning. Gaffky obtained a similar bacillus in 
 cases of gastro-enteritis, following the consumption of meat and 
 sausages, which had been made of horseflesh. 
 
 Bacillus of Choleraic Diarrhoea from Meat-poisoning 
 (Klein). Rods from 3 to 9 p. in longth, 1'3 /x wide, rounded at their 
 extremities, singly or in chains of two. Spore-formation occurs, 
 the spores being 1 ft thick, oval, and situated in the centre or at 
 the end of the rod. 
 
 Feeding mice with the bacilli and inoculation produced positive 
 results. At the autopsy, pneumonia, peritonitis, pleuritis, enlargement 
 of the liver and spleen, and haemorrhages were observed, and bacilli 
 were present in the blood and exudations of these animals. They 
 
372 
 
 INFECTIVE DISEASES. 
 
 occurred in the blood and juices, and especially in the glomeruli of the 
 kidneys, of several fatal cases of choleraic diarrhoea. 
 
 Bacillus enteritidis (Gartner). Short rods in pairs, and short 
 chains. They are motile ; spore-formation not observed. Colonies 
 are granular, and old colonies at the margin have an appearance of 
 
 FIG. 160. TROPICAL DYSENTERY. Mucous membrane of large intestine some 
 months after an acute attack : a,a, representing remains of mucosa ; b,b, inter- 
 vening parts corresponding to the muscularis (HAMILTON). 
 
 powdered glass. On the surface of gelatine a thick greyish- white 
 film develops, which in time becomes wrinkled. In the depth of 
 gelatine a white filament forms. The gelatine is not liquefied. On 
 agar the film is slightly yellowish. On potato it is similar in colour, 
 moist and shining. On blood serum it is very similar, Mice feel 
 
CHOLERAIC DIARRHfKA IN FOWLS. 373 
 
 with the bacilli die in one or two days. Subcutaneous injection i> 
 fatal in guinea-pigs and rabbits in from two to five days. Dogs, 
 cats, and fowls are immune. 
 
 The bacilli were obtained from a cow suffering from a disease 
 associated with diarrhoea, and from the spleen of a man who died 
 twelve hours after partaking of the flesh of this animal. 
 
 DYSENTERY. 
 
 Dysentery is a disease of tropical climates associated with in- 
 flammation and ulceration of the large intestine (Fig. 160). At first 
 the discharge from the bowel is a whitish or brownish mucus, 
 which soon becomes blood-stained; later the evacuations become 
 thin and watery, with altered blood clots, fragments of mucous 
 membrane, and pieces of false membrane ; and in some cases they 
 become purulent. The virus is believed to be in the intestinal 
 discharges, which by contaminating water or soil may give rise to 
 other cases. 
 
 Micrococci have been found in dysentery, but the micro-organ i>m 
 which has received most attention is a protozoon, the Amctba coli, 
 which will be described in another chapter. 
 
 CHOLERAIC DIARRHOZA IN FOWLS. 
 
 Choleraic diarrhoea in fowls, or gastro- enteritis cholerica, is an 
 infectious disease of fowls, occurring in Russia during the summer. 
 The disease Ls very like fowl-cholera. The birds are sleepy, and 
 suffer from diarrhoea, but the temperature is not raist-il, as in 
 fowl-cholera. After death there is usually an abundance of greyish 
 liquid in the small intestine, which is stained with blood. It was 
 investigated by Gamaleia, who found a comma- bacillus, to which he 
 jrives the name Yibrio Metchnikovi. 
 
 Spirillum of Fowl- enteritis (Vibrio Metchnikovi). Curved 
 rods and spirilla; thicker, shorter, and more curved than Koch's 
 commas. They are motile, and possess a single flagellum at one end. 
 They stain with the usual dyes. Spore-formation doubtful. In plate- 
 cultivations minute white colonies appear in from twelve to sixteen 
 hours, and the gelatine is liquefied. The colonies in about three 
 days resemble those of both Fin kler- Prior's and Koch's comma-bacilli, 
 some colonies being more like the one kind, and some like the other. 
 In the depth of gelatine the growth is very much like that of Koch's 
 comma-bacillus, possessing the characteristic air-bubble appearance. 
 
374 INFECTIVE DISEASES. 
 
 On agar a slightl}' yellowish growth is obtained, resembling that 
 of Koch's commas ; on potato a yellowish -brown or chocolate layer 
 develops after incubation at the temperature of the blood, very 
 similar to cultures from Asiatic cholera. Broth becomes turbid, and 
 a wrinkled film forms on the surface ; the addition of sulphuric acid 
 gives the indol test. The spirilla grow in milk, and coagulate it ; 
 the milk becoming strongly acid, and the casein being precipitated. 
 They are pathogenic in chickens, pigeons, and guinea-pigs. Pigeons 
 die in about twelve hours after a subcutaneous injection ; and the 
 spirilla are found abundantly in blood from the heart. Guinea-pigs 
 die from acute septicaemia in about twenty-four hours. The spirilla 
 are found in the blood arid internal organs. Inoculation of pigeons 
 and guinea-pigs with sterilised cultures will produce immunity. 
 
CHAPTER XXVIII. 
 
 TUBERCULOSIS. 
 
 TUBERCULOSIS is a communicable disease of man and animals, charac- 
 terised by the formation of new growths associated with the presence 
 of the tubercle bacillus. Yon Bayle, in 1810, was the first to describe 
 little growths like millet seeds, which were considered to be character- 
 istic of consumption or phthisis. Laennec, in 1834, attached much 
 more importance to the existence of caseous matter and classified 
 miliary tubercle, crude tubercle, granular tubercle, and encysted 
 tubercle, as varieties of tuberculosis. Virchow would not accept 
 all these varieties as tubercular, and only regarded those conditions 
 associated with the presence of miliary tubercles as genuinely 
 tubercular. Laennec's so-called crude tubercle, for example, was 
 simply due to pneumonic caseation. Yillemin threw entirely fresh 
 light upon this controversy by proving that tuberculosis was a 
 communicable disease. Rabbits and guinea-pigs, inoculated with 
 tubercular sputum or caseous tubercle, developed miliary tubercle in 
 a few weeks. Sanderson confirmed these experiments, and pointed 
 out that foreign bodies would produce experimental tubercul<>-i- 
 in rabbits. Cohnheim also confirmed the experiments of Villemin, 
 and maintained that tuberculosis was a specific inoculable disease, 
 and, therefore, everything was .tubercular which, on inoculation, 
 produced tuberculosis. Koch, in 1882, announced the discovery of 
 the tubercle bacillus, and expressed the opinion that without the 
 tubercle bacillus there could be no tuberculosis. Tubercle was 
 defined as tissue containing the tubercle bacillus, whatever might 
 be the clinical manifestations of the case, or the microscopical and 
 naked- eye appearances of the diseased parts. 
 
 A tubercle is a small growth about the size of a millet seed. In 
 the early stage it is circular, hard, grey in colour, and lu-tn>n- : but 
 when it undergoes necrosis and caseation it becomes soft and yellowish. 
 Tn the very early stage it consists of a little collection of round 
 
 375 
 
376 
 
 INFECTIVE DISEASES. 
 
 cells, in which it is possible, though often with extreme difficulty, 
 to demonstrate the tubercle bacillus. The cells originate in the 
 proliferation of endothelial connective tissue and white blood cells. 
 Later on, large oval or circular multi-nucleated cells, or giant cells, 
 make their appearance. The tubercle bacilli are only occasionally 
 found in the interior of human giant cells, whereas in the lower 
 animals, in equine and bovine tuberculosis more especially, the bacilli 
 are often present in great numbers, and very commonly in the form 
 of conspicuous rings, visible under a low power of the microscope. 
 
 KIG. 161. TUBERCLE OF THE LUNG IN A VKKT EARLY STAGE, x 400 : o, An alveolar 
 wall ; b, blood-corpuscles in capillaries of the same ; c, blood-corpuscles 
 extra vasated into the alveolar cavities ; d, alveolar capillaries filled with 
 blood-corpuscles carried forward by the tubercle which is growing into the 
 alveolar cavity ; e, large endothelium-like cells, of which the tubercle in this 
 stage is mainly composed ; /, portion of a branch of the pulmonary artery 
 injected (HAMILTON). 
 
 Whether the absence of blood-vessels or the action of the bacillus is 
 the main factor in producing caseation, is an open question. When 
 suppuration follows caseation, as commonly happens in tuberculosis 
 of the lungs in man, and in experimental tuberculosis in animals, an 
 abscess forms. In cattle there is a remarkable tendency to the 
 formation of calcareous deposit in the caseous masses. 
 
 The tubercle may not degenerate arid die, but live and develop. 
 
TUBERCULOSIS. 
 
 377 
 
 The giant cells, which are more or less central, have been described 
 as sending off processes, which, by dividing and subdividing, and 
 
 FIG. 162. PRIMARY TUBERCLE OF LUNG TWO TO THREE WEEKS OLD, x 50: 
 a, Poi-tion of wall of a branch of the pulmonary artery ; 6,6, giant cella with 
 concentric arrangement of fibrous tissue ; c, centre of tubercle beginning to 
 caseate ; d, small branch of pulmonary artery seen on transverse section ; 
 c, injected capillaries of the alveolar walls (HAMILTON). 
 
 interlacing, form a reticulum, or 
 meshwork. Towards the periphery 
 of the tubercle the reticulum may 
 become arranged in the form of a 
 capsule as the age of the tubercle 
 advances, and the reticular giant 
 cell becomes eventually converted 
 into fibrous tissue. Tbe bacillus has 
 disappeared, and the tubercle has 
 healed. 
 
 Giant cells cannot be relied upon 
 to indicate tuberculosis. They are 
 not always present in tubercu- 
 losis, and they are not peculiar to 
 tubercle, being found, for example, 
 in actinomycosis. The only certain 
 indication of tuberculosis is the pre- 
 sence of the tubercle bacillus, which 
 
 FIG. 163. LA BGK OVAL GIANT CELL 
 KROM TUBERCLE OK LUNG x 300 : 
 a, Granular centre; 6, nucleated 
 jx-riphery forming a mantl--likr 
 sheath; c, proceaaea from the 
 same. 
 
378 INFECTIVE DISEASES. 
 
 can be revealed either by microscopical examination of the suspected 
 tissue, or after inoculation in guinea-pigs. 
 
 Bacillus Tuberculosis (Koch). Rods, 2 to 4 /* and occa- 
 sionally 8 ^ long, very thin, arid rounded at the ends. They are 
 straight or curved, and frequently beaded, and occur singly, in pairs, 
 or in bundles ; there are also involution forms and short branched 
 threads. Spore-formation is observed in old cultures. They are 
 non-motile. Tn the interior of giant cells they are often accompanied 
 by grains which exhibit the same colour reaction. 
 
 The bacilli in tissue sections of bovine tuberculosis are shorter 
 and less granular than those in human tubercular sputum, but in 
 milk they are quite as long, and even longer, and very distinctly 
 granular or beaded, and are thus brought much closer, morpho- 
 logically, to the bacilli in human sputum. Speaking generally, 
 however, the average length of the human bacilli is greater than 
 the average length of the bacilli in cow's milk, but the longest of 
 the bovine bacilli cannot be distinguished in length from the longest 
 human bacilli. There are, however, exceptional cases, for in some 
 preparations of pus from human lungs the bacilli are remarkable, 
 not only for their thinness, and their uniformly beaded character, 
 but more particularly for their extraordinary length. They should 
 be compared with other preparations, in which the bacilli, though in 
 human sputum, are sometimes much more distinctly rod-shaped, 
 much shorter and thicker, with complete absence of any beaded 
 appearance. Neither length nor granularity is a characteristic 
 sufficient to denote any specific difference between human and bovine 
 bacilli. The author has examined minutely the bacilli in tuberculosis 
 of other animals, such as the horse, pig, and cat ; and of birds the 
 fowl, guinea-fowl, pheasant, and ostrich. Here, again, minute 
 morphological differences can be observed. For example, in many 
 cases in fowls the bacilli are conspicuously short and straight. In 
 the liver and lungs of an ostrich, packets of short rod-forms are 
 found, while in other parts of the same sections the bacilli attain 
 a very great length. Many of the long, sinuous forms exhibit a 
 peculiar terminal enlargement. There are also short rods with a 
 similar appearance, and free ovoid bodies, singly and in groups, which, 
 from their connection with the bacilli, and their sharply defined 
 outline in the free state, are similar to spores in old cultures. 
 
 Thus, morphological differences are found under different circum- 
 stances, and within limits the morphology of the tubercle bacillus 
 varies with its environment. 
 
 Koch first succeeded in cultivating the bacillus by employing 
 
DESCRIPTION OF PLATE XI. 
 Bacillus tuberculosis. 
 
 The figures in this plate represent the bacilli of tuberculosis in 
 different animals, examined under the same conditions of amplifica- 
 tion and illumination. x 1200. Lamp-light illumination. 
 
 YiG. 1. Bacilli in pus from the wall of a human tubercular cavity. In 
 this specimen the bacilli are shorter than those in tubercular sputum, 
 and are very markedly beaded. 
 
 F IG> 2. Bacilli in pus from a tubercular cavity from another case in man. 
 They are present in the preparation in enormous numbers. The proto- 
 plasm occupies almost the whole of the sheath, and the bacilli are 
 strikingly thin and long. 
 
 FlG. 3. Bacilli in sputum from an advanced case of phthisis, showing 
 the ordinary appearance of bacilli in sputum ; some beaded, others 
 stained in their entirety ; occurring both singly and in pairs, and 
 in groups resembling Chinese letters. 
 
 FlG. 4. Bacilli in a section from the lung in a case of tuberculosis in man. 
 ^The bacilli in human tuberculosis are found in, and between, the tissue 
 cells ; and sometimes, as in equine and bovine tuberculosis, in the 
 interior of giant cells, but not so commonly. 
 
 FIG. 5. From a cover-glass preparation of the deposit in a sample of milk 
 from a tubercular cow. The bacilli were longer than the average 
 length of bacilli in bovine tissue sections, and many were markedly 
 beaded. 
 
 FIG. 6. From a section of the brain in a case of tubercular meningitis in a 
 calf, showing a giant cell containing bacilli with the characters usually 
 found in sections of bovine tuberculosis. 
 
 FIG. 7. From a section of the liver of a pig with tubercle bacilli at the 
 margin of a caseous nodule. 
 
 FIG. 8. From a cover-glass preparation of a crushed caseous mesenteric 
 gland from a rabbit infected by ingestion of milk from a cow with 
 tuberculosis of the udder. 
 
 FIG. 9. From a section of lung in a case of equine tuberculosis, showing a 
 giant cell crowded with tubercle bacilli. 
 
 FIG. 10. From a section of lung from a case of tuberculosis in the cat, with 
 very numerous tubercle bacilli. 
 
 FIG. 11. From a cover-glass preparation of a crushed caseous nodule from 
 the liver of a fowl, with masses of bacilli. These are for the most part 
 short, straight rods ; but other forms, varying from long rods to mere 
 granules, are also found. 
 
 FIG. 12. From sections of the liver and of the lung in a case of tubercu- 
 losis of a Rhea. Isolated bacilli are found, as well as bacilli packed in 
 large cells, colonies of sinuous bacilli, and very long forms v\ith terminal 
 spore-like bodies and free oval grains. 
 
 The preparations from which these figures were drawn were all 
 >t,.n ed by the Ziehl-Neelsen .method, with the exception of the 
 first, which was stained by Ehrlich's method. 
 
/- 
 
 . 
 
 \3 
 
 v v \ 
 
 ,"- v ^ 
 
 t^-ir m 
 
 k--' 
 
 \\ 
 
 Fig 1. 
 
 Fig 2 
 
 ~ 
 
 Kg 3 
 
 Si^4 
 
 -,\ 
 
 ' N 
 
 ,t 
 
 /I*Z 
 
 Fi^a 
 
 tf&y 
 
 r/^ - 
 
 - \ 
 ^6 
 
 // 
 
 (\ < 
 
 - 1 *tf 
 
 ^^ 
 
 ^ 
 
 % 
 
 Pig 9. 
 
 Pig 30. 
 
 jtf 
 
 i 
 
 
 Fig 12 
 
 BACIT.LUS TUBERCULOSIS. 
 
*^1 
 ^ 
 
 me ~ c * 
 
 TUBERCULOSIS. 379 
 
 blood serum. Solid blood serum, with or without the addition of 
 gt'latine, was employed, and the cultures incubated at 37 C. The 
 jrro \vth takes place very slowly, and only between the temperaturas 
 of 30 C. and 41 C. In about eight or ten days the growth appears 
 as little whitish or yellowish scales and grains. 
 
 The bacillus can also be cultivated in a glass capsule, on blood 
 serum, and the appearances of the growth studied under the 
 microscope. The scales or pellicles were described by Koch as made 
 up of colonies of a perfectly characteristic appearance, which may 
 be still further studied by making a cover-glass impression. They 
 are then seen to be composed of bacilli, arranged more or less 
 with their long axis corresponding with that of the colony itself, 
 and with an appreciable interval between the individual bacilli. 
 The colonies themselves appear as fine curved lines, the smallest 
 being mostly Shaped. Longer colonies have serpentine twistings 
 and bendings, which often 
 recall the curves of fancy 
 lettering. The ends of the 
 lines run to sharp points, 
 but the middle of the 
 growth is spindle- formed. 
 The youngest colonies are 
 extremely delicate and 
 narrow, but the older 
 colonies increase in size, 
 
 are thicker across, and, FlG - 164. - BACILLUS TUBERCULOSIS, FROM 
 ,! ,. .,, TUBERCULAR SPUTUM, x 2500. From Photo- 
 
 blending with each other, graphs 
 
 gradually obliterate the 
 
 characteristic appearances ; a lamellated growth results, which 
 increases, and gives the appearance to the naked eye of the scale 
 or pellicle already described. The blood serum is not liquefied 
 unless putrefactive bacteria contaminate the culture. A fresh tube 
 can be inoculated with one of the little scales, and a new generation 
 started. The scales gradually increase in size, and consist entirely 
 of bacilli. In about three to four weeks the cultivation ceases to 
 increase, and it is then necessary to inoculate a fresh tube. 
 
 In liquid blood serum a film forms on the surface of the liquid, 
 and is easily broken by agitation. In neutralised broth there is 
 very little indication of success. When a triturated culture is added 
 to the broth, a granular, sandy, whitish deposit collects at the 
 bottom of the vessel, with indications of an increase in amount. 
 Koch also tried nutrient agar-agar, which did not prove to be at 
 
380 
 
 INFECTIVE DISEASES. 
 
 BACILLUS TUBERCULOSIS. 
 
 FIG. 165. PURE-CULTIVATIONS ON GLYCERINE-AGAR FROM HUMAN TUBERCULAR 
 SPUTUM, a, After six months' growth. (Fifth sub-culture.) 6 and c, After 
 ten months' growth. (Fourth sub-cultures. ) 
 
TUBERCULOSIS. 
 
 381 
 
 all M favourable medium. Some increase took place, but there 
 was no continuous growth over the inoculated area. 
 
 Glycerine Agar-agar. Nocurd and Roux were among those who 
 worked at the subject and confirmed Koch's 
 observations. Nocard attempted to get 
 cultures of avian tuberculosis on blood serum 
 to which peptone, salt, and cane sugar had 
 been added. The results were more success- 
 ful than with ordinary serum. But he 
 encountered a difficulty in the rapid drying 
 of the surface of the medium, which rendered 
 the tubes unfit for use. It occurred to 
 Nocard and Roux to obviate this by the 
 addition of a hygroscopic agent, and accord- 
 ingly they added sterilised glycerine. The 
 result, which far exceeded their expectation, 
 evidently was not solely attributable to the 
 prevention of desiccation. Following up 
 their discovery, arid 
 being anxious to find 
 a medium more easily 
 prepared than blood 
 serum, they added 
 
 6 to 8 per cent, of glycerine to ordinary 
 nutrient agar-agar. The bacillus grew so 
 abundantly in this mixture that a culture 
 in fifteen days equalled in extent a culture 
 on blood serum which was several weeks old. 
 The bacillus was found to grow abundantly 
 in veal broth, to which glycerine had been 
 added in the proportion of 5 per cent., the 
 bottom of the flask being covered in about 
 three weeks with a flocculent deposit, having 
 some resemblance to anthrax cultivations 
 in liquid media. In beef broth, chicke?i 
 broth, and in Cohn's liquid, cultures were 
 obtained after the addition of glycerine. 
 
 Description of Cultivations on Glycerine 
 Agar-agar. The cultivations on the sloping 
 surface of obliquely solidified glycerine agar- 
 agar begin to appear in from four to six days as very minute white 
 colonies. These steadily increase in size, and either look moist 
 
 FIG. 166. PURE-CULTI- 
 VATION IN GLYCERINE 
 AGAR-AGAR, after ten 
 months' growth. 
 
 FIG. 1G7. -PURE-CULTI- 
 VATION- IN GLYCERINE 
 AGAR-AGAR, A SUB- 
 CULTURE FROM A PURE- 
 CTLTURE ix GLYCERINE- 
 MILK. In two months. 
 
382 INFECTIVE DISEASES. 
 
 smooth, or, even at this early stage, appear dry and crinkled. 
 According to the number of bacilli inoculated, the colonies will 
 either remain isolated or coalesce and form a more or less continuous 
 film. If the nutrient agar-agar has only recently been prepared, 
 there is usually a quantity of liquid present, and the bacillus forms 
 a white coating over the inoculated area and beyond it. The 
 appearances are much more characteristic when this medium is, 
 comparatively speaking, dry. A semi-transparent membranous 
 growth develops, thickens, and assumes a characteristic lichenous 
 appearance. Such a culture, examined with a pocket lens, resembles 
 a model in wax in miniature of the folds of the gastric mucous 
 membrane. In about six weeks to two months the culture has fully 
 developed. In old cultures, especially when the individual colonies 
 remain isolated, the appearance is very characteristic. Some cul- 
 tures in appearance closely resemble cultivations on blood serum. 
 The consistency of the growth depends upon the character of the 
 soil and the age of the culture. When the medium is moist the 
 growth is moist and viscous, but more often it is distinctly tallowy, 
 and in old and dry cultures scaly and friable. 
 
 Cultivations in Glycerine Broth. In a few days minute flakes are 
 visible, which steadily increase in size, and subside to the bottom of 
 the flask, forming in time a very copious deposit. On shaking the 
 flask, this deposit, which is extremely tenacious, rises in stringy 
 masses, and gives an appearance which is more or less character- 
 istic. If the flask is left undisturbed, a delicate veil-like film forms 
 over the surface, which can be readily broken up by gentle agitation, 
 forming flakes which gradually sink in the liquid. If undisturbed 
 for several weeks this film increases in thickness, is irregularly 
 fissured, and has more the appearance of masses of tallow floating 
 on the surface. The growth also may be seen to extend up the side 
 of the flask above the liquid. Pasteur or Erlenmeyer flasks can be 
 employed for these cultures. Solidified egg-albumin added to the 
 glycerine broth seems to increase the amount of growth, which 
 clings to the albumin, and waves to and fro in the liquid when the 
 flask is gently shaken. The author has confirmed the observation 
 of Nocard and Roux, that sub-cultures from glycerine agar-agar, or 
 from glycerine broth, will give cultures in ordinary broth without 
 glycerine. Ordinary broth with egg-albumin, and without glycerine, 
 will also give a good growth when inoculated from previous sub 
 cultures, although the attempt to produce primary cultures in these 
 media has hitherto failed. 
 
 Cultivations in Glycerine- Milk, and other Media. In milk the 
 
TUBERCULOSIS. 383 
 
 author found there was only a slight increase in the number of bacilli 
 inoculated, but milk with glycerine in the proportion of 5 per cent. 
 forms a more favourable medium. The author has also cultivated the 
 bacillus on sterilised urine and glycerine, and ordinary nutrient 
 gelatine with 5 per cent, of glycerine. On potato the growth of 
 the bacillus is extremely slow. Beevor succeeded in obtaining 
 cultures at the ordinary temperature of the room. 
 
 Examination of Cultivations. To examine the bacilli in these 
 various preparations the author prefers to use Neelsen's method, 
 floating the cover-glasses for from five to ten minutes on warm 
 c;irl)olised fuchsine, and passing them through dilute sulphuric acid. 
 In some cultures the bacilli are shorter and thicker than is commonly 
 observed in human sputum, and they are for the most part without 
 the beaded appearance. In old cultures on glycerine agar-agar the 
 number of granular or beaded bacilli increases, and there are also 
 numerous peculiar forms. There are bacilli, sometimes two or 
 three times the length of an ordinary bacillus, provided with a 
 club-shaped enlargement at one or both extremities, and in rare 
 cases with lateral branches. They are no doubt identical with the 
 bacilli with swollen extremities and the branched forms observed 
 by Nocard and Roux. 
 
 In milk the appearance is very striking, many bacilli attaining 
 in old cultures a great length, and all are more uniformly beaded 
 than in any other cultivations. Staining preparations by the method 
 of Gram in aD cases exaggerate this appearance. 
 
 The important part played by the environment is shown by the 
 morphological differences observed in artificial cultivation under 
 varying conditions, and by the fact that by successive cultivation 
 the bacillus can be educated to grow upon a 'medium which is un- 
 suitable for obtaining primary cultures. 
 
 Impression preparations of the growth of the bacillus on the 
 surface of glycerine agar-agar in capsules show a tendency to the 
 formation of serpentine colonies, composed of bundles of more or 
 less parallel bacilli. 
 
 S}wre-farmation. In old cultivations true spore-formation can 
 readily be observed, both in stained and unstained preparations. In 
 the latter case they are recognised in the form of one or two highly 
 refractive bodies in individual bacilli. 
 
 Toxic Products of Cultures. The poisonous substances found in 
 cultures, and the composition and use of tuberculin, have already 
 been described (p. 43). 
 
 Inoculation Experiments. A relatively small portion of a culti- 
 
384 INFECTIVE DISEASES. 
 
 ration inoculated into the subcutaneous tissue, into the peritoneal 
 or pleural cavities, into the anterior chamber of the eye, or directly 
 into the blood stream, produces after three or more weeks artificial 
 tuberculosis in guinea-pigs and rabbits. Dogs and cats can also be 
 infected by experimental inoculation. 
 
 When a trace of tubercular virus is inserted subcutaneously in 
 the thigh of a guinea-pig, in about a week or ten days a chain of 
 enlarged glands will be easily felt in the vicinity of the seat of 
 inoculation. This affords an unfailing test, which can be applied 
 when there is difficulty in ascertaining by the microscope the presence 
 of the bacilli in the material under examination. It also affords a 
 valuable method for testing the effects of antiseptics on tubercular 
 virus. The appearances observed at the autopsy are swollen 
 lymphatic glands, in the neighbourhood of the inoculation, followed 
 by softening and abscess ; enlargement of the spleen and liver, with 
 formation of caseous tubercles ; and tubercular deposits in the lungs, 
 bronchial glands, and peritoneum. 
 
 After inoculation of the eye, grey tubercles appear on the iris, 
 and undergo enlargement and caseation, followed by tuberculosis 
 of the eyeball and organs generally. 
 
 The bacilli appear to be the direct cause of tuberculosis, and 
 the presence of the bacillus in the sputum of patients is a distinctive 
 sign of the existence of this disease. The detection of the bacillus 
 IIMS, consequently, become a test which is constantly applied. 
 
 The bacilli are found in all tubercular growths of man, monkeys, 
 cattle (Perlsucht), birds, and many other animals, and in cases of 
 artificial tuberculosis, in rabbits, guinea-pigs, cats, etc. In man the 
 bacillus can be detected in the tissues, in the sputum, in the blood, 
 and in the urine. 
 
 Tuberculosis may also be produced by inhalation and feeding 
 experiments. The channels of infection in man are also most 
 probably the pulmonary and intestinal mucous membranes. The 
 possibility of inoculation of skin wounds is open to doubt. The 
 bacilli or their spores are inhaled from the air, or taken in with 
 food. Morphologically identical bacilli have also been observed, but 
 very sparsely, in sections of lupus. 
 
 METHODS OF EXAMINING THE TUBERCLE BACILLUS. 
 
 Numerous methods have been recommended for examining the 
 tubercle bacillus. A few of these will be described, as many are 
 only of historical interest, 
 
TUBERCULOSIS. 385 
 
 The Ziehl-Neelsen method is preferred by the author both for 
 
 sections and cover-glass preparation*. 
 
 AW/'x ni'iij'nmt nn'th od Cover-glass preparations or sections are laid 
 in Koch's solution (No. 23, c) for twenty-four hours, or for one hour if 
 the solution is warmed to 40 C. Rinse in water ; immerse in a watery 
 solution of vesuvin for two minutes ; rinse again in water, and examine ; 
 or, after rinsing in water, treat with alcohol, clove-oil, and Canada 
 balsam. 
 
 /,7<r//W/'x mi'tJiotJ. Cover-glass preparations are allowed to float in a 
 watch-glass, containing a solution of gentian-violet or fuchsine, added to 
 aniline water. A saturated alcoholic solution of the dye is added till 
 precipitation commences (10 cc. aniline water, and 10 to 20 drops of the 
 colour solution). The cover-glasses are left in the solution for about 
 half an hour ; then washed for a few seconds in strong nitric acid (one 
 part commercial nitric acid to two of distilled water), and rinsed in 
 distilled water. After-stain with vesuvin or methylene-blue, rinse in 
 water, dry and preserve in Canada balsam. 
 
 Ehrrlich-Koch method. 
 
 Saturated alcoholic solution of methyl- violet or fuchsine 11 
 Aniline water ........ 100 
 
 Absolute alcohol 10 
 
 Preparations are Jeft for twelve hours in this solution (colouring of 
 the cover-glass preparations can be expedited by warming the solution). 
 
 Treat the preparations with (1 to 3) solution of nitric acid a few 
 seconds. 
 
 Wash in alcohol (GO per cent.) for a few minutes (cover-glass prepara- 
 tions need only be rinsed a few times). After-stain with diluted solution 
 of vesuvin or methylene-blue for a few minutes. 
 
 Wash again in 60 per cent, alcohol, dehydrate in absolute alcohol. 
 Clear with cedar- oil, mount in Canada balsam. 
 
 K'nidjli ir/i's method. Prepare a solution composed of 
 
 Saturated alcoholic solution of fuchsine . . 10 drops 
 Aniline water 2 drams. 
 
 Pour it into a watch-glass, and float the cover-glass ; warm the watch- 
 glass over a spirit-lamp until steam rises. Remove it from the flame, 
 and set it aside for five minutes. Take out the cover-glass, and transfer 
 it for a few seconds to acidulated alcohol (two drops of nitric acid in a 
 watch-glass full of alcohol). Wash in distilled water, dry, and preserve 
 in balsam. After-stain, if necessary, with Bismarck-brown, or methylene- 
 blue. 
 
 <;;!,! x' ////-//^/.Cover-glass preparations are placed in Gibbes' double- 
 staining solution which has been warmed in a test-tube, and, as soon as 
 steam rises, poured into a watch-glass. They are allowed to remain for 
 five minutes, and then are washed in methylated spirit till no "more colour 
 comes away, dried in the air or over a spirit-lamp, and mounted in 
 Canada balsam. If the solution is used without warming, the cover-glasses 
 
 25 
 
386 INFECTIVE DISEASES. 
 
 must be left in it for an hour. Sections are treated on the same 
 principles, but must be left in the solution for several hours. The 
 crumpling of the sections by the action of nitric acid is avoided. 
 
 Baumgarten's method. Cover-glass preparations of sputum are made 
 as already described, and immersed in a very dilute solution of potash 
 (1 to 2 drops of a 33 per cent, solution of potash in a watch-glass of dis- 
 tilled water). The cover-glass is pressed down on a slide, and examined 
 with a high power. The bacilli can be thus examined in the unstained 
 condition, and to avoid any mistake from confusion with other species, 
 the cover-glass can be removed, dried, passed through the flame, and 
 stained with a drop of an aqueous solution of fuchsine, or gentian- violet. 
 The putrefactive bacteria are stained, but the tubercle bacilli remain 
 absolutely colourless. 
 
 Baumgarten's new method. A. solution is prepared as follows : Drop 
 4 to 5 drops of concentrated alcoholic methyl-violet solution into a small 
 watch-glass full of water, (a) Stain the sections in this solution, wash 
 them in water, and decolorise in absolute alcohol (five to ten minutes) ; 
 or, before treating with alcohol, immerse the sections for five minutes in 
 a half -saturated solution of carbonate of potash. Pass through clove-oil, 
 and mount in a mixture of Canada balsam, free from chloroform, and 
 clove-oil (equal parts). The object of this process is to differentiate the 
 tubercle bacilli from chance bacteria, inasmuch as the tubercle bacilli 
 are gradually decolorised by the clove-oil. (Z>) Sections stained in the 
 above solution are placed for five minutes in alcohol, and then in a 
 concentrated solution of Bismarck-brown in 1 per cent, solution of acetic 
 acid. The after-treatment may be conducted as already described. 
 
 Ziehl-Neelsen method. Cover-glass preparations may be quickly stained 
 in Neelsen's solution warmed in a watch-glass till steam rises. Sections 
 are left for from five to ten minutes in the solution, and then washed in a 
 watery solution of sulphuric acid (25 per cent.), rinsed in distilled water, 
 and immersed in methylene-blue solution. After two or three minutes 
 they are passed through alcohol and oil of cloves, and mounted in Canada 
 balsam. 
 
 FranlceVs method. Sputum preparations are rapidly double-stained 
 by the following method : Prepare a solution by adding concentrated 
 alcoholic methyl-violet or fuchsine solution, drop by drop, till opalescence 
 arises, to 5 com. of aniline- water heated to 100 C. Float the prepared 
 cover-glasses two minutes in the warmed solution. The process of after- 
 staining and decolorisation is effected by placing the preparation for one 
 to two minutes in one of the following solutions : for fuchsine-stained 
 preparations, a saturated solution of methylene-blue in a mixture of 
 
 Alcohol 50 
 
 Distilled water .30 
 
 Nitric acid 20 
 
 which is filtered before use ; for preparations stained in methyl- violet, a 
 saturated solution of vesuvin may be used in 
 
 Alcohol .70 
 
 Nitric acid . 30 
 
TUBERCULOSIS. 387 
 
 /;/,, 7/r/f'x Method and Eosin.The Author has found that after sections 
 have been stained with methyl-violet and Bismarck-brown by Ehrlich's 
 method, as described by Kooh, they may with advantage be immersed in 
 a weak alcoholic solution of eosin, then rinsed in clean absolute alcohol, 
 clarified with clove-oil, and mounted in Canada balsam. The giant cells 
 are then stained pink, while their nuclei are brown, and the bacilli blue. 
 
 TUBERCULOSIS ix MAN. 
 
 The disease manifests itself in various forms in man, and most 
 frequently in the lungs, producing phthisis or consumption. The 
 sputum contains the bacilli in large numbers, and is extremely 
 virulent. Scrofula and lupus are forms of tuberculosis; they are 
 
 FIG. 168. --SECTION THROUGH A LUPUS NODULE OF THE NOSK. 
 
 probably produced by an attenuated variety of the tubercle bacillus. 
 Lupus can be distinguished from tuberculosis of the skin ; and 
 scrofulous lymphatic glands are distinguished from tubercular glands 
 by the tendency of the latter to produce generalised tuberculosis. 
 This difference in the intensity of the virus in the two cases, 
 Lingard illustrated by the effect upon inoculated guinea-pigs. 
 
 Cavities in the lungs are often thickly lined with bacilli. They 
 are present in great numbers in the caseous matter, though in 
 equine and bovine tuberculosis this is not the case. 
 
 Whether the disease in man is contagious is an open question, 
 though numerous cases of supposed communication between husband 
 and wife, brothers and sisters, have been reported, and Ransome 
 
388 
 
 INFECTIVE DISEASES. 
 
 showed that tubercle bacilli were present in the breath in phthisis. 
 On the other hand, the experience in consumption hospitals does not 
 
 FIG. 169. TUBERCULAR ULCERATION IOF MUCOSA OF HUMAN ILEUM. 
 Between the ulcers there are tubercular lymph -follicles (HAMILTON). 
 
 support this view, there being no evidence of the communication of 
 the disease to nurses and hospital attendants. 
 
TUBERCULOSIS. 
 
 389 
 
 TUBERCULOSIS OF CATTLE. 
 
 In cattle the disease may occur as the result of inhaling bacilli, 
 or of ingestion with food. It is very frequently found in the lungs ; 
 :ind calves may be infected by milk from cows with tubercular 
 udders. Calves may also suffer from congenital tuberculosis, the 
 bacilli having been transmitted from the mother during gestation. 
 
 Breeding in-and-in, over-production of milk, and confinement with 
 insanitary surroundings, predispose to tuberculosis. The disease is 
 known in Germany as ^ Perlsucht"; and in this country the lesions 
 on the pleura are known [as 
 "grapes," and the animals 
 themselves are commonly 
 called " wasters." 
 
 The disease may also exist 
 in the lungs or in other 
 organs, in a limited form, 
 without any indication of 
 ill health. In such cases 
 the disease can be detected 
 by injection of tuberculin, a 
 marked rise of temperature 
 occurring in tubercular 
 animals. 
 
 In advanced cases, the 
 symptoms commonly observed 
 are cough, difficulty in breath- 
 ing, staring coat, wasting, and 
 diarrhoea ; and if the udder is infected, nodules in the gland, and thin 
 bl uish milk. In the lungs, after slaughter, a few small cheesy tubercles 
 may be found in animals apparently in perfect health and in prime 
 condition for the market. In advanced cases, the lungs on section 
 show large yellow masses, containing calcified matter, and the 
 bronchi may be full of yellowish pasty contents. The disease will be 
 found to involve the bronchial glands. The serous membrane may 
 be covered with little warts or grape-like masses. The lymphatic 
 glands may be enlarged to an enormous size. Tubercular ulceration 
 of the intestine is sometimes found, but not commonly. In tubercular 
 disease in the udder, a painless swelling is found which may affect 
 one or more quarters of the gland. 
 
 Transmission of Tuberculosis from Man to Cattle. It is 
 
 FIG. 170. SECTION OF LUPUS OF 
 
 x 700. Giant cell containing a tubercle 
 bacillus (FLUGGE). 
 
390 
 
 INFECTIVE DISEASES. 
 
 for obvious reasons impossible to ascertain by experiment whether 
 tuberculosis can be transmitted from cows to man by milk or other- 
 wise ; but some light may be thrown upon this important question 
 by ascertaining the result of inoculating bovines with human tuber- 
 culosis. If calves can be infected with tuberculosis from a human 
 source by inoculation or ingestion experiments, and especially if 
 the effect of administering human and bovine tubercle to calves 
 by these means is found to be the same, such experiments will 
 not only serve to dispel any doubt there may be as to the identity 
 of the two affections, but they will strengthen the hands of those 
 
 FIG. 171. TUBERCULOSIS OF PLEURA ; ", GRAPE-DISEASE." 
 
 who insist upon the necessity of more thorough inspection of dairy 
 cows, and of power to deal with tubercular animals. 
 
 Inoculation of a Calf with Human Tubercular Sputum.-Th* 
 
 ithor obtained sputum containing numerous bacilli from an 
 
 vanced case of phthisis. The sputum was shaken up with sterilised 
 
 alution and injected into the peritoneal cavity. A few weeks 
 
 if erwards the^calf showed signs of illness. The animal looked dull, 
 
 b feed well, had a slight cough, and showed less inclination to 
 
 bout than usual. These symptoms gradually increased, and 
 
 leath occurred forty-two days after inoculation. Extensive lesions 
 
TUBERCULOSIS. 391 
 
 were discovered at the post-mortem examination. The mesentery 
 was adherent to the abdominal wall, at the seat of the inoculation, 
 and to the rumen ; the liver was adherent to the diaphragm. There 
 was rxten>ive tubercular deposit at the seat of inoculation, and 
 an aWt'ss the size of a walnut. Extending over the mesentery 
 from this point there were hundreds of wartlike, fleshy, new growths, 
 some quite irregular in form, others spherical or button-shaped. 
 There were similar deposits on the tinder surface of the liver, on the 
 spleen, in the gastro- splenic omentum, and on the peritoneal surface 
 of the diaphragm. The spleen was adherent to the rumen, and 
 on dissecting away the adhesions another abscess was opened. The 
 lungs were congested and the pleurae thickened. On microscopical 
 examination of sections extremely minute tubercles were found to 
 be disseminated throughout the whole of the substance of the lungs 
 and liver, and tubercle bacilli were found in these and in the 
 peritoneal deposits. The abscesses contained Streptococcus pyogenes. 
 The calf died of pyaemia, but sufficient time had elapsed for marked 
 local infection leading to generalised miliary tuberculosis. 
 
 TUBERCULOSIS IN RELATION TO THE PUBLIC MILK SUPPLY. 
 
 There is not the slightest doubt that when the udder is involved 
 the milk is highly virulent to the lower animals, and presumably 
 is. therefore, dangerous to man. The virulence of the milk was 
 first insisted upon by Klencke in 1846, and confirmed by Gerlach in 
 1869, and later, by other-. 
 
 This subject was again brought forward with the discovery of 
 the tubercle bacillus, and the demonstration of its existence in the 
 milk in certain cases of bovine tuberculosis. Koch pointed out that 
 the milk only contained bacilli, and was only infective, when the 
 udder itself was tubercular. By this he explained the contradictory 
 results obtained by various experimenters with milk from cows un- 
 doubtedly suffering from " Perlsucht." Koch considered that positive 
 effects were obtained with milk when it happened to contain tubercle 
 bacilli, and negative with milk from which they were absent. Bang 
 in a number of cases verified the presence of tubercle bacilli in milk, 
 and, owing to the contradictory results of previous investigations, 
 r ( -i>eated the ingestion experiments. The milk was found to be 
 virulent both to pigs and rabbits. 
 
 In this country Woodhead and M'Fadyean tested milk for 
 tubercle bacilli. They examined six hundred cows in the Edinburgh 
 dairies, and found thirty-seven suffering from mammitis, but in only 
 
392 INFECTIVE DISEASES. 
 
 six were they able to demonstrate the presence of tubercle bacilli in 
 the milk, and then only in small numbers. 
 
 Hirschberger found in twenty cases of tuberculosis in cattle 
 that the milk of eleven was virulent to guinea-pigs. Three cows out 
 of nine in which the disease was restricted to the lungs gave infected 
 milk. On the other hand, Nocard inoculated milk from eleven 
 tuberculous cows, of which only one had diseased udder, and only 
 this one gave infective milk. Bang injected rabbits with milk from 
 twenty-one cases of tuberculosis, with the udders apparently normal, 
 and the milk was virulent in two. 
 
 The author had two cases of udder tuberculosis under observation, 
 and as no experiments had at the time been made in this country 
 with milk known to contain tubercle bacilli, it was decided to study 
 the effect on rabbits, and test the results obtained by Bang. These 
 cases were both interesting and instructive, and may be referred to 
 in detail. 
 
 One was a case of advanced general tuberculosis. There was extreme 
 emaciation, general apathy, and a peculiar dull expression of countenance. 
 The skin was dry and harsh, the coat staring, and there was loss of hair 
 in patches about the face and neck. There was dulness-on percussion 
 over a large area of the thorax, and the respirations were increased in 
 rapidity. There was also occasional cough and some diarrhoea. But the 
 most interesting condition was observed on examination of the udder. 
 The gland was swollen, especially posteriorly, and distinct induration 
 could be felt on examination. The deposit appeared to be more or less 
 limited to the posterior quarters. The cow evinced no pain during the 
 examination of the udder, not even on the application of firm pressure. 
 
 The author took samples in test-tubes of the milk from all four teats ; 
 when freshly drawn, it differed noticeably from the normal secretion. It 
 was a thin, watery, turbid fluid with whitish flakes in suspension, but it 
 was not gelatinous or muco-purulent in character, and was free from any 
 markedly yellow colour. After being set aside in the laboratory for 
 some hours it separated into a layer of cream and a turbid liquid of a 
 yellowish tint, while at the bottom of the test-tube there was a whitish 
 flocculent deposit, especially in the samples from the posterior quarters. 
 
 There were tubercle bacilli both in the cream and in the deposit. In 
 the cream they were only present in small numbers, and were detected, 
 therefore, only after careful search. But in the deposit they were readily 
 found, as in a cover-glass preparation there were sometimes four or five 
 in the field of the microscope. 
 
 The method adopted for the examination of this deposit was as 
 follows : The whole of the liquid in the test-tube was carefully poured 
 off, and a trace of the sediment spread out on a cover-glass. This was 
 allowed to dry, and passed through the flame, and stained in hot Ziehl- 
 Neelsen solution in the usual manner. 
 
TUBERCULOSIS. 
 
 393 
 
 The other cow was also a case of general tuberculosis, and presented 
 somewhat similar lesions of the udder. The induration of the gland was 
 readily detected, and examination of the milk showed, as in the previous 
 case, the presence of tubercle bacilli. 
 
 It will be observed that in neither of these cases was the disease 
 limited to the udder ; in both the implication of the gland was part of 
 general tuberculosis. 
 
 FIG. 172. TUBERCULAR ULCERATION OF THE INTESTINE OF A Cow. 
 
 The first cow was killed, and the following lesions were found at the 
 post-mortem examination. 
 
 TJini-fix. The lungs and bronchial glands were extensively invaded 
 with tubercular deposit. The glands were greatly enlarged and densely 
 fibrous, in many cases with central, stone-like masses, grating on section 
 against the edge of the knife. In the lung there was every stage, from 
 the early deposit to purulent cavities, cheesy masses, and calcified dibri*. 
 
 Abdvmtn. There were a few caseous nodules in the liver, but none 
 in the spleen. The mesenteric glands formed an almost continuous chain 
 
394 INFECTIVE DISEASES. 
 
 of large tumours, mostly with central cretin" cation. Tubercular deposit 
 in the intestines could be recognised from the outside, and on laying 
 them open the mucous membrane was found to be studded with tuber- 
 cular ulcers. These ulcers were most numerous in the large intestine, 
 and varied in size from a sixpence to a florin. Some were circular, others 
 slightly irregular in form, and others again distinctly oval. In the latter 
 case they were generally situated with their long diameter transversely. 
 The base of the ulcer involved the muscular coat, and was irregularly 
 radiated. The margin was broad, and elevated above the general surface, 
 producing a ring-like appearance. 
 
 Mammary Gland. The udder was infiltrated throughout with tuber- 
 cular new growth, but the invasion was most marked in the posterior 
 quarters. There was apparently very little tendency to caseation. 
 
 Microscopical Examination of the Udder. In order to study the histo- 
 logical characters of the gland, and the distribution of the bacilli, sections 
 were stained with logwood and rubin, and others again with fuchsine 
 and methylene-blue. The tubercular new growth consisted of the usual 
 histological elements, round cells, epithelioid cells, and giant cells. 
 Healthy lobules here and there were sharply marked off from those in 
 which the growth was compressing and obliterating the alveoli in its 
 progress. Bacilli were present in the giant cells, and also distributed in 
 vast numbers throughout the tubercular tissue generally. Bacilli were 
 found in epithelioid cells close to the alveolus, and also between the cells 
 lining the alveoli. In parts also the new growth had involved the milk 
 ducts, and therefore it was easy to account for the presence of the bacilli 
 in the milk. 
 
 The bacilli were found in considerable numbers also in sections of the 
 intestinal ulcers. 
 
 EXPERIMENTAL INFECTION OF RABBITS. 
 
 Ingestion. A rabbit received the contents of a test-tube which 
 had been filled with milk from one of the posterior teats, mixed with 
 a small quantity of bran. In four weeks there was commencing 
 emaciation ; later, diarrhrea set in, and death occurred exactly fifty- 
 eight days after administration of the milk. At the post-mortem 
 examination the mesenteric glands were found to be much enlarged 
 and caseous. A cover-glass preparation from a crushed gland 
 revealed numerous tubercle bacilli. On opening the intestines there 
 was a patch of ulceration, showing the point of access of the bacilli. 
 The intestinal ulceration was a reproduction, to a certain extent, of 
 the condition in the cow which had been the source of the virus. 
 
 Subcutaneous Injection. A second rabbit was injected under the 
 skin of the back by means of a capillary pipette with about ten 
 drops of milk, including some of the deposit from the bottom of the 
 test-tube. The sample of milk had in this case also been taken from 
 
DESCRIPTION OF PLATE XII. 
 Tubercular Mammitis. 
 
 FIG. 1. From a section of the udder of a milch cow. The tubercular deposit 
 is seen to invade the lobules of the gland. Lobules comparatively healthy 
 are marked off, more or less sharply, from the diseased ones in which the 
 new growth in its progress compresses and obliterates the alveoli. Stained 
 by the Ziehl-Neelsen method and with methylene-blue. x 50. 
 
 FIG. 2. Part of the same preparation. On the right of the section part of a 
 healthy lobule is seen. On the left a lobule is invaded by tubercular new 
 growth composed of round cells, epithelioid cells and typical giant cells. 
 Tubercle bacilli can be seen both singly and collected in groups. They 
 are found in and between the cells, and in the interior of giant cells. 
 Bacilli may be seen between the cells lining an alveolus and projecting 
 into its lumen, x 800. 
 
--^ '' ;i 
 
 
 
 Fgl. 
 
 
 : 
 
 
 
 Fig 2. 
 
 TUBERCULAR M/\MMITIS 
 
TUBERCULOSIS. 395 
 
 one of the posterior teats. The rabbit was placed in a separate 
 hutch, and death from general tuberculosis occurred ninety-two days 
 after inoculation. 
 
 The diaphragm and mesentery were studded with tubercles the 
 si/.t* of a pill's head. The kidneys superficially showed whitish 
 rounded nodules projecting above the surface. These were found 
 on section to be continuous, with wedge-shaped deposits in the sub- 
 stance of the kidney. The lungs presented a very striking appear- 
 amv, being, in short, a mass of tubercular deposit; and the bronchial 
 and traclieal glands were similarly affected. In sections of the 
 kidney and lung the bacilli were present, but they were distributed 
 irregularly ; in one part of a section it was difficult to detect a 
 single bacillus, in other parts they were present in large numbers. 
 
 The milk from the two cows, 
 previously to their coming under 
 observation, had been mixed with 
 the general supply of a dairy. There 
 is indeed ample evidence that, both 
 in this and in other countries, the 
 milk of tuberculous animals finds its 
 way into the market. The question 
 which naturally arises is the possi- 
 bility of any manifestation of tuber- 
 culosis in man, arising from the 
 consumption of unboiled milk con- 
 taining tubercle bacilli. We must 
 admit that there is no direct FIG. 173. TUBERCULAR ULCERA- 
 
 evidence of the transmission of TION OF THE IXTESTIXK OF A 
 
 . . . RABBIT. 
 
 tuberculosis by milk from cow to 
 
 man ; but this may arise from the difficulty in tracing such a 
 source of infection, owing to the long time which elapses before 
 symptoms manifest themselves in man. Yet, if milk be a source of 
 infection, we should naturally expect that primary tuberculosis of the 
 intestine would be by no means an uncommon manifestation of the 
 disease; and this in the adult is not in accordance with clinical 
 experience. Such an argument would tend to centra-indicate 
 danger to adults ; but, on the other hand, the possible danger to 
 children has been rightly insisted upon by the earliest writers on 
 this subject. Woodhead has recently stated that, from his experi- 
 ence in two large hospitals, he has been much struck by the fact 
 that, in children who had died from other diseases during the course 
 of tubercular disease of the abdominal glands, there was frequently 
 
396 
 
 INFECTIVE DISEASES. 
 
 not any trace of tubercular disease in other parts ; thus^pointing to 
 the intestine as the channel by which the bacillus made its way into 
 the body. Woodhead also remarks that in a large number of cases 
 
 FIG.: 174. TUBERCULOSIS OF THE LUNGS. 
 
 From a photograph of the lungs of a rabbit which had been injected sub- 
 cutaneously with about ten drops of milk, including in suspension a small 
 quantity of the deposit at the bottom of a sample of milk from a cow with 
 tuberculosis of the udder. Death occurred from general tuberculosis ninety-two 
 days afterwards. The appearance of the lungs was very striking. They were 
 almost completely composed of tubercular deposit. The bronchial glands, 
 as well as the tracheal, of which one is seen in the photograph, were also 
 enlarged and caseous. There were tubercular deposits in the kidneys and 
 other organs, and also at the seat of inoculation. 
 
 of general tuberculosis, where the possibility of infection by the 
 pulmonary passages was evidently excluded, the tubercular process 
 
TUBERCULOSIS. 397 
 
 appeared to have invaded the body by the intestinal canal. These 
 facts, taken in connection with the occasional existence of tubercle 
 bacilli in milk, went far to prove, in his opinion, that milk was a 
 source of tubercular infection, especially to young children. 
 
 From his own experiments and observations the author has 
 drawn the following conclusions : 
 
 1. Cows with tuberculosis of the udder are to be found in dairies 
 
 in this country. 
 
 2. The milk of these cows is, as a rule, mixed with the general 
 
 supply. 
 
 3. The milk in cases of udder tuberculosis contains tubercle 
 
 bacilli. 
 
 4. Rabbits inoculated with, or fed upon, milk containing tubercle 
 
 bacilli contract tuberculosis. 
 
 5. Direct evidence of transmission of tuberculosis by milk to man is 
 
 wanting, but from the effect of such milk on the lower animals 
 it is reasonable to conclude, in the present state of our know- 
 ledge, that there may be danger in using the milk of cows 
 with tubercular udders, and therefore strict inspection of 
 dairies should be enforced ; and boiling of milk before 
 use will, as a rule, be a wise, if not absolutely a necessary 
 precaution. 
 
 Bollinger has shown that the virulence of cow's milk is reduced 
 by dilution with water in the proportion of 1 in 40 and even of 
 1 in 100, and that therefore there would be much less danger 
 in consuming tubercular milk which had been mixed with the 
 milk of healthy cows, than there would be in taking it direct 
 from the infected cow. This is a matter of scientific interest ; but 
 it would be no justification for a dairyman to mix the milk of a 
 tubercular cow with milk of cows known to be healthy. The milk 
 of cows suffering from tuberculosis should undoubtedly be rejected. 
 
 TUBERCULOSIS AND THE PUBLIC MEAT SUPPLY. 
 
 The question of the advisability of allowing the flesh of tuber- 
 cular animals to be sold for food, especially when the disease exists 
 in a very small degree, is a vexed one. Numerous experiments 
 have been made upon the infectivity of the flesh of tubercular 
 animals. Kastner inoculated the juice expressed from the flash of 
 tubercular cows. Sixteen guinea-pigs were unaffected after injection 
 
398 INFECTIVE DISEASES. 
 
 of 1 to 2 cc. into the peritoneal cavity. Nocard injected ten to 
 twenty drops of the muscle juice of the hearts of tubercular cattle, in 
 which the disease was well marked, and none of the guinea-pigs were 
 infected. With juice of the muscles of the thigh derived from ten 
 tubercular cows Nocard inoculated forty guinea-pigs, and one only 
 showed signs of tubercle. Nocard concluded that if there was any 
 danger in the flesh of tuberculous animals, it was the exception and 
 not the rule. On the other hand, Chauveau and Arloing produced 
 tuberculosis in two guinea-pigs out of ten inoculated with muscle 
 juice from a tubercular steer. 
 
 In 1890 a Royal Commission was appointed to investigate this 
 subject, and the report was issued in 1895. Martin, on behalf of 
 the Commission, tested the flesh of twenty-one tubercular cows. In 
 two cases only was evidence obtained of the presence of the bacillus 
 by inoculation of guinea-pigs. The flesh of eight cows affected with 
 mild tuberculosis produced tubercle in one instance by inoculation, 
 but the ingestion experiments were negative. The flesh of five cows 
 severely affected with tubercle gave the disease in four cases, either 
 by feeding or inoculation, but only one gave the disease both ways. 
 Martin thought that some of the results were due to the butcher 
 infecting the meat in the process of dressing the carcase, either by 
 his hands or knives. Woodhead made a series of experiments to test 
 the effects of roasting and boiling on the tubercular virus in meat. 
 It was found that in boiling and roasting experiments, as ordinarily 
 carried out in the kitchen, the temperature, however high it may be 
 on the surface, seldom reaches 60 C. in the centre, except in the 
 case of joints less than about six pounds in weight. Boiling and 
 roasting were found insufficient to destroy tubercular virus enveloped 
 in rolls of meat. 
 
 The following were among the conclusions of the Commissioners : 
 
 We have obtained ample evidence that food derived from tuberculous 
 animals can produce tuberculosis in healthy animals. The proportion of 
 animals contracting tuberculosis after experimental use of such food is 
 different in one and another class of animals ; both carnivora and 
 herbivora are susceptible, and the proportion is high in pigs. In the 
 absence of direct experiments on human subjects, we infer that man also 
 can acquire tuberculosis, by feeding upon materials derived from tuber- 
 culous food-animals. 
 
 The actual amount of tuberculous disease among certain classes of 
 food-animals is so large as to afford to man frequent occasions for 
 contracting tuberculous disease through his food. As to the proportion 
 of tuberculosis acquired by man, through his food or through other means, 
 we can form no definite opinion, but we think it probable that an 
 
TUBERCULOSIS. 399 
 
 appreciable part of the tuberculosis that affects man is obtained through 
 his food. 
 
 The circumstances and conditions with regard to the tuberculosis in 
 the food-animal which lead to the production of tuberculosis in man are, 
 ultimately, the presence of active tuberculous matter in the food taken 
 from the animal, and consumed by the man in a raw or insufficiently 
 cooked state. 
 
 Tuberculous disease is observed most frequently in cattle and in 
 swine. It is found far more frequently in cattle (full grown) than in 
 calves ; and with much greater frequency in cows kept in town cow- 
 houses than in cattle bred for the express purpose of slaughter. Tuber- 
 culous matter is but seldom found in the meat substance of the carcase ; it 
 is principally found in the organs, membranes, and glands. There is 
 reason to believe that tuberculous matter, when present in meat sold to 
 the public, is more commonly due to the contamination of the surface 
 of the meat with material derived from other diseased parts, than to 
 disease of the meat itself. The same matter is found in the milk of cows 
 when the udder has become invaded by tuberculous disease, and seldom 
 or never when the udder is not diseased. Tuberculous matter in milk is 
 exceptionally active in its operation upon animals fed either with the 
 milk or with the dairy produce derived from it. No doubt the largest 
 part of the tuberculosis which man obtains through his food is by means 
 of milk containing tuberculous matter. 
 
 Provided every part that is the seat of tuberculous matter can be 
 avoided and destroyed, and provided care be taken to save from contami- 
 nation by such matter the actual meat substance of a tuberculous 
 animal, a great deal of meat from animals affected by tuberculosis may 
 be eaten without risk to the consumer. 
 
 Ordinary processes of cooking applied to meat which has got con- 
 taminated on its surface are probably sufficient to destroy the harmful 
 quality. They would not avail to render wholesome any piece of meat 
 that contained tuberculous matter in its deeper parts. In regard to milk 
 we are aware of the preference by English people for drinking cow's milk 
 raw a practice attended by danger, on account of possible contamination 
 by pathogenic organisms. The boiling of milk, even for a moment, would 
 probably be sufficient to remove the very dangerous quality of tuber- 
 culous milk, 
 
 TUBERCULOSIS IN EQUINES. 
 
 Tuberculosis is not very common in the horse, but when it does 
 occur, it is frequently mistaken for glanders. There may be miliary 
 tuberculosis in the lungs, or nodules disseminated throughout the 
 lungs, liver, spleen, and bones. In a number of cases investigated 
 by Nocard, the disease commenced in the abdominal organs, and 
 the affection of the lungs appeared to be secondary. The author 
 has examined several cases of equine tuberculosis. In some cases 
 
400 INFECTIVE DISEASES. 
 
 the lungs were affected with the disease in a miliary form. The 
 bacilli could not be distinguished from bacilli in sections of the bovine 
 disease. Giant cells were extraordinarily numerous, and in many 
 cases were densely packed with bacilli, so that they could be recog- 
 nised en masse under a low power. The bacilli were also distributed 
 in the tissue generally, but were much more numerous in the giant 
 cells. 
 
 TUBERCULOSIS IN DOGS. 
 
 Peters described a case of tuberculosis in a pet dog, from eating 
 sputum from a tubercular patient. This is said to be a not 
 uncommon cause of canine tuberculosis. 
 
 TUBERCULOSIS IN CATS. 
 
 Nocard reported a case of tuberculosis in a cat from eating tuber- 
 cular sputum. The abdominal organs were diseased. Bollinger 
 has described two cases of miliary tuberculosis. M'Fadyean also has 
 described a case. The bacilli are very plentiful in the lung. A 
 minute examination of the individual micro-organisms by the author 
 did not reveal any distinctive character. 
 
 TUBERCULOSIS IN SWINE. 
 
 The author examined the tubercular liver of a pig. The pig 
 was about six months old, and after suffering from cough and 
 emaciation, died. 
 
 The liver had caseous nodules scattered throughout its substance, 
 some the size of a pea, and others larger. Tubercle bacilli without 
 distinctive characters were found on examination of sections; but 
 it was in some parts of a preparation difficult to detect any bacilli, 
 and in other parts there were not more than five or six in the 
 field of the microscope. Tuberculosis in swine is said to be very 
 rare in America. 
 
 TUBERCULOSIS IN BIRDS. 
 
 Hdns, guinea-fowls, turkeys, pheasants, and partridges, are sub- 
 ject to tuberculosis, and ostriches and other birds kept in confinement 
 may contract the disease. 
 
 Tuberculosis in fowls appears to be introduced principally with 
 the food, the disease occurring commonly in the intestines and 
 liver. 
 
DESCRIPTION OF PLATE XIII. 
 Tuberculosis in Swine. 
 
 Section of liver of a pig with scattered tubercular nodules. Microscopica 
 sections of the liver showed tubercle bacilli in very small numbers. 
 
Plate XIII 
 
 2 
 ft 
 
 PH 
 O 
 
 i 
 
 I 
 
TUBERCULOSIS. 401 
 
 The author has examined several cases of so-called spontaneous 
 tuberculosis in fowls. Sections of the liver were in one case remark- 
 able on account of the extraordinary invasion of the caseous deposits 
 with bacilli. Cover-glass preparations had been made from the 
 li\vr in the following way for diagnostic purposes : A tubercle was 
 iv.ulilv picked out on the point of a scalpel and crushed between two 
 >liilcs, and the cover-glass preparations stained with the Ziehl- 
 Neelsen solution. The bacilli are for the most part very small. A 
 f.-w attain a considerable length, but the majority are in the form 
 of small, straight rods, with many sizes intervening between these 
 ro Is and isolated granules. 
 
 In July 1888 the author received from Mr. Bland Button the 
 liver and lungs of a Rhea, which had died in the Zoological Gardens. 
 The lung was infiltrated with caseous deposits, and there were 
 scattered caseous nodules in the liver varying in size from a pea 
 to a marble. The naked-eye appearance of a section of the liver 
 through these nodules, at once recalled to mind the naked-eye 
 appearance of the deposits in the pig's liver already described. But 
 win -reas in microscopical preparations of the pig's liver, bacilli were 
 very >cautily present, the sections of the lung and liver of the Rhea 
 contained bacilli in such extraordinary numbers that, under a power 
 of fifty diameters, the collections of bacilli could be recognised as red 
 granular masses. These red masses under a high power were re- 
 solved into dense colonies of bacilli. In their number and their 
 distribution in the tissues, in their varying size, and in the extra- 
 ordinary length of the longest forms, they presented very interesting 
 points for observation. From the naked-eye appearance of the 
 disease and the general microscopical characters, as well as the 
 pr- -fiice of bacilli agreeing in their staining reactions with the 
 classical tubercle bacilli, the author had no hesitation in pronouncing 
 the disease to be avian tuberculosis. 
 
 Klein, who had examined a similar case, alluded to it in a 
 description of leprosy; but this disease is unknown in the lower 
 a niii in Is, and all attempts to infect them from man have been 
 almost, if not entirely, negative. 
 
 The bacilli in the Rhea are principally collected in the caseous 
 parts, but they are also found in the tissue generally, and often 
 collected in large cells. In size they vary to a marked extent. In 
 the cells they often form compact masses of short bacilli, but in 
 other parts, both in collections ami singly, they attain a greater 
 length than is observed in any other form of tuberculosis. Some of 
 the bacilli pre-ent a v ry interesting appearance. They are provided 
 
 26 
 
4()2 INFECTIVE DISEASES. 
 
 terminally with a sharply defined ovoid body. There are also 1 
 collections of short bacilli, many with these spore-like appearances. 
 The author has also seen free ovoid forms, sometimes singly, some- 1 
 times in groups. From their connection with the bacilli and their 
 sharply denned outline they are very suggestive of spores. 
 
 Johne examined the livers of a number of fowls accidentally 
 infected by phthisical sputum. Nocard reported an outbreak in 
 a poultry-yard where the man in charge had consumption. He ! 
 also found the disease amongst fowls fed with the infected organs 
 of tubercular cattle. Subcutaneous inoculation, and feeding of fowls 
 with sputum or bovine virus, will produce the disease. 
 
 Experimental inoculation of tubercular virus from different 
 sources affords an illustration of the different pathogenic effects 
 obtained by varieties of the same species of bacillus. The bacillus 
 of fowl-tuberculosis is a distinct variety. A very small proportion 
 of guinea-pigs, inoculated in the peritoneal cavity with fowl-tubercle, 
 succumb to the disease, though so susceptible to the effects of human 
 or bovine virus. Maffucci maintains that guinea-pigs have an 
 immunity, and that rabbits rarely develop a generalised tuberculosis. 
 Cultures are not identical in appearance with those obtained from 
 man, and on microscopical examination show many long, thick, and 
 branched forms, which are only rarely found in cultures from a 
 human source. 
 
 Stamping-out System. In 1888 a Departmental Committee 
 was appointed to inquire into pleuro-pneumonia and tuberculosis, 
 and they considered that legislation ought to be directed not only 
 to the protection of cattle from tuberculosis, but also to prevent 
 the possibility of the disease being communicated to man. 
 
 The following extracts are from the recommendations of the 
 Committee, which were made on the lines of : 
 
 A. PREVENTION. 
 
 B. EXTIRPATION. 
 
 A. Preventive Measures. 
 
 These should include provision for : 
 
 Improved hygiene of cattle sheds, etc. (especially in the direction of 
 providing proper ventilation, pure water supply, and adequate disinfection 
 of stalls, etc., wherein tubercular animals have been kept). This has 
 been partly met in the Dairy and Milk Shops Order, but its administra- 
 tion by the local health authorities is at present imperfect ; and we would 
 suggest that it should be much more stringently enforced, and that 
 veterinary inspectors should be given more extended powers of entry into 
 all places where animals are kept. 
 
TTHKRCTLOSIS. 403 
 
 Improvement in the hygienic surroundings of animals should include 
 isolation of all suspected cases, precautions against the flesh or milk of 
 diseased animals being given as food to others e.g., to pigs, fowls, etc. 
 and care that fodder, litter, and water should not be taken from one 
 animal or stall and given to another. 
 
 Our attention has been drawn to the frequency with which animals, 
 obviously diseased, sometimes even in the last stage of the malady, are 
 sold in open market. 
 
 Although in England and Ireland, under the provisions of the 
 Nuisances Removal Act as embodied in the Public Health Act, 1885, 
 the medical officer of health or inspector of nuisances may seize such 
 animals, yet such seizure is rarely performed. 
 
 We find the veterinary inspector has no power to prevent such sales, 
 or to seize the beasts for slaughter, since tuberculosis is not included in 
 the Contagious Diseases (Animals) Act of 1878. 
 
 We further find that there is actually a regular trade in such stock 
 infected with tuberculosis, and that they go by the name of ' k wasters " 
 and '' mincers," being frequently slaughtered in the neighbourhood of the 
 larger towns, to which such portions of the meat as are likely to escape 
 the observation of the inspector of nuisances are sent, for the purposes 
 of sale among the poorer inhabitants, and especially for the making of 
 sausages. 
 
 We are, therefore, very strongly of opinion that power should be given 
 to the veterinary inspector to seize all such animals in fairs, markets, or 
 in transit. 
 
 Notwithstanding the uniform prevalence of the disease in Europe and 
 elsewhere, there seems to be no reason to apprehend that, with our 
 present regulations for the slaughter of animals at the port of debarka- 
 tion, and for quarantine of those imported for breeding, there is any 
 special danger of increasing the infection in England by introduction 
 from abroad. The danger, however, exists in regard to the stock brought 
 from countries, which are exempt from slaughter on landing, and sub- 
 jected to the ordinary veterinary inspection during the present period of 
 detention of twelve hours. 
 
 . It is, therefore, evident that the present rules for the prevention of 
 the introduction of disease into the United Kingdom from abroad, are 
 incomplete. 
 
 Since all authorities are agreed that the disease is very marked by 
 heredity, we think it highly desirable that breeders should in their own, 
 a- well as in the public interest, discontinue breeding from tuberculous 
 stock. 
 
 B. Extirpation. 
 
 In order to insure the gradual extirpation of tuberculosis, we are of 
 opinion that it should be included in the Contagious Diseases (Animals) 
 Acts for the purposes of certain sections of those Acts, so as to provide : 
 
 (a) For the slaughter of diseased animals, when found diseased on 
 the owner's premises. 
 
404 INFECTIVE DISEASES. 
 
 (b) For the payment of compensation for the slaughter of such 
 
 animals. 
 
 (c) For the seizure and slaughter of diseased animals exposed in fairs, 
 
 markets, etc., and during transit. 
 
 (d) For the seizure and slaughter of diseased foreign animals at the 
 
 place of landing in this country. 
 
 Notification of this disease should not be compulsory, because it may 
 exist without developing any sufficient outward evidence to enable the 
 owner to detect it, and its growth is so slow, that non-notification of its 
 existence, even in a large number of cases, would do little to nullify the 
 stamping-out effect of the Act of 1878. 
 
 The powers and responsibilities of inspectors in ordering the slaughter 
 -of diseased animals should be the same for tuberculosis as for pleuro- 
 pneumonia, according to section 51 (5) of the Act of 1878. 
 
 Further, tubercle, though hereditary, is nevertheless much less conta- 
 gious than the other diseases included under the Act of 1878, and it is 
 <;lear, therefore, that the immediate slaughter of diseased animals would 
 go far to stamp it out, though, doubtless owing to heredity, this stamping- 
 out process would be gradual in its effect. 
 
 A supplementary report was made by Professor Horsley, in 
 which he expressed the opinion that there ought to be legislation 
 to prevent breeding from diseased animals, and compulsory notifi- 
 -cation : 
 
 1. Breeding. 
 
 Tuberculosis is notorious, even among the laity, as a disease which is 
 transmitted from parent to offspring. This is a fact with which cattle 
 breeders are specially familiar, and which finds strong expression in the 
 evidence attached to this report. Further, this generally received truth 
 has been completely confirmed by the results of scientific investigation, 
 -as is also duly set forth in the report. Considering, therefore, the 
 extreme importance of this point, I think that the act of wittingly breed- 
 ing from animals so affected should be made an indictable offence. The 
 only objection that can be raised to such legislation, which if effected 
 would prevent the dissemination of the disease among cattle in this 
 country, is that, owing to the present state of want of knowledge among 
 cattle owners, and even veterinary surgeons, of the early symptoms, and 
 physical signs on examination, of this disease, prosecutions would occa- 
 sionally occur in cases in which no fault could properly be attributed to 
 the owner, and that, therefore, such prosecutions would be needlessly 
 vexatious. 
 
 Considering, however, the extreme rarity with which such cases would 
 occur, and that, as in the matter of non-notification, each case would be 
 tried before district magistrates on its own merits, this objection is 
 deprived of the force it might have possessed. 
 
TUBERCULOSIS. 405 
 
 2. Notification of the Existence of the Disease. 
 
 This point requires no explanation, since it is clear that, unless the 
 veterinary inspectors or authorities receive information of occurrence of 
 diseases, it is impossible to ensure the thorough carrying out of the 
 provisions of the Contagious Diseases (Animals) Act. 
 
 That deliberate non-notification should be punished cannot be doubted 
 by any one. Objection, however, to legislation in this direction has been 
 put forward, on the same grounds as those upon which the prevention of 
 breeding from diseased animals was contested. As, however, I consider 
 that these objections have been already shown to have no weight, I 
 recommend that both the forbiddance of breeding from diseased animals, 
 and the notification of the disease, should be included in any legislation 
 for tuberculosis. 
 
 The difficulty referred to by the Committee, is presented by 
 cases of the disease which cannot be detected by the ordinary 
 methods of examination, and might possibly be overcome by the 
 use of tuberculin as a diagnostic agent. 
 
CHAPTER XXIX. 
 
 LEPROSY. SYPHILIS. RHINOSCLEROMA. TRACHOMA. 
 
 LEPROSY. 
 
 LEPROSY occurs in three forms tubercular, anaesthetic, and mixed 
 tubercular. It may be classed with the granulomata, as the most 
 common form of the disease is characterised by deposits in the skin, 
 mucous membrane, and internal organs. These deposits are composed 
 of small cells, and large cells resembling giant cells. The cells 
 become deposited in the surrounding tissues, and so the tubercle 
 enlarges, involving the epidermis and developing into an ulcerating 
 sore ; or, after a certain stage of development, beginning to decline, 
 and finally leaving a puffy discoloration. In the anaesthetic form 
 the cells invade the connective tissue of nerves. In the mixed 
 form the varieties occur together,' but the tubercular character 
 predominates. 
 
 Tubercular leprosy commences with the development of an 
 erytheinatous patch, which becomes infiltrated, and finally tubercu- 
 lated, the tubercles varying in size from a millet seed to a marble, 
 or even larger. The eruption on the head and face produces a 
 characteristic leonine expression. The progress of the disease is very 
 slow. After death the following changes may be found in the 
 internal organs : Cirrhosis of the liver and spleen, enlargement of 
 the lymphatic glands, and a condition of the lungs corresponding 
 to cheesy bronchial pneumonia. 
 
 In the anaesthetic form patches develop on the skin, which 
 become anaesthetic ; ulceration follows, and the fingers and toes, or 
 the entire hand and foot, may slough off. 
 
 The disease is undoubtedly communicable, but the infectivity is 
 of a very low type. 
 
 The infectiousness is illustrated by the well-known case of Father 
 Damien. Arning inoculated a man named Keanu, a condemned 
 criminal, and leprosy developed three years afterwards, but this case 
 
 406 
 
LEPROSY. 407 
 
 is not ivirardrd as i-onchiMve, as the man had a family history of 
 leprosy. Tlie disease has never been known to spread from patients 
 in this country, who have contracted the disease abroad. 
 
 The bacilli of leprosy were first observed by Hansen in 1874, and 
 ubeeqnently fully described by him, and his observations confirmed 
 l.v Nasser, in 1879. 
 
 Bacillus Leprae Rods 5 to 6 /A in length and 1 /A in breadth. 
 The bacilli are straight or curved, resembling very closely the 
 tubercle bacilli. They are present in the leprous tubercles of the 
 skin and mucous membrane, in the lymphatic glands, and in the 
 liver, testicles, and kidney; and in the nerves in the anaesthetic 
 variety. They are found between the cells, and in colonies in the 
 cells. They stain readily with the aniline dyes, especially by the 
 Ziehl-Neelsen and Gram's methods. The bacilli are found in extra- 
 ordinary numbers in the skin, and they are rather straighter than 
 tubercle bacilli, and stain more readily. 
 
 Numerous unsuccessful attempts to cultivate the bacillus have 
 been made by many bacteriologists. The author has made repeated 
 inoculations upon glycerine-agar, upon which the tubercle bacillus 
 .iriv\v abundantly, but always with disappointing results. On the 
 other hand, Bordoni-Uffreduzzi showed the author a cultivation 
 which he had obtained from the bone marrow of a leper. The 
 cultivation was made on blood serum and glycerine, and cover - 
 -l,i>- preparations resisted decolorisation with acid. There were 
 slight morphological differences when compared with the appear- 
 ance of bacillus leprse in the tissues, and the results were hardly 
 conclusive. 
 
 The English Leprosy Commission also reported successful cultiva- 
 tion of the leprosy bacillus. The author had the opportunity of 
 examining one of the first cultures received in this country, and 
 found that the bacilli stained deeply in ordinary cover-glass pre- 
 parations, they did not resist decolorisation by the Ziehl-Neelsen 
 method, and they corresponded in culture with one of the varieties 
 of Bacillus subtilis, commonly found on the skin. 
 
 Inoculation of animals has given equally unsatisfactory results. 
 Numerous experiments have been made by Beaven Rake on small 
 animals and birds, with invariably negative results. The blood of 
 leprous patients, tubercles from the living subject, fragments of the 
 skin and of the internal organs after death, have been inoculated by 
 different observers without result. Melcher and Ortmann alone claim 
 to have produced really definite results. Thase observers excised 
 leprous tubercles from the living subject, and inoculated fragments 
 
408 INFECTIVE DISEASES. 
 
 in the anterior chamber of the eye of rabbits. The animals died 
 after some months with extensive deposits in the coecum, lymphatic 
 glands, spleen, and lungs. 
 
 These tubercles varied in size from a pin's head to a millet seed, 
 and contained bacilli, resembling leprosy bacilli in their staining 
 reactions. The question naturally arises whether the lesions were 
 really indicative of leprosy or tuberculosis. Until the experi- 
 ments are independently confirmed, and the result of inoculation 
 differentiated from tuberculosis, it would be rash to accept these 
 experiments as conclusive. 
 
 It has been suggested that tuberculosis and leprosy are identical. 
 There is a similarity in the bacilli and in the lesions of leprosy and 
 tuberculosis, the injection of tuberculin produces a reaction in leprosy 
 nodules, and many lepers die from, tubercular disease of the lung. 
 But while tuberculosis is very readily transmitted to guinea-pigs 
 and rabbits by inoculation of fragments of tubercular tissue, leprosy 
 is inoculable, if at all, in most exceptional instances. The bacilli 
 of tubercle are cultivated with the greatest facility, the bacilli of 
 leprosy, if at all, only with exceptional difficulty; tubercle bacilli 
 are found in giant cells, leprosy bacilli in the so-called leprosy 
 cells. Leprosy bacilli are straighter than human tubercle bacilli,, 
 and differ slightly in their behaviour to staining reagents. On 
 the other hand, the morphological differences are not greater than 
 those existing between different forms of tubercle bacilli obtained 
 from tuberculosis in animals and birds. It would be premature 
 to regard leprosy as a variety of tubercle until cultivations of 
 the bacillus have been obtained, and carefully compared with 
 those of the tubercle bacillus. Differences in morphological details 
 and results of inoculation would then carry less weight as a means 
 of differentiation. 
 
 The tubercular pneumonia of lepers would be regarded, if the 
 bacilli are identical, as a development of leprosy in the lungs, and 
 not, as at present, a result of double infection with tuberculosis. 
 
 METHODS OF EXAMINING THE BACILLUS OF LEPROSY. 
 
 Cover-glass preparations may be made in the ordinary way, or by a 
 special method, which consists in clamping a nodule with a pile clamp 
 until a state of anaemia of the tissue is produced. On pricking with a 
 needle or sharp knife a drop of clear liquid exudes, from which cover- 
 glass preparations may be made, and stained by Neelsen's method. 
 
 For sections the author prefers Neelsen's method and methylene-blue. 
 They can also be stained by Gram's method, which, as a rule, brings out 
 very clearly the beaded appearance of the bacilli. 
 
DESCRIPTION OF PLATE XIV. 
 Bacillus Leprae. 
 
 FIG. 1. From a section of the skin of a leper. The section is, almost in 
 its entirety, stained red, and, with moderate amplification, has a finely 
 granular appearance. Stained by the Ziehl-Neelsen method (carbolised 
 fuchsine and methylene-blue). x 200. 
 
 FIG. 2. Part of the same preparation with high amplification, showing that 
 the appearances described above are due entirely to an invasion of the 
 tissue by the bacilli of leprosy, x 1500. 
 

 
 
 Figl 
 
 Fig 2 
 
 BACILLUS 
 
 
 
LEPROSY. 409 
 
 of Babe*. Preparations are stained in rosaniline hydrochlorate 
 in aniline water, decolorised in 33 per cent, hydrochloric acid, and after- 
 stained with methylene-blue. 
 
 Stamping -out System. The history of leprosy in the British 
 l.-l;tnds during the Middle Ages, and the conditions under which it 
 both increased and declined, have been discussed by several writT>. 
 A large number of institutions of a charitable and ecclesiastical 
 character were established in endemic areas and were occupied by 
 the lepers either voluntarily, or compulsorily by means of the Act 
 De leproso amorendo. These institutions were to a very small extent 
 a means of segregation. According to Dr. Newman the disease, 
 which had reached its zenith about the twelfth or thirteenth century, 
 to decline from that time owing to " a general and extensive 
 improvement in the life of the. people, to a complete change 
 in the poor and insufficient diet (which it is evident consisted far 
 too largely of bad meat, salt, putrid and dried fish, and an almost 
 entire lack of vegetables) and to agricultural advancement, improved 
 sanitation and land drainage." Of all the unfavourable conditions 
 it would appear that food in some way was especially associated with 
 the cause of the disease, either by introducing the bacillus or by 
 rendering the tissues a suitable soil for its reception and development. 
 
 In other countries segregation has been attempted voluntarily 
 or compulsorily, but it has never been completely carried out. There 
 can be very little doubt that the presence of a leper in a healthy 
 community is no greater source of danger than the presence of 
 an individual suffering from tuberculosis, but, for other reasons, 
 voluntary isolation should be carried out as completely as local 
 circumstances will permit. 
 
 The Leprosy Commission in India recommended 
 
 (a) That the sale of articles of food and drink by lepers should 
 
 be prohibited, and that lepers shoidd be prevented from 
 following certain specified occupations. 
 
 (b) That the concentration of lepers in towns should be discouraged. 
 
 (c) That Leper Asylums should be established in which lepers 
 
 might live voluntarily. 
 
 (d) That Leper Farms scattered over the country should be 
 
 encouraged. 
 
 (e) That the few children who are born of lepers should be 
 
 removed to Orphanages. 
 
 They concluded that by means of improved sanitation and good 
 dietetic condition*, a diminution of leprosy will result. 
 
410 
 
 INFECTIVE DISEASES. 
 
 
 
 SYPHILIS. 
 
 Syphilis is a disease peculiar to man, and communicable only by 
 inoculation. The local infection is followed by a period of latency, 
 and by a period during which generalised eruptions appear. One 
 attack confers immunity from future attacks. The virus in its 
 most virulent form is found in the primary seat of inoculation, and 
 
 in the indurated glands which 
 follow. It is also supposed to be 
 present in the blood and secretions. 
 Lustgarten, Eve and Lingard have 
 found bacteria which they believed 
 to be specific. 
 
 Bacillus in Syphilis (Lustgar- 
 ten).- Rods resembling the bacilli 
 of leprosy and tuberculosis, 3 to 4 //, 
 long, -8 fj, thick. Two or more 
 colourless, ovoid points in the course 
 of the rod are visible with a high 
 
 power; it is thought that possibly they are spores. The bacilli 
 are always found in the interior of nucleated cells, which are 
 more than double the size of 
 leucocytes. They have been ob- 
 served in the discharge of the v'f^S '"''n^S--^-^-''^-'?-' 
 primary lesion, and in tertiary vV^Sfe^ / ^ ^.^;. 
 gummata. 
 
 Alvarez and Tavel state that 
 an identical bacillus is found in 
 normal secretions (smegma). Eve 
 and Lingard have described a bacil- 
 lus associated with specific lesions, 
 which differs from the above in its 
 morphology and behaviour towards 
 staining reagents. 
 
 FIG. 175. COVER-GLASS PREPARA- 
 TION OF PUS FROM A CHANCRE, X 
 
 1050 (LUSTGARTEN). 
 
 . - >-?r- N 
 
 '- 
 
 
 
 
 FIG. 176. WANDERING CELL CON- 
 TAINING BACILLI (LDSTGABTEN). 
 
 METHODS OF STAINING THE BACILLUS OF SYPHILIS. 
 
 Method of Lustgarten : 
 
 Sections are placed for twelve to twenty-four hours in the following 
 solution, at the ordinary temperature of the room, and finally the solution 
 is warmed for two hours at 60 C. : 
 
 Concentrated alcoholic solution of gentian-violet 
 Aniline water 
 
 11 
 
 100 
 
SYPHILIS. 411 
 
 The sections are then placed for a few minutes in absolute alcohol, 
 and from this transferred to a 1'5 per cent, solution of permanganate of 
 potash. After ten minutes they are immersed for a moment in a pure 
 concentrated solution of sulphurous acid. If the section is not completely 
 decolorised, immersion in the alcohol and in the acid bath must be 
 repeated three or four times. The sections are finally dehydrated with 
 absolute alcohol, cleared with clove-oil, and mounted in Canada balsam. 
 
 By this method the bacillus is distinguished from many bacteria, but 
 not from the bacilli of tubercle and leprosy which are stained by this 
 process. 
 
 Mi tin nJ i if I)e Giacomi : 
 
 Cover-glass preparations are stained with hot solution of fuchsine 
 containing a few drops of perchloride of iron. They are then decolorised 
 in strong perchloride of iron, and after-stained with vesuvin or Bismarck- 
 brown. 
 
 Method of DoutrtUptoU "nil SrJt/ltz : 
 
 Sections are stained in a weak aqueous solution of gentian-violet and 
 after-stained with safraniu. 
 
 The nature of the contagium in syphilis is unknown. 
 
 Protective Inoculation. Inoculation of the virus, or syphilisa- 
 tion, as a protective measure, was at one time practised and strongly 
 advocated; but ifc is rightly regarded in this country as dangerous 
 and u 11 justifiable. From the experiments of Ricord it would appear 
 that the local results in the vesicular stage resemble the results of 
 the inoculation of virulent vaccinogenic grease or horse-pox. The 
 inoculation goes through the stages of papule, vesicle, ulcer, scab, 
 and scar. The accidental inoculation which occurs in cases of 
 vaccine-syphilis may so closely resemble the results of inoculation 
 with very virulent cow-pox, that it is sometimes difficult to decide 
 as to the exact nature of these cases. 
 
 RHINOSCLEROMA. 
 
 Rhinoscleroma is a rare disease, resembling lupus, and pro- 
 ducing in the nostrils and neighbouring parts nodular swellings, 
 composed of granulation-tissue. The disease is met with in 
 America, Egypt, Austria, and Italy. There are no giant cells, but 
 peculiar large cells, which were first described by Mikulicz. Frisch 
 discovered bacteria in sections, and Cornil and Alvarez pointed out 
 the existence of a capsule. In morphology and cultivation they 
 resemble, according to Dittrich, Friedlander's pneumococcus. They 
 are probably identical with this micro-organism, and Paltauf and 
 Eiselsberg, and others, found that they produced septicaemia in rabbits 
 and guinea-pigs. 
 
 Bacterium of Rhinoscleroma (Bacillus of Ehinosdervma, 
 
412 INFECTIVE DISEASES. 
 
 Cornil and Alvarez). Cocci and short rods, 1*5 to 3 /I in length, 
 5 to *8 //, thick. Deeply coloured points or granules may occur 
 in the course of the rods when stained, but it is very doubtful 
 whether these can be considered as spores. The bacteria are en- 
 capsuled, the capsule being round when enclosing a coccus, and 
 ovoid when enclosing a rod. The capsule is composed of a tough 
 resisting substance; two or more capsules may unite by fusion, 
 enclosing two, three, or a greater number of rods. The bacilli were 
 observed in sections of the tumours, which developed on the lips 
 and in the nasal and pharyngo-laryngeal regions. 
 
 METHOD OF STAINING THE BACILLUS OF RHINO- SCLEROMA. 
 
 Method of Condi and Alvarez : 
 
 Sections are immersed in a solution of methyl-violet (B) for twenty- 
 four to forty-eight hours, with or without the addition of aniline-water ; 
 and are then decolorised after treatment with the solution of iodine in 
 iodide of potassium. If the sections are left to decolorise in alcohol for 
 forty-eight hours, the capsule is rendered visible. 
 
 TRACHOMA. 
 
 Trachoma is a disease of the conjunctiva, common in Egypt. 
 The new growth is composed of round cells, and may be regarded, 
 according to Kartulis, as the chronic stage of either gonorrhceal or 
 Egyptian ophthalmia. Koch failed to find any micro-organisms 
 in the swollen lymph follicles. Sattler asserted that he had culti- 
 vated a micrococcus which produced the disease when inoculated 
 on the conjunctiva. Other observers have found the common 
 pyogenic micrococci in the secretions, especially Staphylococcus 
 pyogenes aureus and albus. 
 
CHAPTER XXX. 
 
 ACTINOMYCOSIS. MADURA DISEASE. 
 ACTINOMYCOSIS. 
 
 ACTIXOMYCOSIS belongs to the class of infective granuloinata. It is 
 a chronic inflammatory affection characterised by the presence of a 
 .-pecinl microphyte, which by irritation produces a neoplasm, composed 
 of round cells, epithelioid cells, giant cells, and fibrous tissue. These 
 neoplasms form nodular tumours of various sizes. In some cases 
 there is a tendency to develop very large tumours, and in others to 
 break down early and suppurate. In cattle, cretification takes 
 place in the fungus tufts. Act ino mycosis closely resembles tuber- 
 culosis in its histological characters. The disease attacks man, 
 horses, cattle, and pigs. 
 
 Many interesting observations have been made upon the origin 
 of this disease in man. Two cases have been recorded in support of 
 the theory of direct infection from the cow. Stelzner described a 
 case of actinomycosis in a man who had had the care of animals, 
 some of which had suppurating glands. Hacker had a case of 
 actinomycosis of the tongue in a man who had charge of cows, one 
 of which had a tumour of the jaw which he had opened. On the 
 -other hand, Mooebrogger found that out of 75 cases, 54 were in men, 
 and 21 in women, including 2 children. In 11 of these men the 
 occupation was not stated. In 33 their occupation did not bring 
 them into contact with diseased animals ; they were, for example, 
 millers, glaziers, tailors, shop people, and students. Only 10 cases 
 occurred among farmers, peasants, and farm -labourers, and in only 
 one case out of the 10, had the patient been brought into contact 
 with diseased animals. 
 
 Out of the 21 women, there were only 4 peasants, and none 
 <f them had been associated with diseased cattle. 
 
 Infection by the flesh of diseased animals has also been clis- 
 <-ued. But there is no evidence of prevalence of the disease 
 
 413 
 
414 INFECTIVE DISEASES. 
 
 among slaughterers and butchers, who would be particularly liable 
 to it, if flesh were a source of infection. The chances of infection 
 by ingestion are minimised by the flesh being almost always cooked. 
 Actinomycosis occurs also in pigs, and pork is very often eaten in an 
 uncooked state ; but Israel has pointed out that this may probably 
 be excluded, as many of the cases occurred among strict Jews. 
 
 The evidence points to the disease originating in man and lower 
 animals from the same source, and there is a very strong suspicion 
 attached to cereals. This view is supported by important obser- 
 vations, with reference to the part played by cereals in inducing 
 the disease in cattle, and it gains additional support from a case 
 described by Soltmann, where the disease resulted from an awn of 
 wall barley. A boy, aged eleven, accidentally swallowed an awn 
 of Hordeum murinum. He became very ill, and suffered great pain 
 behind the sternum, extending to the back. An abscess formed r 
 covering an area extending over six intercostal spaces, and when 
 opened, the awn of this grass was found in the evacuated pus. 
 The pain, however, continued, and fresh deposits occurred, and when 
 the boy was taken to the hospital, the ray-fungus was detected. 
 Possibly the spores of the fungus can be conveyed both by air and 
 water. 
 
 This disease in cattle has long been known in this country, but 
 its various manifestations were either mistaken for other diseases, 
 or simply received popular names. Indeed, the various forms are 
 still familiar to many as wens, clyers or crewels, scrofulous, tuber- 
 cular or strumous abscesses, polypus, lymphoma, cancer of the 
 tongue, scirrhous tongue, indurated tongue, ulcerated tongue, cancer 
 of bone, bone tubercle, osteo-sarcoma, fibroplastic degeneration of 
 bone, spina ventosa, and carcinoma. 
 
 Bovine antinomycosis is especially prevalent in river valleys, 
 marshes, and on land reclaimed from the sea. The disease occurs 
 at all times of the year, but general experience leads to the belief 
 that it occurs more commonly in the winter. 
 
 It is more frequently met with in young animals, and usually 
 occurs between one and three years, but it may be found at almost 
 any age, and probably affects equally both sexes. 
 
 There is little if any evidence to show that the disease is heredi- 
 tary. In numerous cases, the family history has been most carefully 
 inquired into by the author ; and in the case of some imported 
 pedigree animals, the disease was quite unknown on the farm where 
 they had been bred. 
 
 The tongue is so commonly the seat of the disease, that suspicion 
 
ACTINOMYCOSIS. 415 
 
 at once falls on food as the means by which the parasite is conveyed. 
 Skin wounds produced by rubbing against the mangers, posts, or 
 \virt- fencing, may also become infected. 
 
 The evidence is very strong in favour of believing that the micro- 
 organism gain> anvss to the system through wounds or lacerations 
 of tlu- mucous membrane and skin, or through carious teeth. It 
 ha> l>een pointed out that the common occurrence of the disease 
 at the time of the second dentition may be owing to the wound.- 
 produced in the alveolar mucous membrane by the shedding of 
 the teeth. Experience also points to straw being sometimes a 
 fact or in the production of the disease, and it is possible that thistles 
 ami frozen roots also, by wounding the mucous membrane, may 
 afford a way for the entrance of the micro-organism. The disease 
 in the jaws, both in man and in cattle, is very commonly associated 
 with carious teeth. 
 
 The cowsheds, pastures, and chinking tanks may become infected 
 with the discharges from diseased animals. The discharge con- 
 taminates the fodder in the sheds, and falls on thistles and siliceous 
 .Task's in the pasture, which may first wound, and then introduce 
 the micro-organism. The discharge is also coughed out of the 
 mouth, and expelled from the nose, in cases in which a tumour in 
 the pharynx, or the nasal chambers, has undergone suppuration. 
 
 Jensen believed that the disease was produced by different kinds 
 of grain, especially when cultivated on ground reclaimed from the 
 sea. He mentions an instance of a farm, where nearly the whole 
 of the young stock, about thirty in number, had actinomycosis after 
 feeding on mixed forage, grown on a certain field. Two years after- 
 wards the same disease occurred in the same stalls in four animals, 
 after being fed on barley-straw from the same field. According to 
 Jensen, the fungus grows on grain, husks, and straw of different 
 cereals, but most abundantly on barley, which is also the most 
 likely to w^ound the mucous membrane. Johne's observations tend 
 to corroborate this view, for in twenty-two out of twenty-four cases 
 in which he found barley sticking in the tonsils of pigs, he found 
 the beard thickly beset with a fungus very similar to, if not identical 
 with, the ray-fungus. These observations are of great interest in 
 connection with Soltmanii's case. 
 
 Experience points to the belief that the disease is not readily 
 communicable from animal to animal, and it is possible that when 
 it affects a large number of cattle in -a herd, the same causes have 
 been acting to produce the disease in a number, which in another 
 in.-taiice may only produce it in one. At the same time, isolated 
 
416 INFECTIVE DISEASES. 
 
 cases are possibly not quite so common as they are reported to be. 
 It is well known that, as a rule, the services of a veterinary surgeon 
 are not called for except in hopeless or very severe cases. The 
 cowmen themselves, in many districts, treat the cows successfully, 
 and then send them into the market, and thus the existence of 
 previous cases may not have come to the knowledge of the veterinary 
 surgeon. 
 
 Historical. In 1845 Professor von Langenbeck, of Kiel, made 
 notes of a case of vertebral caries in a man, and prepared drawings 
 of peculiar bodies in the pus from an abscess. The drawings were 
 published together with a reference to the case by Israel in 1878. 
 There can be little doubt that these structures were the fungi of 
 actinomycosis. But the first to publish observations was Lebert 
 in 1848. 
 
 Lebert received from M. Louis some pus, of a thick, almost 
 gelatinous consistency, which had been obtained from an abscess of 
 the thoracic wall in a man aged fifty. The patient had been attacked 
 four months previously by a pulmonary affection, which was 
 suspected by M. Louis to be cancerous in nature. The pus contained 
 a very considerable number of little spherical bodies of a slightly 
 greenish-yellow colour, about the size of a pin's head. They 
 could be readily crushed between two strips of glass, and 011 
 examination with a power of fifty diameters two elements could be 
 distinguished : a soft connective substance, and many hard, narrow, 
 wedge-shaped corpuscles, arranged in a radiating manner. Under 
 a high power these bodies were observed to be -^ to ^ of an inch 
 in length, ^ in width at the base, and ^ in width at the 
 apex. Some of these corpuscles were regular, while others showed 
 one or two constrictions, with intermediate flask-shaped swellings. 
 Lebert tested these structures with reagents, with the following 
 results. The bodies were found to remain unaltered by concentrated 
 mineral acids. Acetic acid freed them from foreign elements 
 adhering to their surface. Solution of caustic potash did not affect 
 them if used cold, but a boiling solution reduced the cuneiform 
 structures to a fine greyish powder without dissolving them. Ether, 
 alcohol, and chloroform had no effect upon them when used either 
 hot or cold. Solution of potash, in which these bodies had been 
 heated, mixed with a solution of sulphate of copper and brought to 
 boiling point, did not offer any uniform red colour, which would 
 have been the case if they had contained albumin. Thus, the chief 
 chemical characters of albuminous and fatty substances were 
 wanting, and they resembled chitine in their behaviour to reagents. 
 
ACTINOMYCOSIS. 417 
 
 bore in mind the possible existence of some helminthic debris, 
 of which these bodies might be booklets, but he sought in vain for 
 rrhinococci and cysticerci. 
 
 Actinomycotic pus was later described and figured by Robin. 
 In the illustration accompanying the description, the fungi are most 
 
 FIG. 177. SECTION OF LIVER FROM A CASE OF ACTINOMYCOSIS IN MAN. 
 
 accurately depicted. Robin states that he had found, in two or 
 three cases in the pus of deep-seated chronic abscesses, yellowish grains 
 attaining a diameter of one-tenth of a mm., surrounded by a sort of 
 halo or thin, viscous, finely granular stratum, containing leucocytes. 
 Theee grains were composed of elements 2 to 6 mm. in length, swollen 
 
 27 
 
418 INFECTIVE DISEASES. 
 
 at one end and tapering off at the other, arranged in a regular series, 
 radiating from a common centre which consisted of granular matter. 
 They were highly refractive, possessed a brilliant centre and sharply 
 defined outline ; they were dissolved, or at least rendered indistinct, 
 by acetic acid, and proved insoluble in ammonia and ether. 
 
 The disease in man was next described by Israel in the paper 
 mentioned above. Poiifick was the first to clearly recognise the 
 identity of the disease in man with the disease in cattle, and he 
 described a number of cases in man. Israel subsequently published 
 a work on the subject. The various cases which had been observed 
 up to that date were described, and the disease classified according 
 to the seat of invasion. 
 
 From this time onwards numbers of cases in man have been 
 described, and various important researches published, of which 
 those of Bostrom and Moosbrugger may be especially mentioned. 
 
 In England, Acland recognised a case on examining the liver 
 after death (Fig. 177). H. Taylor was the first, in this country, 
 to detect the fungus during the life of a patient. Shattock found 
 .specimens of the disease in museums. Skerrit, Powell and Godlee, 
 Eve, Delepine, Ransome, Poore, Malcolm Morris and others have 
 published cases. 
 
 In Italy, Perroncito studied the sarcomata of cattle, and claims 
 to have first observed the micro-organism in 1863, In 1875 he 
 described it in the Encyclopaedia Agraria, and, from the negative 
 results obtained by inoculation experiments, was led to regard it, 
 not as the cause, but as a result of the disease. 
 
 Rivolta of Turin also claims to have been the first to have 
 discovered the fungus in actinomycosis bovis. As early as 1868 
 he published a paper on a sarcomatous tumour of the jaw of 
 -an ox. 
 
 Hahn of Munich, in 1870, undoubtedly met with the fungus, for 
 he states that in a case of " wooden- tongue " he found characteristic 
 organised structures, which he provisionally described as a species of 
 mould fungus. 
 
 Bollinger was the first to recognise the nature of this disease in 
 cattle. In 1876 he pointed out that new growths occasionally 
 occurred on the upper and lower jaws of cattle, which either started 
 from the alveoli of the back teeth, or from the spongy tissue of the 
 bone, and by increasing in size loosened the teeth. In their progress 
 they destroyed bone, muscles, mucous membrane, and skin. After 
 some time they frequently broke down, forming ulcers, abscesses, 
 .and fistulas ; but in some cases tumours were formed, which attained 
 
ACTINOMYCOSIS. 419 
 
 the size of a child's head, Bellinger stated that this disease had 
 been known by various names, Osteosarkome, Winddorn (Spina 
 ventosa), Knochenkrebs, Knochenwurm ; in other instances it had been 
 regarded as bone tuberculosis, or mistaken for a simple chronic 
 glossitis. Among breeders of cattle and owners of stock in Germany 
 it had been known under the following names : Ladendruck, Laden- 
 yeschwulst, dicker Backen, Backel, Kinnbeule, Kiefergeschwulst, etc. 
 
 Bellinger pointed out that these swellings consisted of several 
 centres of growth, bound together by connective tissue. They were 
 often as large as a walnut or a hen's egg, and of a pale yellow 
 colour and moist appearance. The cut surface presented yellow ish- 
 white, suppurative foci, while in other cases the growths had a 
 spongy texture, owing to the formation of lacunae or hollow spaces 
 in M fibrous stroma, which contained a turbid, thick, yellow, caseous 
 pulp. 
 
 Microscopical examination of the tumour showed that it had a 
 structure like a sarcoma, while the squeezed-out pulp consisted 
 principally of pus cells, granulation cells, fat granules, and granular 
 detritus. In addition, there were numerous opaque, pale-yellow, and 
 coarsely granular bodies of different sizes, which had a mulberry- 
 like appearance, and were sometimes encrusted with chalk. After 
 careful examination Bollinger found that these bodies were true 
 fungi, and he further maintained, from the constancy of their 
 appearance in all parts of the sarcomatous growth, that they were 
 not accidental, but of pathogenic significance. This was found to 
 be the case, not only in fresh preparations, but in old specimens 
 preserved in the museum. This remarkable form of mycosis was 
 observed by Bollinger, not only in the upper and lower jaws, but 
 also in the tongue. It had long been observed that the tongue was 
 sometimes covered with more or less tubercular growths, scattered 
 abundantly over the surface of the mucous membrane, mostly the size 
 of a millet seed or hemp seed, but often reaching the size of a cherry 
 or walnut, or even larger. In the fresh state these nodules were 
 greyish- white, and semi-transparent, but they soon became cloudy 
 or distinctly puriform in the centre ; they were surrounded externally 
 with a connective tissue capsule. If the nodules were situated on 
 the surface of the tongue, destruction of the mucous membrane very 
 readily followed, leading to the formation of ulcers. The tongue 
 also might become affected with an interstitial glossitis, which often, 
 in spite of the partial atrophy of the muscular fibres, led to a great 
 enlargement and wood-like hardness of the tongue. On account 
 of this peculiar character, such a tongue was long known in South 
 
420 INFECTIVE DISEASES. 
 
 Germany as Holzzunge. In other cases the condition was regarded 
 as " tubercle of the tongue," " chronic sarcoma," " chronic interstitial 
 glossitis," or simply "degeneration of the tongue." 
 
 Bollinger described this disease as occurring in cattle of all ages r 
 developing itself gradually, and being always incurable. As a rule, 
 the animals were slaughtered, because the diminished mobility and 
 enlargement of the tongue interfered with feeding. He also pointed 
 out that this disease of the tongue was by no means rare, as he had 
 had no less than six such tongues from different parts of Bavaria in 
 the space of a year, and he also had been able to prove the existence 
 of the disease in museum specimens. 
 
 On further continuing his researches, Bollinger found the same 
 fungus in tumours which occurred in the pharynx, larynx, and 
 the mucous membrane of the stomach. These tumours were very 
 common in the throat in some parts of North Gernianv, where as 
 many as 5 per cent, of the animals had been known to be affected. 
 The disease frequently occurred in the form of subcutaneous 
 neoplasms, called Lymphonie, Ilohzgeschiviilste, Fibrome, Tuberkel r 
 Tuberkel-scropheln. 
 
 This disease also appeared in the form of abscesses, which were 
 called, in many districts, Schlundbeulen. These growths were found 
 in the neighbourhood of the parotid gland, the larynx, and pharynx, 
 and were similar in every respect to the affection of the jaw. They 
 were described as starting apparently from lymphatic vessels in 
 these parts. Bollinger discovered the fungus in a case of so-called 
 fibroid of the second stomach of a cow, a spongy growth nearly the 
 size of the fist ; and he believed that in another case the disease 
 manifested itself in the form of tubercular ulceration of the 
 intestines. 
 
 Bollinger submitted the fungus to Dr. Harz, a botanist, who 
 described the fungi as mulberry-like masses from -5 to 1 mm. in 
 diameter. They appeared to the naked-eye as opaque, white grains, 
 and when calcified were difficult to recognise. On slight pressure 
 the tufts of the fungi fell apart into segments of unequal size, each 
 of which appeared to correspond to an individual fungus. The 
 latter was described as beginning at the pointed end of the wedge, 
 with a somewhat cone-shaped basal cell, which, in the absence of a 
 mycelium, perhaps took its place, and bore a great number of short 
 linked hyphse. At the ends of the hyphse there were oval, globular, 
 or elongated club-shaped bodies, the reproductive cells or gonidia. 
 
 Cultivation experiments, and inoculation of the tongue of a calf 
 with liquid containing the micro-organism, failed. Harz proposed 
 
ACTINOMYCOSIS. 421 
 
 to call the fungus, from its ray-like appearance, actinomyces ; but 
 what the position of the fungus in nature might be, was difficult to 
 determine. It did not, he believed, belong to the yeast fungi, but to 
 the mould fungi, and might be compared to Botrytis, Monosporium, 
 and Polyartis. 
 
 Bollinger concluded that there could be no doubt that actino- 
 myi'osis occupied an important position in the pathology of cattle 
 diseases. As further evidence of the prevalence of the affection, he 
 remarked that Zippelius of Obernburg had observed in the course of 
 about ten years' practice not less than 254 cases of lymphoma, in 
 the neighbourhood of the larynx and pharynx, besides 157 cases 
 of disease of the jaw; and Bollinger says that he had very little 
 doubt that the greater part of the former, and very likely all the 
 ca>es in the jaw, were due to the fungus which he had discovered. 
 In certain parts of Franconia, according to a communication received 
 from Professor Frank, these tumours of the throat were extremely 
 abundant in cattle. 
 
 Bellinger's researches were followed by those of Siedamgrotzky, 
 and later by a communication from Johne. Johne described the 
 various forms of the disease which had up to that date been 
 recognised, including a description of actinomycosis of the bones 
 of the jaws, of the fauces, of the larynx, of the oesophagus, of the 
 stomach and intestinal canal, and of the udder. He carried out a 
 series of experiments, by which it was clearly established that the 
 disease could be communicated from cattle to cattle. Previously 
 Bollinger, Harz, Perroncito, Ponfick, Siedamgrotzky, and Johne had 
 failed, but subsequently by employing fresh material from the 
 living animal, both Johne and Ponfick succeeded. 
 
 Siedamgrotzky not only confirmed Bellinger's researches, but he 
 described the presence of the fungus in so-called " multiple sarcomas '' 
 of the mucous membrane of the oesophagus. Rabe described the 
 presence of the fungus in tumours known as Winddorn, and pointed 
 out that, in at least one case, he considered that the disease had 
 been carried by the lymphatics. There were eleven subcutaneous 
 tumours in a row on the face, which were connected by swollen, 
 rope-like, lymphatic vessels. They appeared to be secondary to a 
 growth on the nostril, the size of a hen's egg. 
 
 Perroncito described a case of " sarcoma " of the intestines and 
 >tomach, which proved to be actinomycosis. 
 
 Many additional communications were made on the subject of 
 this disease. Ponfick produced it in the lungs by intravenous 
 injection, and subsequently three cases occurring naturally in the 
 
422 INFECTIVE DISEASES. 
 
 practice of veterinary surgeons were published. They not only 
 deserve especial mention, but as this form of the disease appears to 
 be so seldom recognised, they will be given in detail. 
 
 Plug described a case in the lungs. The cow had been out of 
 health for four weeks, did not eat, and had a cough, and two days 
 previous to the visit had become rapidly \vorse. Schmidt found 
 dyspnoea with abdominal respiration ; the nostrils were dilated, the 
 head protruded, and the mouth kept open. There was dulness 011 
 percussion, and crepitation. The animal was killed, and the lungs, 
 which alone were diseased, were sent to Plug. The pleura on exami- 
 nation was normal, but beneath it were numbers of miliary 
 tubercles, many equal in size to a pin's head. On section the lung 
 had a granular appearance from the presence of countless numbers 
 of minute deposits, which all had the appearance of grey tubercles ; 
 in none was there any central softening. They were present in 
 enormous numbers around the bronchi, and in the vessels of the 
 interlobular tissue. Microscopical examination showed, in the middle 
 of most of these nodules, the presence of greenish-yellow, radiating 
 bodies, which under a high power appeared to be undoubtedly 
 actinomycotic granules. In many there were only rudimentary fungi 
 consisting of four or five clubs ; there was only one rosette in each 
 tubercle. The fungus was surrounded by round cells and fibrous 
 tissue. Larger nodules resulted from the agglomeration of several 
 tubercles, or from diffuse infiltration of round cells in the neighbour- 
 hood of a tubercle. 
 
 Hink met with a somewhat similar case. A ten-year-old cow- 
 was slaughtered, and in the middle lobe of the right lung there were 
 yellowish nodules about the size of a pea, scattered over an area 
 the size of the palm of the hand. These nodules were not at 
 first sight distinguishable from ordinary tubercles, but on closer 
 inspection they appeared to be somewhat different, and could be 
 easily shelled out from the thickened lung tissue. On making a 
 section, pus welled up at several points, and contained yellowish, 
 calcareous particles. These particles, on microscopical examination, 
 were found to be strongly calcified tufts of the actinomyces embedded 
 in granulation cells. Addition of hydrochloric acid dissolved the 
 calcareous matter, but had no action on the fungus. 
 
 Pusch described a third case. The lungs of a cow, which had 
 been killed on suspicion of having pleuro-pneumonia, were sent for 
 examination. The front lobe of the left lung was collapsed and 
 firm, the pleura was thickened and opaque ; the larger bronchi were 
 enlarged, filled with pus, and their walls thickened. In the posterior 
 
ACTINOMYCOSIS. 42 & 
 
 ]<>lie of the left lung there was a cavity the size of the fist, which 
 had been opened, and the contents had, for the mo>t part, escaped ; 
 what remained was a greyish, purulent liquid, full of yellowish 
 1 to. lies. By the side of this cavity there was another collection of 
 pus, the size of a walnut. In the lower part of the second lobe of 
 the riiiht lung there was a firm, grey tumour, the size of a hen's 
 ::. <>\er which the pleura was much thickened. On section this 
 was ca vernous, with similar purulent contents, and yellow grain.-. 
 Tlu-f grains under the microscope proved to be ray-fungi. The wall 
 of the cavity consisted of dense connective tissue lined with a soft 
 Lfranulatioii tissue, bathed in pus. There was no disease of any 
 other parts in this case, so that it corresponded in this respect with 
 the two previous ones. Pusch adds that it was difficult to determine 
 whether the organism had gained access to the lungs by the blood- 
 \e-sels. or by the inspired air. In his case he inclined to the latter 
 view, and concludes by saying that the organism is probably very 
 common and attached to the most varied objects, from which it 
 may l>e conveyed by the air. 
 
 Pusch refers in the same paper to an interesting case which 
 ofcurred in the practice of Eggeling. The latter had under his care 
 a cow with extensive paralysis. The spinal cord was compressed 
 by a compact swelling in the neck, consisting of the nodules of 
 actinomycosis. There were no manifestations of disease in any other 
 part of the body. 
 
 J ' i-': faience of the Disease. The author found that the disease 
 was not generally recognised as a common affection of cattle 
 iu this country, in spite of the interest excited by the work of 
 Fleming, to whom is due the credit of first recognising a case in 
 England. In 1887 there was a disease prevailing in Norfolk, and 
 in the following year outbreaks were investigated by the author 
 in Essex, Hertfordshire, Cambridgeshire, and Middlesex. In the 
 Norfolk outbreak the author found on one farm 8 per cent, of the 
 beasts affected with the so-called "wens" or " sitfasts," which 
 proved on microscopical examination to be cases of actinomycosis, 
 Yhr>f growths had previously been described in veterinary text-books 
 as the result of strumous or scrofulous inflammation ; but in all 
 the specimens of wens received from this country and the colonies, 
 the author has been able to demonstrate the presence of the ray- 
 f angu& 
 
 A case of pulmonary actinomycosis, with grape-like growths on 
 the pleura, indicated that wens were not the only manifestation of 
 this disease, which had been lost sight of under the designation of 
 
424 
 
 INFECTIVE DISEASES. 
 
 tuberculosis. Many other cases were examined, and the disease 
 was shown to be prevalent in this country. 
 
 FIG. 178. From a photograph of a Norfolk steer. There is a growth about the 
 size of an orange in front of the throat, an example of a so-called " scrofulous " 
 or "strumous" tumour. This growth was associated with a large polypoid 
 growth in the pharynx which, by interference with deglutition, produced 
 emaciation (Fig. 180). 
 
 In Australia actinomycosis commonly occurs in the form of 
 tumours of the upper and lower jaw, which were attributed to 
 
 ' cancer " or to " scrofulous 
 inflammation." The disease 
 is still commonly known in 
 Australia as " cancer " and 
 " lumpy jaw." 
 
 Reports of the prevalence 
 of actinomycosis in the United 
 States have been published 
 by the Board of Live Stock 
 Commissioners for the State 
 of Illinois. In their Report 
 for 1890 several interesting 
 communications were pub- 
 lished. Mr. Casewell, State 
 Veterinarian, investigated an 
 outbreak of this disease, known also in America as " lumpy jaw,' 
 
 FIG. 179. A NORFOLK HEIFER WITH A 
 LARGE ' WEN " IN THE PAROTID REGION. 
 
ACIINOMYCOSIS. 
 
 425 
 
 on a farm in Yates City, where there were 80 head of cattle, 
 111 were found to be suffering from actinoinycosis. Mr. Case well 
 
 FIG. 180. Photograph of a steer nearly three years old, but about the size of a 
 y. -iii-ling. The emaciation and deplorable aspect recall the appearance of 
 " a piner "or " waster " (tuberculosis). 
 
 reported that the disease was prevalent in nearly every county in 
 that State, and that in his opinion it was spreading. In one 
 instance 109 cases were slaughtered. 
 
 Actinomycosis in Relation to Tuberculosis. When we consider 
 the very high percentage of cases 
 of tuberculosis which has been 
 reported in some localities, the im- 
 portance of differentiating actino- 
 inycosis from tuberculosis cannot 
 be over-estimated. The very 
 great contrast in the appearance 
 of the micro-organisms in the 
 two cases renders this a very 
 matter for the pathologist. 
 But practical veterinarians and 
 breeders of cattle are liable to 
 mistake some manifestations of 
 actinomycosis for tuberculosis. 
 
 FIG. 181. ACTINOMYCOTIC NODULES 
 FROM THE PLEURA. 
 
 It is of the greatest im- 
 1>. nance to bear in mind that 
 wens or elvers are really not tubercular, but actinomycotic ; and 
 
426 INFECTIVE DISEASES. 
 
 that a condition of the lungs may occur as the result of actino- 
 mycosis, which from the naked-eye appearances may be mistaken for 
 "grapes" or " angleberries." It will be well also to remember in 
 connection with the above remarks, that extreme emaciation may 
 result in actinomycosis, producing a condition which, without a 
 post-mortem examination, would probably be attributed to tuber- 
 culosis, the animal being regarded as a " piner " or " waster." 
 If these possible fallacies are taken into account, the excessive per- 
 centage of tubercular cases so commonly reported will be very 
 considerably reduced. 
 
 There is no evidence to show that the flesh of animals suffering 
 from actinomycotic tumours is unfit for human consumption. In 
 very severe cases it is unwholesome, but there is no evidence that 
 it can produce actinomycosis in man. 
 
 MANIFESTATIONS OF ACTINOMYCOSIS IN MAN. 
 
 (I.) Invasion by the Mouth and Pharynx. The fungus may gain 
 access through carious teeth, or wounds or fistulse of the jaw, and 
 very possibly by inflammatory processes in the pharynx and tonsils. 
 
 The disease attacks the 'lower jaw most frequently. The tumour 
 is found in close connection with the bone, or in the sub-maxillary 
 or sub-mental regions, and also in the prse-tracheal region. It occurs, 
 though rarely, in the interior of the bone. 
 
 In a case described by Israel, which occurred in a woman aged 
 forty-six, there was a small tumour about the size of a cherry 
 attached to the external surface of the lower jaw, with an opening 
 through which a probe could be passed into the bone. The tumour 
 was incised and scraped away, and a cavity discovered in the bone, 
 admitting a small sharp-spoon. Later, a further operation was 
 performed : the periosteum was detached, the cavity of the bone 
 enlarged, and the contents scraped out, consisting of granulation 
 tissue, fragments of bone, and the yellowish fungi. At the bottom 
 of the cavity the fang of the canine tooth was found. No return of 
 the growth occurred. 
 
 The first cases of actinomycosis which were observed in America 
 were connected with the jaw. In 1884 Dr. Murphy described two 
 cases at Chicago. The first was that of a woman aged twenty-eight. 
 Two weeks previously she had suffered from severe toothache, with 
 swelling in the throat and great pain in swallowing. It disappeared 
 after poulticing, but she was again attacked with toothache, and a 
 swelling appeared on the angle of the jaw. on the left side. The 
 
ACTINOMYCOSIS. 427 
 
 mouth could not be opened without difficulty; the tonsil was much 
 enlarged, and pus was set free on incision. She still suffered with 
 toothache, and a small swelling now occurred on the left side of the 
 neck below the jaw. She had several carious teeth. The swelling, 
 which was about the size of a walnut, was punctured, and a drainage 
 tube inserted ; a creamy-looking discharge containing yellow granules 
 continued to escape, but the swelling and induration increased. A 
 further operation was decided upon. The carious tooth was removed, 
 and a probe passed into the alveolus showed a communication with 
 the external wound; the angle of the jaw was chiselled away, and 
 the alveolus scraped out. lodoformed gauze was applied, arid the 
 cas" r covered. 
 
 Yh.- second case was a man aged eighteen, who had also suffered 
 with severe toothache and swelling at the angle of the jaw. On 
 examination a carious tooth was noticed. The swelling was well 
 marked, and there was fluctuation ; it was as large as a pigeon's 
 egg. and situated below the jaw. When punctured, thick creamy 
 pu> escaped containing the fungi; the sinus was scraped out, and in 
 ten days the wound was healed. Another swelling appeared, and 
 this was treated as before, and the case recovered. 
 
 The peculiar feature of these growths is their apparent migra- 
 tion. Israel states that in one case a tumour occurred on the alveolar 
 proi-e^s. close to carious teeth, and later was close to the edge of 
 the jaw in the sub-maxillary region. From thence it disappeared, 
 and a large swelling formed below the hyoid bone, and after this had 
 lire:i incised and had healed, an abscess formed above the clavicle. 
 
 Actinomycotic tumours in this region would sometimes appear to 
 correspond very closely with wens or clyers in cattle : they may dis- 
 charge through the skin, and the opening close, or a fistula result ; 
 but they differ, from their tendency to form burrowing abscesses 
 instead of recognisable tumours. In this respect they recall chronic 
 inflammation rather than the sarcoma-like growths in cattle. 
 
 1 'asee in which the upper jaw is attacked are not so frequent as 
 those in the lower jaw. The progress is usually described as slow, 
 and there is a tendency for the deep-seated soft parts to be involved, 
 while in the lower jaw there is a tendency for the tumour to come 
 to the surface. There may be burrowing suppuration, or small 
 tumours, which, after a time, fluctuate and form distinct absc 
 Yh>->e may involve the skin, discharge their contents, and leave 
 h'stulous openings. 
 
 In other cases the disease has been described as extending from 
 th<- alveolar process to the temporal bone, or the base of the skull, 
 
428 INFECTIVE DISEASES. 
 
 destroying bones and even reaching the brain ; or the growth may 
 descend by the spinal column, implicating the vertebrae, and travel- 
 ling and pointing in various directions. 
 
 (II.) Invasion by the Respiratory Tract. In one recorded case 
 the disease existed for seven years, was localised to the bronchi 
 {Bronchitis actinomycotica), and did not extend into the lungs. 
 The sputum was examined, and contained the characteristic fungus. 
 
 If the micro-organisms are inhaled they pass into the bronchioles 
 and alveoli, and produce proliferation of round cells, which undergo 
 fatty degeneration. The resulting patches of peri- bronchitis or 
 pneumonia become yellowish-white ; suppuration and haemorrhage 
 from the capillaries follow, and small cavities result, containing pus 
 cells, fat granules; blood, and the fungi. In the neighbourhood of 
 the new growth there is compression of the alveoli, and ultimately the 
 formation of a dense stratum of connective tissue, separated from the 
 cavities by a lining of granulation tissue containing the character- 
 istic fungus. The symptoms are usually obscure ; but the sputum 
 may contain the fungi, which are often visible to the naked eye. 
 The apices of the lungs are not, as a rule, affected. There is con- 
 siderable clinical resemblance to chronic phthisis : cough, night- 
 sweats, pallor, shortness of breath, and haemoptysis are symptoms 
 common to both. Light may be thrown upon the case by the examina- 
 tion of the sputum. The presence of the actinomyces will be positive 
 evidence as to the nature of the disease. The existence of these 
 symptoms, with absence of tubercle bacilli, would lead to the 
 suspicion of actinomycosis, even failing the discovery of the fungus 
 in the sputum. 
 
 In the second stage the symptoms are more characteristic. The 
 disease spreads to neighbouring parts, and pleurisy commonly super- 
 venes. This extension may involve the peri-pleural tissues. Thus 
 the disease may follow the prse- vertebral tissues, descend behind the 
 insertion of the diaphragm, and point as an ordinary psoas or 
 lumbar abscess ; it may perforate the diaphragm and reach the 
 abdominal cavity. Peritonitis or sub-phrenitic abscess may then 
 result. In some cases adhesions have formed, and the disease has 
 extended to the liver or spleen, or other abdominal organs. The 
 disease may also extend forwards in the direction of the anterior 
 mediastinum and the pericardium. 
 
 The primary affection of the lung becomes of secondary import- 
 ance. Grave symptoms occur, hectic fever, night-sweats, rigors, 
 and marked pallor. In the third stage, the disease comes to the 
 surface, either over the chest, or in the neighbourhood of the dorsal 
 
ACT1NOMYCOSIS. 42 
 
 or lumbar vertebrae ; a swelling appears of a livid colour, and if 
 punctured no fluid escapes, but if allowed to make its own way to the 
 surface, the skin gives way, a muco-purulent discharge mixed with 
 piro-s of the growth escapes, and the fungi can readily be recognised. 
 
 (III.) Invasion of the Digestive Tract. In a case under Chiari, 
 (U-, i tli. with general marasmus, took place at the age of thirty-four, 
 at'trr two \vars' illness. The mucous membrane of the intestines was 
 almost completely covered with whitish patches, raised in the centre, 
 and covered with yellow and brown granules closely adherent to 
 the adjacent tissues. The teeth were carious. 
 
 Small nodules about the size of a pea may be found in the sub- 
 in uc-ous tissue, and in the mucous membrane itself. They soften 
 ami form ulcers with undermined edges, the base reaching the 
 muscular layer. They may undergo cicatrisation, but generally 
 the disease extends through the peritoneum to the abdominal cavity, 
 and perforates the bladder or the intestines, or makes its way through 
 tin abdominal wall. Symptoms are either absent or not character- 
 istic' . The fungus may sometimes be found in the evacuations, or 
 by exploratory puncture. 
 
 (IV.) Cu'lvtei'iiiined. In addition there are a number of recorded 
 cases presenting very varied symptoms and anatomical relations, in 
 which it has not been possible to satisfactorily determine the path 
 i of infection. Delepine has described a most interesting case of an 
 actinomycotic tumour of the brain. 
 
 MANIFESTATIONS OF ACTINOMYCOSIS IN CATTLE. 
 
 (I.) In the Digestive system we find the disease attacking : 
 
 (a) The lips, gums, buccal mucous membrane and palate, and 
 appearing as nodules, wart-like growths, or ulcers. The nodules 
 and ulceration of the palate were well shown in a specimen sent ta 
 the author for examination, under suspicion of being the result of 
 severe foot and mouth disease. 
 
 (b) The upper and lower jaw, where it probably originates in 
 carious teeth, and extending and invading the neighbouring cavities 
 and sinuses destroys the tissues with which it comes in contact,, 
 expanding the bones into thin plates or reducing them to the 
 appearance of pumice-stone. 
 
 (c) The tongue, where we see it most commonly in the form of 
 nodules or wart-like patches under the mucous membrane, with a 
 >p*H-ial tendency to ulcerate, through the irritation of the teeth. These 
 nodules may extend into the deep muscles, and often collect in rows 
 
430 
 
 INFECTIVE DISEASES. 
 
 more or less parallel to the superficial muscular fibres. Complete 
 tran verse sections .of the tongue, double- stained, readily show this 
 arrangement, even to the naked eye. Induration of the tongue 
 results from secondary interstitial glossitis. The author has seen, 
 in one case only, a tumour embedded in the substance of the tongue 
 about the size of a small Tangierine orange, and more or less isolated 
 from any surrounding growth. 
 
 (d) The pharynx, where the disease may occur in the form of 
 polypoid growths producing asphyxia. 
 
 FIG. 182. A NORFOLK STEER WITH EXTENSIVE ACTINOMYCOTIC ULCER ATION OF 
 
 THE SKIN OF THE FLANK. 
 
 (II.) In the Respiratory system we may meet with the disease in : 
 
 () The nasal cavities, originating primarily there or resulting 
 from extension of a growth from the lips, or the pharynx. 
 
 (6) The larynx and trachea, generally in the form of polypoid 
 growths, sessile or pedunculated, which arise primarily or occur 
 secondarily, by extension from the tissues in the neighbourhood. 
 
 (c) The lungs, where the differentiation of the disease is most 
 important, as neoplasms in the lungs, especially in the early 
 stages, and nodular growths on the pleura, may be mistaken for 
 tuberculosis. 
 
 The disease is very rarely found in connection with the 
 Nervous system (III.), but probably does not so rarely attack the 
 Reproductive system (IV.). 
 
ACTINOMYCOSIS. 431 
 
 (Y.) The skin and subcutaneous tissues are a favourite seat of this 
 <li>ease, producing the so-called wens or clyers so commonly sesn in the 
 ten country. A wen is first recognised as a small tumour, the size 
 of a marble or walnut, which increases in size sometimes with great 
 rapidity, and breaks down and discharges its muco-purulent contents 
 through the inflamed and ulcerated skin ; or it may go on increasing, 
 and form a large compact growth, the size of a child's head. These 
 trro \vths when excised, hardened, and cut, have a characteristic 
 honeycombed appearance, produced by the interlacing bands of 
 lil >n>us tissue, which form a spongy structure, from the interstices 
 of which the fungus tufts and thick yellowish pus have for the 
 most part dropped out. 
 
 Actinomyces Hominis. Careful examination of pus from 
 .i case of actinomycosis in man will reveal to the naked eye little 
 vellowish-white or yellow bodies, which a casual observer might 
 mistake for grains of iodoforni. On collecting some of the discharge 
 in a test-tube, and holding it between the light and the eye, the tufts 
 of fungi appeared as brownish or greenish-brown grains, embedded 
 in a muco-purulent matrix. 
 
 On spreading some of the discharge on a glass slip, the largest 
 tufts of the fungus are found to be about the size of a pin's head. 
 They have a distinctly sulphur- yellow colour by reflected tight, but 
 appear of a yellowish or greenish-brown tint by transmitted light. 
 With a sewing needle, or a platinum wire flattened at the end into 
 a miniature spatula, the grains can be readily picked out of the 
 discharge, or taken off the dressing, transferred to a clean slide, and 
 gently covered with a cover-glass. Examined with an inch objective, 
 they have the appearance of more or less spheroidal masses of a 
 pale greenish -yellow colour. On removing the preparation from the 
 microscope, and gently pressing down the cover-glass with the finger, 
 the grains flatten out like specks of tallow ; and on again examining 
 with the same power they are found to have fallen apart into a 
 number of irregular and sometimes wedge-shaped fragments of a 
 faintly brown colour, affording a characteristic appearance. By 
 preparing another specimen, and covering it with a cover-glass 
 without completely flattening out the grains, the spherical, oblong 
 and reniform masses of which the tufts are composed can be 
 recognised with a J- in. objective as rosettes of clubs. By examining 
 the peripheral part of a rosette with a -j^-in., and especially after 
 pressing the grains into a thin layer, with or without the addition 
 of a drop of glycerine, the characteristic clubs are most readily 
 demonstrated, and the most varied shapes observed by carefully 
 
432 INFECTIVE DISEASES. 
 
 examining the form of the individual elements. As in the bovine 
 fungus, every variation in form is found, from single clubs to clubs 
 with lateral offshoots, clubs bifid at the extremity, palmate or 
 fan-shaped groups, and banana-like bunches. In many cases the 
 clubs are divided by transverse fission into two, three, or more 
 segments. As a rule, the clubs are irregular in shape, and of about 
 equal size, while a few are conspicuous by their length. In other 
 parts of the preparation the clubs are replaced by long slender 
 forms, which are sometimes transversely divided into a number of 
 short links. With suitable illumination many clubs are seen to 
 taper off into slender filaments. In addition there are free filaments, 
 which are twisted, branched, and sometimes distinctly spirilliform. 
 Many of the clubs are, composed of layers differing in their refractive 
 power, and many have the appearance of a central channel. There 
 are also in the preparation small, highly refractive bodies, fat 
 granules, granular detritus, round cells, pus cells, and sometimes 
 blood corpuscles. 
 
 The grains differ, as a rule, from those from a bovine source, in 
 the absence of that sensation of grittiness so often transmitted to the 
 finger when pressing the cover-glass upon them, and in the slightly 
 greater tendency of the tufts to retain their compact form. By 
 teasing the grains in a drop of water on a slide, and examining 
 the preparation with a -^ or a ^ objective, the explanation 
 of the latter is forthcoming ; for by this process the clubs are 
 gradually washed away, and a central core remains, which is com- 
 posed entirely of a dense network of filaments. This can readily be 
 observed by using a small diaphragm, and it will be found. that 
 the rosettes of clubs are now replaced by tangled masses, having some 
 resemblance to miniature tufts of cotton-wool. These filaments 
 constitute the delicate network which is seen in sections stained 
 by the method of Gram. This can be readily verified by making 
 a cover-glass preparation of the grains, and staining by that method. 
 The characters of the fungus can readily be studied by proper 
 illumination, without staining. The clubs have a faintly greenish 
 tint, and in form and arrangement are quite characteristic and easily 
 recognisable. Permanent preparations may be made by mounting 
 the fungus in glycerine. 
 
 DESCRIPTION OF STAINED SPECIMENS. 
 
 The fungus may be stained in alcoholic solution of eosin in the 
 manner to be described for the bovine organism, or in orange-rubin, 
 
DESCRIPTION OF PLATES XV. AND XVI. 
 Actinomyces. 
 
 PLATE XV. 
 
 FIG. 1. From a preparation of the grains from an actinomycotic abscess I 
 a boy ; examined . in glycerine. The drawing has been made of a confl 
 plete rosette examined by focussing successively the central and periphera 
 portions. Towards the centre the extremities of the clubs are aloni 
 visible ; they vary in size, and if pressed upon by the cover-glass give th$ 
 appearance of an irregular mosaic. Towards the periphery the clubs aid 
 seen in profile, and their characteristic form recognised. At one pan 
 there are several elongated elements, composed of separate links, x 120<| 
 FIG. 2. Different forms of clubs from preparations in which the rosettes hav* 
 been flattened out by gentle pressure on the cover-glass, x 2500. 
 
 () Single club. (&) Bifid club. (c) Club giving rise to foin 
 secondary clubs, (d) Four clubs connected together, recalling 
 the form of a bunch of bananas, (e) Mature club with a lateral 
 bud. (/) Apparently a further development of the condition 
 represented at (<?). (^) Club with a lateral bud and transversi 
 segmentation, (h) Single club with double tranverse segments 
 tion. (i~) Club with oblique segmentation, (j) Collection dj 
 four clubs, one with lateral gemmation, another with obliqul 
 segmentation, (k) Club with lateral buds on both sides, aul 
 cut off square at the extremity. (1) Club with a daughter cluj 
 which bears at its extremity two still smaller clubs, (m) Clutj 
 divided by transverse segmentation into four distinct elements, 
 O) Elongated club composed of several distinct elements. (0) and 
 (j?) Clubs with terminal gemmation. (q~) Palmate group of clubs, 
 (r) Trilobed club, (s) Club with apparently a central channel 
 () Filament bearing terminally a highly refractiveoval body. 
 
 PLATE XVI. 
 
 FIG. 1. From a section of a portion of the growth removed from a boy 
 during life. The tissue was hardened in alcohol, and cut in celloidin, 
 The section was stained by Gram's method and with orange-rubin. x 50, 
 
 FIG. 2. From the same section. A mass of extremely fine filaments occupies 
 the central part of the rosette. Many of the filaments have a terminal 
 enlargement. The marginal part shows a palisade of clubs stained by the 
 orange-rubin. x 500. 
 
 FIGS. 3 and 4. From cover-glass preparations of the fungus teased out of the 
 new growths produced by inoculation of a calf with pus from a boy 
 suffering from pulmonary actinomycosis. Stained by Gram's method and 
 orange-rubin. The threads are stained blue and the clubs crimson (a] 
 In the younger clubs the thread can be traced into the interior of the 
 club (Z>). In some of the older clubs the central portion takes a yellowish 
 stain, and in others the protoplasm is not continued as a thread, but is 
 collected into a spherical or ovoid or pear-shaped mass. In others, again 
 irregular grains stained blue are scattered throughout the central portion 
 (Fig. 4). x 1200. 
 
 FIG. 5. From a pure-culture on glycerine-agar. (#) branching filaments, (7>) a 
 mass of entangled filaments. Gram's method, x 1200. 
 
 FIG. 6. From a similar but older cultivation, (a) a filament with spores 
 (6) chains of spores simulating streptococci. Gram's method, x 1200. 
 
Figl 
 
 ACTINOMV 
 
Plate XVI. 
 
 ACTIKOMYCOSIS HOMIN1S 
 
ACTIXOMYCOSIS. 43S 
 
 ami in either case mounted in glycerine. But although the fungus 
 can be detected without any staining process, there may sometimes 
 be doubtful appearances, and then cover-glass preparations should be 
 made and stained by the method of Gram with eosin. The filament - 
 c-a ii be readily recognised, and this is of great value, as it form- 
 an additional means for the diagnosis of the disease. In combination 
 with orange-rubin we have a test that is as* characteristic and useful 
 as staining for tubercle bacilli. The discharge, scraping from ;i 
 growth, sputum, or the isolated fungus is squeezed between t\\.> 
 cover-glasses, which are then slid apart ; they are allowed to dry, 
 passed through the flame in the ordinary manner, and then stained. 
 The cover-glasses can be cleared in clove-oil, the excess of clove-oil 
 being removed by gentle pressure between pieces of blotting-paper, 
 and then the preparation can be mounted in balsam and rendered per- 
 manent. On examination of these specimens the masses of filament- 
 will be found to be stained blue, and the tissue elements pink. These 
 filaments vary very much in extent and character in different pre- 
 parations. In some cases there are masses of short threads, which 
 are either straight, sinuous, or twisted, and branched. In other 
 parts the field is occupied by very short, straight, or curved and 
 sometimes spiral fragments ; in others, again, there are comparatively 
 long strands. On examination with a high power, and with careful^ 
 illumination, some filaments will be observed to be moniliform, 
 while others are provided with a terminal oval body. There are 
 also free spherical, and oval, bodies stained blue, which represent th> 
 spores of the organism. When orange-rubin is used instead of 
 eosin, the clubs will be stained and easily recognised. This method 
 enables one to determine the exact relation of the threads to tin- 
 club-shaped bodies ; and this is an interesting point, as it has been 
 suggested that the threads are not connected with the clubs, bir 
 merely an adventitious micro-organism growing in the track ot 
 ray-fungus. The threads are stained blue and the cluh> crh: 
 In theVunger clubs the protoplasm of the thread can U- traced into 
 the interior of the club. In some of the older clubs the cent.,1 
 portion takes a yellowish stain, and in others the pvrfopka 
 continued as a thread, but is collected into Bpherioal, 0* 
 pear-shaped mass. In others again, irregular grain* rtained 
 scattered throughout the central portion. The sheath of 
 is stained pink; and the protoplasm, rtained bh* ill> 
 conosta of small spherical or irregular grains, giving . 
 beaded appearance. 
 
 The effect of various reagents should be tried upon tl 
 
434 INFECTIVE DISEASES. 
 
 grains. The grains are picked out of the pus and transferred to 
 watch-glasses containing strong potash, xylol, and benzol. If 
 returned to a slide and covered with a cover-glass, the clubs are 
 found unaltered. Water or weak potash washes away the clubs, 
 and the filaments become- easily distinguished ; ether and strong 
 acids have no effect upon them. Corallin soda, Hanstem's violet, 
 and iodine zinc-chloride fail to give any particular reaction. 
 Hoffman's blue stains the clubs, but without bringing out any 
 structural details which could not be observed in the unstained 
 specimens. 
 
 Actinomyces bovis. The fungus in cattle may in the same 
 way be detected with the naked eye in the muco-purulent discharge, 
 or in a scraping from the cut surface of a growth. The tufts of the 
 fungus vary in size under different circumstances, from that of a grain 
 of fine sand to that of a pin's head. If the pus or scraping be spread 
 out on a slide and examined against a dark background, the grains 
 appear to be white or yellowish-white in colour ; but if examined by 
 transmitted light, they appear distinctly brownish. On pressing the 
 cover-glass on the slide the grains readily flatten out, being of a 
 soft, tallowy consistency ; or in the process of gently pressing the 
 cover-glass on the slide with slight lateral movement, a distinct 
 gritty sensation is transmitted to the finger, owing to the presence 
 of calcareous matter. On examination with a low power the 
 fungus will be recognised in the form of irregular patches scattered 
 over the field, which might readily be regarded as collections of 
 granular debris of a brownish or yellowish-brown colour, but on 
 careful examination they are observed to have a more or less 
 characteristic appearance. On examining with a higher power, 
 spherical, ovoid, or reniform bodies are to be seen, which are 
 either typical rosettes of clubs or granular masses, with here 
 and there a club-shaped body at the periphery. Pus cells, round 
 cells, fat granules, and minute spherical bodies may also be 
 distinguished. If the grains consist of typical rosettes, and be 
 merely covered with the cover-glass, and examined without being 
 flattened out between the cover-glass and the slide, they will recall 
 to mind, on focussing alternately the centre and the periphery, 
 the appearance of the capitulum of a composite flower. The central 
 portion appears to consist of spherical forms; these are the ex- 
 tremities of the component elements, and as we focus the edge of 
 the rosette these elements are seen laterally, and their characteristic 
 club-form is readily distinguished. The central portion may be 
 flattened against the cover-glass, and as the individual clubs vary 
 
DESCRIPTION OF PLATES XVII. AND XVIII. 
 Actinomycosis Bovis. 
 
 PLATE XVII. 
 
 Section of an actinomycotic tongue stained by the method of 
 Gram and with eosin. 
 
 FIG. 1. This illustrates the appearance which is usually seen under a low 
 power, when a section is stained by Gram's method and with eosin. The 
 central portion of a mass of the fungus is either unstained or tinged with 
 eosin, while the marginal portion is stained blue. The reverse is seen, as a 
 rule, in sections from man ; although under a low power the general appear- 
 ance of sections from these two sources is somewhat similar, x 50. 
 
 FIG. 2. a, I, c, d, represent the earliest recognisable forms of the ray fungus 
 in the interior of leucocytes. In e the club-forms can be recognised. In 
 /"and g there are small stellate groups of clubs, x 500. 
 
 FIG. 3. A part of the section represented in Fig. 1, under a high power. The 
 marginal line of blue observed under a low power is now recognised as the 
 result of the stain being limited to the peripherally arranged clubs. At 
 (a) part of a rosette has undergone calcification ; the clubs are granular, 
 and have not retained the stain. At (&) and close to it there are the 
 remains of rosettes in which the process of calcification is almost complete. 
 x 500. 
 
 PLATE XVIII. 
 
 The figures in this plate are taken from sections of a case of 
 so-called " osteosarcoma," in which the growth of the fungus was 
 remarkably luxuriant. The specimens were stained by Plauts' 
 method. 
 
 FIG. 1. Different forms of clubs in different specimens : x 1200. 
 (a) Very small club-shaped elements. 
 (&) A club with transverse segmentation. 
 (c) A club with lateral daughter clubs. 
 
 (d and e) Clubs with terminal offshoots resembling teleutospores. 
 (/) A club with developing daughter clubs on the left, and on the 
 
 right a mature secondary club. 
 (ff) A segmental club with lateral offshoots. 
 (&) Two clubs undergoing calcification. 
 
 FIG. 2. A very remarkable stellate growth comprised of nine wedge-shaped 
 collections of clubs radiating from a mass of finely granular material. 
 x 500. 
 
 FIG. 3. A rosette undergoing central calcification, and consisting in part of 
 extremely elongated clubs resembling paraphyses. Calcareous matter is 
 also being deposited in the club-shaped structures, x 500. 
 FIG. 4. Part of a rosette with continuation of the club-shaped bodies into 
 transversely segmented branching cells apparently representing short 
 hyphae. x 500. 
 
 FIG. 5. A rosette from another section in which similar appearances are 
 observed as in Fig. 4.- x 500. 
 
ftato H 
 
 to? 
 
 Fig 3. 
 
 ACTINOMYCOSIS BOY IS 
 
 tlnffnt Bv*'."''.- ' 
 
Plate XVIII 
 
 ACTINOMYCOS1S BOV1S 
 
ACTINOMYCOSIS. 435 
 
 considerably ill size, the appearance of an irregular mosaic i> thus 
 produced. 
 
 By pressing upon the cover we break up the rosette, and then 
 the clubs are recognised either singly or in pairs, or attached together 
 in the form of wedge or fan-shaped segments. Calcareous material, 
 if present, may readily be demonstrated by the action of arid,. It 
 will be found that on the addition of dilute hydrochloric, inn 
 acetic acids, the calcareous deposit is dissolved while tin- chilis are 
 not affected, and even with the addition of the strong. M acids the 
 only result will be to dissolve out the calcareous matter and clarify 
 the tufts of the fungus, the form of the clubs being still recognisable. 
 They are not affected by ether or potash, and thus the effect 
 chemical reagents clearly distinguish them from fat crv>tal> ..r 
 calcareous particles. By breaking up the growth into small t 
 ments, we may readily study the shape of the individual club-like 
 elements. By using high-power objectives, and properly arranging 
 the illumination, various forms will be clearly delineated. In some 
 cases the club will be found to be bifid at the extremity ; in other 
 ca>es there are lateral offshoots or daughter-clubs. Here and there 
 will be found clubs closely pressed together like a bunch of bana 
 and in other cases the broken-off pieces have a palmate form. By 
 teasing out the grains in water, and pressing them apart betv 
 the slide and cover-glass, we find that the central portion is com- 
 posed, as a rule, of a structureless core. More rarely there are the 
 delicate filaments which are found in cultures and in the fungus from 
 man. 
 
 If the grains are mounted in glycerine, the appeal am ( of the 
 organism in the fresh state may be preserved. 
 
 The granules may be stained by picking them out with needles 
 and transferring them to a watch-gla.ss containing alcohol, to which a 
 few drops of concentrated alcoholic solution of eosin have been ad 
 They remain in the solution until distinctly stained, and the\ 
 then placed on a glass slide in a drop of glycerine. 
 
 The muco-pus may be spread out into as thin a film M> p<>ihle 
 on a cover-glass, allowed to dry, fixed by warming slight ly offer 
 the flame, and stained by the method of Plaut or of loam. The 
 characters of the fungus can be so readily recognised in the : 
 fresh state, that methods of staining are of secondary import.-. 
 in diagnosis, though there are certain minute joints which can only 
 be satisfactorily determined by means of suitable dyes. 
 
436 INFECTIVE DISEASES. 
 
 CULTIVATION OF ACTINOMYCES. 
 
 Bostrom cultivated actinomyces from five cases in animals, and 
 from one case in man. In all cases he obtained a similar result. 
 The fungi were isolated from pus with sterilised needles, and placed 
 in liquefied nutrient gelatine in which they were teased out, and the 
 gelatine then spread on glass plates. The growth is stated to have 
 become visible in a few days. The fungi were isolated from the 
 plates, crushed between sterilised glass slides arid inoculated on the 
 surface of nutrient agar-agar and blood serum. In this way pure 
 cultivations were obtained. Nutrient gelatine was not liquefied. 
 The cultures in blood serum and agar-agar grew best at from 33 
 to 37 C. The track of inoculation gradually spread out during 
 the first two days, having a finely-granular, whitish appearance. 
 During the next few days small yellowish-red spots appeared in the 
 centre of the inoculated area, while the edge was apparently com- 
 posed of fine processes. The yellowish spots continued to increase 
 for about seven or eight days, and became confluent, and the 
 periphery also was dotted with yellowish-red points. Finally, there 
 were also isolated colonies consisting of a yellowish-red centre with 
 a greyish, downy periphery. The cultivated fungus, if suitably 
 stained, corresponded exactly with that found in human and 
 animal actinomycosis. In the cultures during the first two days 
 threads were found, with true branchings ; later the threads were 
 divided into shorter pieces or rods, and when the yellowish centres 
 appeared there were also a number of very short rods and cocci- 
 like forms. Bostrom also described attenuated club-shaped swell- 
 ings at the end of the threads. He concluded by saying that 
 Actinomyces is not one of the mould fungi, the central threads do 
 not therefore constitute a mycelium ; he was inclined to regard it 
 as a branched Claclothrix, and cultivation seemed to prove this. He 
 suggested that it might be the Streptothrix Forsteri of Cohn. In 
 any case he relegated Actinomyces to the fission fungi or bacteria. 
 
 In 1888 the author made cultures on glycerine- agar from a case of 
 human actinomycosis of the thoracic wall. An abscess was opened, the 
 discharge collected in sterilised tubes, and cultivations prepared with 
 as little delay as possible. Some of the discharge was spread out on 
 a sterilised glass slide, and the grains isolated with sterilised needles 
 and quickly transplanted on the surface of the nutrient medium. 
 The tubes were placed in the incubator at 37 C., and the result 
 watched from day to day. For several days there was to the 
 naked eye no promise of success ; but gradually the grains began 
 
ACTINOMYCOSIS. 
 
 to change, and by the end of a fortnight there wa> an appnviable 
 increase in size. Numerous cover-glas> pqpmtioM u.-iv , n; ,,|,. 
 from what was originally a single grain, an.l on examination b] 
 method of Gram the appearance was very striking. Then- omld be 
 no doubt as to the increase of the mycelial structure. Tin- . 
 masses of filaments covered almast the whole ami of the prepara 
 In parts less thickly covered there were very iiuim-ron- oya ] 
 holies, and rod-like segments with terminal enlargement-. i 
 crocus'' forms corresponded with the appearances previously de- 
 scril>ed as met with in the interior of certain clubs. From t; 
 would appear that some other condition is nere.-sary for the develop- 
 ment of the fully formed club, which is the result of the -heath 
 undergoing some change, possibly mucilaginous, resulting in the 
 formation of a thick investment of the clubbed mass of protopla-m 
 at the end of the thread. 
 
 The.-e cluh-shaped bodies represent organs of fructification, rather 
 than the results of degeneration or death. The difficulty in accept- 
 ing the view of their being entirely lifeless forms lies in the fact that 
 the author has observed daughter-clubs growing from the mature 
 clubs; and, further, in the bovine fungus the author ha> been 
 able to trace the stages in the development of a single dub to a 
 completely formed rosette. 
 
 In the unstained condition, the clubs are found, on the whole, to 
 be very regular in their form and arrangement, and by certain 
 staining methods they can be shown to have a somewhat complex 
 structure. If we take all the characters into account, and par- 
 ticularly the minute structure and the relation to each other of the 
 threads and clubs, we are justified in the opinion that the club in 
 the early stages is an integral part of the living fungus, and that 
 these characters bring the fungus into relation with a higher gi 
 of micro-fungi, Basidiomycetes, although the filam. nled 
 
 by themselves, correspond with the characters of Streptothrix. The 
 life-history of the micro-organism may be summed up thu- : 
 
 The spores sprout into excessively line. straight, or sinuoii.-, and 
 sometimes distinctly spirilliform thre.ids, which branch irr.-_rularly 
 and sometimes dichotomously. The extremities of the branches 
 develop the club-shaped bodies. The clubs are cl<- -1; to- 
 
 gether, so that a more or less globular body is formed, with 
 core composed of a dense mass of threads The tin . be 
 
 differentiated by the method of Gram into ... -nd 
 
 protoplasmic contents. The club-shaped bo.ly externally a; 
 be mucilaginous, while internally it i- c.,ntinuou- with Ui . -m 
 
438 INFECTIVE DISEASES. 
 
 of the thread. It is difficult to say what further changes occur in 
 the club-shaped bodies ; in all probability they represent organs of 
 fructification. If so, the protoplasm in the interior of the club may 
 possibly undergo changes leading to the development of spores, 
 which are ultimately set free; in some cases the terminal segment 
 of a club is separated by transverse fission in the form of a globular 
 body, a process resembling the formation of spores by ab junction, 
 In others, the forms sprouting from the club are suggestive of 
 teleutospores. There are occasionally long, slender forms, very 
 different from the ordinary clubs ; they possibly represent paraphyses 
 or abortive elements. In whatever way they may be formed, there 
 can be little doubt that spores are set free in the vicinity of 
 a rosette, and give rise to fresh individuals; the ultimate result 
 recalling, as has been suggested, the appearance of " fairy rings." 
 There can be little doubt that spores and young fungi are taken 
 up by wandering cells, and conveyed to a distance from the parent 
 fungus, and thus fresh centres of growth are established. 
 
 Appearances of cultures. Bostrom, Wolff, Israel, Paltauf, and 
 others have shown that actinomyces can be cultivated in the 
 ordinary nutrient media. More recently the author has carried on 
 a series of cultivations for some years on glycerine agar, gelatine 
 and milk, broth, bread-paste, and potato, in order to observe the 
 changes which take place, and to study the variations which he 
 found in the appearance of sub-cultures. The actinomyces after a 
 few days on glycerine agar at the temperature of the blood forms 
 little, white, shining, moist colonies, which may remain stationary, or 
 increase and coalesce. In a week or ten days, sometimes earlier, and 
 sometimes after several weeks, the cultures turn a bright yellow colour, 
 but some remain, though white ; others, again, have a tinge of pink, 
 and others are yellowish -brown (Plate XIX). After a time, a powdery 
 efflorescence makes its appearance on the surface of the culture, 
 which may be either yellow or white in colour. The culture may go 
 on increasing, spreading over the surface of the medium, and retain 
 its yellow colour, or it may turn black in parts or completely so, 
 while the agar is coloured brownish-black. Cultures have a peculiar 
 sour smell; the variations in cultures were all proved, by careful 
 testing by sub-cultures, to be due to the growth of the actinomyces 
 under varying conditions of soil, temperature, and the supply of air. 
 The stage of efflorescence corresponds with the breaking up of the 
 filaments into masses of cocci, and chains often closely resembling 
 streptococci. Gelatine is slowly liquefied. 
 
 Wolff and Israel cultivated actinomyces on raw and boiled 
 
DESCRIPTION OF PLATE XIX. 
 Pure-cultivations of Actinomyces. 
 
 These tubes were selected from a great number of cultivations 
 in which there were different appearances. In some instances the 
 growths had a faint tinge of pink. 
 
 FIG. 1. Pure-cultivation on the surface of potato, showing a luxuriant 
 sulphur-yellow growth entirely composed of entangled masses of fila- 
 ments. After three months' growth. 
 
 FIG. 2. Pure-culture from the same series, on glycerine-agar. In this case 
 the culture remained perfectly white. The jelly was coloured reddish- 
 brown. After fifteen months' growth. 
 
 FIG. 3. Pure-culture on glycerine-agar in which the growth was dark- 
 brown, in parts black, and the jelly stained dark-brown. After nearly 
 two years' growth. 
 
PlateJB 
 
 PURE-CULTIVATIONS 
 
 OF 
 ACTINOMYCES. 
 
 - 
 
 
ACTINOMYCOSIS. 439 
 
 eggs, and succeeded, by inoculation in the periton.-.-.l easily, in 
 producing the disease in rabbits and guinea-pigs, in thr form .f 
 tumours of the peritoneum. Sauvageau and R;i :,],., ,i l:i , 
 
 actinomyces should not be included with bacteria, and have sugg< 
 tlif name Oosftora boms. 
 
 PREPARATION AXD EXAMINATION OF TISSUES. 
 
 In order to examine the microscopical appearances, the tissues should 
 be hardened in absolute alcohol and embedded in celloidin. The sections, 
 when stained, are to be dehydrated as a rule iu strong spirit instead of 
 absolute alcohol, as the latter dissolves the celloidin. If the sections are 
 very friable, they can be cleared with clove-oil on the slide. By these 
 means the little fungus tufts, which have a great tendency to fall out of 
 the sections, may be preserved In situ after passing through the various 
 staining processes. To cut the sections, we can use either Jung's m: 
 tome, cutting in alcohol, or the freezing microtome. In the latter case, 
 after the celloidin has hardened, it is necessary to shave off all that 
 surrounds the piece of tissue. It is placed in water until it sinks, and 
 then transferred to gum, and frozen and cut in the ordinary way. 
 
 Stowing Methods. 
 
 There are several methods by which the organism can be stained in 
 the tissues, but it is best to employ for this purpose Gram's method and 
 modifications of Plant's method. 
 
 Gram's Method. By Gram's method the clubs in the bovine disease 
 are distinctly stained, especially if the sections contain the fungus at a 
 suitable stage. Use freshly prepared staining solution. A few drops of 
 nniline-oil are placed in a test-tube, which is filled up with distilled 
 water, the mouth of the tube closed with the thumb, and the mixture 
 shaken up thoroughly. An emulsion forms, which is then filtered, until a 
 perfectly clear solution of aniline-water is obtained. To this is added, 
 drop by drop, an alcoholic solution of gentian-violet until precipitation 
 commences. About fifteen to twenty drops in a small capsule of aniline- 
 water will be sufficient. Sections are floated in this dye for about ten 
 minutes, then transferred to the iodine-potassic-iodide solution until they 
 turn brown like a tea-leaf. They are then decolorised in alcohol ; tl 
 stained in a weak alcoholic solution of eosin, dehydrated in strong com- 
 mercial alcohol, cleared in clove-oil, and mounted in balsam. It will be 
 found that the clubs are stained blue, and that there is a central area, 
 which is, as a rule, tinged by the eosin. There are various modificat 
 of the method, and some of them are extremely successful in affording 
 not only a picture of the fungus, but also the structure of the surrounding 
 tissue. Very instructive results may be obtained by combining the 
 method of Gram with Ehrlich's histological staiu. In this case, after 
 the section has been decolorised in alcohol, it is ready to be transferred 
 to logwood and treated as described below. 
 
 Wei<j> //'* .17, thod. This also gives very beautiful results. The section* 
 
440 INFECTIVE DISEASES. 
 
 are placed for an hour in Wedl's solution of orseille, which is prepared 
 as follows : Add liquid extract of orseille to a mixture of absolute 
 alcohol 20 parts, strong acetic acid 5 parts, distilled water 40 parts, 
 until a dark-red liquid results. This must be filtered before use. The 
 sections are left in this solution for an hour, then just rinsed in alcohol, 
 and transferred to a solution of gentian-violet. Such sections show the 
 nuclei of a violet-blue colour, and the peripheral part of the central core 
 in the larger masses of the fungus also takes a blue colour, while the 
 club-shaped structures are stained a striking wine-red colour. 
 
 Plant's Method and Modifications. This is one of the most valuable 
 methods for staining the clubs. The original method was to float sections 
 for ten minutes in magenta solution warmed to 45 C. This solution 
 consisted of magenta two parts, aniline-oil 3 parts, alcohol of specific 
 gravity 0'830, 20 parts, distilled water 20 parts (Gribbes). The sections 
 were then rinsed in water, stained in concentrated alcoholic solution of 
 picric acid for from five to ten minutes, immersed in water five minutes, 
 50 per cent, alcohol fifteen minutes, passed through absolute alcohol and 
 clove-oil, and preserved in Canada balsam. The clubs are stained a 
 brilliant red and the tissue yellow. Instead of employing the magenta 
 solution, we now use Ziehl-Neelsen's solution. 
 
 By removing the picric acid in Plaut's method by prolonged .immersion 
 in alcohol, and then staining with gentian- violet or methylene-blue, a 
 very successful contrast can be obtained. The most instructive histo- 
 logical picture can be obtained by first staining with Neelsen's solution, 
 removing the stain from the tissue in the way that has been already 
 described, and then transferring the sections to distilled water, and 
 subsequently staining with Ehrlich's histological stain. 
 
 Ehrlick's New Histological Stain, This is a combination of Ehrlich's 
 logwood with orange-rubin. It is of especial value for sections of 
 actinomycosis, and particularly in combination with carbolised fuchsine. 
 It is employed in the following way : The sections must be placed in 
 alcohol or distilled water, and then in Ehrlich's logwood for about half 
 a minute.. From this solution they are transferred to distilled water, 
 washed to remove the excess of stain, and then placed in a large dish of 
 tap-water, where they are left for half an hour or more, until the 
 sections turn blue ; if preferred, they may be left overnight. They are 
 then stained for one or two minutes in a solution of rubin, S, and 
 orange, and washed again in distilled water to remove the excess. They 
 must then be dehydrated in alcohol, cleared in clove-oil, and mounted in 
 balsam. 
 
 Preparation of Large Sections. 
 
 The method of cutting large sections of organs may be employed 
 in studying actinomycosis. The value of these sections depends not 
 only upon their affording an instructive picture of the naked-eye 
 appearances, but' they can also be studied with a pocket lens, or under 
 the microscope with a or -in. objective. By staining the sections, the 
 relation of the morbid to the healthy structures is brought out in greater 
 contrast, and thus the topography of the disease can be studied more 
 
DESCRIPTION OF PLATES XX. AND XXI. 
 Actinomycosis Bovis. 
 
 PLATE XX. 
 
 FIG. 1. From a section of an actinomycotic tongue stained by the triple 
 method (Ziebl-Neelsen, logwood and orange-rubin). In this section the 
 separate centres of growth are clearly shown. Each neoplasm consists of 
 a fungus system, in which the masses of the fungus, situated more or less 
 centrally, are surrounded with round cells, epithelioid cells, sometimes 
 giant cells, and lastly fibrous tissue forming a more or less distinct 
 capsule. In parts the fungi have fallen out of the section. x 50. 
 
 FIG. 2. From a section of a "tubercular" nodule from the lungs of a 
 Norfolk heifer with pulmonary actinomycosis. The nodule is a multiple 
 growth surrounding a bronchus, and is enclosed by a capsule, in the 
 vicinity of which the pulmonary alveoli are compressed. It is composed 
 of a number of separate neoplasms, and each of the latter is composed of 
 secondary centres of growth resembling the giant cell systems of bacillary 
 tuberculosis. The new growth is composed of ray-fungi, large multi- 
 nucleated cells, sometimes distinct giant cells, round cells, epithelioid cells, 
 and, surrounding them, fibrous tissue. On examination of the same 
 specimen with a higher power the typical rosettes of clubs are sometimes 
 surrounded by multinucleated cells, and sometimes small rosettes are 
 found like tubercle bacilli, in the interior of giant cells. From a pre- 
 paration stained by Ziehl-Neelsen, logwood, and orange-rubin. x 50. 
 
 PLATE XXI. 
 
 FlG. 1. () A leucocyte containing the fungus in its earliest recognisable 
 form. (5) A large multinucleated cell containing the fungus in an early 
 stage with the club-form already visible, (c) A leucocyte containing a 
 small stellate fungus, (d) A large cell containing clubs arranged in a 
 small rosette, (e) A multinucleated cell with clubs arranged in a palmate 
 form. All the above are drawn from sections of actinomycotic tongues 
 stained by the triple method, x 500. 
 
 FlG. 2. A giant cell with large vesicular nuclei at the periphery, and in the 
 centre a fully formed rosette of actinomyces with a smaller growth within 
 a " daughter " cell. From a section of the tongue of an ox stained by 
 the triple method, x 500. 
 
 FIG. 3. A very large circular giant cell, with its ring of nuclei at the 
 periphery, enclosing several isolated tufts of actinomyces. From a section 
 of a nodule in the lung. Stained by the triple method, x 500. 
 
 FIG. 4. Three rosettes of actinomyces surrounded by a row of large, some- 
 what angular multinucleated cells. From a section of the tongue of an 
 ox stained by the triple method, x 430. 
 
,_-.., 
 
 Plgl 
 
 
 . 
 
 Fig 2. 
 ACTIN T OMYCOSTS BOVIS 
 

 =* 
 
 
 - . "% 
 
 - * I * ^ * 
 
 I / * 
 
 Fig 3. 
 
 Fig 4 
 
 ACTINO-MYCOSIS BOVIS 
 
 
ACTINOMYCOSIS. HI 
 
 minutely than by simply observing the cut surfaces of organs or growths. 
 And. further, it affords a means of rendering permanent many of tin- 
 instructive appearances observed at the autopsy, without preserving th, 
 whole structure in the form of a museum specimen. V : 
 results can be obtained with material hardened either in spirit or 
 fluid. The fresh material is cut with a large, very sharp knife into -li -,- 
 about a quarter of an inch, or less, in thickness. The- in- placed 
 
 between filter- paper in large porcelain dishes, such as are employed for 
 photographic purposes, and well covered with the hardening solution, 
 which should be frequently changed. By covering the slier \\ ith u small 
 sheet of glass, which is lightly weighted, any curling or turning up of the 
 edges is prevented, and the slice not only kept flat, but hardened with 
 smooth surfaces. Several weeks are required for hardening in I 
 fluid. The slices, after a short time in water, are placed in gum, and 
 then frozen and cut ; the slices which are hardened in alcohol are soaked 
 in water until all trace of the spirit has been removed. A large micro- 
 tome on the Bruce model is used to freeze and cut the sections. But in 
 some cases it will be found better to embed the slices in celloidin, and 
 cut under alcohol with a large microtome of Jung's pattern. The 
 sections are carefully removed from the blade of the knife with a large 
 camel's-hair brush, and in the case of frozen sections floated in w.v 
 
 The next process is to float a section out in spirit, and with the 
 camel's-hair brush to unfold it and spread it out on a sheet of glass. 
 The glass with the section is lifted out and examined, and if the section 
 is sufficiently thin, transferred to the staining solution. In the same 
 way the section is passed through the various stains, as it should be 
 prevented from rolling up or folding in the dye, or it may not be evenly 
 stained throughout. Modifications of this process will suggest them- 
 selves, such as pouring off the dye and leaving the section spread out at 
 the bottom of the dish, and then using the same dish for the next 
 process. The sections are so easily injured, that it is better, as much as 
 possible, to avoid handling them. If the sections are only a few in 
 in diameter, such as transverse sections of the anterior portion of the 
 tongue of an ox, they can readily be transferred from dish to dish l>y 
 means of a large spatula, made by soldering a piece of sheet German 
 silver to thick copper wire. 
 
 To stain them employ carbolised fuchsine and picric acid, or alum 
 cochineal, or logwood and orange-rubin. The proci- 
 precisely the same as with ordinary sections ; but, from their unusual 
 size, experience and practice are required in their manipulation. 
 
 When the section is dehydrated, it is ready to be cleared in clove-oil. 
 The glass on which it is to be permanently mounted shouM be I 
 without scratches or flaws, and thoroughly cleaned and p<> 
 slipped under the section, which is evenly spread out upon 
 lifted out of the dish. The excess of spirit is drained off, t 
 placed on a level surface, and clove-oil poured on the section, 
 until completely clarified : the clove- oil, as much as possible, di 
 and the rest entirely removed by gentle pressure with several thicl 
 
442 INFECTIVE DISEASES. 
 
 of best filter-paper. In this way several large sections can be cleared at 
 the same time. But when only one or two sections are dealt with, they 
 are cleared in clove-oil in a dish, and the mounting glass at this stage 
 passed underneath them as already described. Another plan, which will 
 be found of advantage, is as follows : A piece of clean, thick, filter-paper 
 rather larger than the section, is slipped underneath it, and then raised 
 with the section upon it. After allowing the excess of clove-oil to drain 
 back into the dish, it is carefully laid on the glass with the section down- 
 wards, and gently pressed down. By taking up a corner, the filter-paper 
 is peeled off, and the section left behind on the glass. Any creases or 
 folds are adjusted with needles. After removal of the clove-oil, balsam 
 is run over the section, and a cover-glass gently and dexterously lowered, 
 so as to avoid the presence of air-bubbles. The preparations are set aside 
 to harden in a warm place and on a level surface, and are then ready for 
 fixing in suitable frames. 
 
 Naked-eye Appearances of Large Sections. 
 
 In the sections of an actinomycotic tongue it is at once apparent that 
 the new growth is more or less limited to the periphery of the section. 
 In parts there are dense clusters of little nodular neoplasms, the fungus 
 systems, each having a rounded form, and averaging in size that of a 
 small pea. In other parts small nodules, varying in size from a millet- 
 seed to a hemp-seed, have a linear arrangement between bundles of 
 muscular fibres. The appearance is suggestive of an invasion of the 
 tongue along the lymphatics. 
 
 In many of the nodules the largest tufts of the fungus can be seen. 
 with the naked eye, to occupy a more or less central position. In parts 
 the muscular fibres are replaced by fibrous tissue. 
 
 If now these sections be placed under the microscope, the minute 
 structure may be examined ; but as it is obvious that still better results 
 may be obtained by small sections, any part which it is necessary to 
 examine with high powers can be selected from a corresponding part of 
 the growth, and prepared in the ordinary way. 
 
 In the case of a u wen," the whole growth can be excised with the 
 surrounding tissues, sliced and treated in the way already described, and 
 sections stained by different methods. 
 
 The nature of the growth is at once recognisable as actinomycosis r 
 from the characteristic honeycombed appearance produced by the trabeculse 
 of fibrous tissue which form a spongy structure, from the loculi of which 
 the fungus tufts and caseous matter have for the most part dropped out. 
 In other parts this structure is intact, and the tufts of the fungus can be 
 detected with the naked eye, and readily recognised with a pocket lens. 
 
 A fungus system may be studied more minutely in ordinary sections 
 of the tongue. Each nodule is composed of the actinomyces surrounded 
 by round cells and epitheloid cells, and fibrous tissue which often forms 
 a distinct capsule. In some specimens the fungus is surrounded by a 
 single row of large multinucleated cells, and in other specimens the 
 fungus is found in the interior of large oval giant-cells. 
 
ACTINOMYCOSIS. 443 
 
 TRANSMISSION OF ACTINOMYCOSIS FROM MAN TO THE LOWER ANIMALS. 
 
 Inoculation of cultures has already been referred to. The 
 author successfully inoculated a calf with material direct fn.ni 
 a living patient. 
 
 A calf which had been inoculated in the peritoneal cavity, a ml 
 killed seventy days afterwards, presented the following lesions. The 
 peritoneum of the rumen, in the vicinity of the seat of iim< -ul .,- 
 was studded with hundreds of growths, varying in size from a millet - 
 seed to a pea. The large growths were composed of several small 
 ones collected together. On stripping off the peritoneum, and holding 
 it between the light and the eye, the fungus could be seen with 
 the naked eye in each individual growth. By incising a growth. 
 and examining a scraping under the microscope, the diarart-ri.>tir 
 clubs, and the filaments also, were found to be present. By staining 
 cover -glass preparations with the method of Gram and orange-rubin, 
 the appearances were very striking. The clubs were conspicuoi 
 account of their size, and brilliantly stained. In many, the proto- 
 plasm of the thread was demonstrated in the interior of the club. 
 In sections of the peritoneal nodules stained by Gram's methyl 
 the mycelium was found to be present, and the clubs in part 
 took the stain. With Plant's method the clubs were most dearly 
 demonstrated. 
 
 l>rael and Johne failed to infect a calf by intravenous injec- 
 tion, and Ponfick failed to infect dogs, but Israel succeeded with 
 a rabbit, Israel obtained a small piece of actinomycotic granula- 
 tion tissue from a peri-pleural abscess in a patient with primary 
 disease of the lung, and introduced it into the peritoneal Of 
 The rabbit showed no sign of illness, and was killed ab".,t t.-n 
 weeks afterwards. On examination numbers of tumours were 
 found in the abdominal cavity, varying in si/.e from a hemp-seed 
 to a cherry. Larger ones had a somewhat nodular Mirtac- with 
 yellowish points; others were found on the abdominal wall .n 
 the right side. There was a small growth over the psoas muscle, 
 and one large one attached to an adhesion of th- >1<>>. 
 growths were on the peritoneum, or attached by longer or * 
 adhesions. Some of the larger turnout >h..wed on section a h.ll. 
 space in the centre, which was filial with a pulp, ti 
 fatty degeneration of the traii>plant.-d time, whidi ** 
 differentiated in colour and consistency from the new gr 
 latter consisted of -ranulation fcURM with ahundant 
 fat granules, blood pigment, acicular tat eryttek, and acti 
 
444 INFECTIVE DISEASES. 
 
 grains. From some of the tumours, radiating processes of a yellowish 
 colour penetrated into the retro- peritoneal tissue. 
 
 Rotter inoculated calves, pigs, guinea-pigs, and rabbits. In one 
 case he had a positive result. A piece of actinomycotic growth was 
 introduced into the peritoneal cavity of a rabbit. The animal was 
 killed six months afterwards, and the piece of tissue which had been 
 introduced was found to be encapsuled, and around it were twenty 
 tumours, from the size of a pin's head to that of a hazel-nut, 
 containing the ray-fungus. 
 
 The author also succeeded in transmitting the disease to a rabbit. 
 A small quantity of human pus, containing the yellow grains, was 
 diffused in broth, and injected with a hypodermic syringe into the 
 abdominal cavity of a rabbit. This rabbit died seventy- nine days 
 afterwards. On examination several nodules were found, about the 
 size of a millet-seed, on the peritoneum of the stomach, in the gastro- 
 splenic omentum, and on the peritoneum of the diaphragm. There 
 was a rounded nodule, about the size of a pea, attached to the 
 stomach. There were adhesions between the intestines, and a 
 tumour about the size of a marble attached to an adhesion of 
 the caecum. One of the small nodules was excised and divided, and 
 the scraping from the interior contained typical rosettes of clubs! 
 
 The successful transmission of actinomycosis from man to bovines 
 suggests inter com municability, though the negative evidence as to 
 infection of man from bovines supports the view that the disease 
 is derived from some source which is common to both species. 
 
 TRANSMISSION OF ACTINOMYCOSIS FROM CATTLE TO CATTLE. 
 
 Johne was the first to prove that actinomycosis could be trans- 
 mitted from cattle to cattle, and his results were confirmed and 
 extended by Ponfick. 
 
 A calf was inoculated subcutaneously in the neck and cheek, 
 in the gum, and the abdominal cavity. The animal died in forty 
 days after inoculation with development of actinomycosis. 
 
 A calf was inoculated in the cheek and abdominal cavity. Death 
 occurred 114 days after inoculation. In the peritoneal cavity 
 numerous tumours had formed, and the yellowish grains were visible 
 to the naked eye in sections of the new growth. 
 
 A cow in calf was inoculated in the left posterior quarter of the 
 udder ; phlegmonous mastitis- folio wed, and subsided leaving a small 
 induration, which then increased until the inoculated part of the 
 udder was, in three months, nearly double the normal size, from a 
 
ACTINOMYCOSIS. \ \' t 
 
 deposit resembling a fibroma. The cow was sla tight nvd l:;;j , 
 after inoculation. Typical actinomycosis had heen produced. 
 
 A colt three and a half years old, was inoculated in the jaw 
 and in the forehead after trephining, and also in the trachea. Tin- 
 animal died without any result. 
 
 Ponfick also conducted a series of experiment- which amply 
 confirmed the results which had been obtained by John.-. 
 
 Feeding Experiments. Repeated experiments, made with mnmcc 
 of the growth chopped up, or with isolated grains of the fungus, : 
 negative results. 
 
 Inoculation Experiments. The growth was inoculated in various 
 regions of the body. Small particles of the growth from quite fresh 
 tumours were introduced into the anterior chamber of the eye in 
 rabbits, with negative results. Rabbits were inoculated in the 
 peritoneal cavity from an animal recently slaughtered, but they 
 died of peritonitis. In dogs also the results were negative. 
 
 Seven calves were operated on. In five the abdomen was opened 
 with antiseptic precautions, and in two cases the grout h 
 introduced by injection. In the latter cases the pieces of tumour 
 were suspended in salt solution, but the animals died from peritonitis 
 a few days after the injection. The same result occurred in two out 
 of the five cases in which the abdomen was opened. The three 
 remaining cases gave the following results : 
 
 1. Pieces of tissue, about 1'5 cm. long, were taken from the 
 lower jaw of a recently killed ox. Twelve of these pieces 
 introduced into the peritoneal cavity ; death occurred after 
 days, from exhaustion and recent lobular pneumonia. At the p 
 mortem examination several patches of peritonitis w-n- found, with 
 encystment of the remains of fragments of the inorulati 
 but there was an independent development of several nodules in tin- 
 neighbourhood of the stomach and urinary Madder. Examination 
 of these new formations showed, even to the nal, he\ 
 
 contained yellowish grains, which, on further examination, pi 
 to be the fungi. 
 
 2. Ten pieces of tumour from the jaw of a row pert 
 as before; the calf died suddenly sixty day- atWwanK d 
 injection of fresh pieces of actinomyco-i- into the ju-ul ..r 
 the jJost mortem it was found that various adheoou had occurred, 
 as the result of peritonitis; and in tin* tal>' nn'ml'i 
 sixty-three nodules, varying in si/,-. MkraMOpfcftl 
 showed That all these nodules consisted of typical acti,, 
 formation. 
 
446 INFECTIVE DISEASES. 
 
 3. In this case twelve pieces of growth were introduced into 
 
 the abdomen of a calf eight weeks old. Seven days afterwards fresh 
 pieces were introduced under the skin, in the region of the left lower 
 jaw. A swelling occurred, which was opened. Pus escaped, 
 together with the material which had been used for inoculation, in 
 a state of decomposition. Pieces of tumour were inoculated sub- 
 cutaneously in the neighbourhood of the right side of the neck, 
 ninety-nine days after the first experiment. The animal was bled to 
 death seven months (210 days) after the first experiment. At the 
 post mortem, peritonitis and adhesions were found, with twenty-one 
 large and several small nodules in the mesentery, in the false mem- 
 branes between the viscera, and on and in the serous linings of most 
 of the abdominal organs. There were also several large and many 
 small tumours in the subcutaneous and intermuscular tissue, in the 
 region of the lower jaw and neck on the right side, and numerous 
 large and small nodules in both lungs, some undergoing softening 
 in the centre. 
 
 Isolated fungi were inoculated in dogs, with negative results, 
 nothing remaining after 45 to 80 days, except a thick emulsion. 
 
 Pieces of tumour were introduced in the submucous and sub- 
 cutaneous tissue of dogs, but no change occurred after 600 days. 
 
 In a calf, inoculation under the gum of the upper jaw showed 
 w T hat was possibly only the remains of the growth which had been 
 introduced. At any rate, the experiment was doubtful ; but in 
 a second calf there were numbers of nodules developed around 
 the points of inoculation in the subcutaneous and muscular tissue 
 in the neighbourhood of the lower jaw, and in the region of the 
 neck. The results were observed in these regions in 210 and 110 
 days respectively. 
 
 Inoculation of rabbits and dogs, with isolated fungi, produced 
 no results after 156, 165, and 170 days. Experiments on dogs, 
 with grains from a human source, were also unsuccessful. They 
 were examined after 470 days, and there was no sign of any 
 result. 
 
 After intravenous injection, positive results were discovered m 
 the only calf which survived this operation, on bleeding the animal 
 to death 110 days afterwards. Numerous nodules (27) were dis- 
 covered in the parenchyma of both lungs, without suppuration. 
 
 Two dogs were injected in the jugular vein with isolated fungi 
 mixed with 60 grammes of salt solution. Examined after 45 and 
 80 days respectively, the lungs and all the other organs were found 
 to be free from growths. 
 
ACTINOMYCOSIS. 447 
 
 Ponfick thus summarised these experiments with the Ix.vin,- 
 fungus : 
 
 1. Rabbits and dogs possess a marked immunity from actin.. 
 mycosis, whether pieces of tumour or isolated grains :m . a.lmini>i,.iv.l 
 I >y feeding, or by inoculation in the serous cavities, in the subcutaneous 
 or submucous tissue, or by intravenous injection. 
 
 2. The most common subject of actinmuyr.iMs, the cow, possesses 
 a not less marked susceptibility to the artificial production of the 
 disease. By feeding, an infection was not obtained, pn.hal.lv 
 because the mucous membrane had not been injured; but by 
 inoculation, on the contrary, an independent growth of fn-h 
 neoplasms was produced in the subcutaneous and intermusciilar 
 tissues, occasionally in the submucous tissue, and in a deci<l-<l 
 manner in the abdominal cavity. Clear evidence of this growth 
 is obtained in some cases within a month, or after three or 
 four months. 
 
 3. By intravenous injection, also, it is possible in a few month- 
 to cause typical new growths in the lungs. 
 
 MADURA DISEASE. 
 
 Mycetoma, or Madura foot, is a chronic local disease, attacking 
 <-hiefly the hands and feet, and having considerable resemblance to 
 actinomycosis. It is a disease of tropical climates, and is commonly 
 known as the " fungus-foot disease " of India. A small tumour f< >nn> 
 on the hand or foot, which after a year or two suppurates and 
 bursts, leaving one or more sinuses, from which peculiar black 
 particles, or white or pinkish roe-like bodies, are discharged. 
 
 The disease in the foot may commence in the big toe and spread 
 upwards, involving the leg as far as the knee, and <-\en the thiirh. 
 In a typical case the foot is enlarged and painful, ainl later there 
 are several sinuses from which a purulent and blood-stain*-.! ili>eharge 
 can be expressed, containing the characteristic parti< ! >. 
 
 According to Bocarro all early growths are superficial. Dissec- 
 tion of the growths during an operation, or sections made through 
 the diseased tissues after excision or amputation, show that the 
 disease begins generally in the loose cellular tissue, generally the 
 subcutaneous tissue, and thence extends along the sheath of muscles 
 and tendons to other soft tissues, and finally the bones. 
 
 There are several facts in connection with th- nni>ati..n <>f th- 
 disease, which are of great interest when it is compared to act i 
 mycosis in cattle. Bocarro states that tin- disease originates iu 
 
448 
 
 INFECTIVE DISEASES. 
 
 wounds, sores and pricks of thorns, and that the points of the 
 thorns of the Acacia Arabica have been found embedded in the 
 diseased parts. The disease is common among the agricultural 
 class, and in 90 per cent, of the cases observed in the Hyderabad 
 Civil Hospital it occurred in the hands and feet. 
 
 Vandyke Carter was the first to point out the resemblance to 
 actinomycosis, and he believed that the two varieties of the disease, 
 the black and the white, were the result of the growth of a mycelial 
 fungus, Chionyphe Carteri. 
 
 Kanthack pointed out, that if portions of the growth were 
 placed in ether or chloroform, and afterwards well washed in 
 
 FIG. 183. PART OF HUMAN FOOT WITH MADURA DISEASE. 
 
 caustic potash, small rounded bodies were left, which showed rays 
 under the microscope closely resembling the appearances in actino- 
 mycosis, and that the reaction of the fungus to staining reagents 
 was identical with actinomyces. Hewlett examined sections from 
 the disease in the foot, and also found filaments and clubs. 
 Boyce and Surveyor examined a number of cases, and care- 
 fully studied the fungus in the black and white varieties of 
 the disease. In the black variety the particles were found to 
 vary greatly in size, from that of a grain of gunpowder to that 
 of a marble. If the particles were boiled for from a few minutes 
 to one hour in concentrated caustic potash, and then transferred 
 to distilled water, the brown colouring matter was removed and a 
 
MADURA DISEASE. 449 
 
 mycelial fungus could be seen. If tissue, containing part id.-, was 
 washed for about a minute in eau de Javel and tin -n >t .-lined, the 
 colouring matter was removed, and the relation of the fungus to 
 the tissue could be observed. This fungus consisted of large radiating 
 and branched hyphse, like those of a species of aspergillus, or muc.r. 
 Sections of the white, fish-roe bodies showed, usually in the centre, 
 numerous, small, reniform, deeply-stained masses, surrounded by 
 a radiated zone, with the presence of dwarfed club-like elements 
 resembling actinomyces. 
 
 The author suggested that possibly the presence of the coarser 
 septate mycelium of the black variety might be attributed to a 
 mixed infection. 
 
 Vincent in Algiers succeeded in cultivating the micro-organism, 
 and showed that it was a new species of streptothrix. 
 
 Streptothrix madurae. Vincent found that the streptothrix 
 at first grows scantily in the ordinary culture media, and in such 
 liquids as Cohn's solution. In broth, at the end of about a fortnight, 
 there is a limited growth composed of small, round, greyish masses, 
 and in sub-cultures the growth becomes more abundant. 
 
 The little colonies float in the clear liquid when the tube is 
 shaken, and subside to the bottom when the liquid is at rest, while 
 some adhere to the sides of the tube. They may be very small, or 
 attain the size of a pea ; after two months they acquire a reddish 
 tinge. Later, on the surface of the liquid, there is a white efflores- 
 cence composed of spores. 
 
 The streptothrix grows well in slightly acid infusions of hay or 
 >traw, the proportion of hay to water being 15 grammes to the 
 litre. Vegetable infusions, made with carrots, turnips, and potatoes 
 (20 grammes to 1,000 of water), are suitable media. Th.- 
 streptothrix grows at the temperature of the room, but best at 37 C. 
 and with free access to air. Inoculated in the depth of gelatine th--n- 
 is a scanty growth in the track of the needle and on the surface ; 
 but it grows best in a nutrient medium, composed of infu>in <>t 
 hay or potato 100 cc., gelatine 9 grammes, glycerine 4 grammes, 
 and grape-sugar 4 grammes. Gelatine is not liquefied. 
 
 Ordinary nutrient agar is not a very favourable medium, but 
 on glycerine-agar with grape-sugar there is an abundant ^rmvth -. 
 circular, projecting, shining colonies, slightly yi-llowish-wliit.-. which 
 later become pink or bright red. When the ooboitt aiv numerot 
 they remain small, but isolated colonies increase' rapi-lly ; th. 
 depressed in the centre or umbilicated, and the central part remain! 
 white while the periphery becomes red. Later, the culture loses 
 
 L J 
 
450 INFECTIVE DISEASES. 
 
 colour and becomes a dull white. The growth is very adherent to 
 the surface of the jelly, and so tough that it is almost horny. The 
 Streptothrix can be cultivated in milk, which it slowly pepto- 
 nises. It cannot be grown on blood serum or egg. On potato, on 
 the fifth day at 37 C., there are little prominent colonies, which 
 slowly increase. After a month the growth acquires a pale-rose 
 colour, which gradually increases and changes to orange or dark 
 red. The colour is most intense on acid potato, and after a time 
 an efflorescence, or whitish dust, appears on some of the colonies, 
 consisting of spores. The growth is hard and friable. 
 
 Rabbits, mice, guinea-pigs, and a cat, were inoculated sub- 
 cutaneously with particles from the disease, or with cultures ; but 
 only a local nodule was produced in each case, which, after a slight 
 increase, subsided. Nocard confirmed these results. Intra-peritoneal 
 and intravenous and subcutaneous inoculations in guinea-pigs, rabbits, 
 pigeons, fowls, dogs, and sheep were negative ; and no trace could 
 be found of the cultures in any animal subsequently killed and 
 examined. According to Bocarro, though fresh particles from the 
 disease inoculated in rabbits and dogs gave negative results, inocu- 
 lation of cultures in guinea-pigs, rabbits, monkeys, and rats, pro- 
 duced a local tumour of slow growth, which, on section, had the 
 character of the inoculated material. 
 
 From these experiments we must conclude that the disease has 
 not been shown to be transmissible to the lower animals, by either 
 inoculation of the diseased tissue, or by cultures of the Streptothrix ; 
 and the exact relation of both the Streptothrix and the mycelial 
 fungus to the disease must be considered an open question. 
 
CHAPTER XXXI. 
 
 GLANDERS. 
 
 GLANDERS is a specific inocalable disease of equines, characterised 
 by the formation of nodules and suppurating tumours, with which 
 characteristic bacilli are associated. The disease has been known 
 from very early times. It is described in books of the sixteenth 
 century and in very early treatises on farriery. It attacks horses, 
 n-.-es, and mules. Man and many of the lower animals can be 
 readily inoculated, but cattle and swine have an immunity. The 
 disease is especially prevalent in towns, or wherever li<r>^ may be 
 crowded together without those sanitary arrangements which are 
 .so much attended to in private stables ; and in large establishments 
 fresh horses are being constantly introduced to replace others, and 
 thus the opportunities for the importation of the disease are multi- 
 plied. The disease varies in its virulence. It may occur in a form 
 which proves fatal in a few days, or it may exist for months or 
 years without attracting notice, and yet be capable of being trans- 
 mitted to other animals. The term "farcy" is applied when the 
 disease manifests itself in the form of tumours in tin- .skin. 
 
 Glanders in the horse most commonly produces ulceration of the 
 nostrils and enlargement of the glands. It commences in the form 
 of nodules of the mucous membrane resembling in ilia ry tubercles, 
 and, like them, consisting of a collection of round n-lk Tli.-y 
 .suppurate and coalesce, forming irregular ulcers and raised, congested 
 nodules. The lymphatic glands become enlarged and Mippurate, 
 and the disease extends to the respiratory organs. In the IUII.L' 
 the early stage, the disease is readily mistaken for tubercu! 
 The nodules suppurate, and cavities are formed, but tli.-\ d. 
 tend to caseate. A glairy or muco- purulent discharge from the 
 nostrils should lead to very careful inspection, with every possible 
 precaution ; and it will probably be easy to detect the okmtifla .f 
 the nostrils. In other CUM-S there may be slight discharge from the 
 
452 INFECTIVE DISEASES. 
 
 nostrils and swelling of the glands, and nothing more will be visible 
 until a post mortem examination has been made. 
 
 Glanders in man is found amongst those whose duties bring 
 them into contact with diseased horses, such as stable-men, cavalry 
 soldiers, and veterinary surgeons ; and it is generally the result 
 of accidental inoculation of a wound. An abscess develops, followed 
 by metastatic purulent infiltration of the lungs, liver, spleen, and 
 bones. There may be oedeinatous swelling of the face and ulcers 
 in the nostrils. The joints may become swollen and painful. 
 
 The nodules consist of fibrous tissue and cells, with a tendency to 
 suppuration. In the lungs the disease spreads by the lymphatics. 
 
 *it 
 
 C^ mm 
 
 FIG. 184. BACILLI OF GLANDERS ; section of a glanders nodule, x 700 (FLUGGE). 
 
 The infiltrated patches are necrosed in the centre, which is sur- 
 rounded by leucocytes and fibrous tissue. 
 
 The bacilli were discovered by Lbfner and Schiitz in 1882. They 
 are found in the discharge from the nostrils, in the pus, and in the 
 nodules of animals artificially infected. 
 
 Bacillus mallei. Rods, with rounded ends, shorter and 
 thicker than the tubercle bacillus, occurring singly, or in pairs, 
 and sometimes in filaments. The protoplasm in the rod is broken 
 up in stained preparations,, as in the tubercle bacillus. They stain 
 with the watery aniline dyes, and intensely so with alkaline methylene 
 blue or Neelsen's solution ; they are non- motile and aerobic ; spore 
 formation has been described. They can be cultivated on the usual 
 media, especially on glycerine-agar and on potato ; but they will not 
 grow in infusions of hay, straw, or stable manure. On the surface 
 of glycerine-agar a colourless, transparent growth occurs on either 
 
GLANDERS. 
 
 >id. <>f the track of the needle; on -.'lycerine-agar with milk a 
 whitish layer develops, which gradually chang, .., f n)m 
 
 amber yellow to a reddish-brown. On blood >enim the growth U 
 transparent and yellowish ; on potato it is much more charact. i 
 After two or three days at the temperature of the 1,1, MM!, .-, tilm 
 develops in the vicinity of the inoculated area, which is honey-like, 
 transparent and yellow ; the transparency disappears, and i< 
 week the cultures have become reddish-brown. (Plate II., Fig. '. ) 
 
 The disease has been communicated to man by accid> 
 inoculation with a hypodermic syringe which had been used 
 inoculating cultures. Horses, asses, cats, goats, field mice, and 
 
 <$4t)' 
 
 FIG. 185. SECTION OF A BRANCH OF THE PULMOXAKY AKTEKY .SHOWING 
 GLANDERS BACILLI PENETRATING THE WALL (HAMILTON). 
 
 guinea-pigs, can all be infected with pure culture.-; ra .!> 
 and dogs are slightly susceptible; cattle, swine, and white mice 
 have an immunity. In the guinea-pig a swelling occurs at the seat 
 of inoculation, followed by the formation of a prominent tumour, 
 developing into an abscess. The skin becomes involved, and ..n 
 with indurated margin results. The lymphatic glands also become 
 implicated, and general, infection follows, extending frequently to 
 the testicles or ovaries, and death results after several weeks. 
 rabbits there is generally only a local abscess induced, which termi- 
 nates in a quickly-healing sore. Mice die in three or four days 
 from general infection; glanders nodules are found in the live,- and 
 
454 INFECTIVE DISEASES. 
 
 spleen, closely resembling miliary tubercles. Loffler recommends 
 inoculation of guinea -pigs as the most reliable method of diagnosis. 
 
 Sub-cultures of the bacillus rapidly lose their virulence. The 
 toxic products have been already described (p. 48). Mallein can 
 be prepared from a culture on sterilised potato by extracting 
 with glycerine and water, or from a culture of the bacillus in 
 broth. A virulent culture is obtained from a glandered horse, 
 or from a guinea-pig inoculated with fresh virus. Sub-cultures are 
 prepared in glycerine broth, and incubated at 37 C. for a month 
 or six weeks. If the cultures are found to be pure they are sterilised 
 in the usual way> in the steam steriliser, and by filtering through 
 porcelain a pale, amber- coloured liquid is obtained. To test for 
 glanders, a few drops (2| cc.) are injected underneath the skin, in the 
 middle of the side of the neck. In healthy horses there is no re- 
 action, or a very slight elevation of temperature. In glandered horses 
 there is marked rise of temperature (101 to 105), considerable local 
 swelling at the seat of inoculation, and signs of general disturbance, 
 while the glandered tumours become more swollen and painful. The 
 temperature of the horse to be injected should be taken night and 
 morning for two or three days before the operation ; and in horses 
 suffering from febrile disturbance the test should be delayed. 
 Thomassen made a number of experiments on horses suffering from 
 pleurisy, bronchial catarrh, strangles, and other diseases, without 
 any reaction, except in a glandered horse, used as a control experi- 
 ment. Hunting and M'Fadyean, in this country, have made most 
 careful observations and experiments, and there is no doubt that 
 mallein is a very valuable aid in the diagnosis of glanders. In 
 many cases the reaction has been obtained in horses, and the 
 existence of glanders has been discovered only after a most searching 
 post-mortem examination. With this means of diagnosis it is now 
 possible to determine exactly which are the infected animals in a 
 stable where the disease has broken out. The animals can then 
 be slaughtered, and the disease prevented from spreading. 
 
 Stamping-out System. Whatever may be the stage of the 
 disease or the extent or variety of it, isolation ought to be carried 
 out in its most complete form namely, slaughter. The disease 
 might be completely stamped out, if it were not for the difficult 
 question of compensation. It can undoubtedly be checked by the 
 existing laws. 
 
 In 1869 glanders was included in the list of contagious diseases, 
 and the provisions with regard to giving notice of the disease, the 
 regulation of movement, or exposure, and disinfection, were applied to 
 
GLANDERS. 455 
 
 horses suffering from glanders. Under an earlier Act it was made 
 an offence to expose glandered horses in markets or on common.. 
 In 1878 power to slaughter was incorporated in the Animal* ( )i ,l,-i . 
 
 (1) Where a person having a horse, ass, or mule in his possession, or 
 under his charge, gives notice to a constable that the horse, ass, or mule is 
 affected with glanders, or any person is convicted of an offence against 
 the Act of 1878 by reason of his having failed to give such a notice, then, 
 if at any time thereafter it appears to the Local Authority, on a special 
 report of a Veterinary Inspector, that the horse, ass, or mule is affected 
 with glanders, and the horse, ass, or mule is alive at the end of fourteen daya 
 after the receipt by the Local Authority of that special report, the Local 
 Authority may serve on the owner of the horse, ass, or mule a notice in 
 writing requiring him to slaughter it, or to permit them to slaughter it, 
 within a time specified in the notice. 
 
 (2) If in any case the owner fails to comply with the requisition of 
 the notice of the Local Authority, he shall be deemed guilty of an offence 
 against the Act of 1878, unless he shows to the satisfaction of the court 
 of summary jurisdiction before which he is charged that the horse, ass, 
 or mule, is not affected with glanders, or that the slaughter thereof is 
 for any reason unnecessary or inexpedient. 
 
 (3) The provisions of this Article may be put in force, from time to 
 time, as often as occasion requires, in relation to the same horse, ass, or 
 mule, on a further special report as aforesaid. 
 
 In the order of 1892 it was provided that glanders should i 
 farcy, and power was given to compel slaughter, and to compensate 
 by payment of half the value of a diseased animal, not exceeding 20, 
 and full value in the case of healthy animals. Owing to object 
 urged against the payment of compensation, another order was 
 passed, which came into operation at g the end of 1894; the order to 
 slaughter being amended as under : 
 
 (1) A Local Authority may if they think fit cause to be slaughtered 
 any diseased horse, ass, or mule, provided that if the owner of the horse 
 ass, or mule gives notice in writing to the Local Authority, or their 
 inspector or other officer, that he objects to the horse, ass, or mule being 
 slaughtered, it shall not be lawful for the Local Authority to cause that 
 horse, ass, or mule to be slaughtered except with the further spec 
 authority of the Board of Agriculture first obtained. 
 
 (2) A Local Authority may, if they think fit, cause to be alaugh 
 any suspected horse, ass, or mule, having previously obtained the cent 
 of the owner thereof. 
 
 (3) The Local Authority shall out of the local rate pay oompenaai 
 as follows for any horse, ass, or mule slaughtered under thi 
 
 (a) Where the horse, ass. or mule was diseased the compensation shall 
 
456 INFECTIVE DISEASES. 
 
 be such sum as the Local Authority think expedient, being a 
 minimum in the case of a horse of two pounds, and in the case of 
 an ass or mule of ten shillings ; provided that in no case shall the 
 amount of compensation, if above the said minimum, exceed one- 
 fourth of the value of the animal immediately before it became 
 diseased. 
 
 (5) In every other case the compensation shall be the value of the 
 horse, ass, or mule immediately before it was slaughtered. 
 
CHAPTER XXXII. 
 TETANUS, RABIES, AND LOUPING-ILL. 
 
 TETANUS. 
 
 TETANUS is a communicable disease of man and the lower anim.il>, 
 characterised by spasmodic contraction of the mux-lrs. It is com- 
 monly the result of an injury, and occurs especially after wound* 
 produced by means of splinters of wood or contaminated with earth 
 or dust, and may follow after surgical operations. 
 
 Carle and Rattone first showed, in 1884, that the disease could 
 be communicated from man to other animals. Rabbits inoculated 
 with pus from a case in man developed tetanus, and from these 
 rabbits the disease was conveyed to others. Nicola ier, the following 
 year, found that mice and rabbits inoculated with earth often 
 contracted tetanus, and that the pus which formed at the seat of 
 inoculation contained, amongst other organisms, charact-ri>tic Kaeilli. 
 Pure cultures were first obtained by Kitasato. 
 
 Bacillus of Tetanus. Slender, straight rods, and filamentous 
 forms. Spore formation takes place at the end of a bacillus, giving 
 it a drumstick appearance. They stain with aniline dyes, but best 
 with Neelseii's solution, or by Gram's method. By the Ziehl-Ne*'l>-n 
 method and methylene-blue, the spores can be stained ivd, in 
 contrast to the bacilli, which are stained blue. The bacilli 
 anaerobic, liquefy gelatine, and are slightly motile. They can be 
 grown at the ordinary temperature, but most readily at the t 
 perature of the blood, in an atmosphere of hydrogen, especially 
 the addition of 1 or 2 per cent, of grape sugar to the nutrient 
 medium. The young colonies on plate cultivations some^ 
 resemble those of Bacillus subtilis. They have an opaque <-, 
 and are surrounded by fine rays, extending in all dinvti,,n.- l,k- 
 thistle-down. In the depth of gelatine a ray-like growth QC 
 in the lower part of the track of the needle. The gelatine i> h<jue- 
 tied very slowly, and gas is given off. The cultures 
 
 457 
 
458 
 
 INFECTIVE DISEASES. 
 
 characteristic odour. In slightly alkaline broth, and peptone with 
 alkaline reaction, in an atmosphere of hydrogen, the gas formed 
 will be sufficient to break the flask if it is sealed up. Kitasato 
 obtained his cultures from pus, by taking advantage of the resist- 
 ance of the spores to high temperatures. By raising cultures 
 to 80 C. for three-quarters of an hour, the micrococci and bacilli 
 in the mixed culture were destroyed, while the spores of the 
 tetanus bacillus retained their vitality, and then sub-cultures were 
 obtained in a pure state. The spores are said to be killed by 
 exposure to steam for five minutes. A 5 per 
 cent, solution of carbolic acid with *5 per cent. 
 of hydrochloric, will destroy the spores in two 
 hours. Kitasato and Weyl obtained tetanin 
 from pure cultures of the bacillus, Brieger 
 having previously obtained it from impure 
 cultures. A tetano-toxin, indol and phenol, and 
 butyric acid are also found. Brieger and 
 ITrankel attribute the pathogenic properties to 
 a tox-albumin. These products have been de- 
 scribed more fully in a previous chapter (p. 41). 
 A pure-culture produces tetanus in a mouse 
 in twenty-four hours, and rabbits, guinea-pigs, 
 and rats can also be infected. No pus forms 
 at the seat of inoculation, as after inoculation 
 of earth, but the spasms commence in the 
 muscles nearest to the seat of inoculation. A 
 trace of a broth culture will kill a guinea-pig, 
 the symptoms developing in three days. 
 
 Kitasato succeeded in making animals im- 
 mune to tetanus, and subsequently the discovery 
 was made that the blood in immune animals 
 w ^ produce immunity in other animals, the 
 TURK OF TETANUS explanation being that the toxic principle of 
 
 BACILLI IN GRAPE- t ^ e tetanus bacillus induces the formation of 
 SUGAR GELATINE. , . P ., 
 
 Four days old. tetanus antitoxin; and if equal parts of the 
 
 (FRANK E L AND serum of an immune animal, and a fatal close of 
 tetano-toxin, are together injected into a healthy 
 
 guinea-pig, tetanus will not follow, showing that the virus has 
 been neutralised. Tizzoni and Cattani found that blood from an 
 immunised dog was not only capable of completely neutralising the 
 toxic power of filtered cultures, but that the injection of the blood- 
 serum produced immunity in otherwise susceptible animals, except 
 
 FIG 186 PURE-CUL- 
 
DESCRIPTION OF PLATE XXII. 
 Bacillus tetani. 
 
 FIG. 1. From a cover-glass preparation of a pure-cultivation of the tetanus 
 bacillus in broth ; stained with Neelsen's carbolised fuchsine. / 1200. 
 Lamplight illumination, 
 
 FIG. 2. From a cover-glass preparation from the same source ; stained with 
 Neelsen's solution and methylene blue, x 1200. Lamplight illumination. 
 
i'l.,1. .XXII 
 
 Figl. 
 
 \ 
 
 BACILLUS TETANJ 
 
 
RABIES. 
 
 guinea-pigs or rabbits. This antitoxin is possibly secreted by special 
 .-lands, such as the thymus and thyroid; and, according to Bri 
 and others, extracts of the thymus gland are antitoxic to o( 
 toxins, as well as the tetanus toxin. 
 
 These experiments have resulted in the employment of tetanic 
 antitoxin as a therapeutic agent (p. 63). 
 
 BABIES. 
 
 Rabies, or hydrophobia, is a disease like tetanus, the symptoms 
 being produced by a virus acting upon the nervous system. The 
 specific virus appears to originate in dogs, wolves, and jack;tU ; 
 by wounds inflicted by rabid animals, or by inoculation, the disease 
 may be communicated to man, cattle, sheep, deer, cats, i.il.l.it., an d 
 swine. 
 
 Among the early symptoms observed in the dog are sulkiness 
 and restlessness, depraved appetite, and irritability. A peculiar 
 expression of the countenance maybe noticed, with twitching* of the 
 eyes and face, and the animal's attention appears to be fixed upon 
 some imaginary object. A rabid dog is constantly trying to drink, 
 and at times howls or barks in a peculiar tone, whilst the breathing 
 becomes very irregular. On the fourth day, or later, death follows. 
 After death the glands are enlarged and congested, the tonsils are 
 inflamed, and the vessels of the epiglottis injected. In some cases 
 there is inflammation of the lungs, and the stomach may contain a 
 mass of straw, hair, and horse-dung. The membranes of the brain 
 and the spinal cord may be also congested. All these symptoms 
 may be present, or some only, or they may be entirely absent ; and 
 it is partly for this reason that Pasteur's researches have betM 
 such enormous value. Very little was known of the experimci 
 production of rabies until Pasteur commenced his investigations ; and 
 the test, which can be applied by inoculating rat.l.it-. i> invaluable 
 as a means of diagnosing rabies with absolute certainty. Dogs 
 suffer from symptoms simulating those of rabies; and formerly, \vln-n 
 human beings were bitten, it was impossible in some cases to 
 determine whether the dog had been suffering from rabies or i 
 We are indebted to Pasteur for the only reliable test which can 
 be applied ; and we are now in a position, when a human being is 
 bitten by a, dog supposed to be, but not really, rabiil, to remove 
 all cause for the anxiety which would otherwise remain for months 
 and even years. 
 
 In man the period of incubation lasts from eight days to 
 
460 INFECTIVE DISEASES. 
 
 several months, and in rare cases a much longer period. The wound 
 from 'a bite may have healed, and may again become inflamed, 
 and symptoms follow, owing to the poison affecting the brain, spinal 
 cord, or the peripheral nerves. In the dog the disease appears in 
 two forms raging madness and dumb madness ; and the identity of 
 the virus in the dog and in man is shown by the fact that virus 
 from man can produce both forms of the disease in the dog. The 
 virus may be obtained by inoculation of the saliva of a rabid dog ; 
 but this method is uncertain, as other micro-organisms are present. 
 Pasteur endeavoured to obtain it in a pure state, and was able 
 to demonstrate that the spinal cord, the brain, and the nerves 
 contain the virus. It was also found that by direct inoculation of 
 the nervous system the most certain results followed. 
 
 Bacteria in Rabies. Cocci have been described in connection 
 with hydrophobia by Fol, Babes, and Dowdeswell. The cocci, it is 
 said, were observed in sections of the spinal cord of rabid dogs. The 
 descriptions given by different observers vary considerably, and there 
 is not any particular coccus constantly associated with the disease. 
 Nor have any of the bacteria been isolated from the diseased animal, 
 which were alleged to be present in stained preparations. By many, 
 hydrophobia is believed to be due to the presence of a micro- 
 organism, but at present the nature of the contagium is unknown. 
 
 Protective Inoculation. Pasteur found that a dog inoculated 
 under the dura-mater with virus from the spinal cord of a rabid animal 
 will develop rabies, as a rule, within eighteen days. By trephining 
 rabbits and inoculating the virus, and by, in the same way, 
 transmitting the virus from rabbit to rabbit, the incubation period 
 gradually shortens, until it is reduced to six or seven days. The virus 
 has then reached its maximum virulence in the rabbit, and is " fixed." 
 Pasteur then studied the possibility of producing immunity. The 
 medulla of a rabbit, containing the virulent virus, was suspended in 
 a glass bottle over caustic potash at a temperature of 25 C. If a 
 number of spinal cords were thus treated, and examined from day 
 to day, it was found that they gradually lost their virulence, 
 becoming completely inert in from sixteen to twenty days. A series 
 of cords was thus obtained with diminishing virulence ; by injecting 
 subcutaneously an infusion of rabid spinal cord crushed in broth, 
 and beginning with an inert cord on the first day and using the 
 next in the series on the second day, and so on till a fresh spinal 
 cord could be injected, it . was found that dogs were rendered 
 insusceptible to the strongest virus, administered by inoculation or 
 by exposing them to the bites of rabid dogs. Dogs have usually 
 
RABIES. 1,11 
 
 an incubation period of several weeks, and Pasteur cone. -ivd that it 
 would be possible to anticipate the symptoms, which would naturally 
 follow in a dog which had been bitten or inoculated. I 
 them a mild form of hydrophobia by the injection of att 
 virus of short incubation period. These experiments shou.-d th.it 
 it was possible to do this, and the outcome was the introduction ,,f 
 a system of protective inoculation in the human subject. Pasteur 
 succeeded in giving immunity from hydrophobia to about fifty dogs 
 of every age and breed. 
 
 In 1885 Joseph Meister, a boy nine years of age, bitten badly 
 by a mad dog upon the hands, legs, and thighs, was brought to 
 Pasteur. At a post-mortem examination of the dog, its stomach 
 was found full of bits of hay, straw, and wood, and it had b 
 unquestionably rabid. On July 6th, sixty hours after Meister hail 
 been bitten, a syringe full of marrow from a rabbit which had died 
 on June 21st, and therefore fifteen days old, was injected beneath 
 the skin over the right hypochondriac region. The next morning 
 Meister was inoculated with a spinal cord fourteen days old, and M 
 on every day, till on the sixteenth a cord only one day old was used. 
 So many injections, however, need not have been given. a> it was 
 subsequently found that the spinal marrows injected during th- 
 first five days were inert when tested on rabbits. The marmu 
 the next five days showed an ascending scale of virulency, until, on 
 the last two days of the treatment, Meister had been inoculated 
 with a virus so virulent that it was capable of causing hydrophobia 
 in dogs after ten days' incubation. Meister remained conij>l-t-l\ 
 free from hydrophobia. From that time to the pivx-nt day many 
 thousands of patients have been treated in Pai -i> by >Iii:htly modified 
 methods, and it is very generally believed that a real prophyl. 
 agent has been discovered. 
 
 Pasteur suggested that the rabic virus might consist of two 
 distinct substances a living virus capable of developing in tl,.- 
 nervous system, and a secondary product which, in sutlici.-nt 
 portions, might have the property of hindering the development 
 of the living virus. The nature of tl,i> living viru> i> quite 
 unknown. 
 
 According to a return of the inoculations at the Past 
 stitute, the total number of persons treated in 1895 wi 
 whom five died. In three cases the symptoms of hydro,,!,, 
 occurred within a fortnight of the last inoculation. If these 1 
 cases are omitted, the number of per ***** '* ***** 
 and the deaths to two. The results are shown in the following t 
 
462 
 
 INFECTIVE DISEASES. 
 
 for the nine years previous to 1895, during which Pasteur's method 
 has been in operation : 
 
 Year. 
 
 Number of 
 persons 
 inoculated. 
 
 Number of 
 deaths. 
 
 Eate of 
 Mortality. 
 
 1886 
 
 2,671 
 
 25 
 
 0-94 
 
 1887 
 
 1,770 
 
 14 
 
 0-79 
 
 1888 
 
 1,622 
 
 9 
 
 0-55 
 
 1889 < ... 
 
 1,830 
 
 7 
 
 0-38 
 
 1890 
 
 1,540 
 
 5 
 
 0-32 
 
 1891 
 
 1,559 
 
 4 
 
 0-25 
 
 1892 
 
 1,790 
 
 4 
 
 0-22 
 
 1893 
 
 1,648 
 
 6 
 
 0-36 
 
 1894 
 
 1,387 
 
 7 
 
 0-50 
 
 1895 
 
 1,520 
 
 2 
 
 0-13 
 
 Of the 1,520 persons treated, 156 were bitten on the head or 
 face, 829 upon the hands, and 535 on other parts of the body ; 122 
 were bitten by animals experimentally proved to be rabid, 949 by 
 animals declared by veterinary certificate to be rabid, and 449 by 
 animals supposed to be rabid. 
 
 Babes at Bucharest inoculated 300 persons in one year, and 
 claimed to have reduced the mortality to '4 per cent. 
 
 Stamping-out System. There is every reason to believe that 
 rabies could be stamped out in England in six months, if a general 
 order for muzzling were enforced, and all ownerless dogs were 
 slaughtered. It is the ownerless cur, the vagrant dog, which is 
 mainly responsible for the spread of rabies ; and if a general muzzling 
 order cannot be put into force, it would undoubtedly check the disease 
 if all dogs were compelled to wear a collar with the name and address 
 of the owner, and all dogs without owners were destroyed. 
 
 LOUPING-ILL. 
 
 Louping-ill is regarded by some as an infective disease. It is 
 a disease of sheep, characterised by symptoms due to an affection of 
 the central nervous system. The symptoms consist in contractions 
 
LOUPING-ILL. 463 
 
 of the muscles of the head and limbs, loss of co-ordination and finally 
 complete loss of the power of movement. The name is dn-m-d from 
 the peculiar jumping movements in the early stage. 
 
 Klein and M'Fadyean independently investigated this disease. 
 Klein found bacteria in the cerebral fluid. No micro-organisms 
 were found in the blood. Special attention was drawn to a 
 bacterium which was found by Klein in six out of seventeen cases, 
 and to a micrococcus by M'Fadyean. 
 
 Bacteria in Louping-Ill Klein's Bacterium. Oval cocci a ml 
 rods -6 to 1 /A in length, '2 to '3 /n in breadth. Colonies in gelatin.-, 
 yellowish by reflected light, are brown by transmitted light. On the 
 Mil-face of gelatine the bacteria form a film, which is crenated .it 
 the edge, and thick in the middle, at first grey and later yill\vMi. 
 In the depth of gelatine a filament forms, composed of cl<>>< -ly 
 aggregated minute greyish colonies, and a prominent yellow growth 
 occurs 011 the free surface. The gelatine is not liquefied. The 
 bacteria grow in milk, and broth becomes turbid in two days, and 
 there is a copious flocculent greyish precipitate. 
 
 Injection of broth cultures subcutaneously in rabbits, guinea- 
 pigs, and mice, produced no result, except local swelling at the seat 
 of inoculation, which subsided without causing any constitutional 
 symptoms. The results were equally negative when the cuh 
 were injected subcutaneously in lambs. 
 
 M'Fadyean's Mkrococcus. Cocci -3 p. in diameter. The colon i-> 
 are flat, nearly circular, ahd have a smooth edge. In old col" 
 the centre appears as a dark spot. Gelatine is rapidly liquefied, 
 and a nearly colourless precipitate forms at the bottom of tin- 
 tube. Cultures on the surface of agar have a faint yellow tinge. 
 On potato the colour is deeper but the growth not so well marked. 
 Milk is coagulated. In broth there is an abundant growth rend.-r- 
 ing the liquid turbid and depositing a white precipitate. The mi 
 cocci stain by Gram's method. Inoculated in rabbits or guinea-pigs, 
 they produce suppuration ; in horses and bovines, an inflammatory 
 swelling results without suppuration. They produce abscesw 
 sheep and lambs. The cocci were isolated from abscesses in lamU 
 suffering from louping-ill. Though it is admitted that louping-ill 
 belongs to the class of infective diseases, there can be n> dmil.t from 
 these experiments that the nature of the contagium is unknown. 
 
CHAPTER XXXIII. 
 
 FOOT-ROT. 
 
 SHEEP are subject to several diseases which are classed as foot -rot. 
 There is one form, known as contagious foot-rot, which prevails in 
 certain localities, especially on wet land. Brown describes the 
 disease as primarily a disease of the skin, inducing exfoliation of 
 the cuticle, and exudation of fluid containing epithelial scales. The 
 inflammation extends to the membrane of the foot, leading to exfolia- 
 tion of the hoof, and development of epithelial scales, which form an 
 imperfect horny layer 011 the diseased membrane. In one outbreak 
 investigated by Brown, the disease in the early stage was confined 
 to the skin between the digits of the fore-feet; the surface was 
 red, tumid and pulpy, and white purulent matter existed on the 
 inflamed parts. Later, the hoof grew to an extraordinary length, 
 fungoid growths made their appearance, developing into foot -rot in. 
 an advanced form. 
 
 In France, according to Fleming, the contagious character of 
 this disease has long been recognised. So long ago as 1805 Pictet 
 imported 200 half-bred merino sheep, some of which were infected 
 with foot-rot, and placed them with 200 healthy sheep, and in a 
 short time all the sheep were infected. Several years later Favre 
 and Sorillon carried out investigations which conclusively proved 
 the infective nature of the virus. Among other experiments it 
 was found that when healthy sheep were inoculated in the feet 
 with virus from diseased sheep, the disease was communicated. 
 
 Contagious foot-root may be spread by healthy sheep receiving 
 the virus from infected sheep in fairs and markets. Ships, railway- 
 trucks, and carts in which diseased sheep have been conveyed, unless 
 subsequently thoroughly disinfected, may be the means of trans- 
 mitting the virus to healthy sheep. Healthy sheep turned into 
 pastures quite recently occupied by diseased sheep may be inoculated 
 from the discharges from the feet of the diseased sheep, which 
 contaminate the grass and soil. 
 
 464 
 
FOOT-ROT. 
 
 465 
 
 Law believed that the disease arose from an undiscovered micro- 
 organism, which was probably present in infected pastures. Others 
 in this country have disputed the contagious character of the disease, 
 and considered that the same conditions of the pasture which 
 produced the disease in a flock would produce it again in imported 
 animals, which would account for the apparent contagiousness. 
 
 Brown, in order to test the question of contagion, placed infected 
 sheep in a pen, the bottom of which was covered with straw which 
 was not removed while the experiments were in process. Three 
 healthy sheep, from a locality where foot-rot was unknown, were 
 placed with the infected sheep. At the end of ten days the feet 
 of the sound sheep were still healthy. Subsequently two of the 
 sheep were inoculated, and it was found that the virus introduced 
 
 FIG. 187. FOOT OF SHEEP SHOWING 
 DISEASE OF HORN (BROWN). 
 
 FIG. 188. SECTION THROUGH THR 
 FOOT SHOWING A CRACK BX- 
 
 TKNMM. TllKOOilt TMK W.M.I.. 
 
 subcutaneously in the vicinity of the foot produced the incipient 
 stage of the disease. On making further experiments the c 
 nature of one form of foot-rot was established, but it appea, 
 the contagious property is only developed aft,r long pen 
 exposure, and under certain conditions. On a dry 
 will quickly subside, but on moist land the c<> 
 rot may be communicated by simple contact, in f 
 
 and other experiments Brown has drawn th, 
 
 following conclusions : 
 
 1. That so far as the evidence goes it justifies the state 
 foot-rot is a contagious disease; the infective matter 
 when brought into contact with the skin between the daws, c 
 
466 
 
 INFECTIVE DISEASES. 
 
FOOT-ROT. 
 
 467 
 
 when introduced into the system by inoculation, and probably when 
 taken in by the mouth from contaminated pastures. 
 
 2. That it cannot be produced by long-continued exposure to 
 undraiiied, moist soils, with an abundant, coarse and wet herbage. 
 
 3. That animals exposed to these conditions for many months, 
 and resisting entirely the influences named above, contract foot 
 
 FIG. 191. -DISTORTION OF HOOK is AN ADVANCED FORM OF FOOT-BOT (Bno 
 
 in from fourteen to twenty-one days on being placed among sheep 
 .suffering from the disease. 
 
 4 That sheep affected with foot-rot may improve, a 
 time to time become worse; and finally may recover and ,,n 
 perfectly healthy condition of foot, notwitl.t.u.Un, that 
 been kept the whole period under the conditions irhkh in,h, 
 disease. 
 
468 INFECTIVE DISEASES. 
 
 5. That the contagium of foot-rot remains for some time in 
 the system (ten to twenty days and longer) without any indication 
 of disease appearing in the skin between the claws. An infected 
 sheep may therefore escape detection even by an expert, and may 
 introduce foot-rot into a sound flock. 
 
 One attack does not confer immunity. The disease has been 
 known to recur in sheep which have only recently recovered from 
 an outbreak. 
 
 The disease is no j , communicable to other animals, including 
 man. The flesh is harmless, but as in severe cases the sheep are 
 emaciated, the carcass is in consequence of little, if of any, value 
 as food. 
 
 The nature of the contagium is unknown. 
 
 Stamping-out System. Sheep should not be allowed to be 
 moved from an infected district except for slaughter. When sheep 
 are purchased to add to the stock on a farm, they should be 
 isolated for two or three weeks, and carefully watched before they 
 are allowed to mix with other sheep. If this disease is detected 
 in a flock, every animal should be carefully examined, and any 
 suspicious as well as any diseased sheep should be completely isolated 
 from the rest. Fleming recommends that those which have been 
 in contact, though still apparently quite healthy, should as a pre- 
 cautionary measure, be made to pass through a trough containing 
 a solution of chloride of lime to a 'depth of about four inches. 
 The solution is made by adding one or two pounds of chloride of 
 lime to two buckets of rain-water. If the trough is placed at the 
 entrance to the sheepfold, the sheep will be compelled to traverse 
 it at least twice a day. It is also recommended, when diseased 
 sheep are treated by this plan, that after recovery, all manure 
 in the fold should be removed and destroyed, and the soil dug up 
 to the depth of six inches, or lime freely spread over the surface. 
 Troughs and hurdles must be thoroughly disinfected, and buildings 
 freely ventilated after similar treatment. No locality can be con- 
 sidered free from suspicion until one or two months have elapsed 
 since the recovery of the last case. 
 
CHAPTER XXXIV. 
 
 FOUL-BROOD INFECTIOUS DISEASE OF BEES IN ITALY PEBRINE 
 
 FLACHERIE INFECTIOUS DISEASE OF CATERPILLAR. 
 
 FOUL-BROOD IN BEES. 
 
 FOUL-BROOD is a contagious disease attacking bees and especially the 
 larvae. The larvae rapidly lose their healthy appearance, die anil 
 decompose, turning into a coffee -coloured mass. The cells of the 
 honeycomb are mapped out by the dark-brown cappings of the cells 
 
 FIG. 192. DISEASED COMB (COWAN). 
 
 containing the diseased larvae. The decomposite. , ted with 
 
 the production of an unpleasant odour, which ran be detecte 
 some distance from an infected hive. The disease has been 1 
 from very early times. Preuss investigated it mi-r- 
 attributed it to micrococci." These were really the spoi 
 bacillus, which was first observed by Cohn. Later, Ohfldfr 
 
 IM 
 
470 
 
 INFECTIVE DISEASES. 
 
 FIG. 193. SPOKES OP BACILLUS 
 
 ALVEI. 
 
 Cheyne investigated foul-brood, isolated the bacillus, and fully 
 
 described its morphological and biological characters in nutrient 
 
 media. By removing the cap of 
 one of the diseased cells, the de- 
 composing larva can be withdrawn, 
 and cover-glass preparations will 
 reveal delicate bacilli and large oval 
 spores. The bacilli can be readily 
 isolated and cultivated in nutrient 
 gelatine. 
 
 Bacillus alvei (Cheshire and 
 Cheyne). Rods varying in size, 
 and forming large oval spores. 
 They are motile and possess flagella. 
 Cultivated in nutrient gelatine in 
 
 test-tubes, a delicate ramifying growth appears on the surface, 
 
 and irregular whitish masses arise along the needle track. Pro- 
 cesses shoot out from these masses, and 
 
 extend through the gelatine for long 
 
 distances. They are thickened at points 
 
 in their course, and are clubbed at the 
 
 ends. The gelatine is gradually liquefied, 
 
 and the bacilli form a loose, white, floccu- 
 
 lent deposit at the bottom of the tube. 
 
 The liquid in the tube becomes yellowish 
 
 in colour after a time, and gives off an 
 
 odour of stale, but not amnioniacal, urine. 
 
 On the surface of nutrient gelatine the 
 
 bacilli grow out in chains of rods in single 
 
 file, or of rows of several side by side. 
 
 The processes which .are formed tend to 
 
 curve, and at a short distance from the 
 
 track of the needle form a distinct circle, 
 
 from which another process grows out, 
 
 and a fresh circle is developed. The 
 
 gelatine in the vicinity of the bacilli 
 
 gradually liquefies, and channels are 
 
 formed in the gelatine in which the 
 
 bacilli move backwards and forwards. 
 
 On nutrient agar-agar a whitish layer 
 
 develops, consisting of bacilli arranged side by side, which in a 
 
 few days are replaced by rows of spores similarly arranged. On 
 
 FIG 194. PURE-CULTURE IN 
 NUTRIENT GELATINE, x 4 
 (CHESHIRE AND CHEYNE). 
 
INFECTIOUS DISEASE OF BEES IN ITALY. 471 
 
 potatoes they form a dryish, yellow layer, and in milk a tremulous 
 jelly. A cultivation of the bacillus in milk, sprayed over a honey, 
 comb containing a healthy brood of bee larv*, produced foul-1,, 
 Adult bees fed on material containing bacilli became infected 
 Inoculation of mice and rabbits with the bacillus gave d..nl,tf..l 
 results. 
 
 Stamping-out System.-The infected bees, combs, frames and 
 quilts must be destroyed, and the hives thoroughly disinfected, as 
 this is the only way in which the resistant spores can be got rid at, 
 Cowan believes that if foul-brood were under Government inspection 
 and infected hives were destroyed, the disease could 1* s^nii*-.! 
 
 FIG. 195. CULTIVATION ON THE SURFACE OF GELATINE, x 80 
 (CHESHIRE AND CHEYNK). 
 
 INFECTIOUS DISEASE OF BEES ix ITALY. 
 
 In Italy bees are subject to another infectious malady, and 
 Canestrini has found bacilli in the bees and in the larva;, which are 
 believed to be the cause of the malady. 
 
 Bacillus of Infectious Disease of Bees (('am^ti-iui). Rods 
 2 fj. in width and 4 to 6 /A in length, occurring .-inirly, in j 
 and in chains, sometimes capsulated. They are motile, and spore- 
 formation is present. They liquefy gelatine, colouring tin- liquid 
 pink and forming a white deposit. On agar they form a whit.- 
 growth, and on potato fa claret-coloured layer. ('ultui-> .-in- 
 to be capable of producing the disease in Ix-fs and 1.. 
 
 PEBKINE. 
 
 The silkworm disease known in France as pebrine is characterised 
 by the appearance of black patches on the skin of the won: I 
 was investigated by Cornalia, Nageli, and Pasteur. 
 
472 INFECTIVE DISEASES. 
 
 prophylactic measures have been described in another chapter 
 
 (P 7). 
 
 Panhistophyton ovatum. (Lebert. Nosema bombycis, Micro- 
 coccus ovatus, Corpuscles du ver-a-soie). Shining oval cocci, 2 to 3 /A 
 long, 2 /A wide, singly and in pairs, or masses ; or rods, 2'5 /x, thick, 
 and twice as long. They multiply by subdivision. They were 
 experimentally proved to be the cause of pebrine, gattine, maladie 
 des corpuscles or Flecksucht ; and were discovered in the organs of 
 diseased silkworms, as well as in the pupae, moths, and eggs. 
 
 Metchnikoff believes that these micro-organisms are not bacteria, 
 but psorosperms. 
 
 FLACHERIE. 
 
 Silkworms are also subject to a very destructive disease known 
 as flacherie, flaccidezza, maladie de morts blancs. The worms cease 
 feeding, die and become a putrid mass. The disease is dependent 
 upon bad hygienic conditions, and is very infectious. The cause 
 has not been determined with certainty, but it has been attributed 
 to a streptococcus. 
 
 Streptococcus bombyeis (Mikrozyma bombycis, Bechamp). 
 Oval cocci 1'5 //. diain., singly, in pairs, and in chains. They are 
 said to be present in dust from infected localities. 
 
 DISEASE OF CATERPILLARS. 
 
 Forbes has described an infectious disease of the larvae of a 
 caterpillar (Picris rapce). Cocci which were found singly and in 
 masses, were regarded as the cause of the malady. 
 
PAET III. 
 
 SYSTEMATIC AND DESCRIPTIVE. 
 
 473 
 
CHAPTER XXXV. 
 
 CLASSIFICATION AND DESCRIPTION OF 8PK2EB. 
 
 Ix reviewing the history of the various classifications which h.-.vt- 
 from time to time been proposed, we shall see that the gradual 
 improvements in the means of studying such minute* <>!! 
 the methods of cultivating them artificially, and of studying th<-ii 
 chemistry and physiology, and the ever-increasing revelations of the 
 microscope, have resulted in establishing these microscopic object 
 members of the vegetable kingdom, ranking among the lowest form* 
 of fungi, but with regard to the division into genera and species we 
 are still in a position of doubt and uncertainly. 
 
 Miiller, in 1773, was the first to suggest a classification. I It- 
 established two genera, Monas and Vibrio, and groups! them with 
 the Infusoria. In 1824 Bory de Saint Vincent al>o attcmptr.l 
 a classification; but it was not until Ehrenberg in 1838, ami 
 Dujardin in 1841, worked at the subject, that .1 M-i -ntific distinction 
 of species was attempted. 
 
 Ehrenberg described four genera : 
 
 I. Bacterium . . filaments straight, rigid. 
 
 II. Vibrio . filaments snake-like, flex ihlt 
 
 III. Spirillum . . filaments spiral, rigid. 
 
 IV. Spirochseta . . filaments spiral, flexible. 
 
 Dujardin united Spirillum and Spirocli 1 classed 
 
 thus : 
 
 I. Bacterium . . filaments rigid, vacillating. 
 
 II. Vibrio . filaments flexible, undolatory. 
 
 III. Spirillum . . filaments spiral, rotatory. 
 
 Bacteria were still considered as Ini'ii>ori:.. l.nt in !*"-' Perty 
 maintained that some of the small->t living organism befcoge 
 the animal and others to the vegetable kingdom, an- 1 that Vil.no 
 without question belonged to the latter. In 1853 Robin point, ,i 
 
 475 
 
476 SYSTEMATIC. 
 
 the affinity of the Bacteria and Vibrios to Leptothrix ; and Davaine, 
 in 1859, insisted that the Vibrios were vegetables, and were in 
 fact allied to the Algae. 
 
 Since that time a flood of light has poured in upon this subject 
 through the writings of Hoffmann, Pasteur, Cohn, Rabenhorst, 
 Hallier, Billroth, Warming, Nageli, Magnin, Marchand, Sternberg, 
 Van Tieghem, Koch, Fliigge, De Bary, Zopf, Buchner, Hueppe, 
 Marshall Hall, and many others who have studied the morphology, 
 life-history, and classification of bacteria. 
 
 Of all these writers we are most indebted to Cohn, not only 
 on account of his researches, which extended over many years, but 
 also for his system of classification, which has since been almost 
 universally adopted. 
 
 In his first classification, published in 1872, Cohn considered the 
 Bacteria as a distinct group belonging to the Algse, and divisible 
 into four tribes, including six genera : 
 
 I. Sphserobacteria . . globules (Micrococcus). 
 
 II. Microbacteria . . short rods (Bacterium). 
 
 III. Desmobacteria . . long rods (Bacillus and Yibrio). 
 
 IV. Spirobacteria . . spirals (Sphirochaeta and Spirillum). 
 
 Cohn noted, in spite of placing them with the Algse, that the 
 absence of chlorophyll connected the Bacteria to Fungi, and we find 
 Naegeli subsequently adopting this view, and employing the term 
 Schizomycetes or Fission-fungi. 
 
 Billroth, in 1874, disputed the division into species, and con- 
 sidered that all the forms described by Cohn were but developmental 
 forms of one micro-organism, Coccobacteria septica. In the following 
 year Cohn answered the criticism of Billroth, and produced a second 
 classification, in which he still maintained that distinct genera and 
 species existed. Cohn considered the genera to be distinguished by 
 definite differences in shape, which were adhered to throughout life, 
 while some special feature, as a difference in size or physiological 
 action, or some minute difference in form, determined the various 
 species. 
 
 The second classification of Cohn (1875) only differed from the 
 first in that, instead of keeping the bacteria as a separate group, he 
 placed them, from their close relationship with the Phycochromacae, 
 under a new group, the Schizophytes, and added the genera Lepto- 
 thrix, Beggiatoa, Crenothrix, Sarcina, Ascococcus, Streptococcus, 
 Myconostoc, and Streptothrix. 
 
 Fliigge retained the term Schizomycetes, and divided them thus : 
 
CLASSIFICATION AND DESCRIPTION OK SPECIES. 
 
 -177 
 
 SCHIZOMYCETES (FLtfGGE's ORIGINAL CLASSIFICATION). 
 
 Isolated, or in chains, 
 or united in amor- 
 phous gelatinous 
 families 
 
 Forming gelatinous 
 families of definite 
 form. 
 
 Colonies solid, filled 
 with cells 
 
 Colonies, with simple 
 layer of cells at the 
 periphery . 
 
 In large numbers, 
 in irregular 
 colonies . Aicococcut. 
 
 In small but defi- 
 nite numbers, in 
 regular groups . Sarcina. 
 
 Clathrocyiti*. 
 
 A 
 
 >. 
 
 o 
 
 ID 
 g 
 
 o 
 
 Short. 
 
 Isolated, or in small 
 heaps loosely united, 
 or in irregular gela- 
 tinous families 
 
 . Bacterium. 
 
 Long, x 
 
 
 Short, 
 
 
 
 distinctly 
 
 
 
 jointed 
 
 . Baeillw. 
 
 'Without 
 
 f Straight 
 filaments. 
 
 Long, ^ 
 not dis-1 Thin 
 tinctly j Thick 
 jointed J 
 
 . Leptothrix. 
 . Btggiatoa. 
 
 
 ramifica- 
 
 
 
 
 tions. 
 
 
 
 Threads 
 
 
 
 
 isolated, 
 inter-* 
 
 
 f Short rigid . 
 Wavy, or J 
 
 rio). 
 
 laced, or 
 
 ^ in spirals. I fl PT il e . 
 l J.iOng uexitc 
 
 . Spirillum. 
 
 in bun- 
 
 
 
 dles. 
 
 
 fStreptothrii 
 
 
 Pseudo-ramifications . 
 
 {Clathrotkri 
 
 . Threads in roundish gelatinous masses . 
 
 . ,*** 
 
478 SYSTEMATIC. 
 
 The belief is nevertheless rapidly gaining ground that the lowest 
 forms of vegetable life cannot be divided by a hard-and-fast line 
 into a series with chlorophyll (Algse), and a series without it (Fungi), 
 and the tendency now is to solve the difference of opinion between 
 Colin and Nageli by following the example of Sachs, and amalga- 
 mating the two series into one group, the Thallophytes. 
 
 Researches by competent observers have more recently clearly 
 demonstrated that several micro-organisms in their life cycle exhibit 
 successively the shapes characteristic of the orders of Cohn. 
 
 This doctrine of pleomorphism, now widely accepted, was dis- 
 tinctly foreshadowed in a publication by Lister in 1873, though 
 this memoir contained certain conclusions which have since been 
 abandoned. Lister described forms of cocci, bacteria, bacilli, and 
 streptothrix in milk, which he regarded as phases of the same 
 micro-organism, Bacterium lactis. As a result of his observations, 
 Lister remarks that " any classification of bacteria hitherto made 
 from that of Ehrenberg to that of Cohn based upon absolute mor- 
 phological characters is entirely untrustworthy." To Lankester, 
 however, belongs the credit of having definitely and precisely formu- 
 lated this doctrine. In a paper, also published in 1873, Lankester 
 observed that the series of form- phases which he had discovered in 
 the case of a peach-coloured bacterium led him to suppose that 
 the natural species of these plants were " within the proper limits 
 protean, and that the existence of true species of bacteria must be 
 characterised, not by the simple form-features used by Cohn, but by 
 the ensemble of their morphological and physiological properties as 
 exhibited in their complete life- histories." Lankester inferred that 
 these phase-forms were genetically connected, in that they all pos- 
 sessed the common characteristic of a special pigment, bacterio- 
 purpurin. These conclusions were vigorously opposed by Cohn, and 
 doubt still remains in the minds of some as to whether the different 
 forms are really only stages in the life-history of a single species. 
 Nevertheless the theory of pleomorphism has steadily gained ground 
 ever since. 
 
 Cienkowski and Neelsen worked out the different forms assumed 
 by the bacillus of blue milk ; Zopf has in a similar manner investi- 
 gated Cladothrix, Beggiatoa, and Crenothrix, and traced out various 
 forms (Fig. 196) ; Yan Tieghem has investigated Bacillus amylobacter 
 with a similar result ; Hauser has described bacillar, spirillar, 
 spirulinar, and various other forms in the Proteus mirabilis and 
 Proteus vulgaris. These facts obviously shake the very foundation 
 of Cohn's classification, and we are left without possessing a sound 
 
CLASSIFICATION AND DESCRIPTION OF SIM 
 
 479 
 
 basis for classification into genera or species. The mode of repro- 
 duction is not sufficiently known to afford a better means for 
 distinction than the other morphological appearances taken alone ; 
 nor can we depend upon physiological action, which is hrld ly 
 many to vary with, the change of form, according to altered 
 surroundings. 
 
 FIG. 196. 
 Branched Schizomycete : H Vi>, 
 
 
 - 
 
 (a) Continuous; (6) Composed of long rod 
 
 (Zopf). 
 
 Zopf, who has warmly supported the pleomorphism of barteria 
 hM suggested as a result of his investigate a d,v, 
 8chi Z omycetes, Spaltpihe, or Fission-fungi, mto the 1 
 groups : 
 
480 SYSTEMATIC, 
 
 ZOPFS CLASSIFICATION. 
 
 GROUP I. COCCACE^E. Possessing (so far as our knowledge at present 
 
 reaches) only cocci, and thread-forms resulting from the juxtaposition of 
 
 cocci. The fission occurs in one or more directions. 
 
 Genus I. Streptococcus (Chain-cocci). Division in one or more direc- 
 tions. Individual cocci remain united together to form chains. 
 
 Genus II. Merismopedia (Plate-cocci). Divisions in two directions, 
 forming lamellaB or plates. 
 
 Genus III. Sarcina (Packet-cocci). Division in three directions, form- 
 ing colonies in cubes or packets. 
 
 Genus IV. Micrococcus (Mass-cocci). Division in one direction, cocci 
 after division remain aggregated in irregular clusters, or singly, or 
 in pairs or in chains of three or four elements. 
 
 Genus V. Ascococcus (Pellicle-cocci). Like micrococcus, but the cocci 
 grow in characteristic gelatinous pellicles. 
 
 GROUP II. BACTERIACE^E. Possessing mostly cocci, rods (straight 
 or bent), and thread-forms (straight or spiral). The first may be absent, 
 and the last possess no distinction between base and apex. Division (as 
 far as is known) occurs only in one direction. 
 Genus I. Bacterium. Cocci and rods, or only rods, which are joined 
 
 together to form threads. Spore-formation absent or unknown. 
 Genus II. Spirillum. Threads screw-form, made up of rods (long or 
 
 short) only, or of rods and cocci. Spore-formation absent or 
 
 unknown. 
 Genus III. Leuconostoc. Cocci and rods. Spore-formation present in 
 
 cocci. 
 Genus IV. Bacillus. Cocci and rods, or rods only, forming straight or 
 
 twisted threads. Spore-formation present either in rods or cocci. 
 Genus V. Vibrio. Threads screw-form in long or short links. Spore- 
 formation present. 
 Genus VI. Clostridium.$>&mQ as bacillus, but spore-formation takes 
 
 place in characteristically enlarged rods. 
 
 GROUP III. LEPTOTRICHE.E. Possessing cocci, rods, and thread- 
 forms (which show a distinction between base and apex). The last 
 straight or spiral. 
 Genus I. Crenothrix. Threads articulated ; cells sulphur less ; habitat 
 
 water. 
 Genus II. Beggiatoa. Threads unarticulated ; cells with sulphur 
 
 granules ; habitat water. 
 Genus III. Phragmidiothrix. Threads jointless ; successive subdivision 
 
 of cells is continuous ; cells sulphurless ; habitat water. 
 Genus IV. LeptotJirix. Threads articulated or unarticulated ; successive 
 
 subdivisions of cells not continuous ; cells sulphurless. 
 
 GROUP IV. CLADOTRICHJEE. Possessing cocci, rods, threads, and 
 spirals. Thread-forms provided with false branchings. 
 Genus : Cladothrix. 
 
CLASSIFICATION AND DESCRIPTION OF SPECI1>. IM 
 
 Zopf, however, does not assert that all the fission-fungi cxliil.it 
 this pleoinorphism, nor does he pretend that his classitir.it ion will 
 include all the micro-organisms described. Colin, on the other 
 hand, was ready to admit that all the forms described by him were 
 not truly independent species. De Bary, Hueppe, Baumgarten, and 
 Fliigge have expressed other views with regard to the classification 
 of bacteria. 
 
 De Bary divides them into two great groups bacteria which 
 form endospores, and bacteria which form arthrospores. This 
 affords but little practical assistance, though regarded by 
 botanists, from a scientific standpoint, a* a step in tin- rL'ht 
 direction. 
 
 Hueppe, acknowledging that the fructification must eventually 
 be made the basis for classification, suggests an arranireinent for 
 provisional use in which this view is introduced (p. 482). 
 
 It has already been mentioned that the production of arthrospores 
 is only established in a very few species. Therefore, we are 
 hardly justified in assuming that all bacteria, the spore-formation 
 of which is quite unknown, are to be included with those in which 
 this kind of fructification has been observed, and consequently to 
 distinguish genera on the same grounds may be considered, to 
 the least, somewhat premature. In Baumgarten's <-].iiti -ation the 
 genus bacterium is dispensed with, and the genera divided into two 
 groups, the monomorphic and the pleomorphic. 
 
 GROUP I. MONOMORPHIC. 
 
 Genera. Coccus. 
 Bacillus. 
 Spirillum. 
 
 GROUP II. PLEOMORPHIC. 
 
 Genera. Spirulina. 
 Leptothrix. 
 Cladothrix. 
 
 Fliigge also, in his revised classification, includes the genus 
 bacterium in the genus bacillus. The new clairicati..n differs 
 also from the original one in the grouping together ,,f the dilV- 
 species according to the character and behaviour of tin- ooi 
 in nutrient gelatine. The abolition, in Fliigge'6 and liaum-ar 
 classification, of the genus bacterium is no doubt owini: to OOoft 
 having arisen from the distinction, between a bacterium an 
 bacillus, being made to depend upon length. Observer^ differed 
 as to whether a rod of a certain length ought to be considered a 
 
482 
 
 SYSTEMATIC. 
 
 o 
 
 S 
 o 
 d 
 
 I 
 
 o 
 
 
 
 CB 
 
 .! 
 
 a 
 
 
 
 ' 
 
 IP 
 
 o 
 
 j 
 
 
 
 ^ 
 
 p S 
 
 
 
 & 
 
 
 
 Hi 
 
 S 2 o 
 
 ". ' ' | g 
 . |s 
 
 s 
 
 < 
 
 g|| 
 
 PH >H ^ 
 ^ <1 
 
 'I ^ 
 
 P 03 
 S 
 
 O 
 
 
 ' ' * 
 
 QC 
 
 O 
 
 ^ 
 
 PH 
 
 
 "tj 
 
 -2 .^ 
 
 _2L *^ _JL 
 
 g'SS S 1 
 
 o tC b o 
 
 tH 
 
 p p 
 
 1 If III 
 
CLASSIFICATION AND DESCRIPTION OF SPECIES. 483 
 
 bacterium or a bacillus. To meet this dith'culty a nmi:li-:md 
 ready rule was suggested viz., that a rod le>s than t \\ic-- 
 ln-eadth in length should be considered as a bacterium, and <>tli--r 
 \vi>e a bacillus. But this purely arbitrary division was inadequate, 
 from the fact that a rod at one stage of its growth or under certain 
 conditions might, as far as length went, truly be a bacterium, and 
 under other circumstances be of such a length as to entitle its Innng 
 considered a bacillus (Fig. 197). We should avoid such contusion 
 if we followed Zopf, and acknowledged as a difference U-tween a 
 bacterium and a bacillus the presence or absence of that form of 
 spore- for mat ion now distinguished as endogenous spore-formation. 
 We might then conveniently retain this generic term, to include 
 that group of rod-forms in which this spore-formation is as yet 
 
 FIG. 197. FRIEDLANDKH'S PNKIMOCOOTS, x 1500 
 
 unknown; moreover, we should, by so doing, with one or two 
 exceptions, collect together those short rod-forms which appe. 
 link the simple cocci to the spore-bearing rods or Ixu-ill 
 
 The grouping together of the different specie aocordi 
 character of the colonies in nutrient gelatine is also of quest 
 
 r 
 
 a slignt variation i. - i At the name 
 
 considerably affect the appearances of the colonies. 
 
 case of the comma-bacilli of Finkler and of K 
 
484 SYSTEMATIC. 
 
 The classification of Zopf will lead the investigator to work upon 
 the same lines, and by tracing the life-history of individual forms 
 in pure-cultivations either to extend the work of establishing 
 protean species or to restrict the doctrine of pleomorphism to a few 
 forms. For though the author prefers the classification proposed by 
 Zopf, he is not prepared to accept his views entirely for instance, 
 to regard the bacterium of rabbit septicaemia as a micrococcus. 
 
 Any arrangement at present can only be considered provisional, 
 and therefore the most practical classification must be considered 
 the best. In fact, much more investigation is required before we 
 can arrive at a permanent and thoroughly scientific classification 
 of the known bacteria. Many bacteria have been described by 
 different observers as different species which are really identical. 
 Many micro-organisms have been described and named as new 
 species with very insufficient investigation. The determination of 
 species rests upon the accumulated evidence afforded by a thorough 
 knowledge of their life-history. The morphological appearances 
 under different conditions must be carefully studied, the presence 
 or absence of movement and of spore-formation, and when present 
 the exact character ; the appearances of colonies and of test-tube 
 cultivations in different media and under different circumstances ; 
 the .liquefaction and other changes in nutrient media ; the nature 
 of the chemical products, if any, and the effect on the living animal 
 of the bacterium itself and of its products, in varying doses, must 
 all be taken into account. We must also ascertain whether the 
 bacterium is an aerobe requiring the presence of oxygen, or an 
 anaerobe growing only in the absence of it, or a facultative anaerobe 
 growing equally well with or without it ; and lastly, we must know 
 whether the bacterium is a parasite requiring a living host, or a 
 saprophyte existing on dead animal or vegetable matter, or a 
 facultative parasite capable both of growing in the living animal 
 and of leading a saprophytic existence. Several writers have 
 classified the bacteria which have been described hitherto by taking 
 some of these characters into account, and so preparing a list which 
 is convenient for the purpose of bacteriological diagnosis. A system 
 of this kind is of value in leading investigators to supply information 
 which is wanting in order to verify and amplify the information 
 upon wilich the classification is based, and to identify species which 
 have been described under different names. 
 
CLASSIFICATION OF SPECIES 
 FOR BACTERIOLOGICAL DIAGNOSIS 
 
 COCCI. 
 
 (A) GELATINE NOT LIQUEFIED. 
 
 (a) Chromogenic. 
 
 Micrococcus violaceus 
 Micrococcus carneus . 
 Micrococcus cerasinus siccus 
 Micrococcus cinnabareus . 
 Micrococcus aurantiacus 
 Micrococcus versicolor 
 Micrococcus luteus 
 Micrococcus citreus . 
 Micrococcus ochroleucus . 
 Micrococcus cereus flavus . 
 Micrococcus agilis citreus . 
 Micrococcus flavus tardigradus . 
 Staphylococcus viridis flavescens 
 
 COLOl'R. 
 
 Violet 
 Flesh-red . 
 Cherry-red 
 Cinnabar-red . 
 Orange 
 Yellow . 
 Yellow 
 Yellow . 
 Yellow . 
 Yellow . 
 Yellow . 
 Yellow . 
 Greenish- yellow 
 
 HABITAT. 
 
 Water. 
 
 Water. 
 
 Water. 
 
 Air : 
 
 Water. 
 
 Water. 
 
 Water. 
 
 Water. 
 
 Air. 
 
 Air; water. 
 
 Lymph. 
 
 (b) Non-Chromogenic. 
 
 Micrococcus candicans 
 Micrococcus candidus 
 Micrococcus concentricus . 
 Micrococcus fervidosus 
 Micrococcus cereus albus . 
 Micrococcus aquatilis 
 Micrococcus aquatilis invisibilis 
 Micrococcus cumulatus tenui> . 
 Micrococcus rosettaceus . 
 Micrococcus viticulosus 
 Micrococcus plumosus 
 Micrococcus amylivorus . 
 Micrococcus acidi lactici . 
 Micrococcus urese 
 Micrococcus gingivae pyogenes 
 Micrococcus salivarius septicus 
 Micrococcus of Manfredi . 
 Micrococcus in pleuropneumonia 
 Micrococcus in trachoma . 
 Hsematococcu- bcrria 
 Diplococcus albi<-un> tardttBUnoH 
 
 Air; 
 
 Water. 
 
 Water. 
 
 Water. 
 
 |'u>: water. 
 
 Water. 
 
 Water. 
 
 Nasal mucus. 
 
 Water. 
 
 Air; water. 
 
 Pear-blight. 
 
 Milk. 
 
 Air : urim-. 
 
 1'u- 
 
 - 
 
 S|)UtUIM. 
 
 ph. 
 
 ' ! Vft. 
 
 \ ../ma! ; ' " 
 
 186 
 
486 
 
 CLASSIFICATION OF SPECIES 
 
 HABITAT. 
 
 Diplococcus coryzse Nasal mucus. 
 
 Pseudo-diplococcus pneumoniae Meningitis. 
 
 Micrococcus tetragenus Sputum. 
 
 Micrococcus tetragenus mobilis ventriculi .... Stomach. 
 
 Pediococcus cerevisise Beer ; air. 
 
 Pediococcus acidi lactici Malt ; hay-dust. 
 
 Streptococcus pyogenes Pus. 
 
 Streptococcus brevis Saliva. 
 
 Streptococcus septicus Soil. 
 
 Streptococcus vermiform is Water. 
 
 Streptococcus of mastitis in cows Pus. 
 
 Streptococcus in strangles Pus. 
 
 Streptococcus acidi lactici Milk. 
 
 (B) GELATINE LIQUEFIED. 
 
 (a) Chromogenic. 
 
 COLOUR. 
 
 Micrococcus agilis .... Pink .... Water. 
 
 Micrococcus roseus .... Pink .... Sputum. 
 
 Staphylococcus pyogenes aureus . Orange . . . Pus. 
 
 Micrococcus in pemphigus . . Orange . . . Bullse. 
 
 Staphylococcus salivarius pyogenes . Orange . . . Saliva. 
 
 Micrococcus fuscus .... Brown . . . Water. 
 
 Micrococcus flavus desidens . . Yellowish-brown . Air ; water. 
 
 Micrococcus cremoides . . . Yellowish- white . Water. 
 
 Micrococcus botryogenus . . . Yellowish. . . Equine tumours. 
 
 Micrococcus Finlayensis . . . Pale-yellow . . Yellow fever. 
 
 Staphylococcus pyogenes citreus . Yellow . . . Pus. 
 
 Micrococcus citreus liquefaciens . Yellow . . , Eczema. 
 
 Micrococcus flavus liquefaciens . . Yellow . . . Air ; water. 
 
 Micrococcus tetragenus versatilis . Yellow . . . Blood. 
 
 Diplococcus flavus liquefaciens tardus Yellow . . . Eczema. 
 
 Diplococcus subflavus . . . Yellow . . . Vaginal mucus. 
 
 Diplococcus citreus conglomeratus . Yellow . . . Pus ; air. 
 
 Diplococcus luteus .... Yellow . . . Water. 
 
 Diplococcus roseus .... Pink .... Air. 
 
 Diplococcus fluorescens foetidus . Green . . . Posterior nares. 
 
 (b) Non-Chromogenic. 
 
 Micrococcus albus liquefaciens Nasal mucus. 
 
 Micrococcus aerogenes Intestine. 
 
 Micrococcus radiatus Air ; water. 
 
 Micrococcus fcetidus Nasal mucus. 
 
 Micrococcus in Biskra-button or Pendjeh sore . . . Pus. 
 
 Micrococcus in influenza Blood. 
 
 Micrococcus Freudenreichi Milk. 
 
 Micrococcus in yellow fever Blood. 
 
 Micrococcus lactis viscosus Cream. 
 
 Micrococcus acidi lactici liquefaciens Cheesy butter. 
 
 Micrococcus urese liquefaciens Urine. 
 
 Micrococcus in gangrenous mastitis in sheep .... Milk. 
 
 Staphylococcus pyogenes albus . . . . . . Pus. 
 
 Staphylococcus pyosepticus Pus. 
 
 Diplococcus albicans amplus Vaginal secretion. 
 
FOR BACTERIOLOGICAL DIAG> 
 
 Pediococcus albus 
 Streptococcus liquefaciens 
 Streptococcus septicus liquefaciens 
 Streptococcus albus . 
 Streptococcus of Mannaberg . 
 Streptococcus coli gracilis 
 
 HAHIIAI. 
 
 Blood, 
 
 Water. 
 Urine. 
 
 (c) NO GROWTH IN GELATINE. 
 
 Micrococcus pneumoniae crouposae .... 
 Micrococcus endocarditidis rugatus .... 
 
 Nitromonas of Winogradsky 
 
 Micrococcus in pemphigus ..... 
 
 Micrococcus in influenza 
 
 Micrococcus gonorrhoeas 
 
 Diplococcus intercellularis meningitidis . 
 Micrococcus tetragenus subflavus .... 
 Streptococcus giganteus urethras .... 
 Streptococcus of Bonome 
 
 Saliva. 
 Heart. 
 
 Boa 
 
 Bulls 
 
 Sputum. 
 
 Nasal muciw. 
 Am, 
 
 Meningitis. 
 
 (D) GROWTH IN GELATINE UNDETERMINED. 
 
 Ascococcus Bilrothii 
 
 Leuconostoc mesenteroides 
 
 Streptococcus of progressive tissue necrosis in inir-- 
 
 Micrococcus pyogenes tenuis 
 
 Micrococcus of pyaemia in rabbits .... 
 Micrococcus of progressive abscess formation in mice 
 Micrococcus of Forbes 
 
 Micrococcus in syphilis ... 
 
 Streptococcus Havaniensis 
 
 Streptococcus perniciosus psittacorum 
 Streptococcus bombycis 
 
 :nfiiMun. 
 Beet juice ; mo- 
 
 lasses. 
 
 Putrid blood. 
 Pus. 
 
 infusion. 
 Putrid 
 Cabbage 
 
 pillars. 
 Blood 
 
 Blood of parrot 
 Diseased silk- 
 
 cater- 
 
 PACKET COCCI. 
 
 (A) GELATINE NOT LIQUEFIED, 
 (a) Non-Chromogenic. 
 
 Sarcina pulmonum 
 
 Sarcina ventriculi 
 
 (B) GELATINE LIQUEFIED, 
 (a) Chromogenic. 
 
 Sarcina mqbilis . 
 Sarcina rosea 
 Sarcina flavea 
 Sarcina lutea 
 Sarcina aurantiaca 
 
 COLOUR. 
 Red . . 
 Red . . . 
 
 Yellow . 
 Yellow . 
 Orange-yHlow 
 
 I 'hthihicftl sputum, 
 kch. 
 
 Hui'l. 
 I ...... 
 
488 CLASSIFICATION OF SPECIES 
 
 (b) Non-Chromogenic. 
 
 Sarcina alba 
 
 Sarcina Candida 
 
 Air ; water. 
 Air of breweries. 
 
 RODS. 
 
 (I.) AEROBES OR FACULTATIVE ANAEROBES 
 
 (A) GELATINE NOT LIQUEFIED. 
 (a) Chromogenic. 
 
 (A) SPORE-FORMATION PRESENT. 
 () Motile. 
 
 COLOUR. 
 
 Bacillus cyanogenus 
 Bacillus erythrosporus 
 
 Bacillus brunneus 
 
 Bacillus rubefaciens 
 Bacillus rubescens 
 Bacillus fuscus limbatus 
 Bacillus beroliniensis Indicus 
 Bacillus cyanogenus 
 Bacillus aurantiacus 
 Bacillus fluorescens aureus 
 Bacillus fluorescens longus . 
 Bacillus fluorescens tenuis . 
 Bacillus constrictus 
 Bacillus subflavus 
 Bacillus aureus . 
 Bacillus flavescens 
 Bacillus heminecrobiophilus 
 Bacillus canalis parvus 
 Bacillus fluorescens putidus 
 Bacillus dentalis viridaiis 
 Bacillus virescens 
 
 Bacillus latericeus 
 Bacillus spiniferus 
 Bacillus in purpurc 
 Bacillus uceus 
 Bacillus in cholera in ducks 
 Bacillus flavo-coriaceus 
 Bacillus striatus flavis . 
 Bacillus fuscus 
 
 . . . . Greyish-blue . 
 
 Blue milk. 
 
 Greenish-yellow 
 
 Water. 
 
 iarrhcea (Lesage) Green 
 
 Intestine. 
 
 (ft) Non-Motile. 
 
 
 Brown 
 
 Water. 
 
 (B) SPORE-FORMATION UNKNOWN. 
 
 
 (0) Motile. 
 
 
 Pale-pink 
 
 Water. 
 
 Pale-pink 
 
 Sewers. 
 
 us ... Brown 
 
 Rotten eggs. 
 
 [ndicus . . Indigo-blue 
 
 Water. 
 
 ordaniensis . Bluish 
 
 Sewers. 
 
 Orange 
 
 Water. 
 
 .reus . . Orange 
 
 Water. 
 
 ttgus . . . Greyish-yellow 
 
 Water. 
 
 Quis . . . Greenish -yellow 
 
 Water. 
 
 Pale-yellow 
 
 Water. 
 
 Pale-yellow 
 
 Water. 
 
 Golden-yellow . 
 
 Water; eczema. 
 
 . Yellow . 
 
 Swamp- water. 
 
 philus . . Yellowish 
 
 Caseous glands. 
 
 3 ... Yellowish 
 
 Sewer-water. 
 
 itidus . . Greenish . 
 
 Water. 
 
 aiis . . . Opalescent-green 
 
 Carious dentine. 
 
 Green 
 
 Sputum. 
 
 (i) Non-Motile. 
 
 
 Brick-red 
 
 Water. 
 
 Greyish-yellow 
 
 Eczema. 
 
 emorrhagica . Greyish-yellow 
 
 Blood. 
 
 Orange-yellow . 
 
 Water. 
 
 ducks . . Yellowish 
 
 Blood. 
 
 is . . . Sulphur-yellow 
 
 Water. 
 
 . Sulphur-yellow 
 
 Nasal mucus. 
 
 Deep-yellow . 
 
 Water. 
 
 m-liquefaciens . Greenish -yellow 
 
 Water. 
 
FOR BACTERIOLOGICAL DIAGNOSIS. 
 
 489 
 
 (b) Non-Chromogenic. 
 
 (A) SPORE-FORMATION PI: 
 (a) Motile. 
 
 Bacillus putrificus coli 
 
 Bacillus septicus vesica 
 
 Bacillus in cancer 
 
 1 1 MI,. in fjeces. 
 
 CyHtr 
 
 Stonuu-h. 
 
 (&) Non-Motile. 
 
 Bacillus acidi lactici (Hueppe) . 
 Bacillus coprogenes foetidus 
 Bacillus subtilis similans . 
 Bacillus epidermidis . 
 Bacillus of Colomiatti 
 
 (B) SPORE-FORMATION UNKN"\W 
 (a) Motile. 
 
 Bacillus osdematis aerobicus 
 
 Bacillus of Fulles (I.) 
 
 Bacillus stolonatus . 
 
 Bacillus venenosus brevis . 
 
 Bacillus venenosus 
 
 Bacillus gracilis anaerobiescens 
 
 Bacillus invisibilis 
 
 Bacillus albus . 
 
 Bacillus venenosus invisibilis . 
 
 Bacillus aquatilis sulcatus 
 
 Bacillus argenteo-phosphorescens 
 
 Bacillus gliscrogenus . 
 
 Bacillus cystiformis . 
 
 Bacillus of Guillebeau 
 
 Bacterium Zopfii 
 
 Bacillus ventriculi 
 
 Bacillus coli communis 
 
 Bacillus cavicida 
 
 Bacillus of Utpadel . 
 
 Bacillus aerogenes . 
 
 Helicobacterium aerogenes 
 
 Bacterium aerogenes . 
 
 Bacillus meningitidis purulentae 
 
 Bacillus pyogenes foetidus 
 
 Proteus Zenkrri 
 
 Bacillus enteritklis . 
 
 Bacillus hyarinthi M-pticus 
 
 Proteus lethalis . 
 
 BaciUus endocarditidis griseus . 
 
 BaciUus of Roth (I.) . 
 
 BaciUus Schafferi 
 
 BaciUus of swine-plague 
 
 BaciUus typhi abdominalis 
 
 BaciUus cavicida Havaniensis . 
 
 Bacillus cuniculicida Havaniensis 
 
 ,ilk. 
 
 S\vin- measle*. 
 lluuiiin faeceb. 
 Skin. 
 Cunjunctiva. 
 
 Earth. 
 
 Water. 
 Water. 
 Water. 
 
 Water. 
 
 Sea-water. 
 
 Urine. 
 
 (Trine, 
 
 Milk. 
 
 IIH- ; air. 
 
 St-iniK'ti "f 
 
 ;ii-. 
 mi-. 
 
 Intehtiii'. 
 
 .mi*. 
 
 Kii<l.N-:ipl 
 ol.lrag*. 
 
 
 
 -land*. 
 
 
490 
 
 CLASSIFICATION OF SPECIES 
 
 (&) Non- Motile. 
 
 Bacillus of Okada 
 
 Bacillus pyogenes soli .... 
 Bacillus of Fulles (II.) .... 
 
 Bacillus candicans 
 
 Bacillus septicus agrigenus 
 
 Bacillus scissus 
 
 Bacillus ubiquitus 
 
 Bacillus multipediculus .... 
 Bacillus albus anaerobiescens . 
 
 Bacillus Zurnianus 
 
 Bacillus canalis capsulatus 
 
 Bacillus of Both (II.) .... 
 
 Bacillus tenuis sputigenus 
 
 Bacillus crassus sputigenus 
 
 Bacillus coprogenes parvus 
 
 Bacillus of Fiocca ..... 
 
 Bacillus striatus albus .... 
 
 Bacillus capsulatus mucosus 
 
 Bacillus pseudo-diphtheriticus . 
 
 Bacterium ureae ..... 
 
 Bacillus nodosus parvus .... 
 
 Bacillus oxytocus perniciosus . 
 
 Bacillus lactis pituitosi .... 
 
 Bacillus limbatus acidi lactici . 
 
 Bacillus ovatus minutissimus . 
 
 Bacillus of Belfanti and Pascarola . 
 
 Bacillus of Tommasoli .... 
 
 Bacillus capsulatus 
 
 Bacillus of purpura hsemorrhagica (Babes) 
 Bacillus of purpura hsemorrhagica (Kolb) 
 Bacillus septicaemias haemorrhagicae . 
 Proteus capsulatus septicus 
 Bacillus acidiformans .... 
 
 Bacillus coli similis 
 
 Bacillus hepaticus fortuitus 
 Bacillus filiformis Havaniensis 
 
 Bacillus of Martinez 
 
 Bacillus diphtherias columbrarum . 
 
 Bacillus diphtherias 
 
 Bacillus of Schimmelbusch 
 Bacillus capsulatus Smithii 
 Bacillus erysipelatis suis .... 
 Bacillus in rhinoscleroma .... 
 Bacillus of Friedlander .... 
 Bacillus pneumosepticus .... 
 Bacillus endocarditidis capsulatus . 
 Bacillus septicus keratomalaciae 
 Bacillus of intestinal diphtheria in rabbits 
 Bacillus of acne contagiosa of horses 
 Bacillus pseudo-tuberculosis . . 
 Bacterium tholceideum .... 
 Bacillus gallinarum 
 
 HABITAT. 
 
 Dust. 
 
 Earth. 
 
 Soil. 
 
 Soil. 
 
 Manured soil. 
 
 Soil. 
 
 Air; water. 
 
 Air ; water. 
 
 Water. 
 
 Water. 
 
 Sewer-water. 
 
 Old rags. 
 
 Sputum. 
 
 Sputum. 
 
 Faeces. 
 
 Saliva. 
 
 Nasal mucus. 
 
 Nasal secretion. 
 
 Healthy throat. 
 
 Urine. 
 
 Healthy urethra. 
 
 Milk. 
 
 Milk. 
 
 Milk. 
 
 Eczema. 
 
 Pus. 
 
 Hair with sycosis. 
 
 Blood. 
 
 Blood, 
 
 Blood. 
 
 Blood. 
 
 Blood. 
 
 Liver. 
 
 Liver. 
 
 Liver. 
 
 Liver. 
 
 Liver. 
 
 Diphtheritic de- 
 posit. 
 
 Diphtheritic 
 throat. 
 
 Cancrum oris. 
 
 Intestine. 
 
 Blood. 
 
 Rhinoscleroma. 
 
 Sputum. 
 
 Septic pneumonia. 
 
 Endocarditis. 
 
 Internal organs. 
 
 Intestine. 
 
 Pus. 
 
 Internal organs. 
 
 Intestine. 
 
 Blood. 
 
FOR BACTERIOLOGICAL DIAGNOSIS. 
 
 Bacillus argenteo-phosphorescens 
 1 1; 1 1 i 1 1 us smaragdino-phosphorescei 
 Bacillus phosphorescens gelidus 
 Proteus hominis capsulatus 
 Bacillus of grouse disease. 
 P.at illus lactis aerogenes . 
 
 1TAT. 
 
 Ki>h. 
 Cuttl 
 Blood, 
 Blood, 
 
 Intewtin.-. 
 
 (A) GELATIXE LIQUEFIED, 
 (a) Chromogenic. 
 
 (A) SPORE-FORMATION PRESENT, 
 (a) Motile. 
 
 COLOLL. 
 
 Bacillus' violaceus .... Deep-violet 
 
 Bacillus in disease of bees (Canestrini) Pink . 
 
 Bacillus in " red -cod " . . . Red . 
 
 Bacillus rnesentericus ruber . . Reddish -yellow 
 Bacillus fluorescens h'quefaciens 
 
 minutissimus Greenish-yellow 
 
 \Viit.-r. 
 S;tlt,-<l codfish. 
 
 Potatoes. 
 
 Skin. 
 
 (B) SPORE-FORMATION UNKNOWN. 
 
 (a) Motile. 
 
 Bacillus ianthinus 
 Bacillus violaceus Laurentius . 
 Bacillus lividus . 
 Bacillus carnicolor 
 Bacillus rubidus . 
 Bacillus Indicus .... 
 Bacillus rosaceus metalloides 
 Bacillus ochraceus 
 Bacillus citreus cadaveris . 
 Bacillus buccalis minutus . 
 Bacillus arborescens . 
 Bacillus fulvus . . . . 
 Bacillus plicatilis 
 Bacillus pyocyaneus . 
 Bacillus fluorescens liquefaciens 
 Bacillus cyanofuscus . 
 Bacillus fluorescens nivalia 
 Bacillus chromo-aromaticus 
 Bacillus viscosus. 
 Bacillus pyocyaneus . 
 
 Bluish-violet . 
 Deep-violet 
 Violet-black . 
 Dark flesh-colour 
 Brownish-red . 
 Sealingwax-red 
 Magenta-red . 
 Yellow . 
 Yellow . 
 Yellow . 
 Yellow . 
 Yellow . 
 Yellowish 
 Yellowish-green 
 Greenish-yellow. 
 Greenish-brown 
 Bluish-green . 
 Green or brown 
 Green 
 Green 
 
 (&) No, i- i 
 
 Bacillus coeruleus .... Blue . 
 
 Bacillus glaucus Grey . 
 
 Bacillus membranaceus ainethystinus Violet 
 
 Bacillus lactis erythrogenes . . Red . 
 
 Bacillus mycoides roaeua . . Red 
 
 Water. 
 Water. 
 
 I !!' -'III. . 
 
 Blood, 
 
 Saliva. 
 
 Water. 
 Water. 
 ha 
 
 Water. 
 
 < ' i , , . , 
 
 Water. 
 
 I' ,, 
 
 \\ ..'. i. 
 Milk. 
 
 Boa 
 
 glue. 
 
492 
 
 CLASSIFICATION OF SPECIES 
 
 COLOUR. 
 
 Bacillus prodigiosus .... Blood-red . 
 
 Bacillus hydrophilus fuscus . . Yellow 
 
 Bacillus cuticularis .... Yellow 
 
 Bacillus helvolus .... Yellow 
 
 Bacillus tremelloides .... Yellow 
 
 Ascobacillus citreus .... Yellow 
 Bacillus argenteo-phosphorescens 
 
 liquefaciens Yellowish 
 
 Bacterium ternio (Vignal) . . . Yellowish 
 
 Bacillus smaragdinus fcetidus . . Green 
 
 Air. 
 
 Frog's lymph. 
 
 Water. 
 
 Water. 
 
 Water. 
 
 Skin. 
 
 Sea-water. 
 
 Saliva. 
 
 Ozaena. 
 
 (b) Non-Chromogenic. 
 
 (A) SPORE-FORMATION PRESENT. 
 
 (a) Motile. 
 
 Bacillus inflatus 
 
 Urobacillus Freudenreichi 
 
 Bacillus mycoides 
 
 Bacillus ramosus 
 
 Bacillus gracilis .... 
 
 Bacillus circulans 
 
 Urobacillus Duclauxi 
 
 Urobacillus Maddoxi 
 
 Bacillus limosus .... 
 
 Bacillus butyricus 
 
 Bacillus Hessii .... 
 
 Bacillus lactis albus . 
 
 Bacillus liodermos 
 
 Urobacillus Pasteuri . 
 
 Bacillus mesentericus fuscus 
 
 Bacillus mesentericus vulgatus . 
 Bacillus of potato rot 
 Bacillus maidis .... 
 Bacillus megatherium 
 Bacillus tumescens 
 Bacillus subtilis .... 
 Bacillus subtilis similis 
 Bacillus vacuolosis . 
 Bacillus of Scheurlen 
 Bacillus alvei 
 
 Air. 
 
 Air ; dust ; sewers. 
 
 Soil ; water. 
 
 Soil ; water. 
 
 Water. 
 
 Water. 
 
 Water. 
 
 Water. 
 
 Sea-dredgingK. 
 
 Milk. 
 
 Milk. 
 
 Milk. 
 
 Milk. 
 
 Urine. 
 
 Potato; dust; 
 
 water. 
 
 Potato; water, etc. 
 Rotting potatoes. 
 Maize infusion. 
 Boiled cabbage. 
 Beet-root. 
 Dust ; water ; soil. 
 Liver. 
 Liver. 
 
 Cancerous tissues. 
 Larvae of bees. 
 
 Non-Motile. 
 
 Bacillus aerophilus . 
 Bacillus implexus 
 Bacillus filiformis 
 Bacillus vermicularis 
 Bacillus incanus 
 Bacillus inunctus 
 Bacillus granulosus . 
 Bacillus carotarum 
 
 Air. 
 Water. 
 Water. 
 Water. 
 
 Swamp-water. 
 Swamp- water. 
 Sea-dredgings. 
 Boiled carrot. 
 
FOR BACTERIOLOGICAL DIAGNOSIS. l!;i 
 
 Bacillus brassic* 1TAT - 
 
 Bacillus in gangrene . 
 
 IWillus of Letzerich . " 
 
 I'.acillusanthracis -..'''' ""''' 
 
 () SPOBE-FORMATION UNKNOWN. 
 
 (a) Motile. 
 Bacillus pestifer ...... . . 
 
 Bacillus diffusus . . ^ ,' 
 
 Bacillus gasoformans .... vy. . 
 
 Bacillus h'quidus ..... W t 
 
 Bacillus guttatus ....... ' Water- 
 
 Bacillus liquefaciens ..... AIT, f 
 
 Bacillus radiatus aquatilis ..... Wat-r 
 
 Bacillus nubilis ....... W t 
 
 Bacillus albus putidus ...... ^- . 
 
 Bacillus hyalinus ........ ^y v 
 
 Bacillus vermiculosus ....... -y^v ( . 
 
 Bacillus delicatulus ......... ^\ 
 
 Bacillus punctatus ....... * \yr a . 
 
 Bacillus reticulans ....... \y a . 
 
 Bacillus figurans (Vaughan) ...... \\ 
 
 Urobacillus Schutzenber^i ...... Water. 
 
 Bacillus devorans ......... Water. 
 
 Bacillus venenosus liquefaciens ...... Water. 
 
 Bacillus aquatilis ......... \\ 
 
 Proteus sulphureus .......... Water. 
 
 Bacillus stoloniferus ......... Swamp -v. 
 
 Bacillus phosphorescens Indicus ...... Sea-w:i- 
 
 Bacillus phosphorescens indigenus ...... S<-;t-\\ 
 
 Bacillus cyaneo-phosphorescens ...... Sea-w:i- 
 
 Bacillus litoralis ........ 
 
 Bacillus halophilus ......... SeaKlredgingH. 
 
 Bacillus superficial ......... Sewage. 
 
 Bacillus cloacae . ........ Sewage. 
 
 Proteus microsepticus ........ Vt-rin- . 
 
 Proteus vulgaris .......... I 'utri<l .substance*. 
 
 Proteus mirabilis ......... Putrid substances. 
 
 Proteus septicus ........ . Septiwemia. 
 
 Bacillus foatidus ozaenae ........ Nasal mucus. 
 
 Bacillus septicus ulceris gangraenoai ...... Hlood and organs. 
 
 Bacillus albus cadaveris ........ J'lood. 
 
 Bacillus of Guillebeau ........ Milk. 
 
 Bacillus Ha vaniensis liquefaciens ...... Skin. 
 
 Bacillus carabiformis ........ Stomach. 
 
 Bacillus of Schou ......... KaMiit. 
 
 Bacillus leporis lethalis ........ ' ' 
 
 Bacillus liquefaciens communis ...... I . : . 
 
 Bacillus buccalis fortuitus ........ Saliva. 
 
494 
 
 CLASSIFICATION OF SPECIES 
 
 Bacillus ulna (Vignal) 
 Leptothrix buccalis (Vignal) 
 Bacillus varicosus conjunctive . 
 Bacillus gingivae pyogenes 
 Bacillus pulpse pyogenes . 
 
 Bacillus graveolens . 
 Pneumo-bacillus liquefaciens bovis 
 
 HABITAT. 
 
 Saliva. 
 Mouth. 
 Conjunctiva. 
 Alveolar abscess. 
 Gangrenous tooth- 
 pulp. 
 
 Skin of feet. 
 Lung. 
 
 (c) NO GROWTH IN GELATINE. 
 
 (A) SPORE-FORMATION PRESENT. 
 (a) Motile. 
 
 Bacillus ulna 
 
 Bacillus in putrid bronchitis 
 
 Bacillus mallei . 
 
 White of egg. 
 Sputum. 
 Glandered tissue. 
 
 (&) Non-Motile. 
 
 Bacillus in erythema nodosum Blood. 
 
 Bacillus tuberculosis . Tubercular tissue. 
 
 (B) SPORE-FORMATION ABSENT. 
 
 Bacillus sanguinis typhi . 
 Bacillus septicus acuminatus 
 Bacillus necrophorus 
 
 Blood. 
 
 Septic infection. 
 
 Condyloma. 
 
 (c) SPORE-FORMATION NOT STATED. 
 
 Bacillus allantoides . 
 Bacillus nitrificans . 
 Bacillus in measles . 
 Bacillus in ophthalmia 
 
 Air. 
 
 Soil. 
 
 Blood. 
 
 Conjunctiva. 
 
 (D) GROWTH IN GELATINE NOT STATED. 
 
 Bacillus senilis . 
 Bacillus leptosporus . 
 Bacillus allii 
 Bacillus indigogenus . 
 
 Blood. 
 
 Air. 
 
 Putrefying onions. 
 
 Infusion of indigo. 
 
 II. ANAEROBES. 
 (A) GELATINE LIQUEFIED. 
 
 (A) SPORE-FORMATION PRESENT. 
 
 (a) Motile. 
 Bacillus rubellus 
 
 Bacillus butyricus (Botkin) 
 Clostridium fcetidum 
 
 Dust. 
 
 Milkjwate rjdust 
 
 Earth. 
 
FOR BACTERIOLOGICAL DIAGNOM-. 495 
 
 HABITAT. 
 
 Bacillus radiatus Earth. 
 
 Bacillus liquefaciens magnus Earth. 
 
 Bacillus spinosus Earth. 
 
 Bacillus thalassophilus Sea-dredgingB. 
 
 Bacillus of symptomatic anthrax Tissues (quarter i 1 1 L 
 
 Bacillus oedematis maligni Lymph. 
 
 Bacillus tetani Wounds; earth. 
 
 (ft) Non-Motil. 
 
 Bacillus liquefaciens parvus Earth. 
 
 Bacillus anaerobicus liquefaciens Yellow fever. 
 
 (B) GELATINE NOT LIQUEFIED. 
 
 (A) SPORE-FORMATION PRESEM. 
 
 (a) Motile. 
 
 Bacillus amylozyma 
 
 BaciUus solidus Earth. 
 
 Bacillus polypiformis Earth. 
 
 (b) Non-Motile. 
 Bacillus muscoides 
 
 (B) SPORE-FORMATION AI5- 
 
 (J) Non-Votili. 
 BaciUus aerogenes capsulatus . 
 
 (c) GROWTH IN GELATINE NOT STATED. 
 
 (A) SPORE-FORMATION PRESENT. 
 
 (a) Motile. 
 
 Vegetable infu- 
 Bacillus butyncus .... 
 
 (B) SPORE-FORMATIOX UNKNOWN. 
 Bacillus cadaveris 
 
 CURVED RODS. 
 
 (A) GELATINE NOT LIQUEFIED. 
 
 (a) Chromogenic. 
 
 (a) Motile. 
 
 COL 
 
 Spirillum rubrum . Deep-red . 
 
 (b) Non-Chromogenic. 
 
 Spirillum suis . 
 
496 
 
 CLASSIFICATION OF SPECIES 
 
 Spirillum concentricum 
 Spirillum saprophiles 
 
 (a) Chromogenic. 
 
 Spirillum flavescens 
 Spirillum flavum 
 Spirillum aureum 
 
 Non- Motile. 
 
 Yellowish-green 
 Ochre-yellow 
 Orange-yellow . 
 
 (b) Non-Chromogenic. 
 
 Spirillum linguae 
 Spirillum nasale 
 
 HABITAT. 
 
 Putrefying blood. 
 Hay-infusion; 
 sewage. 
 
 Sewers. 
 Sewers. 
 Sewers. 
 
 Deposit on tongue. 
 Nasal mucus. 
 
 GELATINE LIQUEFIED, 
 (a) Non-Chromogenic. 
 
 (a) Motile. 
 
 Spirillum choleras Asiaticae 
 Spirillum of Tinkler and Prior . 
 Spirillum Metchnikovi 
 Spirillum of Miller . 
 Spirillum of Sanarelli 
 Spirillum tyrogenum 
 Spirillum marinum . 
 
 Intestine. 
 
 Intestine. 
 
 Intestine of fowls. 
 
 Carious teeth. 
 
 Water. 
 
 Old cheese. 
 
 Sea-dredgings. 
 
 NO GROWTH IN GELATINE, OR UNDETERMINED. 
 
 O) Motile. 
 
 Spirillum Obermeieri 
 Spirillum anserum 
 Spirillum undula 
 Spirillum sputigenum 
 Spirillum serpens 
 Spirillum tenue . 
 Spirillum volutans . 
 Spirillum plicatile 
 
 Spirillum dentium 
 Spirillum sanguineum 
 
 (&) Non-Motile. 
 
 Blood. 
 
 Blood of geese. 
 Putrid infusions. 
 Gums. 
 
 Stagnant water. 
 Putrid infusions. 
 Swamp- water. 
 Swamp- water. 
 
 Gums. 
 Brackish water, 
 
 BRANCHING FILAMENTS. 
 
 Streptothrix actinomycbtica. 
 Streptothrix alba. 
 Streptothrix liquefaciens. 
 Streptothrix musculorum suis. 
 Streptothrix Hofmanni. 
 Streptothrix farcinica. 
 Streptothrix asteroides. 
 Streptothrix carnea. 
 
FOR BACTERIOLOGICAL DIAGNOSIS. 1! '7 
 
 Streptothrix aurantiaca. 
 Streptothrix chromogenes. 
 Streptothrix odorifera. 
 Streptothrix viohn-.-a. 
 Streptothrix Forsteri. 
 Streptothrix madurse. 
 
 NOT CLASSIFIED. 
 
 Bacillus indigonacru-. 
 Bacillus prot-u< Hu.r, 
 Beggiatoa altwi. 
 Beggiatoa mirabilis. 
 Beggiatoa roseo-jn-rsit -ina. 
 Cladothrix dichotomy 
 Cladothrix invulnerabilis. 
 Crenothrix Kuhniana. 
 Diplococcus citreus liquefaciens. 
 Leptothrix buccalis. 
 Leptothrix gigantea. 
 
 MILTOCOCCUS aquatilis invi*ihilis 
 
 ^licrococcus crepusculum. 
 
 ^licrococcus foetidus. 
 
 Micrococcus Havanien~i-. 
 
 Micrococcus of septicaemia in ra 
 
 .M.iuas Okenii. 
 
 Mi mas vinosa. 
 
 Monas Warmingii. 
 
 M yconostoc gregarium. 
 
 Rhabdoraouas rosea. 
 
 Sarcina hyalina. 
 
 Sarcina intestinalis. 
 
 Sarcina lit<iralis. 
 
 Sarcina Reitenbachii. 
 
 Sarcina urinse. 
 
 Spliferotilusnatans. 
 
 Spirillum attenuatum. 
 
 Spirillum leucomelaneum. 
 
 Spirillum rosaceum. 
 
 Spirillum Rosenbergii. 
 S})irillum rufum. 
 Spiromonas Cohnii. 
 Spiromonas volubili^. 
 Streptococcus cadav 
 Streptococcus flavus di-i- 
 \'iljrio rugula. 
 
 
DESCRIPTION OF SPECIES ARRANGED FOR 
 REFERENCE IN ALPHABETICAL ORDER. 
 
 Ascococcus Billrothii. Small 
 globular cocci, united into charac- 
 teristic colonies. 
 
 They form on the surface of 
 nourishing fluids a cream-like skin, 
 divisible into an enormous number 
 of globular or oval families. Each 
 family is surrounded by a thick 
 capsule of cartilaginous consistency. 
 In a solution containing acid tar- 
 trate of ammonia the fungi generate 
 butyric acid, and change the origin- 
 ally acid fluid into an alkaline one. 
 
 FIG. 198. Ascococcus BILLKOTHII 
 (Cohn). 
 
 They were first observed on putrid 
 broth, and later on ordinary nourish- 
 ing solutions ; they also readily de- 
 velop upon damp slices of boiled 
 roots, carrots, beetroots, etc. 
 
 Ascobacillus citreus (Tlnna, 
 Tomtnasoli). Rods sometimes 
 curved, 1-3 p. in length, '3 p in width, 
 singly, in pairs, and masses. The 
 
 colonies develop slowly, and are 
 yellowish in colour. 
 
 The cocci inoculated in the depth 
 of gelatine form small colonies in 
 the track of the needle, and a slimy 
 pale-yellow growth on the surface ; 
 liquefaction sets in slowly. 
 
 On agar the growth is gelatinous, 
 and orange in colour, and rapidly 
 extends over the surface. 
 
 On potato the growth is abundant, 
 and pale yellow. 
 
 They were isolated from the skin 
 in eczema seborrhceicum. 
 
 Bacillus acidiformans (Stern- 
 berg). Short rods, 1'5 to 3 p. in 
 length, 1*2 p in width, and filaments 
 5 to 10 p. 
 
 Colonies circular ; iridescent by 
 reflected light. 
 
 Inoculated in the depth of gela- 
 tine they grow freely in the track of 
 the needle, and form a hemispherical 
 mass on the surface. They produce 
 gas bubbles. 
 
 On agar the growth is milk-white, 
 and the jelly becomes strongly acid. 
 On potato the growth is abundant. 
 
 In broth with 5 percent, glycerine 
 they produce opacity and a copious 
 viscid deposit, and the surface is 
 covered with gas bubbles. 
 
 Injected into the peritoneal 
 cavity of rabbits and guinea-pigs, 
 they produce death in twenty-four 
 hours. 
 
 They were isolated from the liver 
 in a fatal case of yellow fever. 
 
 Bacillus acidi lactici (Hueppe). 
 Rods 1 to 2-8 p. long, and '3 to 
 4 p wide, and thread forms. Spore- 
 formation present. In gelatine 
 
 498 
 
DESCRIPTION OF SPECIES. 
 
 499 
 
 cultures the breadth of the rods is 
 diminished. They grow best be- 
 tween 35 and 42 C., and cease 
 under 10 C. Over 45'5 C. they 
 no longer produce acidity. 
 
 Whitish colonies appear on the 
 second day. 
 
 In gelatine a delicate growth 
 appears along the whole track of 
 the needle, with spherical forms 
 here and there. 
 
 In milk they produce lactic acid 
 and the casein is precipitated. 
 
 Bacillus aerogenes (Miller). 
 Small rods varying in length. 
 Colonies white or yellowish- white ; 
 concentric. 
 
 In the depth of gelatine they 
 produce a yellowish filament, and 
 on the surface a grey patch with 
 dentated periphery ; later the fila- 
 ment is brown. 
 
 On potato the growth is yellowish 
 and dry. 
 
 They were isolated from the in- 
 testine in health. 
 
 Bacillus aerogenes capsulatus j 
 (Welch). Rods straight or slightly : 
 curved, 3 to 6 p. ; threads and | 
 chains ; capsulated. 
 
 Colonies on agar greyish- white, 
 with hairy processes. 
 
 They peptonise gelatine and pro- 
 duce gas. Broth becomes turbid, 
 and there is an abundant sediment. 
 Milk is coagulated. Cultures have 
 a faint smell of glue. 
 
 Injected into rabbits they pro- 
 duce gas in the blood and internal 
 organs. 
 
 They were isolated from a patient 
 after death, with blood-vessels full 
 of gas. 
 
 Bacillus aerophilus (Liborius). 
 Rods and filaments. 
 
 Colonies punctifonn ; greyish- 
 yellow. 
 
 Inoculated in the depth of gela- 
 tine the bacilli produce a funnel of 
 liquefied jelly, with flocculi in the 
 lower part. 
 
 On potato they form a smooth 
 yellowish layer. 
 
 They were isolated from con- 
 taminated cultures. 
 
 Bacillus albus (Eisenberg). 
 Rods and chains. 
 
 Colonies circular, white. 
 
 In gelatine the bacilli grow in the 
 track of the needle, ami form a 
 white hemispherical mass on the 
 free surface. 
 
 On agar the growth is pure whit* . 
 and on potato yellowish -whit.-. 
 
 They occur in water. 
 
 Bacillus albus anaerobiescens 
 (Vaughan). Short rods. 
 
 Colonies circular, yellowish- 
 brown. 
 
 Inoculated in the depth of gela- 
 tine they grow in the track of the 
 needle, and on the free sui 
 
 On agar the growth is pure white, 
 and on potato yellowish-white. 
 
 They occur in water. 
 
 Bacillus albus c a d a ve r i s 
 (Straussmann and Strieker). I . 
 J-5 p in length, -7"> /x in width. :in<i 
 filaments. 
 
 Colonies yellowish ; circular, and 
 later radiated. 
 
 Inoculated in the depth of gela- 
 tine they produce a funnel of lique- 
 fied gelatine with a thick dep<> 
 
 On agar there is an abundant 
 white growth. 
 
 On potato the growth is white or 
 yellowish-white, and colours the 
 potato in the vicinity bluish-brown. 
 The cultures have a putrefactive 
 odour. 
 
 Mice inoculated subcutane* 
 die in six hours, and guinea-pigs ID 
 twenty-four. 
 
 They were isolated from putrid 
 human blood. 
 
 Bacillus albus putidus (De 
 Bary). Rods and filam 
 
 Colonies cirnilur ;m<l bro\vni-h. 
 
 Inoculated in the depth of gela- 
 tine they produce rapid lique- 
 faction. 
 
 On agar and potato the growth 
 is slimy. Cultures develop a strong 
 putrefactive odour. 
 
 They occur in water. 
 
 Bacillus allantoides (I- K : 
 
 Rods 2 to 2:.'i ^ in leirj 
 width, and in chains. The rod* 
 develop cocci-forms united by a 
 gelatinous substance into ** 
 masses. They were isolated from 
 a contaminated culture. 
 Bacillus allii ((JriffiUw).-Rod 
 
500 
 
 DESCRIPTION OF SPECIES. 
 
 5 to 7 /z in length, 2 -5 ju, in width, 
 singly, in pairs, and zoogloea. 
 
 On agar they produce a bright 
 green film, and cultures are said to 
 emit traces of sulphuretted hydro- 
 gen. 
 
 They were isolated from putrid 
 onions. 
 
 Bacillus alvei (p. 470). 
 
 Bacillus amylozyma (Perdrix). 
 Rods 2 to 3 ju in length and '5 /n 
 in width, in pairs, and in chains. 
 They are anaerobic. 
 
 Colonies white, and producing gas 
 bubbles. 
 
 On potato in an atmosphere of 
 hydrogen the bacilli partly liquefy 
 it, and there is abundant formation 
 of gas. 
 
 They ferment sugar and starch. 
 
 Bacillus anaerobicus lique- 
 faciens (Sternberg). Slender rods, 
 about "6 //. in diam., in pairs, and 
 in filaments. 
 
 Colonies granular and white ; sur- 
 rounded by liquefied gelatine. 
 
 They grow along the track of 
 the needle when inoculated in the 
 depth of agar. 
 
 They were isolated from the in- 
 testine in a fatal case of yellow 
 fever. 
 
 Bacillus anthracis (p. 192). 
 
 Bacillus aquatilis (Frankland). 
 Rods 2-5 /M in length, and filaments 
 17 /z or longer. They resemble 
 Bacillus arborescens. 
 
 Colonies after liquefaction of the 
 gelatine have a yellowish-brown 
 nucleus from which proceed twisted 
 strands of filaments. 
 
 Inoculated in the depth of gela- 
 tine the growth in the track of the 
 needle is at first almost invisible, 
 later liquefaction occurs. 
 
 On agar the growth is shining 
 and yellowish. 
 
 Broth becomes turbid, and a 
 sediment forms at the bottom of 
 the tube. 
 
 On potato there is a slightly 
 yellowish streak. 
 
 They occur in water. 
 Bacillus aquatilis fluorescens 
 (Lustig). Short thin rods with 
 rounded ends. Non-inotile. 
 
 Colonies fern-like and iridescent. 
 
 Compare Eisenberg's Bacillus fluo- 
 rewen* non-liquefacieris. 
 Bacillus aquatilis graveolens 
 
 (Tataroff). Slender rods 1-3 /u, in 
 length. They, rapidly liquefy gela- 
 tine and produce an odour like that 
 of perspiration from the feet. 
 
 They occur in water. 
 
 Bacillus aquatilis sulcatus 
 (Weichselbaum), No. I. Rods mor- 
 phologically, and in cultures resem- 
 bling Bacillus typhosus. 
 
 Colonies in gelatine exhibit lines 
 and furrows. 
 
 The growth on the surface of 
 gelatine is said to be greater than in 
 cultures of Bacillus typhosus grown 
 for comparison. 
 
 They occur in water. 
 
 No. II. Rods also resembling in 
 morphology and cultivation the 
 Bacillus typhosus. 
 
 Colonies are said to be thicker than 
 those of No. I., and not dentated. 
 
 The growth on potatoes is yel- 
 lowish-brown, and emits a faint 
 odour of urine. 
 
 They occur in water. 
 
 No. III. Very short rods. 
 
 Colonies show lines and farrows, 
 and are yellowish. 
 
 On the surface of gelatine the 
 growth develops as a thin whitish 
 film. 
 
 On agar the growth is white and 
 abundant. 
 
 On potato the growth is yellow. 
 
 They were isolated from water. 
 
 No. IV. Rods and filaments. 
 
 Colonies circular and bluish. 
 
 On the surface of gelatine the 
 growth is greyish-white, and on 
 agar there is a similar appearance. 
 
 They do not grow on potato. 
 
 They occur in water. 
 
 No. Y. Rods rather thicker than 
 those of Bacillus typhosus. 
 
 Colonies similar to those of No. I. 
 
 The growth on the surface of 
 gelatine is yellow. 
 
 On agar the growth is viscid and 
 yellow, and on potato the growth is 
 faintly yellow and the surrounding 
 medium stained bluish-grey. 
 
 They occur in water. 
 
 Bacillus arborescens (Frank- 
 land). Rods 2-5 p in length, and 
 
DESCRIPTION OF SFK< LES. 
 
 5 n in width, singly, in pairs, and 
 in short chains, and filaments. 
 
 Colonies throw out delicate 
 branches with a highly characteristic 
 appearance ; the gelatine slowly 
 liquefies, the nucleus of the colony 
 becomes yellow, and the periphery 
 iridescent. 
 
 Inoculated in the depth of gela- 
 tine the bacilli form a cloudiness in 
 the track of the needle, and an 
 iridescent layer on the surface with 
 central depression of the gelatine 
 and commencing liquefaction. 
 Later the liquefaction produces 
 a funnel, and there is a yellow 
 deposit. 
 
 On the surface of agar the layer 
 is a dirty orange colour. 
 
 On potato the growth is orange 
 red with irregular protuberances, 
 and limited in growth. 
 
 They oc*cur in water. 
 
 Bacillus argenteo -phosphor es- 
 cens (Katz), No. I. Rods slightly 
 curved with pointed ends, 2'5 p. in 
 length, width one-third of their 
 length : singly, in pairs, and long 
 wavy filaments. 
 
 Colonies circular ; at first trans- 
 parent droplets, later yellowish in 
 colour. 
 
 On the surface of gelatine they 
 form a greenish-yellow film. 
 
 In broth they produce turbidity, 
 and later a skin on the surface, and 
 on sterilised fish a pale-yellow sticky 
 growth. 
 
 Cultures are photogenic. 
 
 They were isolated from the sea 
 at Sydney. 
 
 No. II. Rods with rounded ends 
 -21 n in length, '07 n in width, and 
 filaments. 
 
 Colonies on gelatine are circular 
 with sharp contours and greyish- 
 yellow in colour : later they arc 
 irregular and granular. 
 
 Inoculated in gelatine the bacilli 
 form a greyish-white filament in 
 the track of the needle, and a shining 
 patch on the surface. 
 
 On the surface of obliquely solidi- 
 fied gelatine they form a blokh- 
 grey film. 
 
 In broth they produce only 
 turbidity. 
 
 Cultures are photogenic. 
 
 They were isolated from ph<-ph< 
 rescent fi>h. 
 
 No. III. Rods not so thick as 
 those of No. II., sin^lv. in pair-, and 
 filaments. They are motil. 
 
 Colonies are white, scaly, and 
 wrinkled. 
 
 On the surface of gelatine the 
 growth spreads over the im-dium 
 
 On agar the growth 
 
 In broth they produce tin I' 
 and a skin flotfcnfl on the MM- face. 
 
 Cultures are photogenic at* 
 few days' growth. 
 
 They were isolated from a \ > 
 of cuttle-hMi. 
 
 Bacillus argenteo-phosphores 
 cens liquefaciens. 
 or slightly bent, '_' n in length, and 
 in width one-third of their leu. 
 filament >. 
 
 Colonies circular, pale In-own or 
 pale yellow and. after liquefaction, 
 with radiating processes extending 
 into the surrounding gelatine. 
 
 Inoculated in the depth of gela- 
 tine there i- a <_ r n>wth in the track 
 of the needle, and near the surface 
 a cup-shaped area of liquefact: 
 
 In broth they produce tnrl.; 
 and form a skin on the surface. 
 
 On sterilised fish they form a 
 yellow layer. 
 
 They are photogenic but not 
 markedly so. 
 
 Bacillus aurantiacus (Frank 
 land). R 
 in pairs, and in iilain- 
 
 ' lonies are prominent and pale 
 orange in colour. 
 
 Inoculated in th. depth of gela- 
 tine there i- a -liirht u r nwth b 
 track of the needle and an orange 
 patch on the fr. 
 
 On agar and potato the growth 
 i- also orange. 
 
 They occur in wat 
 
 Bacillus aureus (Adan 
 Sl.-nder rods strait . 
 Lent. !: to 4 /* in 1 
 in width : in" pait>. filaments, and 
 They are m 
 
 ( lonies circular or oval and 
 yellow in colour. 
 
 Inoculated in the deptjl of gel 
 tine the growth is very limited m 
 
502 
 
 DESCRIPTION OF SPECIES. 
 
 the track of the needle, while 
 small chrome-yellow hemispherical 
 masses develop on the free surface. 
 
 On potato they form a chrome- 
 yellow growth. 
 
 They occur in water and on the 
 skin. 
 
 Bacillus berolinensis Indicus 
 (Classen). Slender rods, singly, in 
 pairs, and short chains ; capsulated. 
 
 Colonies at first whitish acquire 
 in a few days an indigo-blue colour. 
 
 On the surface of gelatine they 
 form a blue layer, which slowly 
 
 On the surface of agar the indigo- 
 blue colour is very marked. 
 
 On potato they grow abundantly, 
 and develop the same colour. The 
 pigment is insoluble in alcohol, 
 chloroform or water, soluble in 
 strong acids, and decolorised by 
 ammonia. 
 
 They were isolated from river 
 water at Berlin. 
 
 Bacillus brassicse (Pommer). 
 Rods 1-9 to 5-4 p in length, -91 to 
 1*2 /u. in width, and filaments. They 
 form spores. 
 
 Colonies have the appearance of 
 a fine mycelium. 
 
 Inoculated in the depth of gela- 
 tine the growth in the track of the 
 needle sends off fine filaments, and 
 liquefaction quickly follows. 
 
 In the depth of agar white 
 colonies form in the track of the 
 needle, and on the surface the 
 growth is first cloudy and later 
 yellowish. 
 
 They were isolated from infusion 
 of cabbage. 
 
 Bacillus brevis (Mori). Rods 
 25 /u, long and -8 /z broad. Non- 
 motile. 
 
 Colonies pale-yellow ; non-lique- 
 fying. 
 
 Inoculated in the depth of gela- 
 tine small dots appear along the 
 needle track and a pale yellowish 
 growth on the surface. 
 
 On agar at 35 C. a yellowish 
 and on blood serum a greyish 
 growth appears in two or three 
 days. They do not grow on potato. 
 In broth they form a white cloudy 
 -deposit. 
 
 Mice inoculated subcutaneously 
 die in from sixteen to thirty hours. 
 They are also pathogenic in guinea- 
 pigs and rabbits. 
 
 They were found in drain water. 
 
 Bacillus brunneus (Adametz 
 and Wichman). Rods small and 
 slender. Spore formation present. 
 
 Colonies at first white, later 
 brownish. 
 
 Inoculated in the depth of gela- 
 tine the growth occurs along the 
 track of the needle and also on 
 the surface, developing a brownish 
 colour in the surrounding gelatine. 
 
 They occur in water. 
 
 Bacillus buccalis fortuitus 
 (Vignal). Rods 1*4 to 3 ^ in 
 length, singly, and in pairs. 
 
 Colonies circular and liquefying. 
 
 Inoculated in the depth of gela- 
 tine liquefaction occurs slowly, and 
 white flocculi occur in the liquid, 
 and later subside to the bottom. 
 
 In broth they produce turbidity 
 and a skin on the surface. 
 
 They were isolated from saliva. 
 
 Bacillus buccalis maximus 
 (Miller). Rods 2 to 10 ju, in length, 
 1 to 1'3 p in width, and filaments 
 30 to 150 p in length. 
 
 They occur in the mouth. 
 
 Bacillus buccalis minutus (Vig- 
 nal). Rods -5 to 1 /* in length, 
 and slightly less in width. 
 
 Colonies circular and faintly 
 yellow. 
 
 Inoculated in the depth of gela- 
 tine they form a yellowish-white 
 growth in the track of the needle, 
 and a patch of the same colour on 
 the surface ; liquefaction com- 
 mences slowly, and extends down- 
 wards until the gelatine is com- 
 pletely liquefied and a yellow mass 
 collects at the bottom of the tube. 
 In broth there is a similar deposit, 
 and an iridescent pellicle on the 
 surface. 
 
 On potato they form a yellow film. 
 
 They were isolated from saliva. 
 
 Bacillus butyricus, (Praz- 
 mowski. Bacillus ain.ylobacier, Van 
 Tieghem ; Bacillus of butyric c.id 
 fn- mentation}. Rods 3 to 10 p. long, 
 under 1 p wide, often indistinguish- 
 able from Bacillus subtilis. They 
 
DESCRIPTION <>i- si 
 
 
 grow out into long, apparently un- 
 jointed threads. They are mostly 
 actively motile, but also occur in 
 zoogloea. The rods and threads 
 are sometimes slightly bent, like 
 vibrios. They are anaerobic. The 
 shorter rods as a rule swell in the 
 middle, becoming ellipsoidal, lemon, 
 
 FIG. 199. CLOSTRIDIUM BUTYRUT.M. 
 
 A. Active stage, (a, 1) Bent rods (vibrio- 
 
 form) and threads, (c) Short 
 rods, (<Z) Longrods. 
 
 B. Spore-formation. C. Spore-germina- 
 
 tion. (Prazmowski. ) 
 
 or spindle-shaped ; the long rods, 
 and sometimes the short ones, swell 
 at one end : in either case ellipsoidal 
 spores are developed (Fig. 19J); 
 
 Cultivated in nutrient gelatine, 
 the medium is liquefied, and a scum 
 formed on the surface. They grow 
 
 best between 35 J and 40 < Ti,,. 
 spores are widely di-tnl.ntid in 
 nature, and grow readily on fleshy 
 roots, old cheese, etc. They convert 
 the lactic acid in milk into butyric 
 acid, and produce the ripening of 
 
 They occur aUo in solution 
 starch, dextrine, and sugar, and are 
 the active agents in the fermenta- 
 tion of sauerkrautand sour ghei 1. 
 
 Bacillus butyricus (Botkin). 
 
 Rods and tilaiiinits. spmv formation 
 present. They are anaerobic. 
 Colonies consist of felt-like 
 
 Inoculated in the depth of gela- 
 tine with 1-5 per cent, of grape- 
 sugar, the growth commences in 
 the lower part of the needle track 
 with abundant formation of gas 
 bubbles and liquefaction of the 
 
 In milk there is abundant gas 
 formation, which will break the 
 flasks if closed. 
 
 They were isolated from milk, 
 earth, and water. 
 
 Bacillus butyricus 
 (Hueppe). Rods slightly 
 2-1 /* in length, '38 /x in width. 
 and filaments. 
 
 Colonies yellowish; rapidly 
 liquefying. 
 
 Inoculated in the depth of 
 gelatine liquefaction occurs 
 along the track of the needle, and 
 later a wrinkled s-kin floats on 
 the surface. 
 
 On agar the growth is yel- 
 lowish. 
 
 On potato the growi 
 wrinkled and faint! 
 
 They coagulate milk. j>i 
 ting and then dissolving the casein. 
 Thev occur in milk. 
 Bacillus cadaveris (Sternberg 
 -Rods 1-5 to 4 p in length, and 
 1-2 M in width, singly, in pairs, and 
 
 They are anaerobi ><* in 
 
 ( glycenne-agar are irregular, gnu 
 
 <l They produce an acid reaction 
 
 m SuKTneous injection in guinea- 
 | pigs may produce extensive 
 
504 
 
 DESCRIPTION OF SPECIES. 
 
 oedema and death in twenty-four 
 hours. 
 
 They were obtained from the 
 liver in fatal cases of yellow fever. 
 
 Bacillus caeruleus (Smith). 
 Rods 2 to 2'5 IJL in length, and '5 /u, 
 in width, singly and in chains. 
 
 Colonies blue. 
 
 Inoculated in the depth of gela- 
 tine they form a colourless growth 
 in the track of the needle, and a 
 cup-shaped cavity in its upper part, 
 with bluish contents. 
 
 On agar they form a blue layer 
 and a deep-blue growth on potato. 
 
 They occur in water. 
 
 Bacillus canalis capsulatus 
 (Mori). Rods -9 to 1-6 p. in width, 
 capsulated. 
 
 Colonies milk-white. 
 
 The growth in the depth of gela- 
 tine is similar to Friedlander's 
 pneumococcus. 
 
 On agar the growth is viscid, and 
 on potato it is yellowish. 
 
 In broth a skin forms on the 
 surface. 
 
 They are pathogenic in mice. 
 
 They occur in sewage. 
 
 Bacillus canalis parvus (Mori). 
 -Rods 2 to o p. in length, '8 to 1 /A 
 in width. 
 
 Colonies very minute, pale yellow. 
 
 On the surface of gelatine they 
 very slowly form a yellowish film. 
 
 On agar the growth is dry and 
 yellow. 
 
 They are pathogenic in mice and 
 guinea-pigs. 
 
 They occur in sewage. 
 
 Bacillus candicans (Frankland). 
 Very short rods and filaments. 
 
 Colonies pure white. 
 
 Inoculated in the depth of gela- 
 tine they form isolated colonies in 
 the track of the needle, and a white 
 button on the free surface. 
 
 On agar they form a greyish 
 layer, and flourish on potato. 
 
 They occur in soil. 
 
 Bacillus capsulatus (Mori). 
 Oval forms and rods, sometimes 
 encapsuled. Non-motile. Colonies 
 white. 
 
 Inoculated in the depth of gela- 
 tine and agar a nail-shaped growth 
 occurs. 
 
 In broth they form a white- 
 turbidity, and a white pellicle 
 develops on the surface and on the 
 sides of the vessels. rt^Ti 
 
 On potato at 37 C. an abundant 
 moist, yellowish, stringy growth is 
 formed, with production of gas 
 bubbles. They are pathogenic in 
 mice and in rabbits if injected into 
 the pleural cavity. 
 
 They occur in drain water. 
 
 Bacillus capsulatus (Pfeiffer). 
 Rods singly, in pairs, or in chains 
 and in filaments. They have a well- 
 marked capsule. Colonies white. 
 
 Inoculated in gelatine they grow 
 in the track of the needle, and 
 form a white button on the free 
 surface. 
 
 On agar and on potato the growth 
 is also white and very viscid, so 
 that it can be drawn out into long 
 threads. 
 
 They produce a fatal result in 
 mice in two or three days, when 
 inoculated subcutaneously. A 
 minute quantity of a broth culture 
 injected into the peritoneal cavity 
 of guinea-pigs will prove fatal in 
 thirty-six hours. The bacilli are 
 found in the blood which is made 
 viscid. 
 
 They were isolated from a guinea- 
 pig found dead. 
 
 Bacillus capsulatus mucosus 
 (Fasching). Rods 3 to 4 /u. in 
 length, '75 to 1 ^ in width ; capsu- 
 lated. Colonies white. 
 
 Cultures in gelatine resemble 
 Friedlander's pneumococcus. 
 
 They form gas. 
 
 They produce a fatal result in 
 mice in thirty-six hours. 
 
 They were isolated from nasal 
 mucus in cases of influenza. 
 
 Bacillus capsulatus suis 
 (Smith). Rods from 1-2 to 1'8 p 
 in length and '8 to '9 /a in width. 
 There are three varieties of this 
 bacillus having a close resemblance to 
 the pneumococcus of Friedlander. 
 
 They were isolated from the in- 
 testines of swine. 
 
 Bacillus carabiformls (Kac- 
 zynsky). Rods short and slender. 
 
 Colonies develop characteristic 
 processes. 
 
DESCRIPTION OF SI'KMI-X 
 
 505 
 
 Inoculated in the depth of gela- 
 tine, the bacilli produce liquefaction 
 and colour the liquid greenish- 
 yellow. 
 
 On agar they form a yellowish- 
 white layer. 
 
 They were isolated from the 
 stomach of a dog. 
 
 Bacillus carnicolor (Tils).- 
 Rods 2 p. long, and '5 p. broad. 
 Singly actively motile. Spore- 
 formation not observed. 
 
 Colonies are in the form of cup- 
 shaped depressions. 
 
 Inoculated in the depth of gela- 
 tine they grow rapidly along the 
 whole track of the needle, forming 
 a funnel-shaped area of liquefaction 
 at the bottom of which there is a 
 pale pink deposit. 
 
 On potato they form slowly a 
 dark flesh-coloured growth. 
 They occur in water. 
 Bacillus carotarum (A. Koch). 
 Rods -97 to 1-05 p. in length and 
 filaments. 
 
 Colonies white and circular. 
 Inoculated in the depth of gela- 
 tine the bacilli grow slightly in the 
 track of the needle and abundantly 
 on the surface. 
 
 On agar they form a white, and 
 on potato a faintly brown layer. 
 
 They occur on boiled carrot and 
 beet. 
 
 Bacillus cavicida (Brieger,. 
 Rods morphologically and in culti 
 vations similar to Bacillus coli com- 
 munis. 
 
 Cultures are said to be patho- 
 genic in guinea-pigs. 
 
 They were isolated from faeces. 
 Bacillus cavicida Havaniensis 
 (Sternberg). Rods 2 to 3 p. m 
 length, and '7 /* in width, singly 
 and in pairs. 
 
 Colonies are of a pale-straw 
 colour. 
 
 Inoculated in the depth of gela 
 tine the bacilli form small trans- 
 lucent pearl-like spherical colonies, 
 and on the free surface the growth 
 is limited. 
 
 On potato the growth is at t 
 thin and dirty yellow, and latei 
 gamboge yellow. 
 
 Guinea-pigs inoculated subcuta- 
 
 neously die in ten or twelve 
 hours. 
 
 They were isolated from the 
 intestinal contents in a fatal case 
 of yellow fever, by inoculation of 
 guinea-pigs. 
 
 Bacillus chromo-aromaticus. 
 Rods which liquefy gelatine and 
 form a yellowish-white scum on 
 the surface. 
 
 On potato the growth is irides 
 cent and brownish. 
 
 In broth a scum forms on the 
 surface and the broth is ..!. 
 greenish- blue. (Juhmvs have an 
 aromatic odour. 
 
 They are said to produce pneu- 
 monia and pleurisy in rabbits. 
 
 They were isolated from a pig 
 with post-mortem appearances of 
 swine-fever. 
 
 Bacillus circulans (Jordan). 
 Rods 2 to f> p. in length and 1 p. in 
 width, singly and in short ch;i 
 Colonies are brownish. 
 Inoculated in the depth of gela- 
 tine they liquefy the medium in 
 the track of the needle, forming 
 a conical cavity at its upper j art. 
 
 On agar they form a translucent 
 film. 
 
 Milk is slowly coagulated. 
 In broth they produce turbidity 
 and a slimy deposit. 
 They occur in water. 
 Bacillus citreus cadaveris 
 (Strassmann). Rods '.' /* in length, 
 <; p. in width, singly and in chains. 
 Colonies pale yellow. 
 Inoculated in the depth of | 
 tine the bacilli form minute col 
 along the track of the needle, and 
 at its upper part liquefy the gela- 
 tine and produce a yellow depot! 
 
 They were found in the I 
 after death. 
 
 Bacillus cloacae (Jordan) 
 Short rods s fco 1 '.' ,* in length 
 1 ^ in width, nngiy and 
 
 Pa Colonies circular, yellowish. 
 
 Inoculated in the depth of gela- 
 tine liquefaction occurs in the t 
 of the needle, an iridescent 
 forms on the surface, and then 
 an abundant deposit. 
 
 On agar the growth i 
 
.506 
 
 DESCRIPTION OF SPECIES. 
 
 white, and on potato yellowish- 
 white. 
 
 In broth they produce turbidity 
 and a scum on the surface. They 
 reduce nitrates. 
 
 They occur in sewage. 
 
 Bacillus coli communis 
 (Escherich). See p. 344. 
 
 Bacillus coli similis (Stern- 
 berg). Rods 1 to 3 /u in length, -4 to 
 5 p in width ; singly and in pairs. 
 
 Colonies circular and pale brown 
 in colour. 
 
 In the depth of gelatine they 
 form a scanty growth in the track 
 of the needle, and on the free sur- 
 face a translucent film with irregu- 
 lar margins. 
 
 On potato the growth is pale 
 brown or dirty white. 
 
 They were isolated from human 
 liver after death. 
 
 Bacillus constrictus (Zimmer- 
 mann). Rods from 1'5 to G'5 p in 
 length, and '75 /n in width. The 
 rods are segmented. 
 
 Colonies are circular, granular, 
 and greyish-yellow. 
 
 In the depth of gelatine they 
 form a filament in the track of the 
 needle and irregular yellow heaps 
 on the free surface. 
 
 On the surface of agar the growth 
 consists of a yellow shining layer, 
 and on potato the same colour is 
 produced. 
 
 They occur in water. 
 
 Bacillus coprogenes foetidus 
 (Schottelius). Rods about as large 
 as Bacillus subtilis, but shorter. 
 They are non-motile. Spore-for- 
 mation occurs when the bacilli have 
 access to the air, but not in the 
 .animal body. 
 
 In the depth of gelatine a fila- 
 ment forms composed of yellow- 
 ish compact colonies ; and on the 
 surface a fine transparent film ; 
 cultures emit a strong putrefactive 
 odour. 
 
 On potato they form a light grey, 
 dry layer. 
 
 Subcutaneous injection of small 
 <loses had no effect on mice and 
 rabbits, but very large quantities 
 produced a toxic effect in rabbits. 
 Swine are not affected. 
 
 They were found by Schottelius 
 in the intestine in cases of swine 
 erysipelas. 
 
 Bacillus coprogenes parvus 
 (Bienstock). Very short rods. 
 
 On the surface of gelatine they 
 form a very limited, almost in- 
 visible, growth in the track of the 
 needle. 
 
 In mice they produce oedema and 
 death in thirty-six hours, and in 
 rabbits a local rash and death in 
 eight days. 
 
 They were isolated from human 
 evacuations. 
 
 Bacillus crassus aromaticus 
 (Taratoff). Rods 3'5 to 5 /LI long, 
 1-5 ft in width ; constricted in the 
 centre. 
 
 Colonies appear in the form of 
 cup-shaped depressions and pro- 
 duce a fruit-like odour. 
 
 In gelatine they grow in the 
 track of the needle, and later 
 produce a funnel-shaped area of 
 liquefaction. 
 
 They occur in well water. 
 
 Bacillus crassus sputigenus 
 (Kreibohm). Short thick rods, 
 sometimes curved; capsulated. 
 Colonies greyish- white. Cultures 
 in gelatine resemble those of Fried- 
 liinder's pneumococcus. They are 
 pathogenic in small animals. They 
 were isolated from human sputum. 
 
 Bacillus cuniculicida, Banllu* 
 of Rabbit Septicaemia (see p. 228). 
 
 Bacillus cuticularis (Tils). 
 Rods from 2 to 3 p in length, *3 to 
 '5 p. in width, and filaments. 
 
 Colonies are yellow and the gela- 
 tine is liquefied. 
 
 In the depth of gelatine they 
 produce liquefaction, and a skin 
 forms on the surface. 
 
 On potato the growth is slimy 
 and yellow. 
 
 They occur in water. 
 
 Bacillus cuticularis a 1 b u s 
 (Taratoff). Rods 3'2 /* long, con- 
 stricted in the middle. Actively 
 motile. 
 
 Colonies are opalescent and 
 bluish-white. 
 
 Inoculated in the depth of gela- 
 tine they form a white rosette- 
 shaped growth on the surface and 
 
DESCRIPTION OF SIM 
 
 E 
 '."/, 
 
 a shining white growth 
 along the needle track 
 which sends off long 
 rounded processes. 
 
 On agar, glycerine agar 
 and blood serum they 
 produce a luxuriant white 
 shining growth. 
 
 Broth becomes turbid 
 with a whitish deposit and 
 pellicle. 
 
 On potato there is a thick 
 moist brown growth. 
 
 They are found in water. 
 
 Bacillus cyaneo-fuscus 
 
 (Beyerinck). Rods "2 to -6 
 
 H in length, and half their 
 
 length in width. Motile. 
 
 In the depth of gelatine 
 they produce colonies in 
 the track of the needle, 
 which later develop a black 
 pigment. 
 
 In broth with per cent, 
 of peptone they produce a 
 blue colour, changing to 
 brown and finally black. 
 They were isolated from 
 cheese, size and glue. 
 
 Bacillus cyaneo-phosphores- 
 cens (Katz). Rods 2-t; ^ in length, 
 1 p. in width, singly, in pairs, and 
 filaments. Colonies circular and 
 brownish or greyjsh-yellow. 
 
 Inoculated in the depth of gela- 
 tine they form a grey-white fila- 
 ment in the track of the needle and 
 liquefaction follows at the upper 
 part ; later a skin forms on the sur- 
 face and a yellow deposit occurs at 
 the bottom of the liquefied jelly 
 which has a reddish-brown tinge. 
 
 In broth a similar skin floats 
 on the surface and the broth is 
 turbid. 
 
 On sterilised fish the growth is 
 viscid and yellow. 
 
 Cultures are phosphorescent, 
 especially in media containing 
 excess of common salt. 
 
 They were isolated from sea- 
 water at Sydney, and are pos-iUy 
 identical with Bacillus phosphores- 
 c< us of Fischer. 
 
 Bacillus cyanogenus (Hueppe). 
 -Bacterium *ynr>/n//u//t. Bacillut 
 "f Blue Milk. Motile rods, 2'5 to 
 
 \\ 
 
 
 FIG. 200. BACILLUS < , x 050. A 
 
 tive rods. B Rods in xH^ln-a. C. Cliai; 
 short rods. D. Chain of I < '.xjci stage. 
 
 F, G. Spore-forming rods. H. Involi. 
 forms (Xni 
 
 3-5 n in length, and -4 p wide (Fig. 
 200). The rods after division may 
 remain linked together, and form 
 chains. Xon-motile rods occur 
 enveloped in a gelatinous ca; 
 and involution forms. 
 
 Colonies appear after two 
 as small greyish- white points which 
 gradually assume a moist appear- 
 ance. The gelatine becomes steel- 
 grey, throwing the white colonies 
 into strong relief. 
 
 In the depth of gelatine a wli 
 growth appears in the track of the 
 needle near the upper part, an<l on 
 the freo surface, producing also a 
 dark steel-blue discoloration 
 the jelly which spreads down- 
 wards. 
 
 On agar a greyish growth ap- 
 I'Kirs, and the agar is coloured dark 
 brown. 
 
 On potato a yellow i- 
 growth develops, the potato around 
 stained grey-blue. Milk be- 
 comes slightly alkaline and of a 
 slate-grey colour, which on the 
 addition of acid changes to an 
 intense blue. Milk in which the 
 
508 
 
 DESCRIPTION OF SPECIES. 
 
 lactic acid bacillus is growing be- 
 comes sky-blue from the first. 
 
 They are non -pathogenic. 
 
 They are present in blue milk. 
 
 Bacillus cyanogenus (Jordan). 
 Rods 1'3 //. in length, '8 p. in 
 width. Slightly motile. 
 
 Colonies granular and irregular. 
 They colour the surrounding gela- 
 tine brown. 
 
 Inoculated in the depth of gela- 
 tine the bacilli produce a scanty 
 growth in the track of the needle, 
 and a film on the surface with 
 coloration of the gelatine beneath 
 it. 
 
 On agar they form a white layer, 
 and the jelly is coloured brown. 
 
 On potato the growth is brown. 
 
 They were isolated from sewage. 
 
 Bacillus cystiformis (Clado). 
 Short slender rods. Motile. 
 
 Colonies circular, yellowish, gra- 
 nular. 
 
 Inoculated in the depth of gela- 
 tine there is a scanty growth in the 
 track of the needle, and a white 
 patch on the free surface. 
 
 On agar the growth is yellowish- 
 white. 
 
 They were isolated from urine. 
 
 Bacillus delicatulus (Jordan). 
 Rods 2 p. long and 1 p, broad, 
 often in pairs or short chains. 
 Actively motile. Spore-formation 
 not observed. 
 
 Colonies at first whitish with 
 a radiating edge. Later they 
 liquefy the gelatine and the centres 
 become dark. 
 
 Inoculated in the depth of gela- 
 tine they rapidly liquefy it and 
 form a whitish pellicle and a brown 
 deposit. 
 
 On agar a greyish crinkled 
 growth appears, which gradually 
 becomes white and shining. 
 
 On potato there is a grey flat 
 growth. 
 
 Milk is coagulated and becomes 
 strongly acid. 
 
 Broth is made turbid and a white 
 serum and precipitate formed. 
 
 They occur in sewage. 
 
 Bacillus dentalis viridans 
 (Miller). Rods slightly bent, singly 
 and in pairs. 
 
 Colonies circular, yellowish and 
 concentric. 
 
 Inoculated in the depth of gela- 
 tine they grow in the track of the 
 needle and on the free surface, and 
 the jelly is coloured green. 
 
 On agar the growth is colourless 
 or slightly grey. 
 
 Intraperitoneal injections in mice 
 and guinea-pigs produce a fatal re- 
 sult. Subcutaneous injections cause 
 suppuration. 
 
 They were isolated from caries 
 of the teeth. 
 
 Bacillus dentriticus (Bordoni 
 Uffreduzzi and Lustig). Rods '85 
 to 2-8 p. long, and '5 to '85 p. broad, 
 singly and in zoogloea. Motile. 
 Spore-formation not observed. 
 
 Colonies have an arborescent 
 appearance. 
 
 Inoculated in the depth of gela- 
 tine they form a circular raised 
 growth at the point of puncture, 
 and white colonies along the needle 
 track. The jelly is gradually 
 liquefied. 
 
 On agar and blood serum there 
 is a scanty growth on the surface 
 and an abundant growth in the 
 track of the needle. Blood serum 
 is liquefied after some time. 
 
 Broth is rendered turbid ; a 
 white firm pellicle forms. 
 
 On potato there is a thick moist 
 white growth, which later becomes 
 yellow. 
 
 Found in water. 
 
 Bacillus devorans (Zimmer- 
 mann). Rods -99 p. to 1'2 p in 
 length, '74 p. in width ; singly, in 
 pairs and in chains. 
 
 Colonies are circular, granular, 
 and grey, with periphery formed of 
 radiating processes. 
 
 In the depth of gelatine they 
 produce a whitish filament and an 
 excavation at the upper part, which 
 may or may not contain liquefied 
 jelly. 
 
 On agar a greyish film is found. 
 
 They do not grow on potato. 
 
 They occur in water. 
 
 Bacillus diffusus (Frankland). 
 Rods 1-7 /x in length, '5 p. in 
 width ; singly, in pairs, and fila- 
 ments. 
 
DESCRIPTION OF SI 1 ! 
 
 609 
 
 Colonies circular, bluish-green, ! 
 \\ iih a granular nucleus and delicate 
 irregular periphery. 
 
 In the depth of gelatine there is 
 scarcely any growth in the track of 
 the needle, but a shining greenish- 
 yellow film on the surface, and 
 liquefaction below it. 
 
 On agar the growth is faintly 
 yellow. 
 
 In broth they produce turbidity 
 and a yellowish deposit. 
 
 On potato the growth is yellow- 
 ish. 
 
 They occur in earth. 
 Bacillus diphtherias (p. 332). 
 Bacillus diphtheria colum- 
 barum (p. 336). 
 
 Bacillus dysodes(Zopf). Cocci, 
 long and short rods, and spores. 
 
 They were observed in bread, 
 making it greasy and unfit for food, 
 and generating a penetrating odour 
 resembling a mixture of pepper- 
 mint and turpentine. A great loss 
 may result to bakers if the fungus 
 is introduced with the yeast. 
 
 Bacillus endocarditidis cap; 
 sulatus (Weichselbaum). Cocci 
 i. tumbling Fried lander's pneumo- 
 cocci. 
 
 Colonies faintly yellow, with 
 dentated contours. 
 
 In the depth of gelatine the 
 growth produces a filament in the 
 track of the needle, and a patch like 
 stearin on the free surface. 
 
 Large doses injected subcutane- 
 ously, or into the peritoneal cavity 
 prove fatal to rabbits. 
 
 They were isolated from infarcts 
 in a fatal case of endocarditis. 
 Bacillus endocarditidis griseus. 
 Rods motile. 
 
 Colonies granular and brown or 
 yellowish-brown. 
 
 In the depth of gelatine there is 
 a filamentous growth in the track 
 of the needle, and a circular whit- 
 ish patch on the surface. On agar 
 and potato the growth is greyish- 
 brown. 
 
 Cultures cause a fatal result m 
 mice and guinea-pigs. 
 
 They were isolated from a case of 
 endocarditis. 
 Bacillus enteritidis (p. 372). 
 
 Bacillus epidermidis (Bordoni 
 Uffreduzzi). l; ,, ;{ M i n 
 
 length, and -3 p. in breadth. Spore- 
 formation occurs at 25 C. 
 
 They grow very sparingly on 
 gelatine. 
 
 On agar there is a surface growth. 
 
 On potato at 15 the 
 
 growth appears first in the form 
 of drops, which gradually extend 
 and coalesce and form a thin layer 
 
 rll I.I > Kl..l KAN- "N HO 3 
 
 I over the surface. On blood serum 
 they form a thin film. 
 
 Inoculation in rmbbiUmod gumea- 
 ! pigs and on the humai 
 duced no result. 
 
 They were isolat- naket 
 
 of cuticle from between the toes. 
 Bacillus erysipelatis suis 
 
 ' ' " ' 
 
 Bacillus erythrosporus 
 (Ei<l; .-nder rods and 
 
510 
 
 DESCRIPTION OF SPECIES. 
 
 filaments. Motile, 
 tion present. 
 
 Spore-forma- I On potato the growth is brown. 
 They occur in water. 
 
 FIG. 202. PHOTOGRAPH OF PART OF AN IMPRESSION PREPARATION OF 
 BACILLUS FIGURANS ON NUTRIENT GELATINE, x 50. 
 
 Colonies circular, with brown nu- 
 cleus and yellowish-green periphery. 
 In the depth of gelatine they 
 
 Bacillus figurans (Crookshank). 
 Rods, with rounded ends, varying 
 in length. Spore-formation present. 
 
 FIG. 203. PART OF THE SAME SPECIMEN SHOWN IN FIG. 202 x 2(JO. 
 
 grow in the track of the needle 
 and on the free surface, colouring 
 the jelly green by transmitted light, 
 yellow by reflected light. 
 
 In plate-cultivations they cause 
 a cloudy growth, spreading from 
 various points ; if a cover-glas& 
 impression is made, this is found 
 
DESCRIPTION ol SI'KCIES. 
 
 .-.11 
 
 to consist of the regularly-arranged 
 parallel rods. The chains of rods 
 become twisted at intervals into 
 curious convolutions, from which 
 offshoots are continued in various 
 directions. These long shoots or 
 processes are again twisted at 
 intervals into varying shapes and 
 patterns (Figs. 202, 203). Culti- 
 vated in nutrient gelatine, the bacilli 
 form on the surface visible windings, 
 from which fine filaments grow 
 down into the gelatine. They 
 spread out also in almost parallel 
 lines transversely from the needle 
 track. The gelatine is not liquefied. 
 
 On an oblique surface of nutrient 
 agar-agar the filaments spread down- 
 wards into the substance of the 
 jelly, and outwards from the cen- 
 tral streak on the surface, forming 
 a feather-like cultivation (Fig. 201). 
 
 They were obtained from the 
 air, and later were identified by 
 the author with Bacterium Zopfii. 
 
 Bacillus figurans (Vaughau). 
 Rods and threads. 
 
 Colonies composed of curved and 
 
 terlacing lines. 
 
 -In gelatine they grow in the track 
 of the needle, and very slowly 
 liquefy it. 
 
 On agar they form a thin white 
 layer. 
 
 They were found in water. 
 
 Bacillus filiformis (Tils). Rods 
 4 p. in length and 1 /LI in width, 
 singly and in chains. Spore-forma- 
 tion present. 
 
 Colonies are granular, with yel- 
 lowish nucleus. 
 
 Inoculated in the depth of gela- 
 tine there is no growth in the 
 track of the needle, but a whitish 
 growth on the surface, liquefying 
 the jelly slowly. 
 
 In broth they form a skin on 
 the surface. 
 
 On agar the growth is white. 
 
 On potato the growth is dry and 
 after a time brownish. They 
 coagulate milk. 
 
 They occur in water. 
 
 Bacillus filiformis Havaniensis 
 (Sternberg). Long slender rods, 
 3 /z in diam.. and filament-. 
 
 Colonies circular, irregular : deep 
 
 colonies are brownish ; superficial 
 colonies thin and translu 
 
 Inoculated in the depth of gela- 
 tine the growth is scanty in the 
 track of the needle. 
 
 In the depth of agar an opaque 
 branching growth occurs m th. 
 track of the needle, and 
 milk-white growth on the surface. 
 
 In broth they cause opalescence. 
 
 They were isolated from the 
 liver in fatal cases of yellow fever. 
 
 Bacillus flavescens (p<>hl). 
 Rods 2-1 to 2-2 p in length, * p. 
 in width. Slightly motile. 
 
 Colonies yellow and granular. 
 
 Inoculated in the depth of gela- 
 tine the bacilli produce a filament 
 in the track of the needle, and a 
 growth spreads over the free sur- 
 face. 
 
 On the surface of agar the growth 
 is composed of isolated yellow 
 colonies. 
 
 On potato they grow rapidly. 
 forming a shiny yellow layer. 
 
 They occur in marsh water. 
 
 Bacillus flavocoriaceus (Ada 
 metz). Minute rods occurring in 
 zooglcea. 
 
 Colonies circular, sulphur-yellow. 
 Under a low power they show a 
 brownish-yellow nucleus and yellow 
 periphery. 
 
 Inoculated in the depth of gela- 
 tine the growth is granular in the 
 track of the needle and on the free 
 surface. 
 
 They occur in water. 
 
 Bacillus fluorescens aureus 
 i/immermann). Short md-. M ^ 
 in length, '74 /i in width. 
 
 Singly, in airs and in 
 ' 
 
 .- . 
 
 Coloniescircular, granular, yell \*. 
 
 Inoculated in the depth >f gela- 
 tine they form a filament in the 
 track of the needle and a yellow 
 patch on the surface. 
 
 On agar the growth is golden - 
 yellow, and the same on potato. 
 
 They occur in wat 
 
 Bacillus fluorescens lique- 
 faciens ( King- rods with 
 
 rounded (lid- 
 
 Colonies on plates develop an 
 iridescence around them. 
 
DESCRIPTION OF SPECIES. 
 
 In the depth of gelatine a white 
 filament forms in the track of the 
 needle, with liquefaction in the 
 upper part, and an iridescent sheen 
 is produced in the jelly. 
 
 On potato they develop a brown- 
 ish layer. 
 
 They occur in water and in 
 putrid infusions. 
 
 Bacillus fluorescens lique- 
 faciens minutissimus (Unna and 
 Tommasoli). Rods I'd to 2 p. in 
 length, - H p. in width. 
 
 Colonies circular, with brownish 
 nucleus and yellowish marginal 
 zone. 
 
 In the depth of gelatine lique- 
 faction forms in the track of the 
 needle. The liquid is turbid and 
 yellowish, and a white sediment 
 forms at the bottom and a fluores- 
 cent scum on the surface. 
 
 On agar and potato the growth is 
 brownish. 
 
 They were isolated from eczema- 
 tous skin. 
 
 Bacillus fluorescens longus 
 
 -(Zimmermann). Rods varying in 
 length, some curved, T45 to 1'65 p, 
 in length, '83 p. in width, and wavy 
 filaments 14 p. in length. Motile. 
 
 Colonies circular, well-defined, 
 yellowish, with broad twisted 
 markings. 
 
 On the surface of gelatine they 
 produce a layer with a bluish-green 
 fluorescent colour. 
 
 On agar a thin film forms, and 
 the jelly is coloured greenish- 
 yellow. 
 
 On potato the growth is slimy 
 and yellowish. 
 
 They occur in water. 
 
 Bacillus fluorescens nivalis 
 (Schmolck). Rods and chains. 
 Motile. 
 
 Colonies circular, surrounded by 
 liquid fluorescent jelly. 
 
 Inoculated in the depth of gela- 
 tine they produce liquefaction in 
 the track of the needle, and the 
 jelly is coloured green. 
 
 On agar the growth is white and 
 the jelly green. 
 
 On potato the growth is brownish. 
 
 They are probably identical with 
 Bacillus fluorescens liquefaciens. 
 
 They were isolated from snow. 
 
 Bacillus fluorescens non-lique- 
 I faciens (Eisenberg). Short deli- 
 I cate rods. Non-motile. 
 
 Colonies have the appearance 
 of fern-leaves, and are opales- 
 cent. 
 
 Inoculated in the depth of gela- 
 tine the growth is very slight in 
 the track of the needle, and on the 
 surface filmy and fluorescent. 
 
 On agar they form a greenish 
 layer. 
 
 They produce a brownish layer 
 on potato, and bluish-grey dis- 
 coloration. 
 
 They occur in water. 
 
 Bacillus fluorescens putidus 
 (Fliigge). Short rods with rounded 
 ends. Motile ; spore-formation not 
 known. 
 
 They form small dark colonies 
 with a greenish sheen, which have 
 a penetrating odour. 
 
 Inoculated in the depth of gela- 
 tine they produce a pale-grey 
 growth, and after three days colour 
 the medium with a greenish tinge 
 spreading down from above. 
 
 On potato they rapidly develop 
 a brownish layer. 
 
 They occur on decomposing sub- 
 stances, producing a greenish 
 coloration. 
 
 According to Lehmann and Neu- 
 mann the Bacillus fluorescens albus 
 and Bacillus fluorescens longus of 
 Zimmermann and Bacillus fluores- 
 cens non-liquefaciens are merely 
 varieties of Bacillus fluorescens 
 putidus ; and further, cultures of 
 this bacillus on agar and on potato 
 and in milk and in broth cannot 
 be distinguished from those of 
 Bacillus fluorescens liquefaciens. 
 
 Bacillus fluorescens tenuis 
 (Zimmermann). Rods 1 to 1-85 p. 
 in length, -8 p. in width. Singly, 
 in masses, and filaments. Motile. 
 
 Colonies irregular. 
 
 Inoculated in the depth of gela- 
 tine a delicate filament forms in 
 the track of the needle, and on 
 the surface a greyish-white growth 
 spreads and colours the gelatine 
 yellow. 
 
 On agar the growth is shining 
 

 DESCRIPTION OK SI'] 
 
 
 and greenish, and on potato yellow- 
 ish. 
 
 They occur in water. 
 Bacillus foetidus (/>'</, /;,//,* 
 t'.tt'ulnni. Thin). Cocci, short rods, 
 long rods, and leptothrix. The 
 cocci. 1-25 to 1 -4 p. in diam., occur 
 singly or in pairs. Spore-formation 
 present in the rods. 
 
 They were isolated from the 
 exudation in a case of profuse 
 sweating of the feet, and the odour 
 was noticeable in the cultivation 
 (//Wr 1'xn-Uliis saprogene*}. 
 
 Bacillus foetidus ozaenae (Ha- 
 j e k). Short rods, singly, in pairs, 
 and in short chains. Motile. 
 
 Colonies irregular and liquefying. 
 In the depth of gelatine lique- 
 faction occurs along the track of 
 the needle. 
 
 On agar the growth is moist and 
 shiny, and on potato yellowish- 
 brown. 
 
 Cultures emit a disagreeable 
 odour. 
 
 They produce a fatal result m 
 mice and local inflammation in 
 rabbits. 
 
 They were isolated from cases 
 of ozsena. 
 
 Bacillus fulvus (Zimmermann). 
 Rods -88 to 1-3 p in length, 
 77 p in width. Singly, in pairs 
 and in chains. 
 
 Colonies vary in form ; granular, 
 yellowish-grey. 
 
 In the depth of gelatine there is 
 a scanty faintly yellow growth in 
 the track of the needle, and a 
 hemispherical yellow mass on the 
 free surface. 
 
 On agar and potato the growth 
 is yellow and shining. 
 They occur in water. 
 Bacillus fuscus (Zimmermann). 
 Rods -63 p in width, varying m 
 length ; sometimes bent and irregu- 
 lar in form. 
 
 Colonies irregular, granular, and 
 greyish-yellow. 
 
 In the depth of gelatine a 
 hemispherical mass appears on the 
 free surface, which later spreads 
 and forms a yellow wrinkled layer. 
 On agar and potato the layer u 
 similarly coloured. 
 
 They occur in water. 
 
 Bacillus fuscus limbatus 
 (Scheibenzuber). RmK and fila- 
 ments. 
 
 Colonies with brown nucleus and 
 light periphery. 
 
 Inoculated in the depth of gela- 
 tine the growth in the track of the 
 needle is branching and the jelly 
 coloured brown. 
 
 On agar and potato the growth 
 is dark brown. 
 
 They occur in rotten eggs. 
 
 Bacillus gallinarum 
 ofFou-i /;///./;/;.<) <- 
 
 Bacillus gasoformans 
 berg). Rods. 
 
 The colonies are granular, and 
 liquefy the gelatine. 
 
 The bacilli inoculated in the 
 depth of gelatine rapidly produce 
 liquefaction in the track of the 
 needle, and formation of gas 
 bubbles. 
 
 They occur in water. 
 
 Bacillus gingivae pyogenes: 
 vide Bacti rin in <i<m.i"' 
 
 Bacillus glaucus (Maschek). 
 Rods. 
 
 The colonies are well-defined, 
 greyish in colour, and after a time 
 liquefy the gelatine. 
 
 The bacilli inoculated in the 
 depth of gelatine produce a rapid 
 growth in the track of the needle 
 and on the surface, followed by 
 liquefaction and a sediment at the 
 bottom of the liquid. 
 
 On potato and agar they form a 
 greyish layer. 
 
 They occur in water. 
 Bacillus gliscrogenus^ 
 Rods -57 to M4 p in length, '41 p 
 in width. 
 
 Colonies spherical, granular. 
 The bacilli inonilated in the 
 depth of gelatine give rise to a 
 growth in the track of the needle 
 i composed of closely packed disc- 
 i shaped colonies. 
 
 On agar they produce an opal 
 ; cent film. 
 
 On potato they form a vi* 
 yellowish growth 
 They were isolated 
 
 l " Bacillus gracilis 
 
514 
 
 DESCRIPTION OF SPECIES. 
 
 mann). Rods sometimes curved, 
 2'4 to 3'6/Lt in length, '77 p, in width, 
 and filaments. 
 
 The colonies are greyish or 
 yellowish-grey, with concentric 
 markings. 
 
 The bacilli inoculated in the 
 depth of gelatine produce isolated 
 colonies in the track of the needle, 
 and a translucent film on the free 
 surface ; followed after some time 
 by slight liquefaction. 
 
 On agar they produce a bluish- 
 white layer. 
 
 There is scarcely any growth on 
 potato. 
 
 They occur in water. 
 
 Bacillus gracilis anaerobies- 
 cens (Vaughan). Rods. 
 
 The colonies are brownish. 
 
 The bacilli inoculated in the 
 depth of gelatine produce a copious 
 growth in the track of the needle, 
 and gas bubbles, and a film on the 
 surface. 
 
 On agar they produce a thin 
 layer, and on potato a yellowish- 
 white mass. 
 
 They occur in water. 
 
 Bacillus granulosus (Russell). 
 Rods singly, in pairs and in 
 masses, and long filaments. Spore- 
 formation present. 
 
 The colonies have concentric 
 linear markings. 
 
 The bacilli inoculated in the 
 depth of gelatine cause slow lique- 
 faction spreading downwards in 
 the track of the needle. 
 
 On the surface of agar they form 
 more or less isolated whitish or 
 yellowish colonies. 
 
 In broth they produce turbidity, 
 and on potato a thick shining layer, 
 which is at first white and later 
 brownish. 
 
 They were isolated from deep-sea 
 dredgings. 
 
 Bacillus graveolens (Bordoni- 
 Uffreduzzi). Very short rods, - 8 p. 
 in length. 
 
 The colonies are greyish-white, 
 and liquefy the gelatine. 
 
 The bacilli inoculated in the 
 depth of gelatine produce rapid 
 liquefaction in the track of the 
 needle. They colour the gelatine 
 
 greenish-yellow, and produce a foul 
 odour. 
 
 On potato the culture is brown- 
 ish. 
 
 They were isolated from skin 
 from between the toes. 
 
 Bacillus guttatus (Zimmer- 
 mann). Rods 1 to 1'13 p. in length ; 
 93 in width ; singly, in pairs, and 
 in chains. 
 
 Colonies bluish-grey ; granular. 
 
 The bacilli inoculated in the 
 depth of gelatine develop colonies 
 in the track of the needle, and a 
 greyish opalescent film on the 
 surface. 
 
 On agar the growth is greyish- 
 white. On potato it is yellowish 
 and slimy. 
 
 They occur in water. 
 
 Bacillus halophilus (Russell). 
 Rods 1-5 to 3'5 p. in length, -7 p. in 
 width ; singly and in pairs ; and 
 toruloid involution forms. Motile. 
 
 The colonies liquefy gelatine and 
 become frothy from abundant for- 
 mation of gas. 
 
 The bacilli inoculated in the 
 depth of gelatine grow in the track 
 of the needle, and excavate the 
 jelly at the upper part. 
 
 They were isolated from deep- 
 sea dredgings. 
 
 Bacillus Hansenii (Rasmussen). 
 Rods 2-8 to 6 p. long, -6 to -8 p 
 wide. 
 
 Cultivated on sterilised potato, 
 they form in four days a chrome- 
 yellow layer with an agreeable 
 fruit-like smell. Two or three days 
 later the growth dries, and changes 
 to an orange-yellow colour ; later 
 it becomes yellowish or brown, and 
 at the same time spores are formed 
 1-7 p, long, 1-1 p. wide. The colour- 
 ing matter is insoluble in most 
 reagents. 
 
 The bacilli occur as a yellow or 
 whitish skin on nourishing solu- 
 tions, malt infusion, broth, and 
 wine, which have been kept at 
 31 to 33 C. 
 
 Bacillus Havaniensis lique- 
 faciens. Rods "8 p. in width, l - 2 
 to 5 p. in length, singly, and in 
 pairs ; and filaments. Motile. 
 
 The colonies are milky, irregular 
 
DESCRIPTION 01 SPECIES. 
 
 
 in outline, and liquefy the gela- 
 tine. 
 
 The bacilli inoculated in the 
 depth of gelatine cause liquefaction 
 in the track of the needle. 
 
 On agar they form a pale-brown 
 layer. 
 
 They do not grow on potato. 
 
 They were isolated from the skin. 
 
 Bacillus helvolus (Zimmer- 
 mann). Rods 1'5 to 4-5 ^i in length, 
 5 p. in width : in pairs, and in 
 chains. 
 
 Colonies are pale yellow. 
 
 The bacilli form a yellow growth 
 on the surface of gelatine, and pro- 
 duce slow liquefaction. 
 
 On agar the growth is yellow. 
 
 They occur in water. 
 
 Bacillus heminecrobiophilus 
 (Arloing). Rods highly polymor- 
 phic, and filaments 1 to 20 p in 
 length. Slightly motile. 
 
 On the surface of obliquely 
 solidified gelatine they form a 
 yellowish layer. 
 
 On potato the growth is yellowish- 
 white. 
 
 They produce oedema when sub- 
 cutaneously inoculated in the 
 vicinity of wounds. 
 
 They were isolated from a caseous 
 lymphatic gland in a guinea-pig. 
 
 Bacillus hepaticus fortuitus 
 (Sternberg). Rods resembling 
 Bacillus coli communis. 
 
 The colonies, marked with radia- 
 ting striae, are dark brown in colour. 
 
 The bacilli inoculated in the 
 depth of gelatine produce a very 
 slight growth at the upper part of 
 the track of the needle, and a hemi- 
 spherical mass on the free surface. 
 
 On potato they form a creamy 
 white growth. 
 
 They were isolated from the 
 liver in a fatal case of yellow 
 fever. 
 
 Bacillus Hessii (Guillebeau). 
 Rods 3 to 5 n in length, 1'2 p. in 
 width, cocci-forms and filament <. 
 
 The colonies are filamentou-. 
 and liquefy gelatine. 
 
 The bacilli inoculated in the 
 depth of gelatine produce lique- 
 faction, and the liquid jelly is made 
 extremely viscous. 
 
 On potato the growth i*l,m W M M,. 
 
 They coagulate milk. 
 
 Tlu-y wen iM.lat,.,! from milk. 
 
 Bacillus hyacinthi septicus 
 (Heinz). Rods 4 to C> ^ in length, 
 1 p, in width. 
 
 The colonies are transparent and 
 bluish-white. 
 
 The bacilli inoculated in the 
 depth of gelatine produce a fila- 
 ment in the track of the needle 
 and a layer on the surface. 
 
 On potato they produce a slimy. 
 dirty-yellow layer. 
 
 They were isolated from diseased 
 hyacinths. 
 
 Bacillus hyalinus (Jordan). 
 Rods 3'G to 4 |ti in length. 1 ." p in 
 width, and chains. 
 
 The colonies are surrounded by 
 radiating filaments. 
 
 The bacilli inoculated in the 
 depth of gelatine produce liquefac- 
 tion in the track of the nee<l! 
 scum on the surface and a deposit 
 at the bottom. 
 
 On agar they produce a dry, grey 
 growth. 
 
 On potato the growth is greyish- 
 white and tuberculated. 
 
 They coagulate milk. 
 
 In broth they produce turbidity 
 and a pellicle on the surface ; and 
 they are powerful nitrifying 
 agents. 
 
 They occur in \\ 
 
 Bacillus hydrophilus fuscus 
 ( Sanarelli). Rods 1 to tt /* in 
 length, and filaments !."> to I'M p. in 
 length. 
 
 They rapidly produce a funm-1- 
 shaped area of liqm-f:i.-tion when 
 grown in gelatine, followed \>\ 
 complete liquefaction and the 
 formation of a white flocculent 
 deposit 
 
 Inoculated in glycerine agar 
 grow rapidly and product- gas 
 bubbles. On potato th y produce 
 a straw-coloim-l l.-m-r which be- 
 comes distinctly yellow and later 
 brown. 
 
 They are pathogenic in cold- 
 blooded aninnl- 
 blooded animals. (> 
 cnmb in twelve h'. 
 is enlarged and the bn illi arc found 
 
516 
 
 DESCRIPTION OF SPECIES. 
 
 in great numbers in the blood and 
 internal organs. 
 
 They were isolated from the 
 lymph of diseased frogs. 
 
 Bacillus ianthinus (Bacterium 
 ianthinum Zopf, Bacillus violacems). 
 Slender rods, about four times 
 their width in length, with rounded 
 ends. They also form threads, and 
 are actively motile. Spore-forma- 
 tion present in the rods. 
 
 The colonies occur as circum- 
 scribed liquefied areas, in the centre 
 of which is a collection of the 
 coloured growth. 
 
 The bacilli inoculated in the 
 depth of gelatine produce a funnel- 
 shaped liquefaction, and a granular- 
 looking violet mass subsides to the 
 bottom. 
 
 On agar-agar and potato a 
 beautiful violet growth rapidly 
 develops. The colouring matter is 
 soluble in alcohol. 
 
 They were observed on pieces of 
 pigs' bladder floating on the surface 
 of water rich in bacteria. They 
 occurred only on the surface of the 
 bladder exposed to the air, and never 
 on the part under water. They occa- 
 sionally occur in common tap water. 
 
 Bacillus implexus (Zimmer- 
 mann). Rods 2*5 p, in length, 
 1-15 p in width. Non - motile. 
 Spore-formation present. 
 
 Colonies white, granular, develop - 
 ing in three days into masses of 
 interlacing white filaments. 
 
 Inoculated in the depth of gela- 
 tine, a growth develops in the track 
 of the needle and fine filaments 
 penetrate the gelatine. The jelly is 
 liquefied, and a pellicle forms on 
 the surface, and there is a flocculent 
 deposit. 
 
 On agar the growth is white, and 
 on potato yellowish-white. 
 
 They occur in water. 
 
 Bacillus in acne contagiosa in 
 horses (Dieckerhoff and Grawitz). 
 Short rods '2 /u, in diam. 
 
 Inoculated in the depth of 
 nutrient gelatine they form a scanty 
 growth in the track of the needle 
 and a white patch on the free sur- 
 face. They thrive best on blood 
 serum and on agar. 
 
 The bacilli inoculated on the 
 surface of the skin of horses, calves 
 and other animals are said to pro- 
 duce acne pustules. Inoculated 
 subcutaneously in guinea-pigs they 
 produce a fatal result in twenty- 
 four hours. 
 
 They were isolated from pus 
 in cases of acne contaf/iosa in horses. 
 
 Bacillus in cancer (Koubasoff). 
 Rods ; spore-formation present. 
 
 Inoculated in the depth of gela- 
 tine an irregular filament develops 
 in the track of the needle and a 
 transparent growth with central 
 depression on the surface. 
 
 They are said to be pathogenic 
 in small animals, and to produce 
 nodules and ulcers of the mucous 
 membrane of the stomach. 
 
 They were isolated from a case 
 of cancer of the stomach. 
 
 Bacillus in cholera in ducks 
 (Cornil and Toupet), p. 230. 
 
 Bacillus in choleraic diarrhoea 
 (Bovet). Rods 2 to 4 ^ in length, 
 1 to To ju, in width, singly and in 
 pairs, and filaments. 
 
 In the depth of gelatine a filament 
 forms in the track of the needle and 
 a greyish transparent layer on the 
 surface. 
 
 On agar a greyish film is formed. 
 
 On potato the growth is yellowish 
 and abundant. 
 
 Intra-peritoneal injections in 
 guinea-pigs cause peritonitis and 
 death. 
 
 They were isolated from a case 
 of choleraic diarrhoea. 
 
 Bacillus in diphtheritic disease 
 of calves (Bacillus vituloruiii 
 Lofner). Rods about five or six 
 times as long as wide, mostly united 
 in long threads. 
 
 A piece of membrane from a diph- 
 theritic disease in a calf, placed on 
 blood serum developed a white layer 
 composed of the bacteria. Succes- 
 sive generations were not obtainable. 
 
 Mice inoculated directly from the 
 calf died of a characteristic illness, 
 and the same long bacteria were 
 again found in the inoculated 
 animals accompanying widespread 
 infiltration, starting from the point 
 of inoculation. Inoculation of 
 
DESCRIPTION <! si-i:< [ES, 
 
 .'.IT 
 
 guinea-pigs and rabbits gave doubt- 
 ful results. They were found in 
 the deeper stratum of pseudo- 
 diphtheritic patches in calves. 
 
 Bacillus in disease of bees (p. 
 471). 
 
 Bacillus in erythema nodosum 
 (Demme). Rods -2-2 to 2'5 /x in 
 length, ; t. '7 /z in width. They 
 can be cultivated at 37C. 
 
 Colonies on agar are white with 
 radiating lines. 
 
 The bacilli inoculated in the depth 
 of agar grow in the track of the 
 needle, and produce peculiar off- 
 shoots in the surrounding jelly. 
 
 They are said to produce an 
 eruption resembling erythema 
 nodosum when inoculated subcu- 
 taneously in guinea-pigs. 
 
 They were obtained from the 
 eruption and the blood in cases of 
 erythema nodosum. 
 
 Bacillus in fowl enteritis 
 (Klein), p. '2:-'.o. 
 
 Bacillus in gangrene (Tricomi). 
 Rods 3 \L in length, 1 p. in width, 
 singly and in pairs. 
 
 ( olonies circular, granular, dirty- 
 yellow. 
 
 In the depth of gelatine they 
 produce a filament composed of 
 closely aggregated colonies, and at 
 the upper part conical liquefaction 
 of the jelly, beneath a cup- shaped 
 excavation. 
 
 On agar and potato the growth 
 
 is white. 
 
 Injectedsubcutaneouslyin rabbits 
 
 and guinea-pigs they produce gan- 
 grene and death in a few days. 
 
 They were isolated from a case 
 of senile gangrene. 
 
 Bacillus in grouse disease 
 (Klein), p. L': 1 .". 
 
 Bacillus in hog cholera (p. 
 351). 
 
 Bacillus in infantile diarrhoea 
 (Booker). Rods morphologically 
 identical with Bacillus coli com- 
 munis There are seven varieties 
 of this bacillus. They were iso- 
 lated from cases of infantile OUT- 
 rhoea. 
 
 Bacillus in infantile diarrhoea 
 (Lesage). Rods '2'4 /i in length, 
 75 a in width, and filaments. 
 
 Colonies irregular ia contour. 
 colouring the gelatine green. 
 
 On the surface of agar they form 
 a greenish growth, and the gelatine 
 is coloured green. 
 
 Injected intravenously in a rabbit 
 they produced diarrhoea. 
 
 They are said to be identical with 
 Barilla* tluorescens li<jurt':i 
 
 Bacillus in intestinal diph 
 theria in rabbits (Kit 
 Rods 3 to 4 ft in length, 1 to 1 4 ^ 
 in width ; singly, in pairs, and in 
 filaments. 
 
 Colonies greyish ; granular. 
 They produce in gelatine a deli- 
 cate growth in the track of the 
 needle. They are pathogenic. 
 They were isolated from 
 intestine of rabbits suffering t 
 a diphtheritic inflammation of the 
 mucous membrane. 
 
 Bacillus in jequirity infusion 
 (see H(n-;iln. qfSati 
 Bacillus in measles (p. 
 Bacillus in noma (Schinm.. 1 
 busch). Rods singly, in pairs, and 
 filament*. 
 
 Colonies circular, greyish- white, 
 granular, with irregular margins. 
 
 In the depth of gelatine they 
 produce a granular filament anda 
 patch on the sur; 
 
 On agar and potato the growth 
 is greyish-white. 
 
 They are pyogenic in rabbits. 
 They were inoculated from a 
 case of noma. 
 
 Bacillus in ophthalmia ( P 1 - 
 Bacillus in potato rot. 
 2 '> to 4 /i in length. '7 to * M in 
 ! width ; singly, in ch > fila- 
 
 ments. Spore-formation present. 
 
 The bacilli inoculated in the 
 depth of gelatine produce a funnel 
 shaped area of liquefact i 
 
 On agar the growth is composed 
 of greyish-white slimy colonies. 
 
 They were isolated from disease* 
 pot:i' 
 
 Bacillus in purpura hamor- 
 rhaeica '' (| Qto*)- 
 
 _K,d-7.. to i-SpfalHftt, 
 4 M in width, singh 
 in masses. 
 
 Colonies have a greyi*h-yel 
 nucleus and a marginal zone of 1 
 
518 
 
 DESCRIPTION OF SPECIES. 
 
 filaments both in gelatine and agar. 
 Cultures produce a disagreeable 
 odour. 
 
 Subcutaneous in j ections in guinea- 
 pigs and rabbits produce local oedema 
 and death, with haemorrhages in the 
 internal organs. 
 
 They were isolated from the blood 
 in fatal cases of purpura in children. 
 
 Bacillus in putrid bronchitis 
 (Lumnitzer). Rods 1*5 to 2 /z in 
 length, slightly curved. They can 
 be cultivated at 37 C. The colonies 
 on agar are greyish- white. 
 
 The bacilli inoculated on blood 
 serum produce colonies which co- 
 alesce and form a greyish-white 
 film. Cultures have a disagreeable 
 odour. 
 
 Injected into the lungs of rabbits 
 they produce purulent infi amma.tion . 
 
 They were isolated from the 
 sputum in cases of putrid bronchitis. 
 
 Bacillus in "red-cod" (Dantec). 
 Rods similar to Bacillus tetani, 
 with terminal spore-formation. 
 
 Colonies are circular ; reddish. 
 
 On the surface of obliquely 
 solidified gelatine they form a red 
 growth in the track of the needle 
 slowly followed by liquefaction. 
 
 Cultivated on dried cod they 
 produce a red colour. 
 
 They were isolated from red-cod. 
 
 Bacillus in rhinos cleroma (p. 
 411). 
 
 Bacillus in saliva (Fiocca). 
 Very short rods -2 to -33 p in width. 
 
 Colonies circular, granular, and 
 yellowish. 
 
 On the surface of obliquely 
 solidified gelatine they form a 
 growth composed of transparent 
 droplets. 
 
 - On potato they form a transparent 
 film. 
 
 In broth flocculi appear. 
 
 They are pathogenic in rabbits 
 and other small animals, and they 
 are probably a variety of the bacillus 
 of haemorrhagic septicaemia. 
 
 They were isolated from saliva of 
 cats and dogs. 
 
 Bacillus in whooping cough 
 (Afanassiew). Rods (] to 2*2 /* in 
 length, singly, in pairs, and short 
 chains. 
 
 Colonies granular, brownish. 
 
 Inoculated in the depth of gela- 
 tine there is a scanty growth in the 
 track of the needle and a greyish 
 growth on the free surface. 
 
 On agar the growth is greyish. 
 
 On potato the growth is shining 
 and yellowish or brownish. 
 
 They are said to produce symp- 
 toms in rabbits and dogs compar- 
 able to those of whooping cough. 
 
 They were isolated from the 
 throat in cases of whooping cough. 
 
 Bacillus incanus (Pohl). Rods 
 1'2 /j. in length, - 8 p. in width. 
 
 They produce rapid liquefaction 
 in the track of the needle when 
 inoculated in the depth of gelatine. 
 
 On agar a thick white growth 
 develops. 
 
 On potato a whitish growth 
 spreads over the surface. 
 
 They were isolated from the 
 water of marshes. 
 
 Bacillus indicus (Koch). Very 
 short rods with rounded ends. 
 
 FIG. 204. BACILLUS INDICUS COLONIES 
 IN NUTRIENT AGAR, x 60. 
 
 The colonies have a scarlet tint. 
 They are round, ovoid, or spindle 
 shaped, and have granular margins. 
 
 In the track of the needle beneath 
 the surface no pigment is formed. 
 
 Cultivated in nutrient gelatine 
 they liquefy it and colour it crimson, 
 and the growth of a darker crimson 
 hue subsides to the bottom of the 
 tube. 
 
 On the surface of nutrient agar- 
 agar the appearances are very 
 characteristic. In a pure cultiva- 
 tion a brilliant vermilion-coloured 
 reticulated pellicle develops on the 
 surface. (Plate II. Fig. 3.) 
 

 
 They form a vermilion layer on 
 potato. 
 
 They were isolated by Koch in 
 India from the intestinal contents 
 of an ape. 
 
 Bacillus indigogen.es (Alvarez). 
 Rods 3 p. in length and 1-5 /* in 
 width, singly, and in chains : cap>u- 
 lated. 
 
 On agar they produce a yellow Uh- 
 white layer, and are said to develop 
 an indigo-blue colour in infusions 
 of leaves of the indigo plant. 
 
 Intravenous injections in guinea- 
 pigs are said to produce death in a 
 few hours. 
 
 They were isolated from the 
 leaves of the indigo plant. 
 
 Bacillus indigonaceus (Clas- 
 sen). Rods 1'6 to 3 n long, '8 to 
 9 /i wide ; non-motile. 
 
 They form a sky-blue layer on 
 the surface of gelatine. 
 
 On potato the growth is dark- 
 blue, and later has a metallic lustre. 
 
 Bacillus indigoferus, which was 
 found in water at Kiel, is only 
 to be distinguished by its motility. 
 
 Bacillus inflatus (A. Koch). 
 Rods 4'6 to 5 - 5 //. in length, '6 to 
 !* \i in width, and filaments. 
 
 The colonies send out delicate 
 processes. 
 
 The bacilli inoculated in the 
 depth of gelatine send out line 
 filaments in the track of the needle 
 followed by slow liquefaction. 
 
 On agar they form a shining 
 brownish layer. 
 
 In broth a pellicle forms on the 
 surface. 
 
 They occur in the air. 
 
 Bacillus inunctus (Pohl). 
 Rods 3'5 p. in length, '8 to -9 /* in 
 width. 
 
 Inoculated in the depth of gela- 
 tine they grow both in the track of 
 the needle and on the surface ; 
 liquefaction follows in time. 
 
 On agar they form a whitish 
 growth. 
 
 They were isolated from the 
 water of marshes. 
 
 Bacillus invisibilis (Vaughan). 
 Rods ; motile. 
 
 The colonies are irregular and 
 yellowish. 
 
 lno-nl;ited 111 tli.- .1. -;,tl, of 
 
 tine they grow both in tl, 
 the needle and on th. 
 I On agar they form a whit. 
 growth. 
 
 <>n potato they develop an 
 
 visilile l;i\ 
 
 They occur in wai 
 
 Bacillus iridescens 
 Rods from ii . 
 and threads. 8] 
 sent : slightly n,. 
 
 The colon; 
 
 appearance recall in. 
 convolution : tin. 
 
 Inoculated in tin- 
 tine then 
 
 point of puncture. anl 
 like growth 
 track. 
 
 On agar the\ 
 even, moi- 
 cent growth, with a pitted -in: 
 
 Blood s-nim i> li-pi 
 
 On potato t! M.'k. 
 
 dark yellow growth like h< 
 
 Broth : 
 there 
 
 onii.i in v. 
 
 Bacillus lactis aerogenes 
 
 cherich).- ; r:<l thick. 
 
 !. and 1 to 
 with rounded end- : n-iia! 
 
 heaps. Non-m 
 tion not <! 
 
 at :;; 
 
 Colonies on ti 
 moist, >hinini: and p 
 
 UfloW til. 
 
 yellowish iin.-l' 
 
 Inoculat, -rela- 
 
 tine tin- r..d- f..nn 
 nail-shaj.ed gro* 
 cnltnn- i- ''"in; 
 :mil bnbb:- 
 colonie- n -luce 
 
 On blood sei nin there is a raised, 
 
 lU'.i-t, -hllllD'J. "(lit- _'l" 
 
 In milk ' grape ugar 
 
 M.liitinn- th-y prodooi | 
 
 Injc.-t.-d siil- 
 death in : 
 and the bacilli an 
 blood and ii t i rial organs. 
 
520 
 
 DESCRIPTION OF SPECIES. 
 
 They were found in the intestinal 
 tract of animals fed with milk and 
 of infants at the breast. 
 
 Bacillus lactis albus (Loffler). 
 Rods, 3*4 JJL in length, '90 p. in 
 width, and filaments. Spore-forma- 
 tion present. 
 
 Inoculated in the depth of gela- 
 tine they slowly liquefy the upper 
 part, and a white scum forms on the 
 surface. 
 
 On agar they form a white layer. 
 
 On potato the growth is dry and 
 white. They coagulate milk. 
 
 They occur in milk. 
 
 Bacillus lactis erythrogenes 
 (Hueppe).- Short rods, 1 to 1*4 /* 
 in length and - 3 to "5 p in width, 
 and filaments. Colonies small and 
 circular ; greyish-white ; later yel- 
 low and surrounded by liquefied 
 gelatine with a pink tinge. 
 
 In the depth of gelatine the 
 growth in the track of the needle 
 is scanty, but on the surface a 
 whitish patch forms which after- 
 wards turns yellow, and the gela- 
 tine is coloured pink. Later lique- 
 faction sets in, and the liquefied 
 gelatine is turbid and pink. 
 
 On agar a shining yellow layer 
 develops, and the same on potato. 
 
 In broth the bacilli produce tur- 
 bidity, and they coagulate milk. 
 
 They occur in " red milk." 
 
 Bacillus lactis pituitosi (Lof- 
 er). Rods slightly bent. 
 
 Colonies circular, greyish-white. 
 
 On agar and potato they pro- 
 duce a greyish-white layer. 
 
 They render milk viscid. 
 
 They occur in milk. 
 
 Bacillus later iceus (Adametz 
 and Eisenberg). Rods and fila- 
 ments. 
 
 Colonies circular, granular, red- 
 dish-brown. 
 
 In the depth of gelatine there is 
 a scanty growth along the track 
 of the needle and a brick- red 
 growth on the surface. 
 
 On potato the growth is also 
 brick-red. 
 
 They occur in water. 
 
 Bacillus leporis lethalis (Gibier 
 and Sternberg). Rods 1 to 3 /* in 
 length, -5 p. in width. 
 
 Colonies transparent and with 
 the appearance of broken glass. 
 
 In the depth of gelatine there 
 is a growth along the track of the 
 needle with a conical area of lique- 
 faction at the upper part, and a 
 white sediment. 
 
 On agar they form a trans- 
 lucent film. They liquefy blood 
 serum. 
 
 On potato the growth is pale- 
 yellow. 
 
 Cultures injected into the peri- 
 toneal cavity of rabbits are toxic. 
 
 They were isolated from the in- 
 testinal contents in cases of yellow 
 fever. 
 
 Bacillus leprsB (p. 407). 
 
 Bacillus leptosporus (L. Klein). 
 Rods resembling hay-bacilli, 
 singly, in chains and long twisted 
 filaments. 
 
 The spore-membrane is said to 
 form part of the newly grown 
 bacillus, and the filaments are de- 
 scribed as possessing peculiar spas- 
 modic movements. 
 
 They were isolated from a con- 
 taminated culture. 
 
 Bacillus limbatus acidi lactici 
 (Marpmann). Rods short, thick ; 
 singly, in pairs ; capsulated. 
 
 Colonies white. 
 
 In the depth of gelatine they 
 develop slightly in the track of the 
 needle, and produce a white patch 
 on the free surface. 
 
 In milk they produce coagula- 
 tion and lactic acid. 
 
 They occur in milk. 
 
 Bacillus limosus (Russell). 
 Rods 3 to 4 p. in length, 1/25 /u, in 
 width ; singly, in pairs and chains ; 
 spore-formation present. 
 
 Colonies transparent, surrounded 
 by filamentous processes. 
 
 In the 'depth of gelatine pre- 
 pared with sea-water, liquefaction 
 occurs rapidly in the track of the 
 needle, and a deposit forms at 
 the bottom and a thin skin on the 
 surface. 
 
 On agar they form a white layer, 
 and in broth turbidity and a thick 
 scum. 
 
 On potato the growth is greyish- 
 white. 
 
DESCRIPTION OF SIM 
 
 521 
 
 They were isolated from deep- 
 sea dredgings. 
 
 Bacillus liodermos (Fliigge). 
 Small short rods with rounded 
 ends : actively motile. 
 
 Colonies with irregular outlines 
 float on liquefied gelatine in the 
 form of small white flakes. 
 
 Inoculated in gelatine a greyish 
 growth occurs along the track of the 
 needle, but the medium later be- 
 comes liquefied and a greyish-white 
 flocculent deposit settles at the 
 bottom. 
 
 On potato a smooth shining yel- 
 lowish-white layer spreads quickly 
 over the whole surface, and after 
 some days becomes opaque and 
 slightly wrinkled. 
 
 They occur on potato. 
 
 Bacillus liquefaciens (Eisen- 
 berg). Rods short and thick, with 
 rounded ends. Very motile. 
 
 Colonies round, with smooth 
 edges and slimy centres. Lique- 
 faction follows, and a putrefactive 
 odour is noticed. 
 
 In gelatine they make a funnel- 
 shaped whitish growth along the 
 track of the needle. 
 
 On potato the growth is pale 
 yellow. 
 
 They occur in water. 
 
 Bacillus liquefaciens commu- 
 nis (Sternberg). Rods 1 to 2 p in 
 length, and '7 /x in width : singly 
 and in pair-. 
 
 In the depth of gelatine they 
 produce rapid liquefaction in the 
 track of the needle. 
 
 On potato a wrinkled pinkish 
 layer is formed. 
 
 "They were isolated from the eva- 
 cuations of yellow-fever patients. 
 
 Bacillus " liquefaciens magnus 
 (Liideritz). Rods 3 to C> p in 
 length, -8 to 1-1 p. in width, and 
 filaments. They are anaerobic. 
 
 Colonies develop below the sur- 
 face of the gelatine, and liquefaction 
 extends upwards to the surface. 
 
 The bacilli inoculated in the 
 depth of gelatine cause liquefaction 
 in the lower part of the track of 
 the needle. 
 
 In the depth of agar the colonies 
 have delicate branches. 
 
 They liquefy blood-serui 
 produce a putrefactive odour. 
 
 They occur in earth. 
 
 Bacillus liquefaciens parvus 
 (Liideritz). -Rmls 2 to M in 
 length, -5 to 7 M in width. 
 filaments. They are anaei 
 
 Colonies are white, and li , 
 gelatine : but in agar they are 
 spherical or almond-shaped. 
 
 In the depth of gelatine isolated 
 colonies appear in th- track < 
 needle, and in the depth of agar 
 there is gas formation. 
 
 They occur in earth. 
 
 Bacillus liquidus (Frankland). 
 Rods short and flat with rou 
 ends, usually in pair-, the |. 
 of each pair varying from 1 '> n to 
 3-5 /i. They are v 
 size : highly motile 'rma- 
 
 tion not o 
 
 Colonies form cup-shaped excaTa- 
 tions, with almost clear, colon 
 contents. The edg- first 
 
 smooth and circular, but 
 become serrated and granular 
 soon coalesce. 
 
 A broad funnel-shaped 1 
 forms along the who! 
 needle, containing turbid liquid and 
 massesof flocculent matt i ia! Later 
 a thin pellicle form- on the surface, 
 which sinks if the tube is slink 
 
 On agar they grow quickly. B 
 ing a smooth shin in u r i 
 
 On potato a thick ured 
 
 growth a p|" 
 
 Broth is rendered turbid with an 
 abundant sediment, and after a few 
 pellicle fo: 
 
 They are common in unfiltered 
 water. 
 
 Bacillus litoralis (Russell) 
 Colonie- LManular. with regular 
 contour : slowlv liquefying. 
 
 In th* depth of gelat 
 develop a growth in the tra-l 
 the needle, and at the upper part 
 produce a cup-shaped on 
 with th.- culture. The g< 
 tinged with brown in tl 
 
 On agar they produce a gn-\ 
 white film, and in broth turl 
 
 Inoculated in t 
 
 gelatine the bacilli form a 
 funnel-shaped liquefaction along 
 
522 
 
 DESCRIPTION OF SPECIES. 
 
 the track of the needle, and the 
 whole of the gelatine gradually 
 becomes liquid, with a flocculent 
 deposit at the bottom and a greyish 
 wrinkled stain on the surface. 
 
 On potato a thick wrinkled whitish 
 skin forms, which rapidly grows 
 over the whole surface. On at- 
 tempting to raise this skin it will 
 be found to be attached to the 
 potato by a mucous substance 
 which may be drawn out in long 
 threads. According to Hueppe the 
 bacilli cannot form any ropy sub- 
 stances from sugar, but they have 
 an energetic diastatic action. They 
 coagulate the casein in milk in a 
 similar manner to rennet. 
 
 The bacilli are ubiquitous. 
 
 Bacillus lividus (Plagge and 
 Proskauer). Rods. 
 
 Colonies blue-black, liquefying. 
 
 In the depth of gelatine they 
 produce a colourless thread in the 
 track of the needle and a violet 
 layer on the surface followed by 
 gradual liquefaction. 
 
 On agar the growth is blue-black, 
 and on potato violet. 
 
 They were isolated from water. 
 
 They are probably identical with 
 Bacillus ianthinus, or merely a 
 variety. 
 
 Bacillus luteus (Flugge). 
 Short immotile rods. 
 
 Colonies irregular in form, appear 
 brownish under a low power, and 
 yellow to the naked eye. 
 
 In test-tube cultivations they 
 form a 3 7 ellow growth without 
 liquefying the gelatine. 
 
 They occur contaminating plate- 
 cultivations. 
 
 Bacillus maidis (Cuboni). - 
 Rods 2 to 3 /x in length, singly, 
 in pairs, and in chains ; spore- 
 formation present. 
 
 Colonies granular, with wrinkled 
 periphery ; later, liquefying. 
 
 In the depth of gelatine they 
 produce rapid liquefaction in the 
 track of the needle. 
 
 On agar a dry wrinkled white 
 film spreads over the surface. 
 
 On potato the growth is wrinkled, 
 and later yellowish-brown. They 
 liquefy blood serum. 
 
 They were isolated from human 
 evacuations and infusions of maize. 
 
 Bacillus mallei (p. 452). 
 
 Bacillus megatherium (De 
 Bary). Rods 2'5 ^ wide and four 
 to six times as long, with rounded 
 ends and slightly curved, and in 
 short irregular chains. Transverse 
 division occurs, each segment 
 attaining the length of the original 
 rod. In the fresh state they appear 
 non-articulated, but when treated 
 
 FIG. 205. BACILLUS MEGATHERIUM. 
 () A chain of rods x 250, the rest 
 x GOO. (b) Two active rods : d and /, 
 successive stages of germination; h 
 and I, successive stages of germina- 
 tion. (De Bary. ) 
 
 with a dehydrating agent they are 
 i seen to be composed of short seg- 
 i ments with granular contents. They 
 are motile. 
 
 Colonies are small and circular, 
 and the gelatine is liquefied. 
 
 In the depth of gelatine the 
 bacilli grow rapidly, forming a 
 funnel-shaped liquefaction in the 
 upper part. 
 
 On agar they form a whitish 
 layer on the surface, and the jelly 
 ; acquires a dark colour. 
 
 On potato yellowish-white cheesy 
 
 colonies are formed round the point 
 
 I of inoculation. In cultures there 
 
 ' is copious spore-formation. They 
 
 grow best at 20 C. 
 
 They were isolated originally 
 from boiled cabbage. 
 
DESCRIPTION OF SI'K< II :>. 
 
 
 FIG. 206. PURE-CULTURE OF BACILLUS 
 MEGATHERIUM IN GELATINE. 
 
 Bacillus membranaceus ame- 
 thystinus (Eisenberg). Short rods 
 with rounded ends from 1 to 1-4 p. 
 long, and '5 to *8 p. broad. They 
 are grouped together irregularly. 
 Some individual bacilli stain more 
 deeply at the ends than in the 
 middle. Xon-motile. They grow 
 only between 15 and 20 C. Spore- 
 formation uncertain. 
 
 The colonies gradually assume a 
 violet hue, and after liquefying the 
 gelatine float on the surface as violet 
 pellicles, resembling a membrane 
 stained with gentian violet. 
 
 Inoculated in the depth of 
 gelatine a yellowish-white growth 
 appears on the free surface, which 
 after ten days or more becomes 
 violet. Liquefaction takes place 
 gradually, and in about a month 
 a thick violet layer covers the gela- 
 tine which lies beneath the liquid 
 part. 
 
 On agar the growth, which at first 
 has a yellowish milky appearance, 
 becomes violet after eight or ten 
 days. In three or four weeks it 
 has become very much wrinkled, 
 and has a beautiful deep-violet 
 colour with a metallic lustre. The 
 
 jelly is not stain,-,!, an.l the growth 
 can be easily removed from itH 
 surface. 
 
 On potato they gn.w >l,,\vly, :m ,| 
 form a dirty yellow or olive-green 
 colour. 
 
 In broth they grow very slo\ 
 after some weeks a violet d. : 
 and pellicle are formed, and tin- 
 liquid between becom.-.l.uk l.n.un. 
 
 They were found in well water. 
 
 Bacillus meningitidis puru- 
 
 lentae (Neumann an.l Sehaflfer). 
 Rods 2 p. hi length. -0 to '1 p. in 
 width, and filaments. 
 
 Colonies granular, greyish. 
 
 In the depth of gelatine a gre\ 
 yellow filament develops, composed 
 of closely packed colonies, and on 
 the surface a greyish layer. 
 
 On potato the growth is moist 
 and white. 
 
 They are pyogenic in >mall ani- 
 mals and dogs. 
 
 They were isolated from a case 
 of purulent meningitis. 
 
 Bacillus mesentericus fuscus 
 (Fliigge). Rods small and short, 
 singly, in chains of two and i 
 Actively motile. Spore-formation 
 present. 
 
 Colonies are at first roundish and 
 rather white, with a >hai|> outline : 
 later delicate brownish-yellow pro- 
 cesses appear. Liquefaction occurs 
 rapidly. 
 
 Inoculated in the depth of gela- 
 tine a whitish growth forms along 
 the track of the needle, the upper 
 portion of which soon li<|ii< 
 greyish flakes float in the liquefied 
 portion. 
 
 On pott wish 
 
 growth appears on the first 
 but it soon becomes brown ami 
 wrinkled. It remains relatively 
 thin and -uprrii.-ial. ami quickly 
 spreads over the whole surface. 
 
 They are foun-l 1.1 hay <lnst. in 
 the air. on tin- -in face of i>otai 
 anI an- v.-ry wid.-Iy -ii-ti iluted. 
 
 Bacillus mesentericus ruber. 
 Sk-n.lcr rod in pairs, and 
 
 in filaments. 
 
 Colonies are rimii 
 Mi until they come t- th<- -nrface, 
 they produce a network and 
 
524 
 
 DESCRIPTION OF SPECIES. 
 
 liquefy the gelatine. The network 
 disappears and a little deposit occurs 
 at the bottom of the liquefied 
 area. 
 
 Inoculated in the depth of gela- 
 tine a whitish cloudy growth forms 
 along the needle track, liquefaction 
 sets in and extends until the gela- 
 tine is completely liquefied. 
 
 On potato a thin crinkled film is 
 formed, which is yellowish or red- 
 dish-yellow in colour. 
 
 They occur on potato. 
 
 Bacillus mesentericus vulga- 
 tus (Fliigge). Rods large and 
 thick, often forming pseudo-threads. 
 They have an oscillating movement. 
 Spore-formation present. 
 
 The colonies are bluish-white and 
 almost transparent, though the 
 centres become gradually opaque. 
 They sink in the liquefied gelatine, 
 and are granular with irregular 
 contour. 
 
 Bacillus multipediculus 
 (Fliigge). Eods long and slender. 
 Non-motile. 
 
 The colonies consist of a central 
 oval nucleus, from which numerous 
 tapering processes shoot out mostly 
 towards one pole. This form of 
 growth gives a curious resemblance 
 to an insect with feet and antennae. 
 
 Inoculated in the depth of gela- 
 tine a whitish line forms along the 
 track of the needle, from which 
 short processes grow out. 
 
 On potato a rather scanty dirty- 
 yellow growth forms, and the sur- 
 face of the potato becomes dis- 
 coloured around it. 
 
 They are often found as a con- 
 tamination on potato. 
 
 Bacillus muscoides (Liborius) 
 Rods 1 p. thick, sometimes form- 
 ing threads : slightly motile, and 
 with round or oval spores at one 
 end. They are anaerobic. 
 
 The colonies ramify and resemble 
 a delicate moss. 
 
 They were found in the oadema- 
 tous fluid of field mice inoculated 
 with garden earth and stale cheese. 
 
 Bacillus mycoides (Fliigge). 
 Rods rather thick, nearly the size 
 of Bacillus arithracis. Motile, 
 often forming long pseudo-threads. 
 
 Oval and highly refractive spores 
 both in the rods and threads. 
 
 Colonies consist of a whitish tur- 
 bidity in which colourless branched 
 and interwoven processes are seen ; 
 these increase rapidly, and after 
 twelve to twenty hours appear like 
 the mycelium of a fungus. 
 
 Inoculated in the depth of gela- 
 tine they form very fine and closely 
 set hairs extending from the track 
 of the needle. Later liquefaction 
 occurs. 
 
 On potato a whitish layer gradu- 
 ally extends over the surface. 
 
 They occur in earth from the 
 surface of cultivated ground. 
 
 Bacillus mycoides roseus 
 (Scholl). Rods. 
 
 Colonies composed of interlacing 
 filaments. 
 
 Inoculated in the depth of gela- 
 tine they produce liquefaction in 
 the track of the needle ; a reddish 
 scum forms on the surface, and a 
 reddish deposit at the bottom of 
 the liquefied area. 
 
 On agar they produce, in the 
 absence of light, a pink growth. 
 
 Bacillus neapolitanus (Emme- 
 rich). Short rods '9 /M in width. 
 
 FIG. 207. BACILLUS NEAPOLITANUS, x 
 700 (EMMERICH), a, From intes- 
 tinal contents in a case of cholera ; 
 I, From peritoneal fluid of an 
 inoculated guinea-pig. 
 
DESCRIFHON OK SPE< LE8. 
 
 
 Colonies circular, later irregular, 
 granular, strongly refractive and 
 yellowish - brown. They are 
 
 probably identical with Bacillus coli 
 communis. 
 
 They were isolated from cases of 
 cholera at Naples. 
 
 Bacillus necrophorus (Loffler). 
 Rods and filaments. 
 
 They cannot be cultivated on the 
 ordinary media. In rabbit broth 
 they give rise to fluffy masses of 
 filaments. 
 
 Intravenous injection produced 
 in rabbits a pyaemic condition in 
 about a week. The bacilli were 
 found in the pus. 
 
 They were isolated from a rabbit 
 which had been inoculated with 
 fragments of a condyloma. 
 
 Bacillus nitrificans (Wino- 
 gradsky). Yery small rods - 5 /* in 
 in length, singly and in zooglcea. 
 
 Colonies in silica jelly are 
 lenticular, and sub-cultures in liquid 
 media produce a gelatinous de- 
 posit. They are powerful oxidising 
 agents. 
 
 They were isolated from the soil. 
 
 Bacillus nodosus parvus 
 (Lustgarten). Rods 1 '2 to 2*4 p. in 
 length, -4 p in width ; singly and 
 in pairs. 
 
 Inoculated in the depth of agar 
 they produce a white filament in 
 the track of the needle composed 
 of crowded colonies, and on the 
 surface a hemispherical glistening 
 growth. 
 
 They were isolated from the 
 human urethra. 
 
 Bacillus nubilus(Frankland). 
 Slender rods 3 p long and *3 \L wide, 
 and threads. Single bacilli have an 
 active rotatory movement, but the 
 long threads in broth cultun 
 quite motionless. Spore-formation 
 not observed. 
 
 The colonies appear as cloudy 
 undefined patches, which rapidly < 
 liquefy the gelatine. They c<> 
 of a tangled mass of threads. 
 
 Inoculated in the depth of gela- 
 tine they produce along the track 
 of the needle horizontal circular 
 plates, with a delicate cloud-1 ike- 
 appearance, and liquefaction at the 
 
 upper part. Lau-r th,. whole of 
 the gelatine i> li.ju- : 
 
 On agar they tVm thi,, ,)jla- 
 cent blue-violet Him. tin- .-.(- 
 which exhibit la 
 fluorescence. 
 
 On potato there is a slightly 
 yellow growth which is scarcely 
 visible. 
 
 Broth is rendered turbid with a 
 dirty- white d-: u-face 
 
 being covered by a thin pdlirl,-. 
 
 They occur in water. 
 
 Bacillus ochraceus (/ir 
 mann). Rods 1 _'.". IL in 
 
 length; "65 to !', M in width: 
 singly, in pairs, chain-, and fila- 
 ments ; capsulated. 
 
 Colonies circular, granular, yel- 
 low, liquefying. 
 
 Inoculated in the depth of gela- 
 tine they produce liquefaction in 
 the track of tin- nee<l 
 yellow deposit. 
 
 On agar and potato the growth 
 is yellow ochre in colour. 
 
 They occur in \v 
 
 Bacillus oedematis aerobicus 
 (Klein). Rods -8 to -J-4 M in length, 
 ? p. wide, and long filament-. 
 
 Colonies greyish, transparent, 
 with irregular contour. 
 
 In the depth of gelatine a fila- 
 ment occurs in the track of the 
 needle, and gas lu I >lated 
 
 colonies in its lower part, and a 
 transparent patch with irregular 
 margin on the f n 
 
 On the surface of agar they pro- 
 duce a greyish white ! 
 
 In broth there is turbidity with 
 flocculi. 
 
 On potato the growth is yello 
 and v: 
 
 Tl icy give rise to extensive oedema 
 in guinea-pigs, and in a less marked 
 form in rabbits. 
 
 They occur in earth. 
 
 Bacillus cedematis maligni 
 (p. -'-'<>). 
 
 Bacillus of Belfanti and Pas- 
 carola. V 
 
 Colonies circular, granular, yel 
 
 Inoculated in th, -ela- 
 
 rhey produce a filan 
 posed of closely-packed minute 
 
526 
 
 DESCRIPTION OF SPECIES. 
 
 colonies, and on the surface a 
 greyish film. 
 
 On agar they produce a greyish- 
 white growth. 
 
 On potato a transparent whitish 
 film. 
 
 They are fatal to rabbits, guinea- 
 pigs and small birds. 
 
 They are probably identical with 
 Bacillus septicaemias haemorrhagicae. 
 
 They were isolated from pus in 
 a case of tetanus. 
 
 They were isolated from deep-sea 
 dredgings. 
 
 Bacillus of Colomiatti. Minute 
 rods. Spore-formation occurs at 
 the ends of the rods. They can 
 be cultivated at 37 C. 
 
 They form a thin film on agar 
 and on blood serum. 
 
 They were isolated in cases of 
 conjunctivitis. 
 
 Bacillus of Fulles, No. I. Rods 
 1 to 1'2 /u, in length, - 6 p in width. 
 
 Colonies circular, granular, yel- 
 lowish-brown. 
 
 On the surface of gelatine they 
 produce a thin film, and in broth 
 turbidity and flocculi. 
 
 On potato the growth is yel- 
 lowish. 
 
 No. II. Very short rods. 
 
 Colonies circular, granular, yel- 
 lowish. 
 
 In the depth of gelatine the 
 growth resembles Friedlander's 
 pneumococcus. 
 
 On potato the growth is yellowish. 
 
 They were isolated from earth. 
 
 Bacillus of Guillebeau. No. I. 
 Short rods 1 to 2 p in length, 1 /n 
 in width. 
 
 Colonies spherical, granular. 
 
 In gelatine the bacilli produce a 
 growth in the track of the needle 
 and a white patch on the surface. 
 
 On agar the growth is white, and 
 on potato yellowish, viscid, and con- 
 taining gas bubbles. 
 
 They coagulate milk. 
 
 No. II. Rods resembling the 
 above described but distinguished 
 by the production of viscid colonies 
 and, extremely slowly, of liquefac- 
 tion in the jelly. 
 
 No. III. Rods also resembling the 
 above mentioned, but colonies are 
 
 adherent to the jelly and coarsely 
 granular. 
 
 Milk and other liquid culture 
 media are rendered extremely vis- 
 cid. 
 
 Bacillus of Letzerich. Rods 
 sometimes bent, and filaments. 
 
 They rapidly liquefy gelatine. 
 
 They produce purulent peritonitis 
 and death in rabbits. 
 
 They were isolated from urine. 
 
 Bacillus of Martinez ( Stern - 
 berg). Short rods 1 to 1-2 \JL in 
 length and -5 to '8 p. in width ; 
 non-motile. 
 
 Colonies circular and translucent, 
 with a central, nipple-like projec- 
 tion, and the surface covered with 
 mosaic markings. 
 
 In the depth of gelatine the 
 growth consists of large spherical 
 translucent colonies in the track of 
 the needle, and a thin, translucent, 
 scanty growth upon the surface. 
 
 They were isolated from the liver 
 in a fatal case of yellow fever. 
 
 Bacillus 9f Nocard. (Vide Strep- 
 tothrix farcinica.) 
 
 Bacillus of Okada. Short rods 
 rather thicker than the bacilli of 
 mouse-septicaemia, singly, in pairs 
 and in filaments. Spore-formation 
 not observed. 
 
 Colonies granular and brownish. 
 
 Inoculated in the depth of gelatine 
 they form a white filament, and on 
 the surface a milk-white patch. 
 
 Inoculated on agar the growth 
 spreads over the surface forming a 
 milk-white layer. 
 
 In broth they produce cloudiness 
 and a layer floating on the surface. 
 
 They do not grow on potato. 
 
 Cultures produce death in mice, 
 guinea-pigs and rabbits in twenty 
 hours. 
 
 They were isolated from dust. 
 
 Bacillus of Roth. No. 1 and 
 No. 2. Rods. 
 
 Two varieties were isolated from 
 old rags. They appear to be 
 varieties of Bacillus coli communis. 
 
 Bacillus of Sattler. 2 to 4-5 p 
 long and '58 p thick. 
 
 They can be cultivated on nutrient 
 gelatine and blood serum. 
 
 Infusion of jequirity containing 
 
DESCRIPTION Ol sri.i II .-. 
 
 the bacilli, inoculated into the con- 
 junctiva of healthy rabbits, produces 
 severe ophthalmia. The poisonous 
 principle is a chemical ferment 
 ulir'm. Boiling, which does not 
 destroy the spores of the bacillus, 
 destroys the ferment, and cultiva- 
 tions started from these spores, 
 though teeming with jequirity 
 bacilli, are quite harmless (Klein). 
 
 The bacilli occur in infusions 
 of the beans of Abrus precatorius 
 or jequirity. 
 
 Bacillus of Schaffer (Freuden- 
 reich). Rods 2 to 3 p. in length, 
 1 f in width, and long filaments. 
 
 Colonies circular, granular, yel- 
 lowish. 
 
 In the depth of gelatine a growth 
 develops in the track of the needle 
 and a greyish layer on the surface. 
 On agar the growth is greyish 
 and sometimes brownish, and on 
 potato yellowish. 
 
 In broth with peptone and milk 
 sugar there is copious formation of 
 gas-bubbles. 
 
 They closely resemble Bacillus 
 coli communis. 
 
 They were isolated from cheese 
 and potato. 
 
 Bacillus of Scheurlen. Rods 
 1-5 to 2-5 p in length, -5 fi in width. 
 They were isolated from cancerous 
 growths by Scheurlen, and were 
 later identified with Bacillus epider- 
 midis. 
 
 Bacillus of Schou. Short rods 
 and cocci-forms. 
 
 Colonies are spherical, opaque, 
 and granular. 
 
 The bacilli inoculated in gelatine 
 rapidly liquefy it, and a white de- 
 posit forms at the bottom of the 
 liquid. 
 
 Rabbits inoculated in the trachea, 
 or made to inhale pure-cultures, are 
 said to develop fatal pneumonia. 
 They were isolated from rabbits 
 with pneumonia, following section 
 of the vagi. 
 
 Bacillus of swine plague 
 (p. 3ol). 
 
 Bacillus of Tommasoli 
 rods from 1 to 1*8 \i in length, ami 
 25 to :'> p. in width, singly, and in 
 short chains. 
 
 Colonies grey and shiny. 
 
 In the depth of gelatine ti 
 form a filament composed 
 packed colonies. an<l on the surface 
 a shining mass. 
 
 On agar the growth con-i-i 
 greyish patches. 
 
 On potato the growth is granular 
 and yellowish-white. 
 
 Cultures rubbed into the skin 
 are said to produce a vesicular 
 eruption. 
 
 They were isolated from the 
 scalp in a case of sycosis. 
 
 Bacillus of Utpadel. K.uS I-.':* 
 to 1'5 n in length, and '75 to 1 p, 
 in width, singly, in pairs ;unl short 
 chains. 
 
 Colonies milk-white. 
 
 On the surface of gelatine the 
 growth is milk-white, and on agar 
 yellowish-white. 
 
 Injected subcutaneously in cats, 
 guinea-pigs and mice, they produce 
 extensive oedema, and a fatal ter- 
 mination. 
 
 They were isolated from the 
 human intestine. 
 
 Bacillus of Winogradsky. (See 
 Bacillus nitrificans.) 
 
 Bacillus ovatus minutissi- 
 mus (Unna). Short rods with 
 pointed ends '6 to /i in length, 
 4 /x in width, singly, and in 
 masses. 
 
 Colonies are minute, granular an-1 
 yellowish. 
 
 The bacilli inoculated in 
 depth of gelatine form a filament 
 of closely packed gre\ 
 colonies, and on the free surface 
 there is a shiny, greyish 
 layer. 
 
 On agar the growth JH very sum 
 lar, and on potato a! 
 
 They were isolated from tin- akin 
 in eczema seborrlm < 
 
 Bacillus oxytocus perniciosus 
 I Wys-nkowitch). KnU short and 
 thick. 
 
 Ionics circular, granular, yel- 
 lowish. ..r y. ll-.u 
 
 The bacilli inoculated in i 
 depth of g.-latuir produce a gi 
 resembling Friedliwkr'i pneumo 
 coccn> 
 
 '1 1), v coagulate milk. 
 
528 
 
 DESCRIPTION OF SPECIES. 
 
 The products injected intrave- 
 nously produce death in from three 
 to twenty-four hours. 
 
 They occur in sour milk. 
 
 Bacillus pestifer (Frankland). 
 Rods 2 '3 fj. in length, 1 p in 
 width, and filaments. Motile. 
 
 Colonies resemble those of Bacil- 
 lus vermicularis. 
 
 On agar they produce a dentated 
 transparent layer, and on potato a 
 flesh-coloured growth. 
 
 They occur in the air. 
 
 Bacillus phosphorescens geli- 
 dus (Forster). Very short rods. 
 
 Colonies circular, granular, yel- 
 lowish or greenish. 
 
 The bacilli, inoculated in the depth 
 of gelatine, produce very little 
 growth in the track of the needle, 
 and a white film on the surface. 
 
 On agar and potato the growth 
 is whitish. 
 
 Cultures are photogenic. 
 
 They were isolated from phos- 
 phorescent fish. 
 
 Bacillus phosphorescens Indi- 
 CUS (Fischer). Rods singly and in 
 pairs, and filaments. Motile. 
 
 Colonies circular, well-defined, 
 greenish. 
 
 The bacilli, inoculated in the 
 depth of gelatine, produce a greyish 
 filament in the track of the needle, 
 and a hemispherical excavation of 
 the jelly at the upper part. Later 
 the jelly is liquefied, and there is a 
 yellowish scum on the surface. 
 
 On agar and potato the growth 
 is white. 
 
 Cultures are photogenic. 
 
 They were isolated from sea- 
 water. 
 
 Bacillus phosphorescens indi- 
 genus (Fischer). Rods 1-3 to 1-2 
 p in length, 4 to "1 p, in width, 
 singly, in pairs, and filaments. 
 
 Colonies circular, greenish, and 
 later yellowish. 
 
 The bacilli, inoculated in the 
 depth of gelatine, produce a conical 
 excavation in the upper part of the 
 needle track without liquid con- ', 
 tents, but with a dry growth on ; 
 the sides. 
 
 There is no growth on potato. 
 
 Cultures are photogenic. 
 
 They occur in sea-water and on 
 phosphorescent fish. 
 
 Bacillus plicatus (Zimmer- 
 mann). Minute rods, singly, in 
 pairs, and in short chains. 
 
 Colonies yellowish-white. 
 
 The bacilli, inoculated in the 
 depth of gelatine, form minute 
 isolated colonies, and on the surface 
 a wrinkled patch with gradual lique- 
 faction. 
 
 On potato the growth is dry and 
 yellowish. 
 
 They occur in water. 
 
 Bacillus pneumosepticus 
 (Babes). Short rods, '2 /Ltin width. 
 
 Colonies irregular, semi-trans- 
 parent. 
 
 In gelatine, the bacilli grow in 
 the track of the needle. On agar 
 the growth is whitish and shining. 
 
 Rabbits, guinea-pigs and mice 
 die in two or three days of septi- 
 caemia when a culture is injected 
 subcutaneously. 
 
 They were isolated from a fatal 
 case of septic pneumonia. 
 
 Bacillus polypiformis (Libo- 
 rius). Slender rods, spore-forma- 
 tion present. They are anaerobic. 
 
 Colonies composed of peculiar 
 convoluted processes. 
 
 In the depth of blood serum they 
 produce a cloudiness at the lower 
 part of the needle track. 
 
 They occur in soil. 
 
 Bacillus prodigiosus (Micro- 
 coccus prodigiosus : Cohn. Blood 
 rain, Bleeding host). Very short 
 rods with rounded ends, and thread 
 forms '5 to 1 \i in width, forming 
 at first rose-red and then blood-red 
 zooglcea. 
 
 They liquefy gelatine. 
 
 They grow luxuriantly on the 
 sloping surface of nutrient agar- 
 agar, and on sterilised potato, and 
 the colour varies from blood-red to 
 bright-red with sometimes a metal- 
 lic lustre. The cells themselves 
 are colourless. The colouring-matter 
 resembles f uchsine ; it is insoluble 
 in water but soluble in alcohol. 
 The addition of acids changes it to 
 carmine red, and of alkalies to a 
 yellow colour. 
 
 They appear occasionally on 
 
DESCRIPTION 01 M: 
 
 bread, boiled rice, and starch paste, 
 and more rarely on boiled white 
 of egg and meat. Milk sometimes 
 becomes coloured blood-red by the 
 growth of this fungus, an appear- 
 ance formerly attributed to a d 
 of the cow. 
 
 In Paris in 1843 the micro- 
 organism was peculiarly prevalent, 
 attacking especially the bread pro- 
 duced in the military bakehouses. 
 
 Bacillus proteus fluorescens 
 (J;iger). Short thick rods and 
 threads. Actively motile. 
 
 Colonies resemble minute drops 
 of water. 
 
 The rods inoculated in gelatine 
 produce a growth similar to that 
 of Koch's comma-bacilli. 
 
 The jelly becomes greenish, and 
 a pellicle forms on the surface. 
 
 On agar the growth when fully 
 developed is yellowish-white, with 
 a green fluorescence. 
 
 On potato they form a brown 
 layer. 
 
 They are pathogenic in mice. 
 They were isolated from the 
 internal organs of fowls suffering 
 from an epidemic disease. 
 
 Bacillus pseudo-diphtheriticus 
 (p. 335). 
 
 Bacillus pseudo-tuberculosis 
 (Pf eiffer). Rods varying in length. 
 Colonies circular, with dark nu- 
 cleus and transparent zone. 
 
 In the depth of gelatine they 
 produce a filament composed of 
 small colonies, and on the surface 
 a patch with concentric markings. 
 
 They grow on agar, but not readily 
 on potato. 
 
 Inoculated in mice, guinea-pig 
 rabbits, and hares, they produce a 
 fatal result in from six to twenty 
 days. An abscess forms locally, 
 the lymphatic glands enlarge and 
 caseate, and the internal organs 
 contain nodules resembling tubercle 
 
 They were isolated from the i 
 ternal organs of a horse supposed 
 to be glandered. 
 
 Bacillus pulpae pyogenes.- 
 Rods slightly bent and with points 
 ends ; singly, in pairs, and in chains. 
 
 Colonies circular, yel 
 brown. 
 
 Inoculatulin th. -depth of gelatin. 
 liquefaction occurs in the uppi i 
 part of the needle track and ex- 
 tends downward-.. 
 
 Jntraperitoneal injection in mice 
 produces death in from eighteen to 
 thirty-six hours. 
 
 They were isolated from putrid 
 dental pulp. 
 
 Bacillus punctatus (/im- 
 
 maun). Rods I to 1 r, in K-ngth, 
 77 p in width, singly, in pans, and 
 chains. 
 
 Colonies composed of stringy 
 masses in liquefied gelatine. 
 
 The bacilli inoculated in th. 
 depth of gelatine produce rapid 
 liquefaction in the track of th. 
 needle, and a white depo-.it. 
 
 On agar the growth is smooth 
 and shining. 
 
 On potato the growth is browni-h 
 
 They occur in water. 
 
 Bacillus putrificus coli < I 
 stock). Slender, motile rods, \\ p 
 in length, often less, somet 
 
 Fi.;. -'<>*. HACII : 
 
 x in- 
 forming long threads. Spore-for- 
 mation present. 
 
 Cultivations in gelatine are i 
 descent. 
 
 They are constantly present 
 faeces. 
 
 Bacillus pyocyaneus (fiessar^ 
 Slender rods, singly, in two* 
 and : ir mM8W - 
 
 Spore-formation pn 
 
 White colonial . twen 
 
 four hours, which li-i 
 tine. The whole of the median 
 
 :ic,,uirc. :i -n-en.sh - 
 
 ;.. la.illi are cultivated l 
 gelatine, the j-ll.v is 1<|" 
 Soured* .'tedM 
 
 a deep orange! 
 
 On ag r l> * 
 
 :lll ,l odour tin medium a pea-gre 
 
530 
 
 DESCRIPTION OF SPECIES. 
 
 On potato a dry rust-brown 
 growth appears at the seat of 
 inoculation, which becomes green 
 when treated with ammonia. 
 
 The pigment formed by the micro- 
 organism is a definite principle 
 pyocyanin. It can be extracted 
 with chloroform from pus and from 
 washing of bandages ; it is soluble 
 in acidulated water, which it colours 
 red. Inneutralsolutionitbecom.es 
 blue. It crystallises in chloroform 
 in long needles ; and forms some- 
 times lamellae and prisms. 
 
 They cause death in guinea-pigs 
 when injected into the abdominal 
 cavity. Rabbits are not killed by 
 intravenous injection. 
 
 The bacilli are antagonistic to 
 anthrax bacilli. Charrin and others 
 have shown that rabbits inoculated 
 with a. pure-culture of Bacillus 
 pyocyanus after inoculation with 
 Bacillus anthracis will not succumb 
 to anthrax. Woodhead and Wood 
 produced similar results by using 
 sterilised cultures, showing that the 
 results were due to the chemical 
 products of the bacilli. 
 
 The rods occur in the pus of those 
 cases in which the wounds and pus- 
 stained bandages exhibit a greenish- 
 blue colour. 
 
 Bacillus pyogenes feet id us 
 {Passet). Small rods, about T45 ft 
 in length, and '58 ft in width ; 
 
 FIG. 209. BACILLUS PYOGENES 
 FCETIDUS, x 790 (PASSET). 
 
 often in pairs, or linked together 
 in chains. They are motile, and 
 ^pore-formation occurs. 
 
 Colonies like white points appear 
 after twenty -four hours, and de- 
 velop into greyish spots, and these 
 enlarging coalesce into a layer. 
 
 Cultivated in nutrient gelatine 
 
 a greyish, veil-like growth forms 
 on the surface. 
 
 In nutrient agar-agar the culti- 
 vation resembles the growth in 
 gelatine. On blood serum a moder- 
 ately thick greyish-white streak 
 develops, and on sterilised potato 
 an abundant, shining, brownish 
 culture. 
 
 From all these media a putrid 
 odour emanates, but no smell is 
 detected from a cultivation in milk. 
 
 Inoculated into mice and guinea- 
 pigs, abscesses are produced or death 
 from septicaemia results. 
 
 They were isolated from putrid 
 pus. 
 
 Bacillus pyogenes soli(Bolton). 
 Rods resembling Bacillus diph- 
 therias. 
 
 Colonies granular, faintly yellow. 
 
 The bacilli inoculated in the depth 
 of gelatine form colonies in the 
 track of the needle. 
 
 They are pyogenic in mice and 
 rabbits. 
 
 They are present in earth. 
 
 Bacillus radiatus (Luderitz). 
 Rods 4 to 7 ft in length, '8 //, in 
 width, and filaments. Motile. 
 They are anaerobic. Spore-forma- 
 tion present. 
 
 Colonies are composed of delicate 
 interlacing filaments. 
 
 In the depth of gelatine a growth 
 occurs at the lower part of the 
 needle track, from which fine fila- 
 ments are given off in the sur- 
 rounding gelatine, and liquefaction 
 follows. The growth in the depth 
 of agar is also composed of fine 
 filaments. 
 
 In sub-cultures they produce a 
 cloudy liquefaction. 
 
 Cultures have a peculiar odour. 
 
 They occur in earth. 
 
 Bacillus radiatus aquatilis 
 (Zimmermann). Rods 1 to 6'5 ft 
 in length, '65 ft in width. 
 
 Colonies white, with a marginal 
 zone of radiating filaments. 
 
 The bacilli inoculated in the 
 depth of gelatine grow in the track 
 of the needle, and excavate and 
 liquefy the surrounding gelatine, 
 and form on the free surface a 
 wrinkled patch, which later subsides 
 
i>Ksnumox 01 
 
 
 to the bottom of the liquefied 
 .area. 
 
 On agar a smooth slightly 
 brownish layer is formed, and on 
 potato it is yellowish. 
 
 They occur in water. 
 
 Bacillus ramosus (Eisenberg). 
 Rods singly, and in chains ; fila- 
 ments. Spore-formation present. 
 
 Colonies are composed of curi- 
 ously twisted filaments. 
 
 The bacilli inoculated in the depth 
 of gelatine produce delicate fila- 
 ments extending in all directions 
 from the track of the needle, fol- 
 lowed by liquefaction ; later a skin 
 forms on the surface. There is a 
 sediment at the bottom of the tube. 
 
 On agar they form a greyish 
 filamentous layer, and on potato 
 a whitish growth. 
 
 They occur in earth and water. 
 
 Bacillus reticularis (Jordan). 
 Rods 5 /i in length. 1 p. in width ; 
 singly, and in short chains. Motile. 
 
 Colonies are composed of radi- 
 ating filaments, and liquefy the 
 gelatine, forming an excavation with 
 a reticulated lining. 
 
 The bacilli inoculated in the 
 depth of gelatine produce filaments 
 extending from the track of the 
 needle, and at the upper part the 
 jelly is excavated in the form of 
 a cup. On agar they form a dry 
 layer, and on potato a white woolly 
 growth. 
 
 Broth is made turbid, and milk 
 slowly coagulated. 
 
 They occur in water. 
 
 Bacillus rosaceus metalloides 
 < /tiirff-1-tnin rnsacfu.ni 
 Dowdeswell ; Magenta 
 Rods -6 to -8 /* in breadth. 
 
 Colonies in the depth of gelatine 
 are colourless, but superficial ones 
 are prominent and magenta in 
 colour. 
 
 On the surface of obliquely solidi- 
 fied gelatine they form a beautiful 
 magenta band with a metallic 
 lustre. The gelatine is not lique- 
 fied. Similar growths are obtained 
 on agar and potato. 
 
 It is one of the most striking 
 of all the chromogenic bacteria. 
 Cultures have the appearance of 
 
 having been stained with an 
 holic solution of furli-: 
 colour varit-s in siilx-nltnivs f rom 
 magenta to a seal inu-uax red. 
 
 Bacillus rubefaciens < 
 mann). Rods 75 t. llK ,* in 
 length. ;*_' p. in width, singh 
 pairs, and in chainv 
 
 Colonies are faintly red-! 
 yellow. 
 
 The bacilli inoculated in 
 depth of gelatine grow alon^r 
 track of the needle and form ,i 
 greyish layer on the - later 
 
 the jelly acquires a reddish tint. 
 
 On agar the growth is grey 
 abundant. 
 
 On potato the growth is at first 
 grey, later n-ddUh-brown, an<i 
 surface of the potato has a pink 
 discoloration. 
 
 They occur in u 
 
 Bacillus rubellus (Okada). 
 Rods resembling those of malign 
 cedema. They occur singly, in 
 pairs, and filaments: are m- 
 and possess flagella, and are <> 
 capsulated. They are anaerol 
 
 Colonies are whitish, with <>tV- 
 shoots in the surrounding gela 
 which, later, is liquefied, and has a 
 reddish tinge. 
 
 The bacilli inoculated in th" 
 depth of gelatine produce a growth 
 in the lower part <>f the needle 
 track composed of isolated col- 
 with radiating processes. The 
 is liquefied, at first in the 
 corresponding with the gr 
 later completely. The liq" 
 gelatine is o.lomvd red. 
 
 In agar the growth ext 
 below upwards, and th- 
 coloured r <l 
 
 In broth they grow rapidly. 
 
 Th. y were isolated from 
 
 Bacillus ruber < IJn-m.. 
 rouge <! A ' 
 
 . .". p. long, and -7 to '*/* broad. 
 Slightly ni"- 
 
 Colonies b-l.,w tl of 
 
 gelatine are pale yellow, and I 
 ficial ones are blood -red. 
 
 Inocnlat. -I in the depth 
 
 bright red. Th. -n- is aUo forma- 
 tion of gas bnb' 
 
532 
 
 DESCRIPTION OF SPECIES. 
 
 Potato is rapidly covered with a 
 purplish-red growth. Broth be- 
 comes turbid, and pink in colour. 
 
 Milk is coagulated, and a blood- 
 red colour develops on the surface 
 and gradually extends. 
 
 They were found in water. 
 
 Bacillus rubescens (Jordan). 
 Kods 4 p in length, -9 ^ in width, 
 singly, in pairs, and short chains. 
 
 Colonies pure-white. 
 
 Inoculated in the depth of gela- 
 tine there is a little growth in the 
 track of the needle, and a pure- 
 white prominent patch on the free 
 surface. 
 
 On agar the growth is white and 
 shining, and later has a pink tinge. 
 
 On potato the growth is flesh- 
 coloured. 
 
 Broth becomes turbid, and a scum 
 forms 011 the surface. 
 
 Milk after a time acquires a pink- 
 ish colour. 
 
 They occur in sewage. 
 
 Bacillus rubidus (Eisenberg). 
 Rods and filaments. 
 
 Colonies circular, granular, and 
 slightly red. 
 
 The bacilli inoculated in the depth 
 of gelatine produce liquefaction and 
 a brownish-red colour. 
 
 On agar and potato they form a 
 brownish-red growth, and liquefy 
 blood serum. 
 
 They occur in water. 
 
 Bacillus sanguinis t y p h i 
 (Brannan and Cheeseman). Rods 
 1 to 2'5 p. in length, -5 to *8 p in 
 width ; singly, in pairs, and in 
 chains, and involution forms. 
 
 Colonies granular, pale-brown. 
 
 The bacilli inoculated in the 
 depth of glycerine-agar produce a 
 growth in the track of the needle 
 composed of isolated, minute, white 
 colonies. 
 
 Rabbits inoculated die in from 
 two weeks to a month. 
 
 They were isolated from the 
 blood of patients suffering from 
 typhus fever. 
 
 Bacillus saprogenes (Rosen- 
 bach). Three rod-formed organ- 
 isms have been described by 
 Rosenbach as intimately associ- 
 ated with putrefactive processes. 
 
 No. 1. Large rods (Fig. 210) T 
 which form an irregular sinuous 
 streak with a mucilaginous appear- 
 ance when cultivated on nutrient 
 agar-agar. Spore-formation pre- 
 sent. They grow also very readily 
 on blood serum, and all cultivations 
 yield the odour of rotting kitchen 
 refuse. They are not pathogenic. 
 
 No. 2. Rods shorter and thinner 
 than No. 1. They develop very 
 rapidly on agar-agar, forming trans- 
 parent drops, which become grey. 
 The cultivations yield a character- 
 istic odour similar to the last. 
 
 FIG. 210. BACILLUS SAPROGENES, No. 1. 
 (Rosenbach. ) 
 
 They are pathogenic in rabbits. 
 They appear to be identical with 
 Bacillus fcetidus (Bacterium fceti- 
 dum, Thin). They were isolated 
 from a patient suffering from pro- 
 fusely-sweating feet. 
 
 No. 3. See Bacterium sapro- 
 genes. 
 
 Bacillus SCissus (Frankland). 
 Very short rods, 1 to 2 p. in length, 
 and 1 \i in width. 
 
 Colonies -yellowish, opaque in the 
 centre, and periphery 'dentated. 
 
 Inoculated in the depth of gela- 
 tine there is no growth in the 
 track of the needle, but a shining 
 layer forms on the surface, and the 
 jelly is coloured greenish. 
 
 On agar the growth is shining 
 and the jelly coloured green. 
 
 On potato the growth is flesh- 
 coloured. 
 
 They occur in earth. 
 
 Bacillus septicaemia hsemor- 
 rhagicae (p. 231). 
 
 Bacillus septicus (Klein): 
 Rods varying in size. Non-motile. 
 They form threads or leptothrix 
 filaments, and are rounded at the 
 ends. They are anaerobic, and form 
 spores independently of access to air. 
 
DESCRIPTION OF SH 
 
 
 In a nourishing fluid they are 
 overcome by the presence of micro- 
 cocci, Bacterium termo or Bacillus 
 subtilis. 
 
 They occur in the soil, in putrid 
 blood, and many putrid albuminous 
 fluids, and occasionally in the blood- 
 vessels of man and animals after 
 death. 
 
 Bacillus septicus acuminatus 
 (Babes). Rocfe with lancet-shaped 
 ends, about the size of the bacilli 
 of mouse-septicremia. They exhibit 
 polar staining. They can be culti- 
 vated at 37C. 
 
 On agar and blood serum the 
 colonies are circular, transparent, 
 and later coalesce and form a yel- 
 lowish layer. 
 
 They are fatal to rabbits and 
 guinea-pigs in from two to six 
 days. 
 
 They were isolated from an infant 
 after death from septic infection 
 occurring five days after birth. 
 
 Bacillus septicus agrigenus 
 (Nicolaier). Rods resembling Ba- 
 cillus septicaemias haemorrhagicae. 
 
 Colonies circular, granular, with 
 concentric zones of varying tints of 
 brown. 
 
 Intravenous injections are fatal 
 to rabbits in twenty-four to thirty- 
 six hours, and bacilli abound in the 
 blood. 
 
 They occur in recently manured 
 soil. 
 
 Bacillus . septicus keratoma- 
 lacise (Babes). Short thick rods 
 singly, and in pairs : often capsu- 
 lated. 
 
 Colonies white, with dentated 
 contours. 
 
 The bacilli, inoculated in the depth 
 of gelatine, grow in the track of 
 the needle and on the surface ; gas 
 bubbles are developed. 
 
 On agar the growth is arbores- 
 cent and opalescent. 
 
 On blood serum they form a 
 shining, somewhat transparent, 
 dentated film. Cultures have an 
 ammoniacal odour. 
 
 They produce purulent inflam- 
 mation of the cornea. 
 
 They were isolated from the 
 cornea in a case of septicaemia 
 
 following 
 child. 
 
 keratomalacia 
 
 Bacillus septicus ulceris jran- 
 graenosi(Babrs).- -short ro.U 
 ; /x in width. 
 
 Inoculated in the depth of gela- 
 tine they produce li a and 
 gas in the track of the needle. 
 
 On agar greyish-white shining 
 patches are found. 
 
 On potato they develop a trans- 
 parent film. 
 
 They are pyogenic in rabbit - 
 mice. 
 
 They were isolated from the 
 internal organs and blood in a case 
 of septicaemia following gangrene. 
 
 Bacillus septicus vesicae 
 (Clado). Rods 1-6 to 2 p in length, 
 5 /n in width. 
 
 Colonies circular, transparent, 
 yellowish. 
 
 The bacilli inoculated in tin- 
 depth of gelatine form a delicate 
 filament composed of closely-packed 
 colonies, and on the surface th- 
 a filmy growth. 
 
 On agar they form a gri-\ 
 white layer, and on j>tato the 
 growth is dry and brown. 
 
 They are poisonous to rabbi te, 
 guinea-pigs, and mice. 
 
 They were isolated from urine 
 from a case of cy- 
 
 Bacillus sessilis (I- K 
 Rods resembling those of thi 
 bacillus. 
 
 They are said to be distinguished 
 by fission commencing in a newly 
 formed rod before it has been set 
 free from the spore. 
 
 They were isolated from the blood 
 of a cow. 
 
 Bacillus smaragdino-phospho- 
 rescens(Kat/). Rods with pointed 
 c-mN. 'I p in length, 1 j* in width. 
 
 Coloni.- i.iintly yellow, 
 
 with concentric rings. 
 
 Inoculated in the gela- 
 
 tine a white filament forms in the 
 track of tin- m-edle and a greyiah- 
 white patch on the free surface ; 
 and there is son 
 
 In bn.th they produce turbs : 
 
 On potato they produce a thin 
 brownish-yi'llow film. 
 
 They are photogenic, and the 
 
534 
 
 DESCRIPTION OF SPECIES. 
 
 phosphorescence is most marked in 
 cultures containing an excess of 
 salt. 
 
 They were isolated from a phos- 
 phorescent herring. 
 
 Bacillus smaragdinus fcetidus 
 (Reimann). Slender rods slightly 
 bent. 
 
 Colonies on agar irregular, with i 
 a yellowish granular nucleus, and 
 transparent marginal zone. 
 
 The bacilli inoculated in the 
 depth of gelatine produce a growth 
 in the track of the needle, and 
 liquefaction at its upper part, 
 and a greenish coloration. 
 
 In the depth of agar the medium 
 is coloured green. 
 
 On potato the growth is brown. 
 Cultures emit a strong odour. 
 
 Intravenous injection produces 
 death in rabbits in forty-eight ! 
 hours. 
 
 They were isolated from nasal j 
 mucus in oza3na. 
 
 Bacillus solidus (Liideritz). 
 Rods 1 to 10 p. in length, *5 /A in 
 width. 
 
 They are anaerobic. Cultivated in 
 grape-sugar gelatine they produce 
 gas bubbles and a penetrating foul 
 odour. The colonies are spherical, 
 and in agar under a low power j 
 are seen to be composed of fine j 
 filaments like cotton wool. 
 
 In broth with exclusion of oxygen , 
 they produce a copious growth with j 
 abundant formation of foetid gas. 
 They were isolated from earth. 
 Bacillus spiniferus (Unna). 
 Rods sometimes curved, 2 p, in 
 length, -8 to 1 /LI in width, singly, 
 in pairs, and masses. 
 
 Colonies have peculiar spines, and 
 later a radiated marginal zone. 
 
 In the depth of gelatine minute, 
 yellowish, isolated colonies develop 
 in the track of the needle, and a 
 furrowed, yellowish-grey patch on 
 the free surface. 
 
 On agar the same yellowish 
 wrinkled growth appears. 
 
 On potato they form a shining, 
 faintly-yellow layer. 
 
 They were isolated from the skin 
 in eczema. 
 
 Bacillus spinosus (Luderitz). 
 
 Rods sometimes bent, 3 to 8 /M in 
 length, -6 /n in width, and long 
 filaments. Spore-formation present. 
 They are anaerobic. 
 
 Colonies are composed of fine 
 radiating filaments, and liquefy the 
 
 jelly. 
 
 In agar the growth is composed 
 of colonies of matted filaments,, 
 and there is gas-formation. 
 They liquefy blood serum. 
 They occur in. earth. 
 Bacillus stolonatus (Adametz). 
 Rods motile. 
 
 Colonies on gelatine, circular, 
 granular, and whitish, or yellowish- 
 brown. Colonies on agar send off 
 peculiar wavy processes. 
 
 Inoculated in the depth of gela- 
 tine a granular filament develops in 
 the track of the needle, and a white 
 patch on the free surface ; later the 
 gelatine is excavated in the upper 
 part, and the culture lines the 
 cavity. 
 
 On potato the growth is whitish. 
 They occur in water. 
 Bacillus stoloniferus (Pohl). - 
 Rods 1-2 fM in length, '8 /* in width. 
 Motile. 
 
 Inoculated in the depth of gela- 
 tine they produce rapid liquefaction 
 in the track of the needle. 
 
 On the surface of agar they form 
 a white growth. 
 
 On potato they grow abundantly. 
 I but scarcely at all in milk. 
 
 They occur in the water of 
 marshes. 
 
 Bacillus striatus albus (Bes- 
 ser). Rods sometimes bent. 
 
 Colonies on gelatine appear as 
 minute dry points. 
 
 On agar the colonies have a brown 
 ' nucleus and clear marginal zone. 
 
 On the surface of agar the bacilli 
 produce a greyish- white thin layer. 
 On potato the growth is trans- 
 parent and slightly gelatinous. 
 They occur in nasal mucus. 
 Bacillus striatus flavus (Bes- 
 ser). Short rods, straight or 
 | curved ; involution forms. 
 
 Colonies granular, yellowish. 
 On the surface of agar they pro- 
 ; duce a white growth, which later 
 becomes sulphur yellow. 
 
I>I>CKMTIMN 
 
 8PE( IBS. 
 
 585 
 
 On potato a similar colour is 
 produced. 
 
 They were isolated from nasal 
 mucus. 
 
 Bacillus subflavus (Zimrner- 
 niann). Rods 1*5 to H p. in length, 
 77 /i in width, and in chains. Motile. 
 
 Colonies prominent, yellowish- 
 white. 
 
 On the surface of gelatine they 
 form a yellowish-grey layer, and 
 on the surface of agar and potato 
 the growth is yellow. 
 
 They occur in water. 
 
 Bacillus subtilis (Hay bacillus). 
 Cylindrical rods as much as 6 n 
 in length. Single forms grow to 
 double their length, and then 
 undergo division. They also form 
 threads which may be composed of 
 
 FIG. 211. BACILLUS SUBTILI* WITH 
 SPORES (BAUMGAKTKX). 
 
 long rods, short rods, and cocci. 
 They are motile, and provided with 
 a flagellum at each end. If the 
 nourishing medium is impoverished, 
 the multiplication of the rods by 
 division gradually ceases, and spore- 
 formation commences. The rods 
 become motionless, and a dark- spot 
 is visible, either in the middle or 
 towards one end. This gradually 
 develops into a shining spore with 
 a dark outline. The rods swell 
 slightly during this process ; then- 
 contour becomes undefined, and 
 soon disappears entirely : spores 
 being set free in about twenty- 
 four hours. The spores are I"J p. 
 long, and -6 p broad. They develop 
 into rods in the following way :- 
 On one side of the spore a swell] 
 appears, at the summit of which 
 an opening in the spore-meml 
 results, and the germ tin- 
 
 lengthens into a rod, and remains 
 for a time attached to the empty i 
 spore-membrane. 
 
 The 8jx)res are widely distributed, 
 and occur in the air. nil 
 On the excren >rous 
 
 animals the bacilli form ;i 
 efflorescence, and a thirk crumpled 
 skin on liquid manure. 
 
 They flourish equally in liquids 
 and upon damp, solid, noun-hiiii: 
 media. They are aerobic : <K j 
 tion of oxygen causes the growth 
 of the bacilli to cease, and the 
 degenerate. 
 
 In plate-cultivations the colonies 
 are white, and, under a low power, 
 granular and irregular in outline 
 and faintly-green i si i. Liquefaction 
 in, producing <i like 
 
 saucers. The centre i- opaque 
 is surrounded by a network of 
 filaments, which extend into tin 
 gelatine surrounding the colon 
 
 
 Inoculated in the depth f gela- 
 tine, liquefaction 00 
 the tra.-k of the Modi*, MM a him 
 
o36 
 
 DESCRIPTION OF SPECIES. 
 
 floats on the surface. The liquefied 
 gelatine, at first turbid, becomes 
 clear as the bacilli settle at the 
 bottom of the tube. 
 
 On agar a wrinkled film develops, 
 and also on serum. 
 
 FIG. 213. PURE-CULTURE OF BACILLUS 
 SUBTILIS ON THE SURFACE OF Nu- 
 TRIENT AGAR. 
 
 On potato the growth is white, 
 and there is copious spore-forma- 
 tion. 
 
 On ordinary nutrient liquids they 
 develop at first a thin, and subse- 
 quently a thick, dense, crumpled 
 pellicle, with copious spore-forma- 
 tion. 
 
 The simplest way to obtain a 
 culture of the bacillus is to make 
 a decoction of hay. The hay is 
 chopped into small pieces, and 
 boiled with distilled water in a 
 flask for a quarter of an hour. 
 The infusion is then filtered into a 
 beaker, covered with a glass pla.te. 
 and set aside in a warm place. In 
 two or three days the liquid swarms 
 
 with the bacilli, the spores of which 
 exist in great numbers in ordinary 
 hay. A more sure method for 
 obtaining a pure cultivation is as 
 follows : 
 
 (//) Add only a small quantity of 
 water to some finely chopped hay, 
 and set aside for four hours at 
 36 C. 
 
 (6) Pour off the extract, and 
 dilute it to the sp. gr. 1*004. 
 
 (c) Boil gently for one hour in 
 a bulb plugged with cotton wool. 
 
 (d) Set aside 500 com. of the 
 extract at 36 C. 
 
 In about twenty-four hours, as 
 a rule, a pellicle has commenced 
 to develop upon the surface of the 
 liquid. If the reaction is definitely 
 acid, carbonate of soda solution 
 must be added to the decoction. 
 
 METHODS OF STAINING HAY BACILLUS. 
 
 To demonstrate the flagella of the 
 bacilli, they may be stained with 
 hsematoxylin solution (Koch), or by 
 Loffler's method. 
 
 The linking together of cocci, long 
 rods and short rods in the threads, is 
 shown by treating with alcoholic solu- 
 tion or fuchsine, or with iodine solution 
 (Zopf). 
 
 To stain the spores the cover-glass 
 preparations must be heated to a very 
 high temperature (210C.), in the hot- 
 air steriliser for half an hour, or they 
 may be exposed for a few seconds to 
 the action of concentrated sulphuric 
 acid (Biichner), or floated for twenty 
 minutes on hot solution of the dye. 
 
 Bacillus sub tills similans. 
 There are several bacilli closely 
 resembling Bacillus subtilis. 
 
 Two have been isolated froin 
 human faeces by Bienstock which 
 do not liquefy nutrient gelatine. 
 
 No. I. Rods and filaments ; 
 spore-formation present. 
 
 On agar they produce a delicate 
 wrinkled veil. 
 
 No. II. Rods morphologically 
 identical with No. I. 
 
 On agar they produce a smooth, 
 shining layer. 
 
 Bacillus superficialis (Jordan). 
 Rods 2*2 fj. in length, and -1 p. in 
 width ; singly, and in pairs. Motile. 
 
 Colonies have a yellowish-brown 
 
DESCRIPTION 01 SPECIES, 
 
 
 nucleus and transparent marginal 
 zone. 
 
 Inoculated in the depth of gela- 
 tine there is a slight growth in the 
 track of the needle, and after a time 
 liquefaction at the upper part. 
 
 On agar the growth is smooth and 
 shining. 
 
 In broth they produce turbidity. 
 
 They will not grow on potato. 
 
 They occur in sewage. 
 
 Bacillus tennis sputigenus 
 (Pansini). Short rods, singly, and 
 in pairs ; capsulated. 
 
 They produce a whitish growth 
 on the surface of gelatine. 
 
 They coagulate milk. 
 
 They are pathogenic in rabbits. 
 
 They were isolated from sputum. 
 
 Bacillus termo (Mace). Thick 
 rods 1*4 p. long, and - 8 p. wide, 
 usually in pairs, sometimes in chains. 
 Actively motile. 
 
 Colonies whitish, with a grey edge 
 surrounded by liquefied gelatine. 
 
 Inoculated in the depth of gela- 
 tine they form a funnel-shaped 
 area of liquefaction, and later the 
 whole of the jelly is liquefied. 
 
 Broth is rendered turbid and a 
 thin brittle pellicle is formed. 
 
 They are associated with decom- 
 position. 
 
 Bacillus tetani (p. 457). 
 
 Bacillus thalassophilus (Rus- 
 sel). Slender rods varying in 
 length ; and filaments. They are 
 anaerobic. Spore-formation pre- 
 sent. 
 
 Inoculated in the depth of gela- 
 tine the growth appears in the 
 lower part of the track of the 
 needle in the form of cloudy colo- 
 nies, liquefying the jelly and pro- 
 ducing gas-bubbles. Cultures emit 
 a penetrating odour. 
 
 They were isolated from sea-mud. 
 
 Bacillus thermophilus (Mi- 
 quel). Rods varying in size accord- 
 ing to the temperature at which 
 they are cultivated. In broth they 
 grow best between &V and 7< 
 forming a copious deposit. They 
 occur in air. soil, and water. 
 
 Bacillus tremelloides (Tijs). 
 Rods '7o to 1 /i in length, "11) p. in 
 width : and in masses. 
 
 ( 'olonies circular, yellowish- 
 brown. 
 
 The bacilli inoculated in the 
 depth of gelatine produce a growth 
 composed of isolated yellow colo- 
 nies in the track of the needle, 
 and a yellow mass on the surface. 
 They liquefy the gelatine. 
 
 On agar the growth is slimy and 
 golden-yellow. 
 
 On potato they form an abuml 
 ant yellow growth. 
 
 They occur in \\ 
 
 Bacillus tuberculosis (p. :57x). 
 
 Bacillus tuberculosis galli 
 narum (p. 4i-j). 
 
 Bacillus tumescens (/<|.f>. 
 Cocci, long and short rods. They 
 form a jelly-like dine " to 1 ,- m . in 
 diam. on slices of boiled ra 
 with the appearance of a rather 
 tough, crumpled skin of a whitish 
 colour. Examination of 
 licle shows that it is formed of 
 rows of rods lying closely tog. 
 These rods can be observv 
 divide into short rods and < 
 Spore-formation occurs in two 
 stages of development viz., in the 
 cocci and in the short rods. A 
 cultivation is obtained l.\ 
 slices of boiled carrot, slightly 
 moistened, to the air at the tem- 
 perature of the room. 
 
 Bacillus typhi abdominalis 
 
 Bacillus ubiquitus i.Toi 
 Rods 1-1 to _' p. in It-ngth, '1 p. 
 in width : and filaments. 
 
 Colonies granular and well 
 defined. 
 
 The bacilli inoculated in the 
 depth of gelatin.- pro'liu-- 
 resembling tin 
 pneumocoi 
 
 On agar and potato the growth 
 
 Tin v coaL'ulat" milk and reduce 
 nitrates. 
 
 They occur in 
 
 ProbabK 
 candi' 
 
 Bacillus ulna <<ohn 
 ,1,,,,-t I. -'""I thread*, 
 
 of tin- corn I'fi ' 
 
 in both -hort and 
 long r P 1 " - 
 
538 
 
 DESCRIPTION OF SPECIES. 
 
 duced by this bacillus in a nourish- 
 ing liquid. Cloudy masses are 
 found on the surface of the liquid, 
 which later form a thick dry 
 pellicle, consisting of bundles of 
 threads matted together. The for- 
 mation of ellipsoidal spores occurs 
 in the usual way ;. they measure 
 2*5 to 2*8 p long, and more than 
 
 1 p. wide. The bacillus is found 
 in rotting eggs, and can be culti- 
 vated on boiled white of egg. 
 
 Bacillus ulna (Vignal). Rods 
 
 2 p. in length ; singly, and in pairs, 
 and in short chains. 
 
 Colonies composed of concentric 
 zones varying in granularity. 
 
 Inoculated in the depth of gela- 
 tine, liquefaction occurs rapidly in 
 the track of the needle ; later, there 
 is a deposit at the bottom of the 
 liquefied area and a pellicle on the 
 surface. 
 
 On agar they form a white ad- 
 herent layer, and the jelly is tinged 
 with brown. 
 
 In broth a pellicle forms on the 
 surface. 
 
 On potato they form a pellicle 
 with characteristic linear markings. 
 
 They liquefy serum. Cultures 
 produce a putrefactive odour. 
 
 They occur in human saliva. 
 
 Bacillus vacuolosis (Sternberg). 
 Rods 1*5 to 5 p. in length, 1 /x in 
 width, containing vacuolated proto- 
 plasm ; filaments, and involution 
 forms. At times slowly motile. 
 
 Inoculated in the depth of gela- 
 tine, liquefaction occurs slowly at 
 the upper part of the track of 
 the needle, forming a cup-shaped 
 cavity ; the liquefied gelatine is 
 viscid^ and a cream-white layer 
 forms on the surface. 
 
 In agar the development in the 
 track of the needle is scanty ; on 
 the surface a cream-white layer is 
 formed, and the bacilli are united 
 in long jointed filaments. 
 
 On potato a similar growth is 
 produced. 
 
 They were isolated from the in- 
 testine in fatal cases of yellow fever. 
 
 Bacillus varicosus conjunctive 
 (Gombert). Rods 2 to 8 p in length, 
 1 p, in width. 
 
 Inoculated in the depth of gela- 
 tine they produce a greyish-white 
 filament in the track of the needle, 
 and a greyish-white patch on the 
 surface ; liquefaction follows with- 
 out turbidity. 
 
 On the surface of agar a white, 
 dry. adherent film is formed. 
 
 On potato the growth is, at first, 
 white and dry, later, reddish-brown. 
 
 They produce hyperaemia when 
 injected into the conjunctiva. 
 
 They were isolated from the 
 healthy human conjunctiva. 
 
 Bacillus venenosus (Vaughan). 
 Motile rods. 
 
 Colonies circular, whitish. 
 
 Inoculated in the depth of gela- 
 tine there is growth in the track of 
 the needle and on the free surface. 
 
 On agar they form a white film, 
 and on potato a moist brownish 
 layer. 
 
 They are pathogenic in small 
 animals. 
 
 They occur in water. 
 
 Bacillus venenosus brevis 
 (Vaughan). Rods short and thick. 
 
 Colonies are yellow and composed 
 of concentric rings. 
 
 Inoculated in the depth of gela- 
 tine they grow in the track of the 
 needle and over the free surface. 
 
 On agar they produce a white 
 film. 
 
 On potato the growth is brownish. 
 
 They are pathogenic in small 
 animals. 
 
 They occur in water. 
 
 Bacillus venenosus invisibilis. 
 Slender rods. 
 
 Colonies irregular, granular. 
 
 Inoculated in the depth of gela- 
 tine the growth is extremely slow 
 both in the track of the needle and 
 on the surface. 
 
 On agar there is a whitish film, 
 and on potatoes a brownish layer. 
 
 They are pathogenic in small 
 animals. 
 
 They occur in water. 
 
 Bacillus venenosus lique- 
 faciens (Vaughan). Rods. 
 
 Colonies circular, granular, yel- 
 lowish. 
 
 Inoculated in the depth of gela- 
 tine they grow in the track of 
 
INSCRIPTION OF SI I 
 
 589 
 
 the needle and on the surface, and 
 liquefaction occurs after some 
 weeks. 
 
 On agar they produce a white 
 growth, and on potato it is brown- 
 ish or yellowish. 
 
 They are pathogenic in small 
 animals. 
 
 They occur in water. 
 
 Bacillus ventriculi (Raczyn- 
 sky). Rods 1-5 to 3 p. in length, 
 1 fi in width, singly, in pairs, and 
 in short chains. 
 
 Colonies have a dark nucleus and 
 transparent periphery. 
 
 On agar they form a white layer. 
 
 They were isolated from the 
 digestive tract of dogs. 
 
 Bacillus vermicularis (Frank- 
 land). Large bacilli 2 to 3 /A in 
 length, 1 p. in width, and long 
 threads. Spore-formation present. 
 
 Colonies are irregular in contour, 
 the irregularity increasing as the 
 colony comes to the surface. The 
 peripheral part is composed of 
 closely packed, wavy bands of bacilli, 
 and the centre is irregular and 
 wrinkled. 
 
 The bacilli inoculated in the 
 depth of gelatine form a flattened 
 band in the track of the needle, and 
 a grey layer on the surface ; lique- 
 faction slowly follows. 
 
 On agar they produce a smooth, 
 shining, grey layer, and on potato 
 a thick, irregular, flesh-coloured 
 growth. 
 
 They reduce nitrates. 
 
 They occur in water. Probably 
 identical with Bacillus vermicu- 
 losus. 
 
 Bacillus vermiculosus (Zimmer- 
 mann). Rods 1-5/i in length, *;"> ^ 
 in width, singly, in pairs, very short 
 chains and long filaments. They 
 are slowly motile. 
 
 Colonies irregular ; grey, granu- 
 lar. 
 
 Inoculated in the depth of gela- 
 tine they produce, after four days, 
 liquefaction in the upper part of 
 the needle track, which spreads 
 downwards, and a reddish-grey sedi- 
 ment collects at the bottom of the 
 liquefied area. 
 
 On agar the growth is smooth and 
 
 shining, and on potato yellov. 
 grey. 
 
 They occur in water. 
 
 Bacillus violaceus ( >><!< Bacil- 
 lus ianthiniis). 
 
 Bacillus violaceus Laurentius 
 (Jordan). Rods ; to 3*6 p in 1,-ngth, 
 7 /i in width. 
 
 Colonies violet, surrounded \>\ 
 liquefied gelatine. 
 
 Inoculated in the depth of gela- 
 tine liquefaction occurs HI th,- track 
 of the needle, and a violet s,.<linient 
 collects at the bottom. 
 
 On agar the growth is violet, later 
 black. 
 
 On potato there is a copious 
 growth, changing in colour from 
 violet to black. 
 
 In broth a violet colour i- pp. 
 duced in the presence of nitrates. 
 
 They coagulate milk, and render 
 it bluish-violet. 
 
 They occur in water. Probably 
 identical with Bacillus ianthiniis 
 (Zopf). 
 
 Bacillus virescens (Frick). 
 Rods and filaments. 
 
 Colonies irregular, granular, 
 green. 
 
 On the surface of gelatine they 
 colour the medium green. 
 
 They grow on agar. 
 
 < )n potato they form a brownish 
 growth. 
 
 In broth a pellicle is formed on 
 the surface, and beneath it the 
 liquid is coloured green. 
 
 They were isolated from green 
 sputum. 
 
 Bacillus viscosus (I'r.mkl 
 Rods |-."i to -2 n in length, singly 
 and in pairs. 
 
 Colonies granular, with hairlike 
 processes extending into the gela- 
 tine, which i- l:qut Tied and has a 
 green colour. 
 
 Inoculated in 
 
 tine they produce liquefaction and 
 a green fluorescence. 
 
 On agar they form a greet 
 white layer, and colour th.- j.-llv 
 green. 
 
 On potato the growth is bn> 
 
 Probably identical with Ba< 
 fluoreseens liquefari' 
 
 Bacillus Zurnianus < i 
 
DESCRIPTION OF SPECIES. 
 
 Rods 1-2 to 1-5 fi in length, -6 to -8 ft 
 in width. Colonies greyish-white, 
 viscid. 
 
 The bacilli inoculated in the 
 depth of gelatine develop slightly 
 in the track of the needle, and pro- 
 duce a prominent grape-like growth 
 on the free surface. 
 
 On potato the growth is grey or 
 tinged with yellow. 
 
 They occur in water. 
 
 Bacterium aerogenes (Miller). 
 Short rods, singly and in pairs. 
 Motile. 
 
 The colonies are circular, well 
 denned, and yellowish. 
 
 Inoculated in the depth of gela- 
 tine the growth in the track of the 
 needle is brownish-yellow, and a 
 flat greyish button forms on the 
 free surface. 
 
 On agar a pulpy layer develops. 
 
 On potato the growth is pulpy 
 and yellowish-white. 
 
 The bacteria possess great power 
 of resisting the effect of acids. 
 
 They were isolated from the diges- 
 tive tract. 
 
 Bacterium brunneum 
 (Schroter).- Motile rods, produ- 
 cing a brown colour. 
 
 They were observed on a rotting 
 infusion of maize. 
 
 Bacterium decalvans (Thin). 
 Cocci, singly or in pairs, 1*6 ft in 
 length. 
 
 They were observed in the roots 
 of the hair in cases of Alopecia 
 weata. 
 
 Bacterium fusiforme (Warm- 
 ing). Rods spindle-shaped, with 
 pointed ends, 2-5 p. long, and *5 to 
 8 fi thick. They were described 
 .as forming a spongy Ikyer on the 
 surface of sea-water. 
 
 Bacterium gingivae pyogenes 
 (Miller). Short rods. 
 
 The colonies are circular and 
 rapidly liquefy gelatine. 
 
 The bacteria inoculated in the 
 depth of gelatine produce rapid 
 liquefaction in the track of the 
 needle and a white sediment. 
 
 On agar they produce a moist 
 white growth. 
 
 They are pyogenic when inocu- 
 lated subcutaneously in small ani- 
 
 mals, and cause a fatal result when 
 injected to the peritoneal cavity. 
 
 They occur in the deposit on the 
 teeth. 
 
 Bacterium hyacinthi (Wakker). 
 Cocci resembling Bacterium 
 termo. 
 
 They were observed in the yellow 
 slime of diseased hyacinth bulbs. 
 
 Bacterium hydrosulfureum 
 ponticum (Zelinsky). Long 
 motile rods. 
 
 On agar a dark coffee-coloured 
 pigment is produced, which turns 
 black when exposed to air. 
 
 Cultures give off sulphuretted 
 hydrogen. 
 
 They were isolated fromdredgings 
 in the Black Sea. 
 
 Bacterium litor eum (Warming). 
 Cocci ellipsoidal, 2 to 6 ft long, 
 1*2 to 2'4 ft wide ; singly, never 
 as chains or zoogloea. 
 
 They occur in sea-water. 
 
 Bacterium luteum (List). 
 Rods from I'l to 1*3 /j. long. Non- 
 motile. The colonies are slimy, with 
 orange centres. 
 
 Inoculated in the depth of 
 gelatine an orange growth occurs, 
 principally at the point of puncture. 
 
 Milk is coagulated. 
 
 They occur in water. 
 
 Bacterium merismopedioides 
 (Zopf). Threads 1 to 1*5 u in 
 thickness ; these subdivide into 
 long rods, short rods, and finally 
 into cocci. The cocci divide first 
 in one and subsequently in two 
 directions, forming characteristic 
 groups, which appear like merismo- 
 pedia. These groups may eventually 
 consist of 64 by 64 cells or more, 
 and ultimately form zoogloea. The 
 cocci develop again into rods and 
 threads. 
 
 They were observed in water 
 containing putrefying substances 
 (River Panke, Berlin). 
 
 Bacterium navicula (Reinke 
 and Berthold). Cocci spindle-form 
 or ellipsoidal, including motile and 
 non-motile forms. They have one 
 or more dark spots, which may be 
 coloured blue by iodine. 
 
 They have been observed in rot- 
 ting potatoes. 
 
DESCRIPTION OF SPK< 1 1 -. 
 
 .'.I! 
 
 Bacterium photometricum(En- 
 gelmann). Rods slightly reddish in 
 colour : motile. 
 
 The movements are stated to 
 depend on light. 
 
 Bacterium synxanthum (Ehren- 
 berg : Bacterium .mntti'minn : Bnc- 
 
 tiTtUiu <>f y< l/otr milk). Cocci '7 to 
 
 1 p, in length, and rod-forms. 
 They produce a yellow colour in 
 boiled milk, which at first becomes 
 acid, and then strongly alkaline. 
 They also occur on boiled potatoes, 
 carrots, etc., where they form small 
 lemon-yellow masses. 
 
 The colouring-matter is soluble 
 in water, insoluble in ether and 
 alcohol, unchanged by alkalies, de- 
 colorised by acids. It is similar 
 to yellow aniline colours, both 
 spectroscopically and in ordinary 
 reactions. 
 
 Bacterium termo (Vignal). 
 Rods 1-5 to 2 p. in length, '5 to *7 /* 
 in width. 
 
 Colonies white, surrounded by 
 liquefied gelatine. 
 
 The bacilli inoculated in the 
 depth of gelatine produce a funnel- 
 shaped area of liquefaction ; later, 
 the jelly is completely liquefied and 
 coloured green. Cultures have a 
 strong putrefactive odour. 
 
 In broth they form a white 
 deposit and colour the medium 
 green. 
 
 They were isolated from human 
 saliva. 
 
 Bacterium tholoeideum (Gess- 
 ner). Rods similar to Bacillus 
 lactis aerogenes. 
 
 Pathogenic in small animals. 
 
 They were isolated from healthy 
 human evacuations. 
 
 Bacterium ureae (Cohn). Cocci 
 1'25 to 2 p. in diam., singly or in 
 chains, and rods. The rods split 
 up by division into chains of cocci, 
 which after a time are set free. The 
 cocci increase further by subdivi- 
 sion 1 , and a jelly-like membrane 
 develops around them. Masses of 
 cocci exist m the form of irregular 
 or roundish lumps. They are 
 aerobic. 
 
 Cultivations, after twenty-four 
 hours, consist exclusively of rods ; 
 
 after forty-eight hours, of cocci 
 chains ; and in fourteen days, of 
 zopgloea : the cocci transplanted 
 into fresh nourishing solution again 
 grow into rods. These observations 
 point to the existence of a \ 
 morphic specie 
 and the former nomenclati. ; 
 
 M/v, must be regarded as 
 
 untenable. 
 
 In urine they set up ammoniacal 
 fermentation, converting urea 
 carbonate of ammonia. Rods. 
 long and 1 p. wide, have been iso- 
 lated from stale urine (Bacilli^ 
 urese, Leube), which also mo*t 
 energetically cause the ammon 
 fermentation of urine. 
 
 Bacterium ureae (Jaksch). 
 Rods 2 pin length, 1 p. in width. 
 
 Colonies on gelatine semi-trans- 
 parent. 
 
 Inoculated in the depth of gela- 
 tine the i>;u-illi form a delicate 
 branching growth in the track o 
 the needle. 
 
 They convert urea into carbonate 
 of ammonia, ami cultures smell of 
 herring brine. 
 
 They occur in ammoniacal urine. 
 
 Bacterium yiolaceum (Bt-rgon- 
 zini). Rods similar to Bacterium 
 termo, - G to 1 p. thick long. 
 
 They occur on white of egg, 
 forming a violet pigment. 
 
 Bacterium Zopfii (Kurth). 
 
 Cocci, 1 to l'2f> p. in iliani. : rods 
 and threads. Cultivated in a streak 
 on nutrient gelatine spread out on 
 a glass slide, a peculiar develop- 
 ment takes place. In t\v 
 hours after inoculation threads 
 have developed; in forty eight 
 hours windings .>f tin- threads 
 are observed, and in six day- 
 threads ha vi- broken up into cocci. 
 They were observed in the ink 
 of fowls, especially in the contents 
 of the vermiform appcn 
 oculation of rabbits was followed 
 byn-_ lite. Identical with 
 
 Bacillus ftf 
 
 BeggiatoaalbaiViiui i 
 
 1-o.ls, spiraN :inl tlir.-aoS i 
 Tin- ' 
 
 laU.l 
 
 less; their protoplasm contains 
 
-542 
 
 DESCRIPTION OF SPECIES. 
 
 numerous strongly refractive gran- 
 ules consisting of sulphur. They 
 occur as greyish or chalk-white 
 gelatinous threads, 3 to 3 '5 p 
 thick, in sulphur springs and 
 marshes. 
 
 Beggiatoa mirabilis (Cohn). 
 Threads distinguished by their 
 breadth, which may reach 30 /j. 
 They are motile, bent and curled 
 in various ways, and rounded at 
 the ends. Around the threads, 
 isolated cells have been observed, 
 
 families, bound together by gela- 
 tinous substance. Later they be- 
 come larger, globular or ovoid in 
 shape, and hollow, containing 
 watery fluid in their interior. The 
 families reach a diameter of 660 /z, 
 in which the cocci form simply a 
 peripheral layer. The hollow fami- 
 lies or' vesicles are often perforated, 
 presenting a delicate reticulated ap- 
 pearance, which finally may become 
 broken up into irregular structures. 
 The red colouring-matter can be 
 
 FIG. 214. BACTERIUM ZOPFII. SUCCESSIVE CHANGES IN THE SAME THREAD, 
 x 740. , A thread form ; b, breaking up into rod forms ; c, into cocci (Kurth). ' 
 
 macrococci, but spiral forms are as 
 yet unknown. The threads are 
 filled with sulphur granules. They 
 occur in sea-water, forming a white 
 gelatinous scum on decomposing 
 algae. 
 Beggiatoa roseo-p ersicina 
 
 {Colinia rowo-jH-rx/ciiHt. Bacterium 
 rubescens, or Peach-coloured bac- 
 terium, Lankester). Cocci, rods, 
 spirals, and threads (Fig. 21 6). The 
 cocci, globular or oval, reach 2'5 jj. 
 in diam. They form at first solid 
 
 distinguished from other red pig- 
 ments, and it is designated by the 
 name bacterio-piu-purm. It is quite 
 distinct from the pigment produced 
 by Micrococcus prodigiosus, being 
 peach-blossom red, and insoluble 
 in water, alcohol, etc. Examined 
 spectroscopically, it shows a strong 
 absorption in the yellow, and a 
 weaker band in the green and blue, 
 as well as a darkening in the more 
 refrangible half of the spectrum. 
 In the cocci, especially of the older 
 
DESCRIPTION 01- SPEl IKS. 
 
 , dark granules are to be 
 seen, which consist of sulphur. 
 The micro-organisms occur on the 
 surface of marshes, or on water in 
 which algae are rotting. They 
 form a rose-red, blood-red, violet- 
 red, or violet-brown scum ; and 
 .sometimes in such quantity that 
 
 Cladothrix dichotoma (Cohn). 
 Threads resembling those of lep- 
 tothrix ; slender, colourless, not 
 articulated, straight or slightly 
 undulated, and in places twisted 
 in irregular spirals with pseudo- 
 branchings. The development can 
 be traced from the cocci to rods and 
 
 Kit;. i'i:>.-HK..i;iATOA ALBA. 
 
 
 
 whole marshes and ponds may be 
 coloured blood-red by them. 
 
 Spirillum sanguineum, 
 violaceum,mona^vin.-uun<H>kenii. 
 
 and Rhabdomonas rosea are 
 <ibly phase-forms of Beggiatoi 
 i oseo-persicina. 
 
 jreftds; imniou 
 
 x .M 
 
 threads. The latter are at the 
 beginning simple threads, wni 
 were formerly described as Lepto 
 if coloured 
 
 mu.iv.j.Kition with imn. 
 
 thrix ' 
 
 false branches by single rod. turning 
 
544 
 
 DESCRIPTION OF SPECIES. 
 
 aside, which by repeated division 
 lengthen into threads. A thread 
 appears to be first composed of 
 long rods, then of short rods, and 
 lastly of cocci. The iodine reaction 
 must be applied to distinguish these 
 forms, especially when the sheath 
 of the threads has a yellow, rust- 
 red, olive-green, or dark brown 
 coloration. The cocci may grow 
 into rods while still in the sheath, 
 and finally become leptothrix 
 threads, surrounded by a delicate 
 gelatinous sheath, from which the 
 
 media small tufts, about 1 to 3 /u r 
 and floating masses. 
 
 Cladothrix Forsteri (vide Strqi- 
 tothrix Forsteri, Cohn). 
 
 Cladothrix intricata (Russell). 
 Rods and filaments. 
 
 Colonies are composed of a net- 
 work of twisted threads. 
 
 Inoculated in the depth of gela- 
 tine fine filaments spread out from 
 the track of the needle, and the 
 gelatine is liquefied. 
 
 Grown on agar the filaments 
 penetrate the jelly. 
 
 FIG. 216. PHASE-FORMS OF BEGGIATOA ROSEO-PERSICINA (WARMING). 
 
 false branching proceeds. Frag- 
 ments may break off, which are 
 actively motile, and appear as 
 vibrios, spirilla, and spirochseta- 
 forms. They may also occur in 
 zoogloea (Fig. 217). 
 
 They are the commonest of all 
 bacteria in both still and running 
 water, in which organic substances 
 are present. They are observed 
 also in the waste water of certain 
 manufactures, such as sugar. Arti- 
 ficially they can be cultivated on 
 infusions of rotting algae and ani- 
 mal substances, forming on these 
 
 In broth the growth is abundant. 
 
 They were isolated from sea 
 dredgings. 
 
 Cladothrix invulnerabilis (Ac- 
 osta, y Grande Rossi).- Filaments 
 which produce in gelatine a white 
 thread, and liquefy it very slowly. 
 
 On potato the growth is abundant 
 and chalky in appearance. 
 
 In milk they form a firm yel- 
 lowish pellicle ; and in broth and in 
 water the growth is abundant. 
 
 They occur in water. 
 
 Clostridium butyricum (vidr 
 Bacillus bntyriciis). 
 
DESCRIPTIOX OF SPECIES. 
 
 546 
 
 Clostridium foetidum (Libo- 
 nus). Rods 1 p. in width, singly and 
 in filaments. Spore-formation re- 
 sembles that of Bacillus butyricus. 
 They are anaerobic. 
 
 Colonies rapidly liquefy gelatine. 
 
 gas-formation with unpleasant smell 
 and splitting up of ti 
 They were iv-ht,.,! ( ,,,;, 
 Crenothrix Kuhniana (Raben- 
 norst). Cocci, rods, and thread- 
 forms. Ti,, cocci are globular, 
 
 FIG. 217. CLADOTHRIX DICHOTOMA. 
 
 A. Branching schizomycete : (a) Vibrio-form ; (b) Spirillum-form [slightly mag- 
 nified J. 
 
 B. A screw-form with () Spirillum-form; (//) Vil.rio-f.irm. 
 < . Long spirochaeta-form. 
 
 !> Fragment with spirillum-f on i. n at tli- i>tln-r. 
 
 K. Screw-forms: (a) cont in : <.mposedofi 
 I . .^pirochaeta-form: (a)continuo 'tig rocfs; ( 
 
 (d) cocci (Zopf). 
 
 On agar the colonies form branch- 
 ing processes resembling colonies of 
 Bacillus oedematis maligni. 
 
 Inoculated in the depth of gela- 
 tine liquefaction spreads from be- 
 low upwards. There is abundant 
 
 1 to 6 ^ in dim I threads are 
 
 colourless. 1 - 1<> .'. ^ tin. L 
 
 shaped at the 
 
 a diam. <>; 
 
 form colonies with a 1 
 
 green, or dark-brown to brown-black 
 
546 
 
 DESCRIPTION OF SPECIES. 
 
 coloration, caused by impreg- 
 nation with oxide of iron. The 
 threads are distinctly articulated, 
 and ensheathed. The segments are 
 
 set free when the sheath bursts, 
 and develop into new threads. In 
 other cases the segments remain 
 enclosed, and subdivide into discs, 
 
 FIG. 218. CRENOTHRIX KUHNIANA. 
 
 a, 6, c, d, e. Cocci in various stages of fission, x 600. 
 
 /. Zoogloea of cocci, x 600. 
 
 rj. Various forms of zoogleea, natural size. 
 
 h. Colony of threads composed of rods grown out of a zoogloea of cocci. 
 
 i r. Thread-forms ; some straight, others spiral, with more or less differentia- 
 tion between base and apex, (r) is composed of short rods at the base, and above 
 these of cylindrical segments, and at the apex these segments have divided into 
 cocci, x 600 (Zopf). 
 
DESCRIPTION OF SPE< IBS. 
 
 .-.17 
 
 which, by vertical fission, break up 
 into globular forms (cocci). These 
 again develop into new threads, 
 either within the sheath, eventually 
 penetrating it, or after they are set 
 free. 
 
 The micro-organism appears in 
 little whitish or brownish tufts in 
 wells and drain-pipes, and it not 
 only renders drinking-water foul, 
 but may stop up the narrower 
 pipes. 
 
 Diplococcus albicans ampins 
 (Bumm). Cocci resembling gono- 
 cocci but much larger, singly and 
 in tetrads. 
 
 Colonies are prominent and 
 greyish-white. 
 
 In the depth of gelatine they 
 produce a greyish-white growth in 
 the track of the needle and on the 
 free surface. They slowly liquefy 
 the gelatine. 
 
 They were obtained from the 
 vaginal mucous membrane. 
 
 Diplococcus albicans tardis- 
 simns (Bumm). Cocci morpho- 
 logically identical with gonococci. 
 They grow extraordinarily slowly 
 on gelatine. 
 
 They form very minute colonies, 
 which are opaque and greenish- 
 brown in colour. 
 
 Inoculated in the depth of gela- 
 tine isolated greyish-white colonies 
 develop in the track of the needle, 
 and on the free surface a thin, 
 white, waxy film with dentated 
 edge. 
 
 On agar the growth is very 
 similar. 
 
 They were isolated from the 
 vaginal mucous membrane. 
 
 Diplococcus citrens conglome- 
 ratns (Bumm). Cocci in pairs 
 resembling gonococci, 1 '5 \i in diam., 
 in tetrads and in masses. 
 
 Colonies lemon-yellow ; irregulai 
 in form. 
 
 Inoculated in the depth of gela- 
 tine the cocci develop in the t 
 of the needle, and liquefaction com- 
 mences at its upper part. 
 
 The growth on the free surface 
 is yellow, and floats on the liquefied 
 gelatine or subsides to the bottom 
 of the liquefied area. 
 
 They are present in gonorrhoaal 
 
 ms in air ami in 
 
 Diplococcus citrens liqne- 
 faciens (Unna). Oval ooooi 4 to 
 1 p, in liain.. in ; radn, 
 
 short chains, and masses. 
 
 Colonies appear in the form of 
 circular discs, at first greyish-white, 
 later lemon-yellow. They are finely 
 granular, and have sharply defined 
 contours. 
 
 Inoculated in the depth of gela- 
 tine, at the end of a week the 
 growth is found on the free 
 surface, forming a shining yellow 
 layer; in two weeks liquefaction 
 commences, and the growth floats 
 on the liquefied gelatine which is 
 also yellowish and turbid. 
 
 On the surface of agar a yellow- 
 ish-! >rown layer is rapidly formed. 
 The appearance is similar on 
 potato. 
 
 They were isolated in cases of 
 eczema seborrhoeicum. 
 
 Diplococcus coryzae (Hajek).- 
 Large diplococci. 
 Colonies are white, prominent. 
 Inoculated in the depth of gela- 
 tine the growth resembles the 
 pneumococcus. 
 
 On agar a white layer is formed. 
 They are probably identical with 
 Friedliinder's pneumococci. 
 
 They were isolated from the 
 mucus in acute nasal catarrh. 
 
 Diplococcus flavns liquefaciens 
 tardns. Cocci reMnbnog gono- 
 cocci. 
 
 Colonies are circular, tuning, 
 and chrome-yellow in colour. 
 
 Inoculated in the depth of gela- 
 tine a yellowish growth o< 
 along the needle track, and also 
 on the free surface. In 
 the surface is depressed, bvt 
 ge la t liquefied until about 
 
 two months have elapsed. 
 
 On agar a yellow i-h white layer 
 is formed, and on potato the colour 
 is more pronounced. 
 
 They were isolated from the 
 skin in eczema selwirhceicum. 
 
 Diplococcus fluorescens fceti- 
 dus (klainann ). Cocci in pairsand 
 
 OoiooiM circular, forming a 
 
548 
 
 DESCRIPTION OF SPECIES. 
 
 brownish deposit surrounded by 
 liquefied gelatine which has a violet 
 or greenish tinge. 
 
 Inoculated in the depth of gela- 
 tine they produce liquefaction along 
 the track of the needle, with a 
 hemispherical excavation of the 
 gelatine at the upper part. An 
 iridescent film floats on the surface 
 and a greenish sediment forms at 
 the bottom of the liquefied area. 
 
 On agar the layer is brownish. 
 
 On potato the growth is granular, 
 and the potato in the vicinity has 
 a bluish colour. 
 
 They were cultivated from the 
 nasal mucus. 
 
 Diplocqccus inter cellularis 
 meningitidis (Weichselbaum). 
 Cocci singly, in pairs, tetrads and 
 masses. They grow at 37C. 
 
 Colonies on agar are granular and 
 yellowish-brown. 
 
 On the surface of agar they form 
 a greyish-white viscid growth. In 
 the depth of agar the growth only 
 occurs in the upper part of the 
 needle track. 
 
 On blood serum and broth there 
 is very little growth, and none on 
 potato. 
 
 Cultures quickly lose their 
 vitality. 
 
 They are pathogenic in mice, 
 guinea-pigs, rabbits, and dogs. 
 
 They were isolated from the exu- 
 dation in cases of cerebro-spinal 
 meningitis, and were observed in 
 the interior of pus cells. 
 
 Diplpcoccus luteus (Adametz). 
 Cocci 1*2 to 1'3/M in diam., singly 
 and in chains. Motile. 
 
 Colonies are circular and slightly 
 yellow, and granular. Old colonies 
 are bright yellow. 
 
 On the surface of gelatine a 
 growth occurs in concentric circles 
 of a lemon-yellow colour, and the 
 gelatine is coloured reddish-brown. 
 After several weeks liquefaction 
 sets in. 
 
 On agar a yellow layer forms, and 
 the jelly is coloured reddish-brown. 
 
 On potato the growth changes 
 from yellow to brown. Milk is 
 coagulated. 
 
 They were obtained from water. 
 
 Diplococcus of pneumonia in 
 horses (Schutz). Oval cocci, singly 
 or in pairs, capsulated. 
 
 Colonies small and white. 
 
 In the depth of gelatine a row of 
 colonies develops in the track of the 
 needle. 
 
 On agar the growth is composed 
 of transparent droplets. 
 
 Injection into the lung is said to 
 produce pneumonia, ending fatally 
 in eight or nine days. 
 
 They are pathogenic in rabbits, 
 guinea-pigs, and mice. 
 
 They were isolated from the 
 lungs of a horse suffering from 
 pneumonia. 
 
 Diplococcus roseus (Bumm). 
 Cocci identical in description with 
 gonococci. 
 
 Colonies are pink, granular, and 
 irregular in form. 
 
 Inoculated in the depth of gela- 
 tine the cocci grow freely in the 
 track of the needle and on the 
 surface, developing a pink colour 
 and slowly producing liquefaction. 
 
 They are present in the air. 
 
 Diplococcus subflavus (Bumm). 
 Diplococci similar to gonococci. 
 
 Colonies greyish-white, later 
 yellow. 
 
 They grow in gelatine and on 
 blood serum, and liquefy broth. 
 
 They produce suppuration when 
 injected subcutaneously in man. 
 
 They were isolated from lochial 
 discharges, the vesicles of pem- 
 phigus, and from the secretion in 
 colpitis in children. 
 
 They stain by Gram's method. 
 
 Haematococcus bovis (Babes). 
 Cocci oval, singly, in pairs, and in 
 masses. 
 
 Inoculated in the depth of gela- 
 tine minute colonies develop in the 
 track of the needle. 
 
 On agar the growth is composed 
 of transparent droplets. 
 
 On potato a yellowish shining 
 film is formed. 
 
 On blood serum the growth is 
 similar to that on agar. 
 
 They produce a fatal result in 
 rabbits and guinea-pigs in a week 
 or ten days. 
 
 They were isolated from the 
 
DESCRIPTION OF SPECIES. 
 
 549 
 
 blood and organs of cattle which 
 died of an epidemic disease asso- 
 ciated with hfemoglobinuria. 
 
 Helicobacterium aerogenes 
 (Miller). Bacilli singly, in chains 
 and long wavy filaments. Motile. 
 
 Colonies whitish, varying in form. 
 
 Inoculated in the depth of gela- 
 tine the bacilli give rise to a faintly 
 yellow growth in the track of the 
 needle, and an almost invisible, 
 rapidly growing layer on the 
 surface. 
 
 On potato the growth is dry and 
 brownish. 
 
 They were isolated from the 
 healthy intestinal tract. 
 
 Leptothrix buccalis (Robin). 
 Long, thin threads, -7 to 1 /x broad, 
 colourless, often united in thick 
 bundles or felted together. Masses 
 of cocci occur with the threads, 
 and the threads themselves are com- 
 posed of long rods, short rods, and 
 cocci. The threads may break up 
 into spiral, vibrio, and spirochaeta 
 forms. The last-named occur in 
 large numbers in the mouth, and 
 have been named Sj>ti-m-lifrfn &///- 
 calls. Leptothrix buccalis is found 
 in teeth slime, and is believed 
 to be intimately connected with 
 dental caries. The threads pene- 
 trate the tissue of the teeth, after 
 the enamel has been acted upon by 
 acids generated by the fermentation 
 of food. The short rods, long rods, 
 cocci, leptothrix-forms, and screw- 
 forms are found in the dental 
 canals. 
 
 The threads of Leptothrix buc- 
 calis have a special staining reaction 
 (Leber). They become coloured if 
 placed in an acid medium with 
 iodine ; if the medium be alkaline, 
 it must first be acidified with very 
 dilute hydrochloric acid or acetic 
 acid. The contents are stained 
 violet, and contrast with the sheath 
 and septa, which remain uncoloured. 
 
 Leptothrix buccalis (Vignal). 
 Rods 1 to 1-5 n in width, 1-6 to 
 30 fi in length. 
 
 Colonies greyish-white, promi- 
 nent and furrowed. 
 
 Inoculated in the depth of gela- 
 tine a filament forms in the track 
 
 of the needle, and a growth occurs 
 on the free surface. Liquefa* 
 
 " at the upper part, forming 
 a cup-shaped cavity, and a I.I 
 skin float- upon the liquid, 
 liquefaction gradually extends to 
 the side of the tube, and a deposit 
 is formed at the bottom of the 
 liquefied gelatine. 
 
 On agar the layer is white, 
 wrinkled and transparent, and later 
 yellowish. 
 
 In broth there is turbidity, but 
 no skin on the surface. 
 
 On potato the growth is greyish- 
 white. 
 
 They are occasionally present 
 in the mouth in health, and 
 are possibly identical with lepto- 
 thrix buccalis ( Robin). 
 Leptothrix gigante i 
 Long rods, short rods and cocci 
 can be observed in the same thread. 
 There are also screw-threads, which 
 may take the form of spirals, 
 vibrios, or spirochsetae. The threads 
 increase in diameter from base 
 to apex ; corresponding with the 
 thickness of the threads, the rods 
 and cocci show different dimensions. 
 They have been observed in the 
 diseased teeth of dogs, sheep, oats 
 and other animals. 
 
 Leuconostoc inesenteroides, 
 Cienkowski ('/ 
 
 FrnscJihiifli/"!'.. < : /'). 
 
 Cocci and rods singly, in chains, 
 and in zooglcea, surrounded bv a 
 thick gelatinous envelope. The I 
 history has been very thoroughly 
 investigated. The spore*. 
 J /* in diam., are >: 1 or 
 
 ellipsoidal form, with thick mem- 
 brane and shining contents. The 
 outer membrane-layer bursts, and a 
 middle lamella oozes out, and forms 
 a thick gelatinous envelope, v 
 
 remains adherent 
 to the pla c spore- 
 
 germination leads to the forma 
 of a coccus with a gelatinous 
 velope. The coccus then elongates 
 into a short rod-form, and the 
 lope becomes tffip 
 soidal The rod divides into two 
 cocci, and hese lengthens 
 
 into a rodand divide*. Byrepet: 
 
550 
 
 DESCRIPTION OF SPECIES. 
 
 of this process a chain of cocci 
 results, encased in a cylindrical 
 or ellipsoidal envelope. The chains 
 increase in length, become twisted 
 up, and eventually fall apart into 
 pieces of various lengths. 
 
 In nourishing liquids a great 
 number of little masses are formed, 
 which adhere together, and produce 
 pseudo-parenchymatous structures. 
 These latter may join together, 
 forming still larger agglomerations. 
 
 This micro-organism occurs occa- 
 sionally in beet-root juice and the 
 molasses of sugar-makers, forming 
 large gelatinous masses resembling 
 frog-spawn. The vegetation is so 
 rapid that forty-nine hectolitres of 
 molasses, containing 10 per cent, 
 of sugar, were converted within 
 twelve hours into a gelatinous 
 mass ; consequently, it is a for- 
 midable enemy of the sugar manu- 
 facturers. 
 
 FIG. 219. LEUCONOSTOC MESENTEROIDES. 
 
 1. Spores. 2. Spores after germination, showing gelatinous envelope. 3, 4, 5, 6. 
 Increase by division. 7. Glomerular form of zooglcea. 8. Section of an old 
 mass of zooglcea. 9. .Cocci chains with arthrospores (Tieghem and Cien- 
 kowski). 
 
 The masses of zooglcea are of \ 
 
 almost a cartilaginous consistency, I 
 
 and admit of sections being made | 
 
 with a razor. After a long time ! 
 
 the envelope liquefies, and the cocci ! 
 
 are set free ; the latter introduced j 
 
 into fresh nourishing media develop j 
 
 new colonies. In the chains some | 
 
 of the cocci become enlarged with- i 
 
 out changing their form. These j 
 acquire the properties of spores, 
 and 
 
 Micrococcus acidi lactici 
 
 (Marpmann). Large cocci, singly 
 and in pairs. 
 
 Colonies yellowish-white. 
 
 On the surface of gelatine the 
 cocci produce a yellow layer. 
 
 They grow in milk, producing a 
 reddish colour, and coagulation due 
 to the formation of lactic acid. 
 
 They were isolated from milk. 
 
 Micrococcus acidi lactici lique 
 faciens (Kreuger). Cocci oval, 1 
 
DESCRIPTION OF SIM 
 
 551 
 
 to l-r fj. in diam.. in pairs and in 
 tetrads. 
 
 Colonies white. 
 
 Inoculated in the depth of gela- 
 tine the cocci produce a granular 
 filament, changing in two or three 
 lays to liquefaction in the form of 
 a funnel ; later, a wrinkled mem- 
 brane floats on the surface of the 
 liquid. 
 
 In milk they produce lactic acid. 
 
 They were isolated from butter 
 which had turned cheesy. 
 
 Micrococcus aerogenes (Miller). 
 Oval cocci. 
 
 Colonies are dark and regular in 
 contour, but have a peculiar spotted 
 appearance. 
 
 Inoculated in the depth of gela- 
 tine a brownish-yellow growth 
 occurs along the track of the needle, 
 and on the free surface a white 
 button-like elevation. After a time 
 the gelatine is slowly liquefied. 
 
 On agar a yellowish-white pulpy 
 layer forms, and a similar growth 
 appears on potato. 
 
 They resist the action of acids, so 
 that the presence of gastric juice 
 does not impede their develop- 
 ment. 
 
 They were obtained from the 
 intestine. 
 
 Micrococcus agilis (Ali-Cohen). 
 Cocci 1 AI in diam.. singly, in 
 pairs, tetrads, and in chains. They 
 are motile, and possess flagella. 
 
 Inoculated in the depth of gela- 
 tine they grow in the track of the 
 needle, and produce, after two or 
 three weeks, liquefaction or excava- 
 tion of the jelly. 
 
 On agar and potato the growth 
 is pink. 
 
 They occur in water. 
 
 Micrococcus agilis citreus 
 (Menge). Cocci in pairs, chains 
 and masses. They are motile, and 
 each coccus possesses a single fla- 
 gellum. 
 
 Colonies appear surrounded by 
 clouded gelatine. 
 
 Inoculated in the depth of gela- 
 tine there is a scanty growth in the 
 track of the needle, and on the sur- 
 face a bright yellow patch. 
 
 On agar they form a yellow layer, 
 
 which is viscid, and may be drawn 
 out in long threads. 
 
 In broth they produce cloudiness 
 and a viscous deposit. 
 
 The growth on potato is bright 
 yellow. 
 
 Milk is not coagulated. 
 
 They were isolated from an in- 
 fusion of peas. 
 
 Micrococcus albus liquefaciens 
 (Besser). Large cocci in chains and 
 in masses. They are anaerobic. 
 
 Colonies on agar exhibit concen- 
 tric rings of different shades of 
 brown. 
 
 Inoculated in the depth of gela- 
 tine they produce liquefaction in 
 the track of the needle. 
 
 They occur in mucus from the 
 nose. 
 
 Micrococcus amylivorus (Bur 
 rill). Oval cocci, 1 to 1-4 /i long, 
 7 p. broad, singly, in pairs, and rarely 
 in fours, never in chains, are found 
 embedded in an abundant mucilage 
 which is very soluble in water. 
 
 They have been described as pro- 
 ducing the so-called "fire blight" 
 of the pear tree and other plants. 
 
 Micrococcus aquatilis < Bolton). 
 Small cocci in masses. 
 
 Colonies circular, prominent, and 
 pure-white. 
 
 Inoculated in the depth of gela- 
 tine, there is a white growth in the 
 track of the needle and also on the 
 free surface. 
 
 On agar the growth is white. 
 
 They occur in \\ 
 
 Micrococcus aquatilis invisi- 
 bilis ( Vaughan). Cocci oval. 
 
 Colonies brown. 
 
 In gelatine there is a slight 
 growth in tin- track of the needle, 
 and a more abundant growth on 
 the free surface. 
 
 On agar they form a white film. 
 
 On potato the growth is in- 
 visible. 
 
 They occur in water. 
 
 Micrococcus aurantiacus 
 (Cohn). Coeci spherical or o 
 1*3 to \-itp in diam., singly, in pairs, 
 aii'l in groups. 
 
 Colonies orange-yellow. 
 
 Inoculated in gelatin- ih -y form 
 minute colonies in the track of the 
 
552 
 
 DESCRIPTION OF SPECIES. 
 
 needle, and a prominent hemi- 
 spherical yellow growth on the free 
 surface. 
 
 On agar the growth is orange- 
 yellow, and on potato yellow and 
 slimy. 
 
 They occur in water. 
 
 Micrococcus botryogenus 
 (Johne, Rabe). Cocci 1 to 1-5 /x in 
 diam., in wavy chains. 
 
 Colonies circular, sharply defined. 
 At first silver-grey, later yellowish- 
 grey with metallic lustre, they 
 produce an odour like that of 
 strawberries. 
 
 Inoculated in the depth of gela- 
 tine a greyish-white filament de- 
 velops, with slight liquefaction of 
 the gelatine ; later, it becomes 
 milk-white, and at its upper part a 
 characteristic bubble appears. 
 
 They make hardly any growth 
 on agar. 
 
 On potato they grow very abun- 
 dantly, forming a yellowish layer 
 with the same odour as the colonies 
 on plate cultivations. 
 
 Inoculated guinea-pigs die of 
 septicaemia ; in sheep and goats 
 severe inflammation spreads from 
 the point of inoculation. Mice are 
 immune. In horses an inflamma- 
 toTj oedema is at first set up, 
 followed in four to six weeks by 
 the formation of new growths, 
 which sometimes suppurate and 
 contain large numbers of micro- 
 cocci. 
 
 They were found in "tumours of 
 the spermatic cord and of the 
 connective tissue in other parts in 
 horses. 
 
 Micrococcus candicans 
 (Fliigge). Cocci which collect in 
 masses. 
 
 In plate-cultivations they form 
 in two or three days milk-white 
 colonies ; while those below the 
 surface of the gelatine are yellow- 
 ish. Under a low power the deep 
 colonies are quite circular, with 
 smooth margins, of a blackish-brown 
 colour, and very slightly granular ; 
 the superficial colonies are quite 
 irregular in outline, and are finely 
 granular. 
 
 Cultivated in test-tubes they form 
 
 a white nail-shaped cultivation. 
 They were isolated from contami- 
 nated plate-cultivations. 
 
 They occur in the air. 
 
 Micrococcus candidus(Cohn). 
 Cocci forming snow-white points 
 and spots upon slices of cooked 
 potato. 
 
 Micrococcus carneus (Zimmer- 
 mann). Cocci '8 /* in diam., occur- 
 ring in masses. 
 
 Colonies circular, greyish-white, 
 with the centre tinged with red. 
 
 Inoculated in the depth of gela- 
 tine they form a white, granular 
 filament in the track of the needle, 
 and a pale pink layer on the free 
 surface. 
 
 On the surface of oblique gela- 
 tine a flesh-coloured layer develops, 
 which later assumes a violet colour. 
 
 On agar the growth is similar. 
 
 On potato the growth is abundant 
 and red in colour. 
 
 They were isolated from water. 
 
 Micrococcus cerasinus siccus 
 (List). Cocci -25 to '12 ^ in diam., 
 singly and in pairs. They can best 
 be cultivated at 37 C. 
 
 On agar they form a cherry-red 
 layer, and a similar growth on 
 potato. 
 
 The pigment is insoluble in alco- 
 hol, ether, and water, and is not 
 destroyed by acids or alkalies. 
 
 They occur in water. 
 
 Micrococcus cereus albus 
 (p. 178). 
 
 Micrococcus cereus flavus 
 (p. 178). 
 
 Micrococcus cinnabareus 
 (Fliigge). Large cocci occurring in 
 twos, threes, and fours. 
 
 Colonies develop very slowly, and 
 are punctiform, and bright red at 
 first, and afterwards reddish-brown. 
 
 The cocci inoculated on the sur- 
 face of gelatine form a heaped-up, 
 red-coloured growth. 
 
 They were found contaminating 
 old cultivations. 
 
 Micrococcus citreus (List). 
 Cocci 1'5 to 2 - 2 p. in diam., singly, 
 in pairs and chains. 
 
 Colonies are irregular in form, 
 moist and shining, and yellowish in 
 colour. 
 
DESCRIPTION OF >ll < DBS, 
 
 
 In the depth of gelatine the 
 growth is very scanty. 
 
 On the surface of agar the growth 
 is yellowish. 
 
 On potato the growth is similar 
 but more abundant. 
 
 Micrococcus concentricus 
 (Zimmermann). Cocci -9/iin diam., 
 in masses. 
 
 Colonies bluish-grey. 
 
 Inoculated in the depth of gela- 
 tine there is no growth in the track 
 of the needle, but concentric rings 
 form on the free surface. 
 
 On agar the growth is greyish- 
 white and smooth. 
 
 On potato yellowish and slimy. 
 
 They occur in water. 
 
 Micrococcus cremoides (Zim- 
 mermann). Cocci -8 p in diam., 
 occurring in masses. 
 
 Colonies are spherical, granular 
 and yellowish. The margins are 
 dentated or irregular, and processes 
 extend into the surroundinggelatine. 
 
 Inoculated in the depth of gela- 
 tine liquefaction occurs in the 
 track of the needle in a few days. 
 A yellow growth floats on the 
 liquefied gelatine, and a yellowish 
 mass subsides to the bottom of the 
 liquid. 
 
 On agar a smooth shining layer 
 is formed, and on potato the growth 
 is abundant. 
 
 They occur in water. 
 
 Micrococcus crepusculum 
 (Cohn. Mowu crcpwcuktm, Ehren- 
 berg. Mibrokolcke* in nmlenden 
 *<ib*tr<itf>n, Fliigge). Round or 
 short oval cells, scarcely 2 /* in 
 diam. : singly or in zoogloea. 
 
 They occur in various infusions 
 and putrefying fluids in company 
 with Bacterium termo. 
 
 Micrococcus cumulatus tenuis 
 (Besser). Large cocci, oval : in 
 masses. 
 
 Colonies on agar have a brown 
 nucleus. 
 
 Inoculated in the depth of gela- 
 tine they form a white filament, 
 and on the surface a transparent 
 laver. 
 
 "in broth there is an abundant 
 deposit, and the supernatant liquid 
 is clear. 
 
 They occur in mucus from tin- 
 nose. 
 
 Micrococcus endocarditidis ru 
 gatus (Wrirhsrlbaum). Cocci re- 
 sembling pyogenic staphyloco. 
 
 ColoMH-v h;ivr ;i In-own or M'lloW- 
 
 i-h brown nucleus. 
 
 In the depth of agar there 
 slight growth in the track of th.- 
 needle, and a wrinki layer 
 
 on the surface. 
 
 On potato the growth is dry and 
 brownish. 
 
 On blood serum th growth i 
 colourless and adher> 
 
 Injectedsubcutan.-ou-ly.thry pro- 
 duce, in rabbits, local swelling and 
 redness, and suppuration in guinea- 
 pigs. Injected into the veins I 
 injury to the aortic valves, they 
 produce endocarditis 
 
 They were isolated from a case 
 of ulcerative endocarditis 
 
 Micrococcus fervidosus (Ada- 
 metz). Cocci '6 p. in diam., in pairs 
 and in 
 
 The deep colonies are pale-yellow, 
 and look like watery droplets 
 superficial colonies are granular and 
 irregular with jagged edges. 
 
 Inoculated in the depth <>f gela- 
 tine a granular filament develoi 
 the track of the needle, and on the 
 surface a circular patch with den- 
 tated margin. 
 
 On agar the growth is whit, 
 slimy, and on potato greyish - 
 white. 
 
 They occur in 
 
 Micrococcus Finlayensis 
 nberg). Cocci ~< to 7 ^ in 
 diam., singly, in pairs, tetrads, and 
 in masses. 
 
 In the depth of gelatine they 
 produce a growth in th 
 the needle, with 1 
 the upper part with a pale-yellow 
 deposit. 
 
 < )n agar the growth is pale-yellow. 
 
 'I' hey were isolated from the liver 
 in a fatal case of yellow f- 
 
 Micrococcus flavus desidens 
 (Flii'_ in P*>r, 
 
 i sofa few elein 
 
 Colonies yellowish-^ 
 
 Inoculated in th. gela- 
 
 tine they grow along the track of 
 
554 
 
 DESCRIPTION OF SPECIES. 
 
 the needle, and form a yellowish- 
 brown layer at the point of punc- 
 ture. 
 
 Later liquefaction sets in, and a 
 deposit forms at the bottom of the 
 turbid liquid. 
 
 They occur in air and in water. 
 
 Micrococcus flavus lique- 
 faciens (Flligge). Cocci mostly in 
 twos and threes, also in masses. 
 
 Small yellow colonies appear after 
 two or three days, which have a 
 shallow depressed zone surrounding 
 them. Under a low power they 
 are granular and yellowish-brown, 
 with lines radiating from the centre 
 to the circumference. Later they 
 liquefy the gelatine, and coalesce. 
 
 Inoculated in the depth of gelatine 
 the cocci produce spherical yellow 
 colonies in two days along the track 
 of the needle. These become con- 
 fluent, and at the end of eight days 
 the whole of the jelly has become 
 liquid ; later the upper part becomes 
 clear, and a yellow mass subsides to 
 the bottom of the tube. 
 
 They occur in air and in water. 
 
 Micrococcus flavus tardigra- 
 dus (Fliigge). Large cocci showing 
 at times peculiar dark poles ; gener- 
 ally arranged in masses. 
 
 Colonies develop slowly ; the 
 superficial ones have a smooth wax- 
 like surface with projecting centre ; 
 those below the surface are of a 
 dark chrome-yellow colour, and are 
 round or oval. 
 
 Inoculated in gelatine the cocci 
 develop slowly along the track of 
 the needle, forming small isolated 
 colonies ; the gelatine is not 
 liquefied. 
 
 They occur in air and in water. 
 
 Micrococcus fcetidus (Klamann). 
 Cocci singly, in pairs, and short 
 chains and masses. 
 
 Colonies circular or oval, white. 
 
 Inoculated in the depth of gela- 
 tine a pure white, shining growth 
 forms in concentric circles at the 
 point of puncture, and develops a 
 brownish colour ; and liquefaction 
 occurs after a time, and extends 
 along the needle track. 
 
 A white layer spreads over the 
 surface of agar. 
 
 On potato the growth is slimy 
 and grey in colour, with a red tinge. 
 
 Cultures produce an odour like 
 that of ozaena. 
 
 They were isolated from the 
 nose. 
 
 Micrococcus foetidus (Rosen- 
 bach). Small oval cocci. 
 
 Cultivated in agar-agar they 
 develop gas-bubbles and a foetid 
 odour. They were isolated from 
 carious teeth. 
 
 Micrococcus Fr eudenr eichi 
 i (Guillebeau). Large cocci, singly 
 I and in chains. 
 
 Colonies are granular and puncti- 
 j form. 
 
 In broth turbidity is produced, 
 j and, later, a flocculent deposit. 
 
 On potato a shining film develops, 
 yellowish or brownish-yellow in 
 colour. 
 
 In milk the cultures become 
 viscous, and can be drawn out into 
 threads several yards in length. 
 
 They were isolated from milk 
 with viscous fermentation. 
 
 Micrococcus fuscus (Maschek). 
 Cocci oval. 
 
 Colonies pale-brown or black. 
 
 Inoculated in the depth of gela- 
 tine there is a slight growth along 
 the track of the needle, and a brown 
 layer forms on the surface followed 
 by liquefaction. 
 
 On potato the growth is brown 
 or brownish-black and slimy. 
 
 Cultures give off an odour of 
 putrefaction. 
 
 They occur in water. 
 
 Micrococcus gingiyae pyogenes 
 (Miller). Large cocci, singly and 
 in pairs. 
 
 Colonies spherical, with sharp 
 contours. 
 
 Inoculated in the depth of gela- 
 tine there is an abundant growth 
 along the track of the needle and 
 on the free surface. 
 
 On agar a thick film develops, 
 with a faint tinge of purple by 
 transmitted light. 
 
 Injected into mice subcutaneously 
 
 I they produce local suppuration, and 
 
 sometimes death. Injected into 
 
 the peritoneal cavity they produce 
 
 peritonitis and death. 
 
ffnoH o 
 
 565 
 
 They were isolated from an 
 abscess of the gums. 
 Micrococcus gonorrhceae 
 
 (p. I'.IO). 
 
 Micrococcus havaniensis 
 (Sternberg). Cocci 4-5 p. in diam. 
 
 The colonies are circular and of 
 a blood-red colour. 
 
 The cocci inoculated in the depth 
 of gelatine produce a colourless 
 growth in the track of the needle 
 and a carmine patch on the surface. 
 
 On agar and on potato they form 
 a thick irregular carmine layer. 
 
 Micrococcus in Biskra-button 
 ( Hey denreich). Cocci in pairs, 
 *'i to 1 p. in length, occasionally 
 tetrads : capsulated. 
 
 Inoculated in the depth of gela- 
 tine they form a greyish-white fila- 
 ment composed of closely packed 
 colonies, and a yellowish-white film 
 on the free surface. Liquefaction 
 commences at the upper part of the 
 needle track in a few days, forming 
 a funnel which extends until, in 
 two weeks, the gelatine is com- 
 pletely liquefied. 
 
 On the surface of agar a shining 
 white or yellowish-white layer de- 
 velops in twenty-four hours. 
 
 On potato the growth is similar. 
 
 Inoculations are said to produce 
 in rabbits, dogs, fowls, sheep and 
 horses a morbid condition of the 
 skin similar to the disease known as 
 Biskra-button or Pendjeh sore. In 
 man they produce suppuration when 
 rubbed on the skin. 
 
 They were isolated from the 
 disease known as Pendjeh sore, 
 Biskra-button or cl<> <!'- />/'*/./. 
 
 Micrococcus in gangrenous 
 mastitis in sheep. Cocci singly, 
 in pairs, and in masses. 
 
 Colonies are spherical, white, and 
 under a low power have a brown 
 nucleus and transparent margin. 
 
 The cocci inoculated in the depth 
 of gelatine produce a conical area 
 of liquefied jelly with a copious 
 white deposit. 
 
 On agar they produce a white 
 layer, which later turns yellowish 
 in colour. 
 
 On potato they form a greyish 
 growth. 
 
 Injected into the mania ry . 
 of sheep they produce inflammatory 
 oedema, and a fatal 
 twenty-four to forty-eight hours. 
 
 1 n rabbits they are pyogenic. 
 
 They were isolated from the milk 
 in cases of gangrenous masti; 
 
 Micrococcus in infectious 
 p e u r o-pneumonia (IWN ami 
 Nolen) p. _'}_!. 
 
 Micrococcus in influenza 
 chel). Cocci from 1 to 1 _>.', M in 
 diam., singly, in pairs, and chains. 
 
 Extremely minute colonies appear 
 in three days. 
 
 Inoculated in the depth of gela- 
 tine a milk-white filament forms 
 along the track of the needle. L \- 
 faction commences in four 
 at the upper part, and extends 
 slowly. 
 
 On agar the colonies are pure- 
 white. 
 
 On potato the growth is yellowish- 
 white. 
 
 They do not grow on blood serum 
 or in milk. 
 
 Intravenous injection in dogs is 
 said to produce symptoms like 
 dvtemper. 
 
 They were obtained from the 
 blood in cases of influenza. 
 
 Micrococcus in influenza 
 (Kircbner). Cocci in pairs and 
 chains ; capsulated. They grow at 
 37 C. 
 
 The colonies are transparent, 
 whitish. 
 
 On the surface of agar thti 
 an abundant growth, but it is 
 limited in the depth ol 
 
 Inoculation experiment*! were 
 inconclusive. 
 
 They were obtained from the 
 sputum in cases of \\\\\\\- 
 
 Micrococcus in pemphigus 
 
 . Cocci > kO 1 M in 
 
 diam., singly and in pairs ; id- 
 eal with Staphylococcus pyogenas 
 
 The cocc '1* 
 
 are said to have produced bull. 
 
 They were isolated from pem- 
 phipoid bulUe in children. 
 
 Micrococcus in pemphigus 
 (Demme). Cocci -x to 1-4 M in 
 
556 
 
 DESCRIPTION OF SPECIES. 
 
 diam., singly, in pairs, and in 
 masses. They can be cultivated 
 at 37 C. 
 
 The colonies on agar are milk- 
 white and prominent. Later, off- 
 shoots occur from the margin, 
 producing a resetted appearance. 
 
 Inoculated in the depth of 
 gelatine the cocci form clubbed or 
 stalactitic out-growths from the 
 filament which develops in the 
 track of the needle. 
 
 On the surface of agar a creamy 
 layer is formed with similar off- 
 shoots. 
 
 Injected into the lungs of guinea- 
 pigs they are said to produce 
 broncho-pneumonia. 
 
 They were obtained from the 
 bullae in acute pemphigus. 
 
 Micrococcus in pneumonia 
 (Manfredi). Oval cocci -6 to 1 /z 
 in width, 1 to 1*5 p. in length, singly, 
 in pairs, and short chains. 
 
 Colonies on gelatine are circular, 
 whitish, and later spread out and 
 become bluish by transmitted light, 
 and of a pearly lustre by reflected 
 light. 
 
 Inoculated in the depth of gela- 
 tine there is a limited growth along 
 the track of the needle. 
 
 On blood serum they form a 
 shining, granular, faintly greenish- 
 yellow layer. 
 
 They also can be cultivated on 
 potato and in broth. 
 
 They are pathogenic in dogs, 
 rabbits, guinea-pigs, mice and birds. 
 Birds die in a few days ; mammals 
 in from one to three weeks. After 
 death new growths composed of 
 granulation tissue are found in the 
 internal organs, varying in size 
 from a millet seed to a pea. They 
 were obtained from the sputum of 
 pneumonia complicating measles. 
 
 Micrococcus in progressive 
 abscess formation in rabbits 
 (Koch). Cocci only about '15 ju, in 
 diam., principally in thick zoogloea. 
 The disease was induced by the in- 
 jection into rabbits of decomposing 
 blood. At the place of injection a 
 spreading abscess formed, which was 
 fatal to the animal in about twelve 
 days. No bacteria were observed 
 
 in the blood, but in the walls of the 
 abscess thick masses of cocci were 
 found. The pus is infectious, 
 causing the same disease in healthy 
 rabbits. 
 
 Micrococcus in pyaemia in 
 rabbits (Koch). Round cocci and 
 diplococci -25 p in diam. 
 
 The disease was produced by the 
 subcutaneous injection, in a rabbit, 
 of distilled water in which the skin 
 of a mouse had been macerated. 
 At the autopsy there were found 
 great infiltration around the site of 
 injection, peritonitis, and accumu- 
 lations in the liver and lungs ; in 
 short, the appearances of pyaemia. 
 
 FIG. 220. MICROCOCCUS IN PY.EMIA 
 IN RABBITS : VESSEL FKOM THE 
 CORTEX OF THE KIDNEY x 700. 
 a, Nuclei of the vascular wall ; 
 f, , Masses of raicrococci adherent 
 to the wall and enclosing blood- 
 corpuscles (Koch). 
 
 In the capillaries of the organs 
 examined, masses of cocci were 
 observed enclosing blood-corpuscles. 
 Fresh inoculations in rabbits with 
 exudation-fluid, or blood from the 
 heart, reproduced the same disease. 
 Micrococcus in septicaemia in 
 rabbits. Ellipsoidal cocci '8 to 1 
 p. in largest diam. The disease 
 was produced by the injection of 
 putrid meat infusion. After death 
 slight oedema was noted at the site 
 of injection, slight extravasation of 
 blood, and great enlargement of 
 the spleen. No emboli or peri- 
 tonitis resulted. Masses of cocci 
 were found in the capillaries of 
 
DESCRIPTION "i -1 
 
 different organs, especially in the 
 srlomeruli of the kidneys. Rabbits 
 and mice inoculated with blood 
 from the heart proved susceptible 
 to the disease. 
 
 Micrococcus in syphilis (Ditse 
 and Taguchi). Cocci and diplo 
 cocci. 
 
 They produce a greyish-white 
 growth in nutrient media. 
 
 They are said to produce inflam- 
 matory changes in the internal 
 organs and disease of the blood- 
 vessels when inoculated in dogs, 
 rabbits and sheep. 
 
 They were obtained from the 
 blood in cases of syphilis. 
 
 Micrococcus in trachoma 
 (p. 190). 
 
 Micrococcus in yellow fever 
 (p. -260). 
 
 Micrococcus lactis viscosus 
 (Conn). Cocci in pairs and chains. 
 Colonies circular and granular. 
 Inoculated in the depth of gela- 
 tine liquefaction begins at the upper 
 part of the needle track, and extends 
 until the gelatine is completely 
 liquefied. The liquefied gelatine is 
 viscous, and may be drawn out i 
 long threads. 
 
 On the surface of agar they form 
 a white, shining layer. 
 
 In broth there is an abundant 
 growth, and a film on the surface. 
 " They coagulate milk, producing 
 butyric acid, and giving it a b 
 taste. They were obtained 
 bitter cream. 
 
 Micrococcus luteus (Cohn).- 
 Oval cocci 1 to 1-2 p. in diam. 
 
 Colonies yellow, with irregular 
 contours, and granular. 
 
 Inoculated in the depth of gel 
 tine a granular filament develops in 
 the track of the needle, and on the 
 free surface a yellow patch 
 
 On agar the growth is slimy and 
 
 elliptical, with highly refractive o-ll 
 contents. 
 
 They form yellow drop* of I 
 'A mm. diam. on boiled potato : and 
 a thick, wrinkled, yellow skin on 
 nutrient liquids. 
 
 The colouring-matter in insoluble 
 in water, and unchanged by 
 phuric acid or alkalies. 
 
 Micrococcus ochroleucus 
 (Prove). Cocci '5 to '8 p in diam., 
 singly, in pairs. ;ui<l ^hort chains. 
 
 Colonies minute and colourlett, 
 with crenated margin, from whit -h. 
 later, processes extend into the gela- 
 tine, while the centre of the colony 
 becomes yellow. 
 
 On the surface of gelatine a film 
 develops, which in a few days turna 
 yellow. Old cultures have a p 
 liar smell. 
 
 The yellow pigment can be ex- 
 tracted with alcohol, i iblf 
 in water, and decolorised by a 
 
 They were obtained from human 
 urine. 
 Micrococcus of Forbes 
 
 'Micrococcusplumosus (Hi 
 gam). Cocci '8 M i diam., i 
 masses. 
 
 Colonies yellowish-white. 
 Inoculated in the depth of gel 
 tine long delicate acicular procea* 
 shoot out from the needle track 
 and on the free surface. 
 
 On potato the growth is simila 
 They were isolated from watei 
 Micrococcus pneumonia crc 
 
 v,u puu^ the growth is yellow, 
 and after a time wrinkl. 
 
 The pigment is insoluble in water, 
 ether and g alcohol, and not destroyed 
 by acids or alkali^ 
 
 They occur in water. 
 
 Micrococcus luteus <Mn;- 
 -Cocci similar m size to the above, 
 
 tenuis 
 irre^- 
 somewhat larger than staphvlocooci. 
 
 Si. 11 !?** 5-H 
 
 deeply, leaving a clear space m the 
 
 ^Intulatedinthedepthof^la- 
 tine a slightly opaque gro 
 
 f rmed agar a thin depomt yipmn 
 ^e needle track, *1 
 
 M-enfowi't 
 in three out of tbil 
 
558 
 
 DESCRIPTION OF SPECIES. 
 
 Micrococcus rosettaceus (Zim- 
 mermann). Cocci '7 to 1 p in diam., 
 singly, and in masses. 
 
 Colonies circular, whitish, or 
 greyish-yellow. 
 
 Inoculated in the depth of gela- 
 tine the growth is very scanty in 
 the track of the needle, but 
 spreads over the surface as a grey 
 rosette. 
 
 On agar a smooth layer with den- 
 tated margin is formed. 
 
 On potato the growth is faintly 
 yellowish. 
 
 They occur in water. 
 
 Micrococcus roseus (Eisen- 
 berg). Cocci forming pink colonies, 
 and a rose-coloured growth on the 
 surface of nutrient agar-agar. 
 
 They were found in sputum in a 
 case of influenza. 
 
 Micrococcus salivarius septi- 
 CUS (Biondi). Oval cocci, diplo- 
 cocci, in chains and small masses ; 
 capsulated. 
 
 They grow best on acid gelatine, 
 or in an atmosphere of carbonic 
 acid. 
 
 Colonies are small and circular, 
 with an opalescent centre and a 
 transparent margin, with sharply 
 defined outline. In the interior of 
 the colonies there is an appearance 
 of a network. 
 
 Inoculated in the depth of gela- 
 tine the cocci form a delicate fila- 
 ment and white dots on the free 
 surface. 
 
 On agar the cultures should be 
 made direct from the blood. The 
 growth appears on the surface and 
 resembles dewdrops. 
 
 Broth cultures remain clear. 
 
 They are fatal to mice in twenty- 
 four to seventy-two hours, and to 
 rabbits in fifteen to thirty days, 
 producing septicaemia. Attenuated 
 cultures are said to give immunity. 
 
 They were found in the saliva of 
 healthy and diseased persons. 
 
 Micrococcus stellatus (Mas- 
 chek). Cocci singly. 
 
 Colonies stellate. 
 
 Inoculated in the depth of gela- 
 tine a branching growth appears in 
 the track of the needle. The jelly 
 becomes brownish. 
 
 On potato the growth is brownish- 
 yellow and shining. 
 
 They occur in water. 
 
 Micrococcus tetragenus 
 (Gaffkey). Cocci about 1 //, in 
 diam., in tetrads, and surrounded 
 by a hyaline capsule. 
 
 Colonies form in twenty-four to 
 forty-eight hours as small white 
 dots, which are finely granular, and 
 have a vitreous lustre ; when they 
 reach the surface they form thick 
 raised masses. 
 
 Inoculated in the depth of gela- 
 tine, the cocci form an irregular 
 white growth, especially in the 
 upper part of the track of the 
 needle. 
 
 On agar the colonies occur along 
 the needle track, and are white, 
 round and circumscribed. 
 
 On potato they form a thick, 
 slimy, viscous layer. 
 
 White mice inoculated with a 
 minute quantity of a pure-cultiva- 
 tion die in from two to ten days, 
 and the groups of the characteristic 
 tetrads may be found in the capil- 
 laries throughout the body, especi- 
 ally in the spleen, lung and kidney. 
 
 Double infection can be produced 
 by inoculating a mouse with a 
 pure cultivation of Bacillus an- 
 thracis two or three days after 
 inoculation with Micrococcus tetra- 
 genus. On examination after 
 death, the capillaries of the lungs, 
 liver and kidney are filled with 
 both anthrax bacilli and masses of 
 tetrads (Plate V., Fig. 3). 
 
 Micrococcus tetragenus 
 mobilis ventriculi (Mendoza). 
 Cocci in tetrads ; capsulated ; 
 motile. 
 
 Colonies circular, whitish and 
 granular. 
 
 Inoculated in the depth of gela- 
 tine they grow on the free surface 
 only, and give off a peculiar odour. 
 
 They were isolated from the 
 stomach. 
 
 Micrococcus tetragenus sub- 
 flavus (Von Besser). Cocci singly, 
 and in tetrads. 
 
 They do not grow on gelatine. 
 
 Colonies on agar are brown and 
 irregular in contour. 
 
DESCRIPTION OF >!!: DBS, 
 
 559 
 
 On the surface of agar the cocci 
 form a greyish-white band, which 
 turns brown at the periphery, and 
 later is all dark or orange-yellow. 
 
 On potato the growth is brown. 
 
 They occur in nasal mucus. 
 
 Micrococcus tetragenus ver- 
 satilis (Sternberg and Finlay). 
 Cocci varying in size from '5 to l*o 
 f, in tetrads and irregular groups. 
 
 Colonies are circular and lemon- 
 yellow in colour. 
 
 Inoculated in the depth of gela- 
 tine there is very scanty develop- 
 ment along the line of puncture, 
 and the gelatine is liquefied in the 
 form of a cup near the surface. 
 At the bottom of the liquefied 
 gelatine a viscid, pale-yellow mass 
 accumulates. 
 
 On the surface of agar a thick, 
 viscid, yellow layer is formed along 
 the line of inoculation, which gradu- 
 ally extends over the entire sur- 
 face. The colour varies from cream- 
 yellow to lemon-yellow, and the 
 surface is moist and shining.- 
 
 On potato there is a similar 
 growth. 
 
 They were isolated from the 
 skin of patients suffering from 
 yellow fever, from mosquitoes after 
 attacking these patients, and from 
 the air. 
 
 Micrococcus ureae liquefaciens 
 (Fliigge). Cocci spherical, l"-~> t<> 
 2 p. in diam., singly, or in chains 
 of three to ten elements, or in 
 irregular groups. 
 
 Colonies appear in two days 
 as small white points. They have 
 sharply defined edges and a granular 
 surface. The gelatine gradually 
 liquefies, and the edges of the colo- 
 nies become irregular. 
 
 The cocci inoculated in the depth 
 of gelatine produce a continuous 
 white line along the track of 
 the needle. Finally, the whole of 
 the gelatine liquefies, and appears 
 as a whitish-turbid fluid with a 
 thick whitish-yellow deposit at the 
 bottom. 
 
 They were obtained from urine. 
 
 Micrococcus versicolor 
 (Fliigge). Small cocci, in pairs 
 and in masses. 
 
 White colonies develop in twenty- 
 four hours : after two days th. 
 JCtyOWUh, with .sharp contour* of 
 yellowish-green colour, an.l finely 
 granular. The sii 1 ,rrii.-i.-il colonies 
 form flat deposits, j r, mm . in size, 
 increasing to 10 mm. after four or 
 five days. 
 
 On the surface of gelatine the 
 cocci form a shining layer \v: 
 greenish or bluish ihimiMR lik. 
 mother-of-pearl. 
 
 Inoculated in h o f gela- 
 
 tine the growth is composed of 
 spherical yellowish colonies, ami 
 on the free sir \ form an 
 
 iridescent film. 
 
 They occur in the air. 
 
 Micrococcus violaceus 
 ter). Cocci or elliptical 
 scribed as uniting into violet-Mm 
 gelatinous spots, which again ; 
 to form larger j.it 1 
 
 The colonies on gelatine are v ; 
 in colour. 
 
 Inoculated in the depth of gela- 
 tine the growth is scanty in 
 track of the necdl.- 
 
 On the surface of gelatine they 
 form a bluish-violet layer, and the 
 same on agar and potato. 
 
 They were observed on boiled 
 potatoes exposed to the air, and are 
 also found in water. 
 
 Micrococcus yiticulosus (Kate). 
 Oval cocci 1 n in width. and \"2 n 
 in length, in masses, but without 
 formation of murli gelatinous ma- 
 terial. 
 
 The superficial colonies are ( 
 different in appearance from the 
 deep colonies. From the deep 
 colonies fine hairlike tendrils are 
 thrown off from a niing 
 
 a very delicate an- 1 .\tonsive net- 
 work. The threads are found to 
 consist of zoogloea masses, irregular 
 in size, arranged like HtringB of 
 beads. The colonies which are ex- 
 posed to the air form :i thin layer 
 of muddy-white gelatinous 
 stance, which rapidly -pn-ada, some- 
 times sending on prooeMW 
 into the depth of the gela- 
 
 In..culatcd in th- !. pfl. of gela- 
 
 ddicatr feather-lito growth 
 
 occurs along the track of the needle, 
 
560 
 
 DESCRIPTION OF SPECIES. 
 
 and on the free surface they pro- 
 duce the appearance which has been 
 described in colonies. This micro- 
 organism is exceedingly rare. It 
 was obtained from a contaminated 
 culture. 
 
 Monas Okenii. Short cylindrical 
 cells, 5 p. wide, 8 to 16 p, long, with 
 rounded ends. They exhibit lively 
 movements, each end being provided 
 with a flagellum twice as long as 
 the cell itself. They have pale-red 
 cell-substance, with dark grains. 
 
 They occur in stagnant water. 
 
 Monas vinosa. Round or oval 
 cells of about 2*5 p, in diam., often 
 united in pairs. Their motion is 
 slow and tremulous, and the cell- 
 substance is pale-red with dark 
 grains interspersed. Flagella have 
 not been observed. 
 
 They were found in water with 
 decaying vegetable matter. 
 
 Monas Warmingii. Cylindrical 
 cells, rounded at the ends, 15 p. long, 
 5 to 8 p. broad. They are possessed 
 of a flagellum at each end, and 
 exhibit rapid, irregular movements. 
 The cell-substance is pale-red, inter- 
 spersed at the ends with dark-red 
 grains. 
 
 Myconostoc gregarium (Cohn). 
 The threads are very thin, colour- 
 less, unarticulated, but fall apart 
 into short cylindrical links when 
 dried. 
 
 They form gelatinous masses, 
 10 to 17 p, in diam., singly or 
 heaped into slimy drops on water 
 in which algse are decomposing. 
 
 Nitromonas ofWinogradsky. 
 Very short rods, '9 to 1 p. in width, 
 I'l to 1'8 p. in length. Singly, in 
 masses, and in very short chains. 
 
 They can be cultivated in silica- 
 jelly. 
 
 They are active agents of nitrifi- 
 cation. 
 
 They were obtained from the soil. 
 
 Pedio coccus acidi lactici 
 (Lindner). Cocci '6 to 1 p. in diam., 
 singly, in pairs, and tetrads. 
 
 Colonies colourless. 
 
 On the surface of agar the cocci 
 form a colourless layer. 
 
 On potato the growth is almost 
 invisible. 
 
 The cocci produce lactic acid in 
 solutions containing sugar. 
 
 They occur in hay infusion and 
 malt. 
 
 Pediococcus cerevisise (Balcke). 
 Cocci singly, in pairs, and tetrads. 
 
 Colonies at first colourless, later 
 yellowish-brown. 
 
 Inoculated in the depth of gela- 
 tine a greyish-white filament 
 occurs in the track of the needle, 
 and a white layer on the free 
 surface. 
 
 On agar the growth is transparent 
 and iridescent, and on potato almost 
 invisible. 
 
 They, were isolated from the air 
 of a brewery. 
 
 Pneumobacillus liquefaciens 
 bovis (p. 242). 
 
 Proteus capsulatus septicus 
 (Banti). Rods isolated from a case 
 of septicaemia, and identical with 
 Proteus hominis capsulatus. 
 
 Proteus hominis capsulatus 
 (p. 224). 
 
 Proteus in gangrene of the 
 lung (Babes). Rods -8 to 1'5 p. 
 thick, irregular in form, and fila- 
 ments with irregular enlargements. 
 
 Colonies whitish and transparent, 
 with ramifications extending over 
 the gelatine. 
 
 In the depth of gelatine a growth 
 occurs along the track of the 
 needle, and a ramifying growth on 
 the free surface. 
 
 On agar the growth is slightly 
 yellowish. 
 
 On potato the growth is brownish. 
 
 They are extremely pathogenic 
 in mice and guinea-pigs. 
 
 They were isolated from a case 
 of gangrene of the lung. 
 
 Proteus microsepticus (Kar- 
 linski). Cocci, rods and filaments 
 in morphology, and cultures re- 
 sembling Proteus vulgaris. 
 
 Inoculated in the depth of gela- 
 tine liquefaction occurs in the 
 track of the needle, forming a 
 funnel with cloudy contents, and 
 in a few days the whole of the 
 gelatine is liquid. 
 
 They produce a general infection 
 in mice, and death in twenty-four 
 hours, and occasionally death in 
 
DESCRIPTION OF SPK< IBS. 
 
 50] 
 
 ral>1its. and local suppuration in 
 guinea-pigs and white rats. 
 
 They were isolated from pus in 
 a fatal case of puerperal pyaemia. 
 
 FIG. 221. PROTEUS MIRABILIS : SWARM IN-. 
 I >i.AM>> ox THE SURFACE OF GELATINE, x 285 
 
 (H.u - 
 
 Proteus mirabilis. Cocci -4 ^ 
 to -9 M- They occur singly and in 
 zoogloaa, and sometimes in tetrads, 
 pairs, chains, or as short rods in 
 twos resembling Bacterium termo 
 
 concentric circles, which in time 
 liquefies the medium. Similar 
 movements are observed in capsule- 
 cultivations as in Proteus vulgaris. 
 
 They were isolated t 
 putrid meat infusion. 
 
 Proteus septicus 
 (Babes). Rods -4 p in 
 width, and filamentous 
 forms. 
 
 Colonies rapidly liquefy 
 the gelatine. 
 
 Inoculated in the depth 
 of gelatine the bacilli form 
 a turbid funnel, or com- 
 pletely liquefy the gelatine 
 in twenty-four hours. 
 
 On agar the growth is 
 reticulated. 
 
 On potato brownish- 
 white. 
 
 Cultures have an un- 
 pleasant odour. 
 
 They are pathogenic in 
 mice. 
 
 They were isolated from 
 the organs in a case of 
 human septicaemia 
 
 Proteus sulfureus 
 denborn). Rods '8 M 
 width, varying in length, 
 and long filaments. 
 
 They correspend in mor- 
 phology and cultures with 
 Proteus vulgaris. 
 
 They produce sulphuretted hy- 
 drogen in cultures. 
 
 They were isolated from water. 
 Proteus vulgaris (Hauser).- 
 
 H- 
 
 *>"~T- ' 2/^g^\ 
 
 *- .3 wC^Jf *?&:> 
 
 !X i* *^ / 5^x 4 
 
 **^ ' -^ 
 
 FIG. 222.-P*' *''"" lNv".n'N KOMW, 
 
 in fact, in all conceivable transition 
 
 forms. , 
 
 Cultivated on nutrient gelatine as r , 
 
 they form a thick whitish layer m ^ 
 
 M broad as long, and others vary 
 
562 
 
 DESCRIPTION OF SPECIES. 
 
 63 /n wide. They are actively 
 motile. 
 
 Cultivated on nutrient gelatine 
 they convert it into a turbid, grey- 
 ish-white liquid. If cultivated in 
 a capsule containing 5 per cent, of 
 nutrient gelatine, a few hours after 
 inoculation the most characteristic 
 movements of the individual bacilli 
 
 FIG. 223. PROTEUS VULGARIS, FROM 
 SURFACE OF NUTRIENT GELATINE, > 
 (HAUSER). 
 
 are observed on the surface of the 
 nutrient gelatine, although at this 
 early stage no superficial liquefac- 
 tion can be detected. Probably the 
 movements depend upon the exist- 
 ence of a thin liquid layer, as they 
 are not observed if the nutrient 
 medium contains 10 per cent, of 
 gelatine. 
 
 They were isolated from putrid 
 meat infusion. 
 
 Proteus Zenkeri. Cocci -4 ^ in 
 twos like Bacterium termo, and 
 short rods 1 '65 p. long. 
 
 Cultivated on nutrient gelatine 
 no liquefaction results, but a thick 
 whitish-grey layer is formed. The 
 
 bacilli are motile, and the same 
 phenomena are observed on the 
 solid medium as in Proteus vulgaris. 
 In cover-glass impressions most 
 varied groupings of the bacilli are 
 seen, and also developmental and 
 involution-forms. 
 
 They were isolated from putrid 
 meat infusion. 
 
 Pseudo-diphtheritic bacil- 
 lus (p. 335). 
 
 Pseudo-diplococcus pneu- 
 moniae (Bonome). Oval cocci 
 in pairs and short chains ; cap- 
 sulated. 
 
 Inoculated in the depth of 
 gelatine small colonies develop 
 in the track of the needle in 
 twenty-four hours. 
 
 On agar there is a scanty, 
 moist growth. 
 
 On potato an almost in- 
 visible film. 
 
 In broth the cocci grow 
 rapidly, and the cultures give 
 off a peculiar odour. 
 
 They produce septicaemia in 
 mice, guinea-pigs and rabbits. 
 This micro-organism is 
 probably a variety of the 
 pneumococcus. 
 
 They were isolated from a 
 fatal case of cerebro-spinal 
 meningitis. 
 
 Ehabdomonas rosea. 
 Spindle-form rods, 3*8 to 5 p 
 broad, 20 to 30 /z long. They 
 exhibit slow, trembling move- 
 ments, having at each end of 
 the cell a flagellum. The 
 cell-substance is very pale, with 
 dark grains interspersed. 
 
 They occur in brackish water. 
 S arc in a alba. Small cocci. 
 They form small white colonies on 
 nutrient gelatine. 
 
 Inoculated in the depth of gela- 
 tine they grow slightly along the 
 needle track, but are heaped up on 
 the surface without liquefying the 
 gelatine. 
 
 They are present in the air. 
 S a r c i n a aurantiaca, Cocci 
 singly, in pairs, in tetrads, and in 
 packets. 
 
 Colonies orange-yellow. 
 Inoculated in the depth of 
 
 THE 
 
 285 
 
DESCRIPTION OF SPECIES. 
 
 563 
 
 gelatine they slowly liquefy it along 
 the whole needle track, and form 
 on the surface an orange-yellow 
 growth. On potatoes they slowly 
 develop the same pigment. 
 
 Sarcina Candida (Reinke). 
 Cocci 1*5 to 1*7 /x in diam., singly, 
 in pairs, and in tetrads. 
 
 Colonies are circular and shining, 
 white, and later yellowish. 
 
 Inoculated in the depth of gela- 
 tine liquefaction quickly takes place 
 along the track of the needle. 
 
 On the surface of agar a white, 
 moist layer develops. 
 
 They were found in the air of 
 breweries. 
 
 Sarcina flava (De Bary). Small 
 cocci in packets. 
 
 Inoculated in the depth of gela- 
 tine they produce liquefaction. 
 
 On agar they form a yellow 
 layer. 
 
 On potato the growth is limited 
 and yellow. 
 
 They were isolated from beer. 
 
 Sarcina hyalina (Kutzing). 
 Cocci round, 2'5 p. in diam., almost 
 colourless. United in families of 
 4 to 24 cells, reaching 15 p. in diam. 
 
 They occur in marshes. 
 
 Sarcina intestinalis (Zopf). 
 Cocci in groups of four or eight. 
 Yery regular in form ; never in 
 the large packets which occur in 
 Sarcina ventriculi. 
 
 They are found in the intestinal 
 canal, especially the ca3cum, of 
 poultry, particularly fowls and 
 turkeys. 
 
 Sarcina litoralis (Oersted). 
 Cocci 1"2 to 2 \JL in diam., bound 
 together in 4 to 8 families, which, 
 in their turn, may unite and in- 
 clude as many as 64 tetrads. 
 Plasma colourless ; in each cell 
 1 to 4 sulphur granules. 
 
 They were found in sea water 
 containing putrefying matter. 
 
 Sarcina lutea (Schroter). 
 Cocci singly, in pairs, tetrads and 
 packets. A single individual in a 
 tetrad may be divided into two, or 
 into four, so that a tetrad within a 
 tetrad results. 
 
 Colonies are round, slightly 
 granular in appearance, and yellow. 
 
 Inoculated in the depth of 
 gelatine they grow rapi.lly : the 
 gelatine becomes liquefied, and the 
 yellow growth sinks to the bottom 
 of the tube. 
 
 FlO. 221. SABCIXA X 600 (FLt'GOK. 
 
 Cultivated in agar they form a 
 colourless growth along the track 
 of the needle, and a bright canary- 
 yellow layer upon the surface. 
 
 On potato they form a yellow 
 layer. 
 
 They are present in air. 
 
 Sarcina mobilis ( Maurea). 
 Cocci 1-5 ^ in diam., in pairs, and 
 in tetrads. They are motile. 
 
 Colonies, at first white, become 
 brick-red. 
 
 Inoculated in the depth of gela- 
 tine, there is, after several days, a 
 slight growth along the track of 
 the needle, and a patch of growth 
 on the free surface which gradually 
 turns red. In about two weeks 
 liquefaction produces a fin, 
 shaped appearance ; later the lique- 
 faction extends to the sides of the 
 test-tube. 
 
 In broth turbidity is produ 
 and a yellowish-red depo- 
 
 On agar the growth, at firet w 
 changes to a brick -n,l colour. 
 
 There is no growth on potato, 
 ami milk i< not OOOgolftted. 
 
 They were isolated from a.- 
 Boid 
 
 Sarcina pulmonum ( Mauser). 
 Cocci from 1 to l'"> /i in liam . in 
 tetrads and packets. 
 
 Colonies white and small. They 
 are coarsely granular. 
 
 Inoculated in the depth of gela- 
 tine the growth is scanty in the 
 track of the needle, but on the free 
 surface there is a circular, well- 
 defined, translucent patch, which 
 
564 
 
 DESCRIPTION OF SPECIES. 
 
 later becomes greyish -brown, shin- 
 ing, wrinkled and irregular. 
 
 On potato the growth is very 
 slight and limited. 
 
 They cause ammoniacal decom- 
 position of urine. 
 
 They were isolated from phthisi- 
 cal sputum. 
 
 Sarcina Reitenbachii (Caspary). 
 Cocci about 1-5 to 2'5 /M in diam., 
 at the time of division lengthened 
 to 4 p. Mostly united together 
 from 4 to 8 in number ; occasion- 
 ally 16 or more. Colourless cell- 
 wall, lined with rose-red layer of 
 plasma. 
 
 They were found on rotting 
 water-plants. 
 
 Sarcina rosea (Schroter). 
 Large cocci, in packets. 
 
 Inoculated in the depth of gela- 
 tine, liquefaction quickly takes 
 place, and cultures after a time 
 have a reddish colour. 
 
 On agar the growth is slow and 
 limited. 
 
 On potato the growth is abundant 
 and of a bright-red colour. 
 
 In broth they produce turbidity, 
 and a red deposit. 
 
 They occur in the air. 
 Sarcina urinae (Welcker). Very 
 small cocci, 1/2 /z in diam., united 
 in families of 8 to 64. They were 
 observed in urine. 
 
 Sarcina yentriculi (Goodsir). 
 Cocci reaching 4 p. in diam., united 
 in groups of four, or multiples of 
 four, producing cubes or packets 
 with rounded-off corners. Contents 
 of the cells are greenish or yellow- 
 ish-red. 
 
 Colonies are round and yellowish. 
 A yellowish growth forms on the 
 surface of oblique gelatine without 
 liquefaction. 
 
 On potato they form a yellow 
 growth, and on serum also. 
 
 They grow well in hay-infusion, 
 forming brownish scales and a simi- 
 larly-coloured deposit. 
 
 They occur in the stomach of man 
 and animals in health and disease, 
 and were first detected in vomit. 
 
 Sphaerotilus natans. Cells 4 to 
 9 p. long, and 3 //, thick, united in 
 a gelatinous sheath to form threads. 
 
 The cells comprise rods and cocci- 
 forms ; the cocci are set free, and 
 develop into rods, which again form 
 threads. In the last a false branch- 
 ing has been observed. The plasma 
 of the cells break up into minute, 
 strongly-refractive portions, which 
 develop into round spores, at first 
 of a red and afterwards a brown 
 colour. 
 
 They occur in stagnant and flow- 
 ing water contaminated with or- 
 tanic matter, and form floating 
 akes of a white, yellow, rust-red, 
 or yellow-brown colour. 
 
 Spirillum amyliferum (Van 
 Tieghem). Filaments 6 n in length 
 and 1'4 to 1/5 n in width ; with 
 from 2 to 4 screw curves. 
 
 They act as a strong ferment in 
 the absence of air. 
 They occur in water. 
 Spirillum anserum (Sakharoff). 
 Spirilla resembling the Spiro- 
 cha3 ta O bermeieri. Extremely 
 motile. 
 
 They have not been cultivated 
 artificially. 
 
 Blood from diseased geese, con- 
 taining the spirilla, produces the 
 disease when inoculated in healthy 
 birds. The geese suffer from 
 diarrhoea, and die in about a week. 
 They were found in the blood of 
 geese suffering from an epidemic 
 form of septica3mia prevailing at 
 some of the stations on the Trans- 
 caucasian Railway. 
 
 Spirillum attenuatum (Warm- 
 ing). Threads much attenuated at 
 the ends, which consist usually of 
 three spirals. The middle spiral is 
 about 11 p high and 6 p. in diam., 
 and the end ones 10 /* high and 2 p 
 in diam. 
 
 They are found in brackish water. 
 Spirillum aureum (Weibel). 
 Curved rods with blunt ends, spi- 
 rilla and spirilliform filaments, and 
 involution forms. 
 
 Colonies are circular and golden- 
 yellow. 
 
 Inoculated in the depth of gela- 
 tine, a finely granular growth forms 
 in the track of the needle, and a 
 yellow-ochre prominent mass on 
 the free surface. 
 
DESCRIPTION OF SPKi IBS. 
 
 565 
 
 On agar a greyish growth ex- 
 tends over the surface, and later 
 prominent yellowish heaps make 
 their appearance. 
 
 On potato there is an abundant 
 growth of a golden-yellow colour. 
 
 They were found in sewage 
 mud. 
 
 Spirillum choleras Asiaticae 
 
 (P. 3in. 
 
 Spirillum choleroides (Bu j wid). 
 Curved rods very similar mor- 
 phologically and in cultures to 
 Koch's comma-bacilli. 
 
 They were isolated from river 
 water. 
 
 Spirillum choleroides (Orlow- 
 ski). Curved rods very similar to 
 Koch's comma- bacilli. 
 
 They were found in well water. 
 Spirillum concentricum 
 (Kitasato). Short spirilla, and 
 spirilliform filaments. 
 
 Colonies are circular, and com- 
 posed of concentric rings alternately 
 opaque and transparent. 
 
 Inoculated in the depth of gela- 
 tine there is only a little growth 
 in the track of the needle, and a 
 cloudy growth on the surface 
 extending into the jelly. 
 
 On agar the growth is extremely 
 adherent. 
 
 In broth they produce turbidity, 
 which disappears after a time ; and 
 there is a slimy deposit at the 
 bottom of the tube. 
 
 They were found in putrefying 
 blood. 
 
 Spirillum dentium (Miller).- 
 Spirals 10 to 20 /* in length, pomte< 
 at the ends. 
 
 They have not been cultivated. 
 They occur in the deposit on tt 
 teeth, and in company with Lepto- 
 thrix buccalis in carious teeth. 
 
 Spirillum flavescens (Weibel) 
 Commas thicker than those f oun< 
 in Asiatic cholera, spirilla, an< 
 spirilliform filaments. 
 Colonies yellowish. 
 Inoculated in the depth of gela 
 tine a finely granular 
 develops in the track of the need e 
 and on the free surface a pal 
 yellow patch. 
 
 On agar the growth, at 
 
 greyish- white, becomes yellow, and 
 forms a thick layer. 
 
 On potato -the growth is abun- 
 dant, and similar in colour. 
 
 They were fmnxl in sewage 
 mud. 
 
 Spirillum ftavum 
 Spirilla morphologically idem 
 with spirillum aureum. 
 
 Colonies on gelatine are pale- 
 yellow, and later the colour is more 
 intense. 
 
 On agar and potato tin y forma 
 ayer the colour of yellow-* > 
 
 They were isolated from sewage 
 mud. 
 
 Spirillum leucomelaneum 
 (Koch). Dark un^'l ilike spaces 
 alternate in the spirillum, resulting 
 from a regular arrangement of 
 dark granular contents. A rare 
 :orm observed in water covering 
 rotting algae. 
 
 Spirillum lingiise (Weibel). 
 Curved rods, spirilla, ami sj.inlli 
 form filaments, and involir 
 forms. 
 
 Colonies are composed of mtei 
 lacing filaments, and offshoots 
 extend into the surrounding gela- 
 
 Inoculated in the depth of gela- 
 tine a delicate growth occurs in the 
 needle track. 
 
 On agar the growth is whiti-h 
 and granular. 
 
 In broth a cloudiness is produced, 
 as well as a flocculent deposit. 
 
 They are especially .listiu^uisbed 
 from other spirilla described 
 Weibel by their staining by C 
 method. 
 
 They were isolated from 1 
 
 ^Spirillum marinum (Russell). 
 -Curved rods, and spiral filament*. 
 
 Colonies circular, granular and 
 striated ; later, flocculent ma** 
 float in liquefied areas. 
 
 Inoculatld in the depth of gela- 
 tine liM".-fa.-tion occurs I 
 track of the needle, and a mem- 
 brane form^on the surface of the 
 
 ie growth is yellowish 
 
DESCRIPTION OF SPECIES. 
 
 develops, and extends over the 
 surface. 
 
 Broth made with sea- water be- 
 comes rabidly turbid. 
 
 They were obtained from sea- 
 water. 
 
 Spirillum Metchnikovi (p. 373). 
 
 Spirillum nasale (Weibel). 
 Large curved rods and spirilliform 
 filaments. Non-motile. 
 
 Colonies circular, finely granular, 
 and brownish-yellow. 
 
 Inoculated in the depth of gela- 
 tine a delicate growth develops in 
 the track of the needle. 
 
 On the surface of agar they form 
 a whitish slimy film. 
 
 They occur in nasal mucus. 
 
 Spirillum Obermeieri (p. 258). 
 
 Spirillum of Finkler and 
 Prior (p. 258). 
 
 Spirillum of Griinther (Vibrio 
 aquatilis). Curved rods, very simi- 
 lar to Koch's comma-bacilli, but 
 there is no growth on potato. 
 
 They occur in water. 
 
 Spirillum of Miller. Curved 
 rods, singly, in pairs, and spiral 
 filaments. 
 
 They liquefy gelatine. 
 
 They were isolated from carious 
 teeth. 
 
 Spirillum of Neisser (Vibrio 
 berolinensis). Similar to Koch's 
 comma-bacillus, but smaller. 
 
 Colonies colourless, granular and 
 transparent, liquefying the gelatine 
 much more slowly than Koch's 
 com ma-bacillus. 
 
 Inoculated in the depth of gela- 
 tine they produce a growth similar 
 to that of Koch's comma-bacilli, 
 but much more slowly in milk. 
 
 In broth they grow abundantly. 
 
 They were found in water. 
 
 Spirillum of Renon. Curved 
 rods longer and broader than the 
 comma-bacilli of Koch. 
 
 Colonies yellowish, with dark 
 nucleus. 
 
 In gelatine the cultures resemble 
 those of Koch's comma-bacillus. 
 
 On agar the growth is white and 
 abundant. 
 
 They cause turbidity in broth. 
 
 They were isolated from impure 
 water from a well. 
 
 Spirillum of Smith (vide 
 Spirillum suis). 
 
 Spirillum of Weib-el. Curved 
 rods resembling Koch's cooima- 
 bacilli, morphologically, and in 
 cultures on jelly. The gelatine is 
 more quickly liquefied. 
 
 There is no growth on potato. 
 
 They occur in well water. 
 
 Spirillum plicatile (Ehrenberg). 
 Thin threads 2*25 /A in breadth, 
 110 to 125 ju- long, occurring also in 
 spirulinar forms. The threads have 
 primary and secondary windings ; 
 the former are in each example of 
 equal size, but the latter are often 
 
 FIG 225. SPIROCH^ETA PLICATILE. 
 
 irregular. Their ends are cut off 
 bluntly, and they exhibit rapid 
 movement. 
 
 They occur abundantly in marsh- 
 water in summer, and can be ob- 
 tained by allowing algae to decom- 
 pose in water. On cultivation the 
 threads break up into long rods, 
 short rods, and, finally, cocci. This 
 change is rendered visible by making 
 cover-glass preparations, and stain- 
 ing with aniline dyes. 
 
 Spirillum rosaceum (Klein). 
 Resembles Spirillum undula, but is 
 reddish in colour ; the colouring- 
 matter is insoluble in water, alcohol 
 or chloroform. 
 
 Spirillum Rosenbergii. 
 Threads with 1 to 1^ windings, 4 
 
DESCRIPTION OF SPECIES. 
 
 567 
 
 to 12 /* long, 1-5 to 2-t ; M thick. 
 They are colourless, but the con- 
 tents include strongly refractive 
 sulphur granules. Also spirals 6 to 
 7 :. /z in height, which are actively 
 motile. 
 
 They were found in brackish 
 water. 
 
 Spirillum rubrum (Esmarck). 
 Curved rods, spirilla and spiro- 
 chetae. They are actively motile. 
 
 The growth on artificial media is 
 extremely slow. 
 
 Inoculated in the depth of gela- 
 tine they grow along the track of 
 the needle, forming a filament of a 
 wine-red colour, without causing 
 liquefaction ; and on the free sur- 
 face the growth is colourless. 
 
 In broth long spirillar threads 
 are formed. 
 
 They were isolated from the 
 putrid tissues of a mouse. 
 
 Spirillum rufum (Perty). 
 Filaments from 8 to 16 /* in length, 
 with 1 to 4 screw curves ; non- 
 segmented ; chiefly motile ; tinged 
 with red. 
 
 They form rose or dark red spots 
 on the sides of wells. 
 
 Spirillum sanguineum (Cohn). 
 Threads 3 p. and more in thick- 
 ness, with 2 to 2 spirals, each 9 to 
 12 p. high. The ends are provided 
 with flagella. Their colour is due 
 to the presence of reddish granules 
 contained in the cells. 
 
 They were observed in brackish 
 water containing putrefying sub- 
 stances. (Vide Beggiatoa roseo- 
 persicina.) 
 
 Spirillum saprophiles. (I.) 
 Curved rods with pointed ends, *6 p 
 in width, 3 /* in length : spirilla, 
 spirilliform filaments, and involu- 
 tion forms. 
 
 Colonies yellowish or greenish- 
 yellow. 
 
 Inoculated in the depth of gela- 
 tine a white growth forms in the 
 track of the needle, later becoming 
 yellowish ; and on the free surface 
 there is a white growth, and 
 beyond this a transparent film 
 spreads over the jelly. 
 
 On agar the growthi s creamy, 
 and the jelly clouded beneath it. 
 
 On potato the growth is slimy 
 and yellowish or dark-brown in 
 colour. 
 
 They were obtained from sewage 
 
 FIG. 226. COMMA 1', \. i i. u IN WATER 
 
 CONTAMINATKI) WITH J 
 
 mud and decomposing hay infu- 
 sion. 
 
 (II.) Curved rods about 2 /* in 
 length, with blunt ends and in pain. 
 Extremely motile. 
 
 Colonies circular and yellowish- 
 brown. 
 
 Inoculated in the depth of gela- 
 tine a white growth develops in 
 the track of the needle, and later 
 becomes yellowish-red : on the free 
 surface a white patch forms, -un- 
 rounded by a transparent film. 
 
 In the depth of agar there is no 
 growth in the track of the needle, 
 but a yellowish-white patch on 
 the free surface adherent to the 
 jelly. 
 
 On potato the growth is also 
 adherent, and in appearance shining 
 and brownish-green. 
 
 They were isolated from decom- 
 posing hay infusion. 
 
 (III.) Curved rods, spirilla, and 
 spirilliform filaments, and involu- 
 tion forms. 
 
 Colonies are circular, granular, 
 and with irregular margin ; yellow 
 in the centre, and white at the 
 periphery. 
 
 Inoculated in the depth of gela- 
 tine a white growth develops in 
 the track of the needle, and on the 
 surface, without producing lique- 
 faction. 
 
 On the surface of agar the gr 
 is white. 
 
 On potato the growth is distinctly 
 brown. 
 
568 
 
 DESCRIPTION OF SPECIES. 
 
 They were isolated from sewage 
 mud. 
 
 Spirillum serpens (Miiller). 
 Long spirilliform filaments ; often 
 collected in masses. 
 
 They were observed in vegetable 
 infusions and stagnant water. 
 
 Spirillum sputigenum (Lewis). 
 Curved rods, very similar to the 
 comma-bacilli of Koch ; but many 
 observers having failed in repeated 
 
 
 FIG. 227. COMMA-BACILLI OF THE 
 MOUTH, x 700 (VAN ERMENGEM). 
 
 attempts to cultivate them, main- 
 tain that they are biologically dis- 
 tinct from those associated with 
 Asiatic cholera. Klein asserts that 
 they can be cultivated in an acid 
 gelatine, and that they are iden- 
 tical with Koch's comma-bacilli in 
 their mode of growth. They 
 occur with other bacteria in saliva 
 and in scrapings from carious 
 teeth. 
 
 Spirillum suis (Smith). Com- 
 mas and spirilla. 
 
 Colonies in gelatine are circular, 
 granular and brownish, and later 
 appear to be composed of concen- 
 tric rings. The gelatine is not 
 liquefied. 
 
 Broth with 1 per cent, of peptone 
 becomes in a few days clouded. 
 
 On potato they develop a thin 
 yellowish layer. 
 
 The commas are said to be slightly 
 larger than those obtained from 
 Asiatic cholera, and are not 
 pathogenic. 
 
 They were obtained from the 
 large intestine in swine. 
 
 Spirillum tenue. Very thin 
 threads, with at least 1, usually 
 2 to 5 spirals. Height of a single 
 screw is 2 to 3 /, and the length of 
 spiral therefore 4 to 15 p. They 
 are very swiftly motile. 
 
 They often occur in dense felted 
 swarms in vegetable infusions. 
 
 Spirillum tyrogenum (Deneke). 
 Curved rods, slightly smaller 
 than Koch's comma-bacilli, with 
 a great tendency to form long 
 spirillar threads (Fig. 228). 
 
 FIG. 228. DENEKE'S COMMA-BACILLI 
 FROM CHEESE, x 700 (FLI'GGE). 
 
 Colonies on plate-cultivations 
 are sharply denned and of a 
 greenish-brown colour. After a 
 time they liquefy the gelatine, but 
 the liquefaction is much more 
 marked than in colonies of Koch's 
 commas of the same age, though 
 not so rapid as in the case of the 
 commas of cholera nostras. 
 
 Inoculated in the depth of nu- 
 trient gelatine a turbid liquefaction 
 occurs along the needle track, and 
 on the surface of nutrient agar-agar 
 a yellowish-white layer develops. 
 
 Inoculation of potatoes gives no 
 result. 
 
 Administration of the bacilli by 
 the mouth, in the manner employed 
 for testing the pathogenic effect of 
 Koch's bacilli, produced a fatal 
 result in a few cases ; on the other 
 hand, injection into the duodenum 
 failed entirely. The pathogenic 
 properties may be therefore con- 
 sidered as not yet established. 
 
 They were isolated from old 
 cheese. 
 
 Spirillum undula. Threads 
 1-1 to 1-4 p thick, 9 to 12 p long ; 
 spirals 4'5 p. high ; each thread has 
 1| to 3 spirals. They are actively 
 motile, and possess a flagellum at 
 each end. 
 
 They occur in various infusions. 
 
 Spirillum v olutans (Eh r enberg) . 
 Threads 1-5 to 2 p thick, 25 to 
 30 p. long, tapering towards their 
 
DESCRIPTION OF SPECIES. 
 
 extremities, which are rounded off. 
 They possess dark granular con- 
 tents. Each thread has 2 to 3$ 
 windings or spirals, whose height is 
 1) to 13 ^. They have a flagellum 
 at each end, and are sometimes 
 motile, sometimes not. 
 
 They are found in the water of 
 marshes and in various infusions. 
 
 Spiromonas Cohnii. Colourless 
 cells, consisting of 1 J spirals, with 
 both ends acutely pointed and pro- 
 vided with a flagellum. Breadth 
 of the cells, 1-2 to 4 p. 
 
 They occur in water containing 
 decomposing matter. 
 
 Spiromonas volubilis (Perty). 
 Colourless, transparent cells, 15 
 to 18 /A long. Rapidly motile, and 
 revolving round a longitudinal 
 axis. 
 
 They occur in marsh-water and 
 putrefying infusions. 
 
 Staphylococcus pyogenesalbus 
 (p. 178). 
 
 Staphylococcus pyogenes 
 aureus (p. 176). 
 
 Staphylococcus pyogenes cit- 
 reus (p. 178). 
 
 Staphylococcus pyosepticus 
 (Heucourt and Ricnet). Cocci 
 identical with Staphylococcus pyo- 
 genes aureus. 
 
 Subcutaneous injection causes in 
 rabbits intense oedema, and death 
 in twenty-four hours. 
 
 They were isolated from pus 
 from an abscess in a dog. 
 
 Staphylococcus saliyarius pyo- 
 genes (Biondi). Cocci '3 to ."> n 
 in diam., singly and in masses. 
 
 Colonies white and opalescent, 
 producing liquefaction. 
 
 Inoculated in the depth of gela- 
 tine the growth appears in the 
 track of the needle, and is followed 
 by liquefaction. 
 
 On agar the growth is orange- 
 yellow. 
 
 The cocci produce local suppura- 
 tion when inoculated in animals. 
 
 They were isolated from an 
 abscess in a guinea-pig following 
 subcutaneous injection of saliva. 
 
 This coccus is probably identical 
 with Staphylococcus pyogenes 
 aureus. 
 
 Staphylococcus viridis flaves 
 cens (Guttmann).--Cocci singh. in 
 pairs and masses ; morphologically 
 agreeing with Staphylococcus pyo- 
 genes aureus. 
 
 Colonies are greenish-yellow. 
 
 Inoculated in the depth of gela- 
 tine a filament forms composed of 
 greyish col< 
 
 On agar the growth is greenish- 
 yellow. 
 
 They grow well on potato. 
 
 They were isolated from the 
 vesicles of chicken- \ 
 
 Streptococcus acidi lactici 
 (Grotenfeld). Oval cocci > t< 1 ^ 
 long, -3 to -6 /x in width, and long 
 chains. They are partially anaerobic. 
 
 Colonies are circular and wli 
 
 Inoculated in the depth of gela- 
 tine a growth occurs only ii 
 track of the needle. 
 
 Milk is coagulated. 
 
 They were isolated from coagu- 
 lated milk. 
 
 Streptococcus albus (Tils). 
 Cocci forming motile chains. 
 
 Colonies are flat and circular. 
 with white periphery and dark 
 nucleus, rapidly liquefying. 
 
 Inoculated in the depth of gela- 
 tine there is rapid liquefaction in 
 the track of the needle, and a 
 white deposit. 
 
 On potato they form a white 
 slimy layer. 
 
 They were found in water. 
 
 Streptococcus bomby c i 
 
 Streptococcus brevis (LingeU- 
 heim). Cocci singly, in pain 
 and chains, of eight to ten 
 elements. 
 
 Colonies on gelatine are circular 
 and very minute. 
 
 Inoculatfl in the depth of gela- 
 tine there is a funnel-shaped cavity 
 near the surface, and below this, 
 in the track of the needle, Mnall 
 isolated colonies. 
 
 On agar a yellowish-grey 
 develops along the line of inocula- 
 tion. 
 
 On potato there is a copious white 
 growth in forty-eight hours. 
 
 Broth is made turbi-1 
 
 They were isolated from healthy 
 saliva. 
 
570 
 
 DESCRIPTION OF SPECIES. 
 
 Streptococcus cadaveris (Stern- 
 berg). The description corresponds 
 with that of Streptococcus pyo- 
 genes. 
 
 Inoculated in the depth of gela- 
 tine the colonies are said to be 
 larger and more opaque. 
 
 On the surface of agar they form 
 a thin translucent layer. 
 
 In broth little flocculi develop, 
 composed of chains in which in 
 some cases the elements varied 
 considerably in size. 
 
 They were obtained from the 
 liver in a fatal case of yellow fever. 
 
 Streptococcus coli gracilis 
 (Escherich). Cocci from "2 to '4 p, 
 in diam., forming chains composed 
 of from six to twenty elements. 
 Some elements in a chain are irre- 
 gular in form, and show transverse 
 fission. 
 
 The colonies are spherical and 
 sink down in the liquefied gela- 
 tine. 
 
 Inoculated in the depth of gela- 
 tine liquefaction occurs in the track 
 of the needle on the second day, 
 and a white deposit forms at the 
 bottom of the liquid. In about a 
 week the gelatine is completely 
 liquefied. 
 
 On agar there is a very slight 
 growth. 
 
 On blood-serum small scales 
 develop. 
 
 On potato the growth is com- 
 posed of small white prominent 
 colonies. 
 
 Milk is coagulated. 
 
 They were found in the evacua- 
 tions of healthy infants. 
 
 Streptococcus conglomeratus 
 (Kurth). Cocci and chains, identi- 
 cal with Streptococcus pyogenes. 
 
 They form an adherent film at 
 the bottom of the tube, which is 
 not broken up by agitation. This 
 is observed in other varieties of 
 Streptococcus pyogenes, and is not 
 sufficient to distinguish it. 
 
 They are pathogenic in mice. 
 
 They were isolated from cases of 
 scarlet fever. 
 
 Streptococcus flavus desidens 
 (Fliigge). Cocci, diplococci, and 
 short chains. They form yellowish- 
 
 white colonies, which gradually 
 sink down in the gelatine. 
 
 Inoculated in the depth of gela- 
 tine the cocci form china-white, 
 confluent masses in the track of 
 the needle, and on the surface a 
 yellowish-brown slimy layer. 
 
 They occur in air and in water, 
 and were originally isolated from 
 contaminated cultures. 
 
 Streptococcus giganteus 
 urethrge (Lustgarten). Cocci '8 
 to 1 /n in diam., forming chains 
 composed of several hundred 
 elements. In description they 
 correspond with Streptococcus 
 pyogenes. 
 
 They do not grow at the tem- 
 perature of the room. 
 
 Colonies on agar are transparent 
 and iridescent. 
 
 They were isolated from the 
 healthy urethra. 
 
 Streptococcus Havaniensis 
 (Sternberg). Cocci '6 to *9 p. in 
 diam., forming long chains, com- 
 posed of cocci, in pairs, and oval 
 elements showing transverse divi- 
 sion. 
 
 This streptococcus is probably 
 a variety of Streptococcus pyo- 
 
 They were found in the acid 
 vomit of a yellow-fever patient. 
 
 Streptococcus in contagious 
 mammitis in cows (Nocard and 
 Mollereau). Cocci spherical or 
 oval, united in long chains. 
 
 Colonies are spherical, granular, 
 pale-yellow, or brownish by trans- 
 mitted light. 
 
 The cocci inoculated in the depth 
 of gelatine produce a granular 
 filament in the track of the 
 needle. 
 
 On the surface of nutrient gela- 
 tine minute spherical colonies are 
 formed, which are bluish by re- 
 flected light. 
 
 Injected into the mammary gland 
 of cows and goats they produce 
 mastitis. 
 
 They were isolated from the milk 
 of cows suffering from contagious 
 mammitis. 
 
 From the description this strepto- 
 coccus appears to be closely related 
 
DESCRIPTION OF SIM 
 
 571 
 
 to, if not identical with, Strepto 
 coccus pyogenes bovis(Crookshank) 
 Streptococcus in progressiv 
 tissue necrosis in mice. Koch 
 produced a disease in mice by sub 
 cutaneous injection of putrid blood 
 In tissue sections a chain coccu 
 was found which was similar to 
 Streptococcus pyogenes. 
 
 FIG. 229. STREPTOCOCCUS ix PRO- 
 GRESSIVK Tisst'K XKCHOMS ix Mi< K. 
 ". Necrotic cartilage cells, and (6) 
 chains in masses ; c, isolated chains. 
 (Koch.) 
 
 ^Streptococcus in Strangles 
 (Schutz). Cocci forming long 
 chains, which, it is said, do not grow 
 on nutrient gelatine or agar, but 
 form a transparent iridescent cul- 
 ture on blood serum. Cultures in 
 broth produced the disease in horses 
 and mice. Rabbits, guinea-pigs, 
 and pigeons are not aifected. 
 
 Strangles is a disease of the horse, 
 associated with suppuration of the 
 glands of the head and neck, prin- 
 cipally in the sub-maxillary, sub- 
 parotideal, and retro-pharyngeal 
 regions, gchutz found that the pus 
 contains streptococci and produces 
 a fatal disease in mice. 
 
 Streptococcus liquefaciens 
 (Sternberg). Spherical and oval 
 
 ingh 
 
 cocci, -4 to -6 M in 
 pairs and short rha 
 
 Inoculated in the depth of gela- 
 tine liquefaction occurs rapidly in 
 the track of the needle, and in a 
 week the gelatine is comi.l 
 liquefied, slightly opalescent, and a 
 scanty deposit forms at the bo 
 of the tube. 
 
 In the depth of ngar a filament 
 is formed compose 
 crowded colonies. 
 
 On potato a thin and limited 
 dry white layer is formed along the 
 line of inoculation in four to five 
 days. 
 
 They are non-pathogenic. 
 They were isolated from the liver 
 and intestines of fatal cases of 
 yellow fever. 
 
 Streptococcus mirabilis (Ros- 
 cpe and Lunt). Cocci 4 M in .li.tm.. 
 singly, and in long chains. 
 
 The growth on nutrient media is 
 very scanty. 
 
 In broth the growth is composed 
 of a mass of delicate filaments 
 which collect at the bottom of the 
 liquid. 
 
 They were isolated from sewage. 
 Streptococcus of Bonome.- 
 Cocci forming chains They corre- 
 spond in description with 
 coccus pyogenes, but. it is said, 
 they do not grow in gelatin- 
 on blood-serum, and they are said 
 to be distinguished by the characters 
 of the colonies on agar plates. 
 They are pathogenic. 
 Inoculated in rabbits and white 
 mice they produce symptoms 
 similar to those produc 
 oculations of the pneumococcus. 
 Sub-cultures rapidly lose their 
 virulence. 
 
 They were isolated from rusoa 
 of cerebro-spinal menin^ 
 
 Streptococcus of Manneberg 
 Cocci ft in diam., singly, in pairs, 
 M<l :n BU 
 
 noculat. 1 in tin- depth of gela- 
 ine a white filament forms along 
 he track of the needle componed 
 of minute colonies. In about a 
 10 nth the filament is replaced by 
 funnel of semi-liquefied gelati 
 On the surface of agar the 
 
572 
 
 DESCRIPTION OF SPECIES. 
 
 growth resembles Streptococcus 
 pyogenes. 
 
 "On potato they form a slimy 
 layer. 
 
 Milk is rapidly coagulated. 
 
 They are pyogenic in dogs and 
 rabbits. Injected into the veins 
 they produce inflammation of the 
 kidneys. 
 
 They were isolated from the 
 urine in a case of Bright's disease. 
 
 Streptococcus perniciosus 
 psittacorum (Parrot disease). 
 Cocci, singly, in chains, and in 
 zoogloea, have been described in 
 connection with a disease of the 
 grey parrot (Psittacus erithacus). 
 This disease is fatal to about 80 per 
 cent, of these parrots imported to 
 Europe. They suffer from di arrhoea 
 and general weakness ; their feathers 
 are ruffled ; their wings hang 
 loosely, and their eyelids close ; 
 convulsions set in, and death fol- 
 lows. At the autopsy greyish 
 nodules are found in the lungs, liver, 
 spleen and kidney. In and around 
 the capillaries of these nodules, and 
 in the blood of the heart, the cocci 
 are found in great numbers in 
 zoogloea, and more rarely in chains. 
 Inflammatory change in the sur- 
 rounding tissue is absent. 
 
 Streptococcus pyogenes (p. 178). 
 
 Streptococcus radiatus 
 (Flligge). Cocci less than 1 p in 
 diam., singly, in small masses, and 
 sometimes in short chains. 
 
 Colonies appear in twenty-four 
 hours. They are white, with a 
 yellowish-green sheen ; later they 
 liquefy the gelatine and develop a 
 circlet of rays. 
 
 Inoculated in gelatine, isolated 
 centres form along the track of the 
 needle which throw out horizontal 
 rays. At the same time a funnel- 
 shaped area of liquefaction forms 
 very slowly in the upper part. 
 
 On potato the growth is yel- 
 lowish-brown. 
 
 They occur in air and in water. 
 
 Streptococcus septicus(Flugge). 
 Cocci in chains, indistinguishable 
 microscopically from Streptococcus 
 pyogenes. 
 
 Colonies on gelatine grow more 
 
 slowly than those of most strepto- 
 cocci. 
 
 They are pathogenic. Mice die 
 in forty-eight to seventy-two hours 
 after subcutaneous inoculation of 
 a minute quantity of a cultivation. 
 During the last twenty- four hours 
 there is a distinct motor and sensory 
 paralysis of the hind legs. In 
 rabbits inoculation of the ear pro- 
 duces local redness, then a general 
 disease, and death in two or three 
 days. 
 
 They were found by Nicolaier, and 
 independently by Guarneri, in earth. 
 
 Streptococcus septicus lique- 
 faciens (Babes). Cocci -3 to -4 ^, 
 in pairs, in short chains. 
 
 Inoculated in the depth of gela- 
 tine a granular filament forms in 
 twenty-four hours along the track 
 of the needle, followed by lique- 
 faction of the gelatine forming a 
 funnel in which the gelatine is 
 clouded ; flat, whitish deposits 
 form on the side of the funnel. 
 
 On the surface of agar minute 
 shining, transparent colonies are 
 formed. 
 
 On blood-serum the growth is 
 almost invisible. 
 
 They are pathogenic. Subcu- 
 taneous injection in mice and 
 rabbits produces local inflammation 
 and oedema, followed by death in 
 about a week. 
 
 They were found in the blood 
 and organs of a child which had 
 died of septicsemia complicating 
 scarlet fever. 
 
 Streptococcus yermiformis 
 (Tils). Cocci forming motile 
 chains. 
 
 Colonies are yellowish-white, the 
 central portion finely granular, 
 the periphery radiated. 
 
 Inoculated in the depth of gela- 
 tine there is rapid liquefaction, 
 and a yellowish deposit at the 
 bottom of the liquid. 
 
 On potato the culture forms a 
 dirty-yellow layer. 
 
 They were found in water. 
 
 Streptothrix actinomycotica 
 (p. 431). 
 
 Streptothrix alba (Gaspanm). 
 A variety of Actinomyces bovis. 
 
DESCRIPTION OF SPECIES. 
 
 
 Streptothrix asteroides 
 { Oospora <i*tt-r'i' l> .--, S a u v a g e a u 
 and Radais ; Cladothrix asteroid?*, 
 Eppinger). Branching filaments 
 which form on the surface of 
 grape-sugar-agar a whitish growth, 
 which is later of a brownish-yellow 
 colour. 
 
 Broth remains clear, and small 
 pellicles float on the surface re- 
 sembling drops of stearin. 
 
 On potato they form snow-white 
 points, which turn brick-red in 
 colour, and are later covered with 
 a delicate white efflorescence. 
 
 The streptothrix is pathogenic 
 in rabbits and guinea-pigs. 
 
 It was isolated from pus. 
 
 Streptothrix anrantiaca 
 {Oospora mtrantiaca, Sauvageau and 
 Radais, and Doria). Similar to 
 Streptothrix asteroides. 
 
 Streptothrix carnea (Doria: 
 Oospora carnea, Sauvageau and 
 Radais). Similar to Streptothrix 
 asteroides, but the cultures on 
 gelatine are pink. 
 
 They are not pathogenic. 
 
 Streptothrix chromogenes 
 (Gasparini ; Oospora chromogenes, 
 Lehmann and Neumann). Culti- 
 vated on the surface of gelatine the 
 filaments produce a chalky growth, 
 and the jelly is coloured brown, and 
 is slowly liquefied. 
 
 \ On potato the growth is yellowish 
 or brown, and the potato itself is 
 coloured dark brown or black. 
 
 The streptothrix has been isolated 
 from air and water and the con- 
 tents of the stomach. 
 
 Streptothrix farcmica (Hticille 
 n de bozuf, Nocard : o,,x//o/v 
 ti. Sauvageau and Radais). 
 
 Inoculated on the surface of 
 gelatine there is in about two weeks 
 a very scanty granular growth. 
 
 In broth greyish pellicles develop 
 with a dusty surface. They are 
 pathogenic in cattle, guinea-pigs, 
 and sheep. 
 
 They were isolated from the 
 disease known as farcin de bnuf. 
 
 Streptothrix Forsteri (Cohn). 
 Cocci rods, and leptothrix threads. 
 The threads are twisted in irregular 
 spirals, and branch sparingly and 
 
 irregularly. Screw-forms are pro- 
 duced by the threads breaking 
 into small pieces. 
 
 Colonies slowly Ii,,u,-fy k <eh 
 
 On agar they form a wh 
 growth. 
 
 In broth they form shining 
 masses, floating in clear li.jui.i. 
 
 They occur in the lachrymal 
 canals of the human eye, in the 
 form of closely felted masses, and 
 in the air, and in fresh- and sea- 
 water. 
 
 Streptothrix Hofmanni i '/ 
 myct* i 
 
 Huti'ii'iinni, Sauvageau and Radais). 
 Thefilaments flourish in tin* n! 
 ary culture media with tin- a<M 
 of sugar, but they do not grow on 
 potato. 
 
 They produce suppuration in 
 rabbits. 
 
 They were isolated from the air. 
 
 Streptothrix liquefaciens 
 (f'ltnl,, //<//> //y (rarten). 
 
 A variety of Actinomyces I 
 
 Streptothrix madurae (p. 4 
 
 Streptothrix muacnlorum 
 suis (- 1 /;//"/// //* not, 1 )unker). 
 A variety of actinomyces found 
 in the muscles - 
 
 Streptothrix odorifera 
 odnriftf". Kiillinanii). Probably 
 identical with ( ><m]><>ra hromogenes. 
 
 Streptothrix violacea 
 i-;,,/.n:,i. Sauvageau and Radais, 
 and Doria. I 
 
 liquefies gelatine, and gives it a 
 pale wine-red colour. 
 
 Agar is coloured a violet 
 and potato becomes a reddish - 
 brown. 
 
 Urobacillus Duclauxi 
 (Miquel). Rods -I*. pi in 
 
 diam., and filaments 2 to 10 p in 
 length. Motile. Spore-formation 
 present. 
 
 In gelatine containing ammonia 
 
 or urea they develop in the track 
 
 of the needle and cause liquefac- 
 
 The liquefied gelatine is 
 
 viscid. 
 
 Broth containing ammonia be- 
 comes turbid, a sediment forms, and 
 the liquid gives off an unpleasant 
 odour. 
 
 They occur in sewage. 
 
574 
 
 DESCRIPTION OF SPECIES. 
 
 Urobacillus Freudenreichi 
 (Miquel). Rods 1 to 1-3 /LI in width, 
 and filaments 5 to 6 /u, in length. 
 
 Colonies circular, white. 
 
 Inoculated in the depth of gela- 
 tine growth occurs in the track of 
 the needle, and a pure white growth 
 on the surface, followed by slow 
 liquefaction. 
 
 In broth they produce turbidity. 
 
 They decompose urea. 
 
 They occur in air, sewage and 
 dust. 
 
 Urobacillus Maddoxi (Miquel). 
 Rods 1 n in width, 3 to 6 /u, in 
 length, and involution forms. 
 
 Inoculated in the depth of gela- 
 tine containing urea they produce 
 white colonies and crystals. 
 
 In broth they produce turbidity. 
 
 They decompose urea. 
 
 They occur in sewage. 
 
 Urobacillus Pasteuri (Miquel). 
 Rods attaining 1-2 ju. in width, 
 and 4 to 6 p in length, singly and 
 in pairs. Spore-formation present. 
 
 They grow in ammoniacal gela- 
 tine, slowly liquefying it and form- 
 ing crystals. The liquefied gelatine 
 is viscid. 
 
 They ferment urine, producing 
 a copious deposit of crystals. 
 
 They were isolated from decom- 
 posing urine. 
 
 Urobacillus Schutzenbergi 
 (Miquel). Short rods "5/i in width, 
 1 ^ in length. 
 
 They rapidly liquefy gelatine. 
 
 On agar they form a white layer. 
 
 They grow readily in broth, 
 especially after the addition of 
 urea. The liquid is made cloudy, 
 but after a few days it becomes 
 clear again. 
 
 They occur in water. 
 
 Vibrio rugula (Miiller). Rods 
 and threads, 6 to 16 p long, about 
 5 to 2-5 [i thick. The rods are 
 either simply bowed, or possessed 
 of one shallow spiral (Fig. 230). 
 
 They bear a flagellum at each end. 
 The rods form swarms when caus- 
 ing decomposition, and then, or 
 after, grow out into threads, curved 
 in a screw-like manner. In the 
 
 FIG. 230. VIBRIO RUGULA, x 1020. 
 A. Bowed threads. B. Slightly- 
 curved rods. C. Rods swollen pre- 
 paratory to spore-formation. I). 
 Rods swollen at the spore-forming 
 end. E. Various stages of the 
 developing spores. (Prazmowski. ) 
 
 next stage of development the rods 
 cease to move, and become swollen 
 with granular contents. One ex- 
 tremity develops an enlargement, 
 giving the rod the appearance of 
 a pin. The spore formed by 
 the contraction of the plasma in 
 the swollen end finally becomes 
 globular. 
 
 The vibrios appear in vegetable 
 infusions, causing fermentation of 
 cellulose. 
 
APPENDICES. 
 
APPENDIX I. 
 
 YEASTS AND MOULDS. 
 
 Yeast- fungi and mould-fungi, like bacteria or fosion-fungi, are- 
 achlorophyllous Thallophytes. They belong to two separate orders 
 the Saccharomycetes and Uyphomycetes which are intimately related 
 to each other, but quite distinct from bacteria. Their germs occur 
 widely distributed in air, soil and water, and are constant I v 
 encountered in bacteriological investigations. In addition, many 
 species are of hygienic and pathological interest and importance in 
 being either accidentally associated with, or the cause <>f \ 
 morbid processes and fermentations. For a complete account of 
 all the described species and full details of the various form- <>t 
 development, reference must be made to botanical and oth-i 
 works.* A description of certain species is appended here, an 
 afford some useful information to the worker in a bacteriological 
 laboratory. 
 
 YEAST-FUNGI OR SACCII.MSM.MY* i:n>. 
 
 Saccharomyces cerevisise (Tonda cere/v.s/</).- ' 
 oval, 8 to 9 /A long, singly or united in Mn.-ill dud Sj,.,!-.^ 
 
 occur three or four together in a mother- c 11. I t. '> // in diam. 
 S. cerevisi*, S. pastorianus and S. ellip>oi 1,-ns an- ;n-tiv ftk 
 ferments. According to Jorgensen they will produce in fourteen 
 days in beer-wort from 4 to 6 per cent., by volume, of alc-h-.l. 
 
 Saccharomyces ellipsoideus (Il.m-n). I. Klliptira: 
 mostly 6 ^ long, singly or unit.-l in littl- br 
 four spores found in a mother-cell, 3 to 3'5 p in diam. 
 on the surface of wort -gelatine th-y pro,lu<-- i" etovn to fourteen 
 days, at 25 C., a net-like growth l.y wl.irh they can be recogDiBed 
 
 * Sachs, Text-book of Botany ; Jorgensen, Micr*-0rgariim* and Fermen- 
 tation. 
 
.578 APPENDICES. 
 
 with the naked eye. II. Hound, oval, and rarely elongated 
 cells. They produce yeast-turbidity. There are two so-called 
 disease-yeasts allied to this species. The colonies of one kind form 
 -a network. This yeast causes turbidity in beer, and a bitter after- 
 taste. In the other kind the colonies are sharply defined. It 
 produces a disagreeable aromatic taste to beer, and an astringent 
 after- taste. It is widely distributed, and is the principal agent in 
 accidental fermentation. 
 
 Saecharomyces conglomeratus (Reess). Cells round, 5 to 
 6 fj, in diam., united in clusters, consisting of numerous cells 
 produced by budding from one or a few mother- cells. There are 
 2 to 4 spores in each mother-cell. They occur on rotting grapes 
 and in wine at the commencement of fermentation. 
 
 Saecharomyces exigims (Reess). Conical or top-shaped 
 cells, 5 fji long, and reaching 2*5 //, in thickness, in slightly 
 branching colonies. Spore-forming cells are isolated, each contain- 
 ing 2 or 3 spores in a row. They occur in the after-fermentation 
 of beer ; but, according to Hansen, they do not produce disease 
 in beer. 
 
 Saecharomyces Jorgensenii (Lasche). Cells small, round 
 or oval. On the surface of wort-gelatine the culture is greyish- 
 white, and the gelatine is slowly liquefied. They ferment saccharose 
 -and dextrose, but not maltose. When grown in wort with other 
 yeasts they are rapidly crowded out. 
 
 Saecharomyces pastorianus, I. Cells oval or club-shaped. 
 Colonies consist of primary club-shaped links, 18 to 22 /x long, 
 which build lateral, secondary, round or oval daughter-cells, 5 to 
 6 fji long. Spores 2 to 4. They occur in the after- fermentation 
 of wine, fruit- wines, or fermenting beer, and in the air of breweries. 
 They produce a bitter taste and unpleasant odour and turbidity 
 in beer. II. Cells mostly elongated, but also oval or round. 
 Cultivated on the surface of gelatine and yeast- water a growth is 
 produced with smooth edges, by which it can be differentiated 
 from No. III. They occur in the air of breweries, but do not 
 produce disease in beer. III. They produce yeast-turbidity in 
 beer. On the surface of yeast -water gelatine the cultures, after 
 sixteen days, have hairy edges. 
 
 Saecharomyces apiculatus. Cells lemon -shaped, both ends 
 bluntly pointed, 6 to 8 /w, long, 2 to 3 //, wide. Budding occurs only 
 at the pointed ends. Rarely united in colonies. Spores unknown. 
 They occur with other yeasts in various accidental fermentations 
 .and in ripe fruits. 
 
YEASTS AM> M>ru>>. .">7! 
 
 Saccharomyces sphaericus. ( Vlls varying in form; the 
 basal ones of a colony oblong or cylindrical, 10 to 15 p. long, 
 5 /x thick ; the others, round, 5 to 6 /t in diam. Unit-d in ramified 
 families. Spores unknown. 
 
 Saccharomyces anomalus (Hansen). Cells small, oval, and 
 sometimes elongated. Spores are hemispherical, with j trojecting rims 
 at the base. They were found in impure brewery yeast. 
 
 Saccharomyces mycoderma (Mycoclerma cereri*ln- et vini). 
 Cells oval, elliptical, or cylindrical, 6 to 7 p lonjr. - t" ." /t thick, 
 united in richly-branching chains. Spore-forming cells may be 
 20 /A long. Spores 1 to 4 in each mother-cell. The colonies in 
 gelatine are greyish and filmy. They form the so-called "mould" 
 on fermented liquids, and develop on the surface without 
 fermentation. When forced to grow submerged, a little alcohol is 
 produced, but the fungus soon dies. They occur on wine, beer, fruit 
 juices and sauerkraut. 
 
 Saccharomyces albicans (Oidium albicans, Fungus of thrush). 
 Cells round, oval, or cylindrical, 3'5 to 5 /t thick ; the cylindr 
 cells 10 to 20 times as long as they are thick. The bud-colonies 
 mostly consist of rows of cylindrical cells, from the end< of which 
 oval or round cells shoot out. Spores form singly in roundish celb 
 In plate-cultivations the colonies are pure whitv. In tin- depth 
 of gelatine a filament is formed composed of white colonies, some 
 with ray-like processes extending into the gelatine. On potato the 
 fungus forms a rapid white growth, and on bread also, 
 can be easily cultivated in a nutrient solution containing sugar 
 and ammonic tartrate. The cells germinate acc-.nlin- to the rie 
 ness of the fluid in sugar; they either grow into long 
 or, in a very strongly saccharine solution, many daughi 
 formed and bud out in various directions. According t- 
 the thrush-fungus is pathogenic in rabbit* death tak 
 twenty-four to forty-eight hours after an intrav,nou 
 a pure-culture. Long mycelial threads are found in the i 
 organs. They occur on the mucous membrane 
 especially of infants, in greyish-white patches which n 
 epithelium, bacteria, yeast* and the mycelin -us m, 
 
 Saccharomyces pyriformis (Marshall \Sard)- 
 They convert saccharine solutions contain,,,,' 
 beer. They occur with other micro-organisms . 1 
 
 glutini, O,K 
 vl in <le,s, 5 to 11 , long, 4 , -1, to***, -f 
 
580 APPENDICES. 
 
 Cell- membrane and contents are colourless in the fresh state, but 
 when dried and re-moistened possess a pale-reddish nucleus in the 
 middle. Spore-formation unknown. They form rose-coloured, slimy 
 spots on starch paste, and on sterilised potatoes. The colouring 
 matter is not changed by acids or alkalies. 
 
 Saceharomyees ilicis (Grb'nlund). Cells spherical. Spore- 
 formation present without vacuoles. Cultures on the surface of 
 gelatine have a powdery appearance. They produce about 2 '8 
 per cent., by volume, of alcohol in beer- wort, and cause a disagree- 
 able, bitter taste. They were obtained from the fruit of Ilex 
 aquifolium. 
 
 Saceharomyees aquifolii (Gronlund). Cells large and 
 spherical. Spores contain vacuoles. Cultures on gelatine are 
 variable, smooth and shining, or powdery. They produce about 
 3 '7 per cent, alcohol in beer- wort, and cause a sweet taste with 
 bitter after-taste. They also were obtained from the fruit of Ilex 
 aquifolium. 
 
 Saecharomyces Marxianus (Hansen). Cells elongated. 
 They develop a mycelial growth on solid nutrient media. They 
 occur on grapes. 
 
 Saceharomyees membranaefaciens (Hansen). Cells elon- 
 gated and vacuolated. Spore -formation abundant. Cultivated on 
 wort-gelatine they produce circular, flattened and wrinkled colonies, 
 greyish, and sometimes with a reddish tinge. The gelatine is slowly 
 liquefied. They occur in the slimy secretion of the roots of the 
 elm, and were also isolated from well-water. 
 
 Saceharomyees Hansenii (Zopf). Cells with small spherical 
 spores. They set up alcoholic fermentation in solutions containing 
 sugar. They were found in cotton- seed flour. 
 
 Saceharomyees Ludwigii. Cells irregular in form, oval, 
 bottle-shaped, lemon-shaped, and elongated, and mycelial filaments. 
 On wort-gelatine the growth is greyish or yellowish. 
 
 Saceharomyees acidi lactici (Grotenfelt). Cells oval, 2 to 
 4-35 \L in length, and 1*5 to 2-9 /x in width. Colonies on nutrient 
 gelatine are porcelain-white. They coagulate milk. 
 
 Saceharomyees minor (Engel). Cells spherical. Spore- 
 formation present. They are said to be the most active ferment 
 in the fermentation of bread. 
 
 Saceharomyees rosaceus (Pink Torula}. Cells 9 to 10 /x 
 in diam. They form a coral-pink growth in nutrient gelatine, 
 nutrient agar-agar, or on sterilised potatoes. They are present 
 in the air. 
 
YEASTS AM. Mi iri.I,.-. 
 
 Saccharomyces niger (Black 7W)._<vli, 
 
 MOII.D-KI XGI OR HYPHOMYCETES. 
 
 The mould-fungi have been divid.-d into 
 five orders : Hypodermii, Phycomycetes, Asco- 
 mycetes, Basidiomycetes and Myxomycete. 
 The following species, with the orders to 
 which they belong, are of especial inter, - 
 
 HYPODERMII. 
 
 Ustilago carbo (Milh.r, Smut). Spores 
 brown, circular ; episporium smooth ; sporidia. 
 ovoid cells. The spores or conidia occur as 
 a black powder in the ears and panicl* > 
 wheat, barley and oats. 
 
 Tilletia caries. Spores round, pale 
 brown ; episporium with reticulated thicken- 
 ings. In germinating, the sporidia grow 
 out radially from the end of the promyce- 
 lium ; these, at their lower part, conjugate 
 
 I i i , r '' 231. Bi ' 
 
 by a cross branch and separate from the , 
 
 promycelium, and at some point of the pair .v POTATO. 
 
 a hypha grows out, on which abundant 
 
 secondary sporidia develop. The latter are long, oval o-lN. \\hich 
 
 can in turn germinate. The fungus occurs in the form of a 
 
 stinking powder in grains of wheat, which renders i im- 
 
 pure, and gives it a disagreeable smell. 
 
 Urocystis occulta. The spores consist of several cells united 
 together; partly, large dark-brown evils in the interior, and 
 side, several flat, semicircular, colourless cells. Tin- pn.inyr.'Iiuin 
 germinates as in Tittetia, but the cylindrical evil- pn-lm-.- a li\ ; 
 without, as a rule, previous conjugation. T :r as a M 
 
 powder in rye-straw in long disintegrated stripes, which are at i 
 greyish. The affected plant produces abort i\- I 
 
 Empusa muscse. A spore or conidiirra of thi^ huigus 
 alighting upon the white area of the un-l.-i -mt'an- .t' the body 
 of the house-fly germinates into a hypha. The latter, penetrating 
 the skin, forms toruloid cells, which multiply by germ 
 are disseminated in the blood throughout the body of the fly. 
 
582 APPENDICES. 
 
 These cells again grow into hyphse, which penetrate the skin, each 
 forming a conidium, which is cast off with considerable force. The 
 parasite is fatal to flies, especially in the autumn. They are often 
 observed attached to the walls or window-panes, surrounded by a 
 powdery substance, consisting of the extruded conidia. 
 
 Empusa radicans. The spores form long hyphse, which pierce 
 the transparent skin of the caterpillar of the cabbage white butter- 
 fly. The terminal cells ramify, and fill the body of the caterpillar 
 with a network of mycelial filaments. The caterpillars attacked 
 become restless, then motionless, and death ensues. 
 
 Tarichium megaspermum. The spores are black in colour, 
 and provided with a thickened episporium. They occur at the 
 sides and ends of mycelial threads, attacking caterpillars (A gratis 
 segetum). 
 
 PHYCOMYCETES. 
 
 Saprolegnia. Colourless threads, forming dense radiating tufts, 
 occur on living and dead animal and vegetable matter in fresh 
 water. The filaments penetrate into the substratum, and branch 
 more or less in the surrounding water. The cylindrical ends of the 
 threads are shut off by a septum -forming zoosporangia, or mother- 
 cells, in the interior of which a number of spherical zoospores 
 develop. These are set free through an apical opening in the 
 thread, and after a time coming to rest, give rise to new plants. 
 In the sexual mode of reproduction a spherical bud, the oogonium, 
 develops at the end of a mycelial thread; from the thread small 
 processes or antheridia sprout out laterally towards the oogonium 
 and blend with its protoplasm. The latter breaks up into a number 
 of oospores, which clothe themselves with a membrane while still 
 within the mother-cell, and, eventually being set free, grow into 
 fresh mycelial filaments. The fungus attacks fish and tritons, 
 and produces a diseased condition of the skin, which may be 
 ultimately fatal. In salmon it produces the common " disease of 
 salmon." 
 
 Peronospora infestans. The conidia -bearers of this fungus 
 have as many as five branches, each bearing an egg-shaped 
 conidium. The contents of the conidia falling off and reaching a 
 drop of moisture, break up into a number of swarming zoogonidia, 
 which in turn develop upon plants. Fixing themselves to the 
 cuticle of the host, they throw a germinating filament into an 
 epidermal cell ; after piercing first its outer wall, and then its inner 
 
YEASTS AND MOULDS. 
 
 wall, the filament reaches an intercellular space, where the myiM-lium 
 develops. This continues to grow and spivad thn.u.irh.mt tin- plant. 
 In tubers it can hibernate and develop in the young shoots in tin- 
 following spring. The fungus appears in the form of brown 
 patches on the green parts of the plants, especially the leaN 
 The attacked parts wither and turn yellow or brown in od 
 If the under surface of a diseased leaf is examined, a corresponding 
 .lark spot may be observed, accompanied with a faint greyish-wh 
 bloom, which covers it. The latter consists of the conidia-bear 
 
 branches. 
 
 Pilobolus. The fruit-hyphse possess spherical 
 containing conidia. When ripe the receptacles with th-ir tronidia 
 are detached at their bases, and spring by their elasticity 1 
 distance. The fungus occurs as glassy tufts on the 
 cows, horses, etc. A cultivation can generally be obtain 
 keeping fresh horse-dung under a bell-glass. 
 
 Mucor mucedo. Hyphse colourless, simple or brui 
 rania yellowish-brown or black; spores ovoid. Tl,,v t 
 familiar white mould on fruits, bread, potatoes and 
 penetrate into the interior of nuts and .ftta, A i 
 fibrils develops in the substance of nutrient gelatine v 
 turn of sporangia on the free Burfcu* Th, mmrfoa d 
 spores and development into hyph* can b ' * ' 
 
 ^urs if the Z ^ cultivated n Uta <* ***-*?* 
 
 Mucor racemosus.-Hyph. short; spcW 
 " , und By continm-d cultivatu.n in hqdi 
 
 Mucor Phycomyces U, IK ^ 
 
 *"' Mucor fusiger (Li,l,,heim).-0void por,. 
 
584 APPENDICES. 
 
 Mucor mellittophorus (Lichtheim). Spores elliptical. Found 
 in the stomach of bees. 
 
 Mucor corymbifer (Lichtheim). This fungus forms branched 
 fruit-hyphse. The sporangia have a smooth membrane. It has 
 been found in the external auditory meatus, and on bread it forms 
 a dense snow-white growth. Pathogenic in rabbits. 
 
 Mucor rhizopodiformis (Lichtheim). The spores of Mucor 
 rhizopodiformis and Mucor corymbifer, when introduced into the 
 vascular system of rabbits, can germinate in the tissues, especially 
 in the kidneys, where they set up hsemorrhagic inflammation. 
 Dogs are immune, and only artificial mycosis is known. It occurs 
 on bread. 
 
 Mucor erectus. Resembles Mucor racemosus. It occurs on 
 rotting potatoes. 
 
 Mucor circinelloides. Mycelium much branched, and 
 sporangium carrier is curved. 
 
 Mucor spinosus. Sporangia chocolate. Columella has short 
 processes or spines. 
 
 ASCOMYCETES. 
 
 Oidium Tuckeri. Fruit-hyphse bearing single ovoid conidia. 
 Observed in the form of brown patches, covered with a white mildew- 
 like layer on the leaves, branches and young fruit of the vine, 
 producing " grape disease." 
 
 Oidium lactis. Fruit-hyphse simple, erect and colourless, 
 bearing at their ends a series or chain of conidia. In some cases, 
 the fruit-hypha branches beneath the chain of spores. Spores are 
 short cylinders. The conidia germinate into filaments of varying 
 length, which by subdivision form septate mycelial hyphse ; these 
 and their branches give rise in turn to spores or conidia. The 
 fungus is deeply stained by the ordinary aniline dyes. In a plate- 
 cultivation the colonies appear as white points, and develop into 
 delicate stellate colonies which ultimately coalesce and form a fine 
 mycelial network covering the surface of the gelatine. The gela- 
 tine is not liquefied. The growth on the surface of agar is similar 
 to that on gelatine. The fungus occurs in sour milk. 
 
 Achorion Schonleinii (Fungus of favus). Threads branching 
 at right angles. Favus in man forms yellow crusts on the hairy parts 
 of the body. The crusts are composed of epidermis and mycelial 
 filaments and spores. In plate-cultivations whitish colonies are 
 formed surrounded by liquefied gelatine. Cultivated on the surface 
 
AM, MOULD* 585 
 
 gelatine the growth resembles that of T, i 
 
 
 Fn;. 232.--IlKAi> AND XK< K OF C.U.K \M i -WORM 
 
 and the fungus forms a mi-mi rane mi tin- liijunl j.-lly \\hidi i- white 
 
 above and \v]]mv beneath. 1i 
 
 In man the disease varies in ajiiearanre in difb 
 
 body. Cattle, horses and dogs al>< Mitlei ;n ; but >heep 
 
 and pigs rarely, if ever. Th- (!i>-.i-- ; 
 
 Sometimes a small jiortimi of tin- -kin i- d&MMl ; nr mif, 
 
 the head, neck, che>t and alulm: Mink, m.-i;. 
 
 :-"l with >eah> or ern>t-. 'l'ii-i-.- i- often loss of hair in patches, 
 and the skin may be covered with >riirf. The disease is tranxmiK 
 
586 APPENDICES. 
 
 to the human subject. In one case, according to Brown, seven 
 grooms were infected on the arms from a grey pony which was 
 suffering from the disease in an aggravated form. 
 
 Fungus of fowl-scab. Fowls are liable to a disease similar 
 to favus. According to Schiitz this disease is characterised by 
 greyish-white patches on the comb and wattles of fowls, which may 
 extend over the neck and body. On nutrient gelatine a white 
 mycelium is formed ; and the gelatine is liquefied, and acquires a 
 reddish tint. The fungus can be readily cultivated on bread-paste, 
 agar-agar and potato. Cultures inoculated in fowls produce the 
 disease, but have no effect on mice and rabbits. 
 
 Fungus of mouse-favus. Mice suffer from a form of favus 
 which can be communicated to healthy mice by inoculation of scabs 
 or infected skin (Nicolaier). On nutrient agar the fungus forms a 
 thick mycelium, at first white, and later of a red or reddish-brown 
 colour. Mice can be infected with cultures. 
 
 Microsporon furfur This fungus occurs in Pityriasis 
 versicolor. Grawitz regarded it as identical with Oidium lactis, and 
 it is very closely related. Cultivated on gelatine the jelly is hollowed 
 out and the mycelial growth sinks down, and is yellowish in colour. 
 
 Oidium albicans. Vide Saccharomyces albicans. 
 
 Aspergillus glaucus (Eurotium aspergillus glaucus}. Mycelium 
 at first whitish, becoming grey-green or yellow-green. Spores 
 grey-green, thick -walled. It is found on various substances, chiefly 
 cooked fruit, and is non-pathogenic. 
 
 Aspergillus repens (Eurotium repens, De Bary). Fruit -heads 
 fewer than in the above, which are at first pale and then blue -green 
 to dark-green in colour. Conidia mostly oval, smooth, colourless 
 or pale to grey- green. 
 
 Aspergillus flavus. Gold-yellow, greenish and brown tufts. 
 Fruit-heads round, yellow, olive-green or brown. Conidia round, 
 seldom oval, sulphur-yellow to brown in colour. Saprophytic in 
 man, pathogenic in rabbits. 
 
 Aspergillus fumigatus. Greenish, bluish or grey tufts. 
 Fruit-heads long and conical. Conidia round, and rarely oval, 
 smooth, mostly pale and colourless. This fungus occurs on bread, 
 and has been found in the human lungs, external auditory 
 meat us and middle ear, and in the lungs of birds. The spores 
 introduced into the vascular system of rabbits, or into the peritoneal 
 cavity, establish metastatic foci in the kidneys, liver, intestines, 
 lungs, muscles, and sometimes in the spleen, bones, lymphatic 
 glands, nervous system and skin. 
 
YEASTS AND MOULDS. 587 
 
 Aspergillus niger (Enroth (Kf er^ ,, De 
 
 Dark chocolate-brown tufts. Conidia ,,,!. l.|, 1( * ,,, OWJI> o 
 brown when ripe. This mould can be cultivate,! , ..... lih on 
 moistened with vinegar, on slices of lemon, and on ari.l 'lYuus and 
 liquids It flourishes best of all, accorfing to H,mli,,, in 
 or the tollowing composition : _ 
 
 Oraminw. 
 
 LM 
 
 Sugar-candy . - (| . 
 
 Tartaric acid . . . . } . 
 
 Nitrate of ammonia . . i. 
 
 Phosphate ... .,; 
 
 Carbonate of potassium ... .5 
 
 magnesium .... 
 
 Sulphate of ammonia ... 25 
 
 zinc ..... 
 
 iron ...... .07 
 
 Silicate of potassium ... -07 
 
 It was also found that the fungus grew best when tin- 
 was spread out in a layer 2 or 3 cm. in depth in a .-shallow dj 
 and a temperature of 35 C. proved to be the most fa\ 
 The abstraction of zinc from the nutritive liquid reduced the weight 
 of a crop from 25 (the average) to 2 grammes, and the presence 
 f T?OOOOO P art f Citrate of silver, or 50 ooo P art f corr " 
 subHmate, stopped the growth altogether. It is saprophytir in 
 living body. 
 
 METHOD OF EXAMINING ASPERGILLUS NIGER. 
 
 Species of aspergillus stain intensely with rannine. fuchsine or methyl- 
 violet : but to examine Aspergillus niger with a high power ah 
 special technique is employed, as follows : A drop of glycerine is placed 
 on a clean slide, and a drop of alcohol on a cover-glass. With a fine i 
 of forceps a few of the fruit-hyphae with their hlack heads are immersed 
 in the alcohol. The cover-glass is then turned over on to the drop of 
 glycerine, and the slide held in the flame of a Bunsen burner till the 
 spores or conidia are dispersed. To make a permanent preparat 
 remove the cover-glass, and transfer the fruit-hyphie so treated to a 
 mixture of glycerine and water (1 to .".) : a drop may b< 
 placed ready on a slide provided with :i ring of Canada balsam. The 
 specimen is then permanently mounted by .-4 a circular cover- 
 
 glass, and surrounding it with a ring of cement in th- IIMKI! 
 
.588 APPENDICES. 
 
 Aspergillus ochraceus. At first flesh-coloured, and then 
 ochre-yellow heads. 
 
 Aspergillus albus. Pure-white fruit-heads. 
 
 Aspergillus clavatus. Club-shaped fruit-heads on long stems. 
 
 Aspergillus nidulans. Bread and potatoes acquire a reddish- 
 brown colour. Pathogenic in rabbits. Occurs on bread. 
 
 Aspergillus subfuscus. The growth is olive-yellow in colour. 
 Pathogenic in rabbits. Occurs on bread. 
 
 Aspergillus flavescens. The growth is yellowish-green. 
 Pathogenic in dogs and rabbits. Occurs on bread. 
 
 Penieillium glaueum. Occurs as a white, and later a blue 
 green, mould, on which dew -like drops of liquid may appear. Its 
 spores are present in large numbers in the air, and are liable 
 to contaminate cultivations. The fruit-hypha bears terminally a 
 number of branched cylindrical cells, from which chains of greenish 
 <3onidia are developed. It is the commonest of all moulds. 
 
 Botrytis Bassiana. Hyphae and spores colourless. Hyphse 
 usually simple, but sometimes united in arborescent stems. It is 
 the cause of muscardine, a fatal disease of silkworms, and occurs 
 also in various other caterpillars and insects. 
 
 Chionyphe Carteri. Mycelial filaments observed by Carter 
 in Madura disease. 
 
APPENDIX II. 
 
 H^EMATOZOA. 
 
 H^MATOZOA IN MAN, BIRDS AND TURTLES. HAJMATOZOA 
 EQUINES, CAMELS, RATS AND FISH. ILEMATOZOA IN FRO<.-. 
 
 HJGMATOZOA IN MAN (MALARIA). 
 
 IN 1880 Laveran, in Algiers, noticed the existence of peculiar 
 structures in the blood of a patient suffering from malaria, and 
 his researches were communicated to the Academy of Medicine in 
 Paris in 1881 and 1882, and subsequently published in extenso in a 
 treatise on the subject. 
 
 Laveran described various bodies which he was led to regard as 
 different stages in the life-history of the same micro- parasite. ] 
 most striking forms were cylindrical elements with pointed azl 
 ruities. They were crescent-shaped and pigmental in tin* middle. 
 There were other forms, more frequently found, which were either 
 free in the serum or in contact with the red blood-c< 
 They were more or less spherical, pigmented, and endowed with 
 amceboid movement. Other forms, again, were provide! with motile 
 filaments three or four times as long as the diameter of a r.-d hlood- 
 corpuscle. And, lastly, there were little masse- of hyaline material, 
 which Laveran regarded as dead forms. 
 
 These observations at first attracted little ai hut t hex- 
 
 have since been confirmed and extended by Richard. < 'minrilman and 
 Abbot, Marchiafava and Celli, Golgi, Sternberg, < Me- khor, 
 
 Vandyke Carter, Manson, and other>. and their imjurtance fully 
 recognised. 
 
 The different forms assumed by the hematoznnn of malaria 
 be described in two groups: those xvi thin the red hi, ^-corpuscles, 
 and those free in the serum. 
 
 Intra-corpuscular bodies. These are of three 
 
590 
 
 APPENDICES. 
 
 structureless protoplasmic bodies much smaller than, and within 
 or attached to, the red blood-corpuscles (Fig. 233). These rapidly 
 change their shape, exhibiting amoeboid movement. They were 
 first described by Marchiafava and Celli, and possibly represent the 
 first stage in the life-history of the hsematozoon. Marchiafava and 
 Celli suggested the name Plasmodium malarise. Second, minute 
 
 FIG. 233. NON-PIGMENTED AMEBOID FORMS (Marchiafava and Celli). 
 
 masses of finely granular or of hyaline protoplasm enclosing granules 
 of pigment (Fig. 234). These forms are sometimes present in large 
 numbers, and at other times can be found only with difficulty. 
 They are more or less spherical, but exhibit amoeboid movement, 
 and rapidly change their form. The pigment granules are also in 
 active movement. There may be one or more of these amoeboid 
 
 FIG. 234. PIGMENTS n AMOEBOID FOEMS (Golgi). 
 
 bodies to a blood-corpuscle, and they vary in size ; one may occupy 
 the whole of the corpuscle. In cases of pernicious malaria, similar 
 bodies may be seen, in tissue sections, in the corpuscles filling the 
 capillaries. Third, forms which appear like isolated grains, and 
 larger homogeneous bodies surrounded by clear spaces which change 
 in outline. 
 
 Extra-corpuscular bodies. These are the most striking, and 
 
 FIG. 235. SEMI-LUNAR BODIES OF LAVEKAN (Golgi). 
 
 perhaps the most interesting, forms. First, the semi-lunar bodies 
 of Laveran. These are crescent-shaped bodies, sometimes pointed 
 
ANIMAL MICRO- PARASITES. 
 
 501 
 
 at the extremities, but more usually round*'. 1 off (Fig. -_>;J5). They 
 are not always curved; some, indeed, are almost spheric -a I, and 
 others sausage-shaped. They are motionless. In IIM mens 
 
 a delicate line is visible on the concave side of the crescent connect- 
 ing the extremities. On careful examination this is found to be 
 
 ' 
 
 & 
 
 Fi.;. 236. ROSETTE FORMS WITH SEGMENTATION (Golgi). 
 
 the edge of a very delicate membrane. The body is composed of 
 homogeneous protoplasm. Centrally placed is a collection of pigment 
 granules, which on careful examination can be distinctly seen to be 
 in movement. The semi-lunar bodies vary in number in different 
 cases. Sometimes several can be seen in the field at the same time, 
 and in other cases they are only observed after a long ami ji.n 
 search. They are, as a rule, free in the serum ; but they have aU, 
 been seen within the red blood-cells. Second, finely granular masses 
 of protoplasm, which arise, according to Golgi, from the ii 
 corpuscular pigmented bodies. The pigment is collected in a ro>. 
 and the protoplasm by segmentation gives rise to a number of small 
 
 FIG. 237. FLAGELLATED FORMS (Vandyke Carter). 
 
 1. A flagellated spherule ; a, the same in t! "f a phagocyte; 6, free 
 
 motile filaments. 
 
 spherical forms, which are ultimately set f 
 
 believes that these changes occur in definite relation to the 
 
 ment of the paroxysm. Thifl. spherical, pear-shaped, or 
 
 bodies rather smaller than the red bl ..... 
 
 with one or more actively motile flagell, ff> These flagella 
 
592 APPENDICES. 
 
 are long lash-like filaments, which by their activity set the neigh- 
 bouring blood- corpuscles in motion. Free filaments in active move- 
 ment have also been observed. Fourth, small spherical pigmentecl 
 bodies about one-quarter the size of a red blood-corpuscle, which 
 exhibit amoeboid movement. 
 
 Inoculation experiments. Marchiafava and Celli assert that 
 inoculation of a healthy subject with blood containing the parasites 
 will produce a paroxysm of ague with development of the hsematozoa. 
 The pathogenic power of these parasites, however, has not been 
 established. There has been no cultivation of the parasite outside 
 the animal body, and reproduction of the disease with a pure culti- 
 vation. In favour of its being a pathogenic organism, Laveran 
 points out its invariable presence in some form or other in cases of 
 malaria ; the marked changes it effects in the red blood-cells ; the 
 increase in the number of the parasites in proportion to the severity 
 of the attack ; and, lastly, their disappearance after the admini- 
 stration of quinine. Others, again, have doubted the parasitic 
 nature of these bodies, and have looked upon them as representing 
 pathological changes in the blood-cells. 
 
 Laveran first of all suggested the name Oscillaria malarise ; but 
 subsequently he recognised that these bodies belonged to the animal, 
 not to the vegetable, kingdom. Osier has suggested that, tempo- 
 rarily at any rate, the organism should be placed in the genus 
 Hsematomonas of Mitrophanow, thus : " Genus, Hsematomonas ; 
 species, Hsematomonas malarise. Definition Body plastic ; ovoid 
 or globose ; no differentiation of protoplasm, which contains pigment 
 grains ; flagella variable, from one to four ; highly polymorphic, 
 occurring in (1) amoeboid form, (2) crescents, encysted form, (3) 
 sporocysts, (4) cellular free pigmented bodies." 
 
 EXAMINATION OF THE H^MATOZOA OF LAVERAN. 
 
 In the Living Condition. Select a patient by preference who has had 
 several attacks of malaria, and is markedly anaemic. Examine before the 
 invasion of the febrile paroxysm. Take two perfectly clean cover-glasses 
 and two clean slides ; wash one of the fingers of the patient with soap 
 and water, and then cleanse with alcohol ; apply a ligature, and with a 
 clean needle puncture the thin skin near the root of the nail ; touch the 
 drop of blood which collects, with a clean slide ; cover quickly with a 
 cover-glass, and gently press it if the layer of blood be too thick. 
 Examine with a T V - * 
 
 In Stained Preparations. Puncture the finger again if necessary ; 
 touch the droplet of blood with a clean cover-glass ; apply another cover- 
 glass ; press them gently together, and then slide them apart ; stain with 
 
AMMAI. - .-J 9S 
 
 two or three drops of alcoholic solution of methylene-l.lue ; 
 
 wwh off 
 
 H KMATOZOA OF 
 
 Aeeording to Danilewsky, birds suffer f,,, m malaria in Uth - m 
 acute and a chronic form. The haematozoa ,, 
 those found in malaria in , llalif ailll :inv ,1,,,,, ,, in -. IVIKV may ^ 
 attributed to the different character of the blood in birds. Graari 
 and Feletti have described two kinds of malarial ha-mato/,, 
 1-irds, one kind belonging to the genii* ffmamcA* and th- o( 
 to the genus 
 
 H.EMATOZOA OF Tl'KT! 
 
 Danilewsky has also minutely described and figured hsemato/- 
 the blood of turtles, which in some stages of their life hist 
 do>ely resemble those found by Laveran in man. 
 
 H^MATOZOA OF EQUIXES AND CAMELS (SURRA DISEASE.) 
 
 Surra is a blood disease occurring in hoi-so, mnl. 
 i-haracterised by fever accompanied by jaundice, petechiae of mucous 
 membranes, great prostration, and rapid wasting terminating in 
 death. The average duration of the disease is about tu.> m.-ntl^. 
 No organic lesions are found after death, but a para in 
 
 the blood during life. By means of subcutaneous inoculation. 
 by the introduction into the stomach of blood containing 
 parasite, the disease, according to Evans, can be ti. to 
 
 healthy animals. The importance of this disM>- may be reali 
 from the fact that on one occasion in India tin- :!nl Punjab <'a\. 
 lo>t no less than three hundred horses from it. 
 
 The disease has not been observed to be contagious 
 in the ordinary sense, but the possibility of its conveyance by means 
 of large brown flies has been sugg->t.-d. These flies attack ; 
 horses so vehemently that the blood frequently streams from tin- 
 bit*->; and the opinion that they propagate th- .li>.M>- i- | 
 among the natives. At the same time it has been j.articularls 
 noted that where the disease ha- broken mit the water * 
 Impure, 
 
 Kv.-m> discovered a hiemato/.,N,n, in Issn, in all 1 1,,- -liM-ased howes- 
 and mules examined ; in all di.--. b, with a 
 
 in the dogs which had Wn >ubj.-- 
 
 38 
 
094 APPENDICES. 
 
 Evans stated that when he first discovered the parasite he 
 thought it was a spirillum, but very speedily on closer examination 
 arrived at an opposite opinion. 
 
 To him the organism presented the appearance, when fresh and 
 active, of an apparently round body, tapering in front to form a 
 neck and terminating in a blunt head. Posteriorly he described a 
 tapering tail, from which there extended a long slender lash. At 
 the head end there appeared in one or two cases a circlet of 
 pseudopods, and as the body slowly died in serum it gave the 
 appearance of flattening out. After watching very closely all its 
 changes of form and movements, Evans came to the conclusion that 
 there, existed on either side of the body two fin-like papillae, one 
 near where the neck began and the other close to where the tail 
 began. In only very few instances he was able to see the four at 
 once. He suggested that these processes were of the nature of 
 pseudopods. 
 
 The parasite he described as extremely active in its movements, 
 with an undulatory, eel-like motion, progressing for the most part 
 head-end foremost, but occasionally moving in the direction of the 
 lash when tugging at a corpuscle. In fresh blood these organisms 
 resembled spermatozoa in colour ; but their peculiar characteristic 
 was the power they possessed of attacking and disintegrating the 
 red corpuscles. 
 
 Occasionally two were observed to unite and swim off as one 
 body ; but the mode of union was a disputed point. Evans thought 
 that they joined with their respective heads and tails in the same 
 direction, overlapping each other; but others to whom they were 
 shown were of opinion that they fastened with their tails in 
 opposite directions. 
 
 The parasites were not always present in the blood, but were 
 observed to come and go in successive broods. Evans referred the 
 organism to Lewis for his opinion as to its nature. Lewis arrived 
 at the conclusion that the parasite was " more nearly related to 
 that which he found in the blood of rats than to any other " ; but 
 he was of opinion at the time that they did not appear exactly 
 the same. 
 
 Five years later Surra broke out in British Burma, and Steel 
 was deputed to investigate the outbreak. Steel confirmed the 
 communicability of the disease to dogs, horses and mules by 
 ingestion and inoculation, but he considerably supplemented Evans' 
 views as to the nature of the disease by careful thermometric 
 observations : these finally led him to regard the disease as a true 
 
ANIMAL MICRO-PARASITES. .")!>.") 
 
 relapsing fever, closely resembling relapsing fever in 111:111. At the 
 same time it is worth recording that until Steel observed the 
 presence of the parasite described by Evans he regarded the <> 
 break as malarious in origin, and provisionally tcnnrl it ;M~ 
 typhoid. In the Burma outbreak, as in the Punjab epidemic, 
 considerable evidence was adduced in favour of regarding the disease 
 as being due to bad water supply. 
 
 Steel succeeded in staining the organism with aniline dyes, hut 
 his description of the parasite in the fresh state differs very 
 materially from that given by Evans. 
 
 Steel failed to recognise the round body tapering in front to a 
 neck. To him the bodies appeared thick in the middle, gradually 
 diminishing in size in either direction, with a blunt ami rigid 
 extremity at one end. The opposite end he described as tapering 
 in such a way as to produce a subspiral prolongation, which was 
 
 uncurled and lashed about freely like a whip. This tail was 
 described as slender in relation to the general size of the parasite ; 
 
 but under the highest power available the presence of a col 
 
 fiagsllum could not be detected, nor, he adds, did the movements of 
 
 the blood-constituents indicate its existence. 
 
 Steel also failed to see the slightest sign of the two fin-like p;. 
 
 on each side as described by Evans an opinion in which he was 
 
 supported bv Lewis. 
 
 These two observers, Evans and Steel, also differed as I 
 
 the movement could be called spiral. Steel felt con 
 
 movement was as much of that nature at times as can 
 
 from organisms with so open a corkscrew shape; whil, 
 
 maintained an opposite view. In the dried and stained , 
 
 Steel observed that they retained their subspn-ai 
 
 markedly spiral form of tail. 
 
 Steel found that the disease could be column 
 
 and to the monkey, and then the resembl 
 
 to the spirillum of n-lap>in- f,-vrr in 
 
 From the different apr.n.n,^ pr nted l.y < h- ,,- - 
 
 in the living state and when ,lri,l.,.,.l st ,i.,. 3 
 
 therewas^ahlya.illel , r^-hla,,,. , H, 1, 
 
 anaMa......!........,, 
 
 r.= 
 
 , 
 
 pointing out the 1 f F ** ' 
 
596 APPENDICES. 
 
 called into question ; and had Steel studied photographs of 
 spirilla he would not have regarded the corkscrew appearance as 
 imaginary. 
 
 Steel found the parasite in all cases, and further observed that 
 it appeared as the temperature rose and disappeared during the 
 apyrexial periods. 
 
 From all these observations Steel concluded as follows : That 
 relapsing fever of mules is an invariably fatal disorder, characterised 
 by the periodical occurrence of attacks of high fever, during which a 
 special organism closely resembling the spirillum of relapsing fever 
 in man is found in the blood. This organism is one-sixth the size of 
 a red corpuscle in width and three to six times in length. It is 
 eel-like, and, when dried and stained, presents a thick portion the 
 body and a spiral tail. The latter takes less of the dye than the 
 former, and commences as a sudden narrowing of the body, termi- 
 nating by a fine point. This, he insisted, had nothing of the nature 
 of an infusorian flagellum. The thick portion tapers in either 
 direction from its centre, and terminates in front abruptly in a rigid 
 process, with probably some holdfast organ. The sharpness of the 
 head end varies in different animals. The body portion he described 
 as spiral, and so closely in general appearances to resemble the 
 spirillum of relapsing fever that he concluded that the organism 
 was undoubtedly a spiral bacterium and named it after its discoverer 
 Spirochceta Evansi. This view, however, would not be accepted by 
 Evans, who maintained that, whatever it might be, it was not a 
 member of the family of bacteria. 
 
 In the face of these conflicting opinions Evans, in 1885, submitted 
 to the author preparations of the organism in the blood as well 
 as material from the lungs and intestines of a camel that had 
 succumbed to the disease. 
 
 On examining a stained preparation the author found that 
 with a power of 200 diameters a number of the parasites could 
 be distinguished in the field of the microscope, and with T V and 
 y 1 ^ o. i. objectives the individual characteristics were clearly brought 
 out. These were quite sufficient at once to dispel the idea of its being 
 a spirillum. It was obvious that it was a more highly organized 
 micro- parasite, presenting very peculiar and distinctive structural 
 appearances. 
 
 The author came to the following conclusions : 
 
 The somewhat tapering central portion, or body, of the parasite 
 is continuous at one end with a whip-like lash, and at the other end 
 terminates in an acutely-pointed stiff filament or spine-like process. 
 
ANIMAL Mint. '-PARASITES. .~>17 
 
 I lore and there. po>silily from injury >r want <!' l-\r!. .jnn.-nt . the 
 spine-like process appears to be blunted or absent. By very caret u I 
 focussing on the upper edge of the central portion, the author 
 discovered. the existence much more markedly in M.HM 
 parasites than in others of a longitudinal incin n .-itli.-r 
 
 a straight or undulating margin. The membrane is attarl, 
 the body, arising from the base of the rigid filament, and I:.V..UMB 
 directly continuous at the opposite end with the flagellum. In some 
 caaefl the edge only is deeply stained, giving the appearance <>' 
 thread continuous with the flagellum, so that one might be en- 
 led to overlook the membrane, and imagine that the flagellum aroae 
 from the opposite end of the body, at the base of th- -pin-like 
 
 process. 
 
 Close to the base of the spine-like process a clear un^am 
 is in many parasites easily distinguished ; and at the opposite < 
 there is, in some, the appearance of the deeply-stained protoplasmic 
 contents having contracted within the faintly-stained me 
 investment. When the longitudinal membrane has a 
 the undulations are much more marked in some cases, than i 
 Here and there the wavy outline appears iir>t OH th. 
 the central portion and then on the other; but there 
 waving outline on both sides of the same part of the I 
 was explained by a careful examination, which showed 
 somewhat ribbon-like parasite had become doubled on 11 
 discovery of this undulating membrane at once sugge* 
 author "an explanation of the lateral pHeudopodia < 
 Evan, If we imagine that we are looking down upon U 
 with the edge of the membrane towards us, one can co 
 
 , , ..... ....... 
 
 - 
 
 was about the same length a> th- 
 
 * 
 
598 
 
 APPENDICES. 
 
 sites. But the union obviously took place by the non- flagellated 
 ends, for the two flagella were frequently turned in the same 
 direction, so that the fused parasites resembled, as Evans sub- 
 sequently suggested, a trophy of buffalo horns. Here and there 
 more than two parasites had united, forming a stellate group ; and 
 in one case the author noticed that the individuals had apparently 
 united with their non-flagellated ends just overlapping, so that the 
 unstained spot in one was just situated in a line with the unstained 
 spot of the other. 
 
 In Evans's Report, Lewis's opinion is given that these parasites 
 differed slightly, but still were closely allied to certain flagellated 
 organisms which had been observed by him in rats in India. On 
 
 FIG. 238. "SUKRA" PARASITES OCCURRING SINGLY AND FUSED. 
 (From preparations stained with magenta, x 1200. Lent by Dr. Evans.) 
 
 referring to his original memoir, it will be found that his description 
 and woodcut differed very materially from the Surra parasite as 
 just described, though a microphotograph which Lewis had appended 
 to the memoir after it was written, indicated a great similarity 
 to this organism. To the author the organisms appeared not only 
 closely allied, but, as far as one can judge from figures and descrip- 
 tions, morphologically identical with the parasites described by 
 Mitrophanow in the carp, and as a matter of fact, instead of a 
 mere resemblance, the rat and the Surra parasites, when stained, 
 are found to be morphologically identical. 
 
ANIMAL MICRO-PARASITES. 
 
 H.EMATOZOA OF RATS. 
 
 Iii describing these organisms, Lewis remark.-! (hat it wa 
 strange that they had not occupied attention before, and suggested 
 as an explanation that possibly European rats did net hai-Uur these 
 parasites. The author examined a few white rats, but \\iihout 
 success, and then proceeded to examine the blood ..f (-nun..,, 
 rats, trapped from the London sewers, and discovered that these 
 organisms are to be found in no less than 25 per cent, of apparently 
 healthy animals. The first question which naturally arose wa 
 whether these organisms in European rats were identical with those 
 described by Lewis in Indian r.r 
 
 If we refer to the description given 1>\ L \\i>. \\e lind that he 
 states that when he first noticed them he thought they were vibrios 
 or spirilla. The drop of blood 
 under examination appeared to 
 quiver with life ; and on diluting 
 the blood, motile filaments could 
 be seen rushing through the 
 serum and tossing the blood- 
 corpuscles about in all directions. 
 
 The filaments were pale and 
 translucent, without any trace 
 of visible structure or granu- 
 larity, and they were more un- 
 dulatory in movement than , Bux)n 
 
 spirilla. A corpuscle might be OF RATS (Lewis). 
 
 observed to quiver, and thi> 
 
 could be distinctly traced to be due to the existence of a flagell.m., 
 apparently a posterior flageUum, as the organisms seemed generally 
 to move with the thicker end forward; no flap-Hum ..iM 
 detected at the opposite end. The greater numb i 
 in the woodcut (Fig. 239) are dex-ril-d as representing these 
 organisms a few hours after the blood had been obtained, when 
 their movements are not so rapid, and the flagellum becomes more 
 ea>ily recognisable. 
 
 This observation led Kent, who named the organism Uer t 
 Lewiri, to remark that if, as L-wi> H inclined to 
 organ "propels instead of draws the animalcule . 
 habited serum, we have presented a struct ,,,-,,1 tnd t 
 without parallel among the other represents 
 flaffeUata, the recognition of which would demand .- 
 
1300 APPENDICES. 
 
 distinct generic and family group for the reception of these singular 
 organisms." In his later paper, however, Lewis came to the con- 
 clusion that, like the generality of flagellated organisms, the rat 
 parasites moved with the lash in front. 
 
 On careful examination the plasma which constituted the thicker 
 portion of their substance was observed to suddenly swell out so as 
 to divide the body into two parts, as seen in the centre of the figure ; 
 at other times two or three such constrictions or dilatations were 
 detected, and at other times the body assumed an arrow shape, as 
 depicted at the lower part of the figure. When dried, and stained 
 with a little weak solution of aniline-blue, the body presented a very 
 different appearance. It was found to have contracted irregularly, 
 and to manifest a somewhat granular and shreddy appearance, 
 suggestive of a coagulated fibro-albuminous substance. The body 
 portion became flattened towards its middle to double its original 
 width, and both ends almost acutely pointed, while the flagellum 
 was only partly visible. After fixing with osmic acid they measured 
 0*8 to 1 /u, in width, and 20 to 30 /u, in length ; the flagellum was about 
 .as long as the body : so that the total length of the organism was 
 .about 50 //. Lewis detected these parasites in 29 per cent, of the 
 species Mus decumanus and Mus rufescens, but failed to find them 
 in mice. He considered that they had many features in common 
 with motile organisms of vegetable origin ; but they appeared to 
 .approach much more closely to the Protozoa, more particularly 
 several of the species of Dujardin's Cercomonas. He points out 
 that many, however, believe that these organisms are zoospores and 
 not animalcules. To him they also seemed to be not unlike the 
 flagellated parasite described by Butschli. 
 
 The latter observer detected flagellated organisms (Leptomonas 
 Biitschlii) in the intestinal canal of a free nematode (Trilobus 
 gracilis). They, too, form stellate colonies, like the Surra parasite, 
 owing to their being attached by their non-flagellated ends. When 
 detached from these colonies they presented a somewhat spindle- 
 shaped body about 11 JJL in length, with a somewhat thick flagellum 
 about double this length, so that the total length of the protozoon 
 would be 33 /x, or, as Lewis states, about half the length of the 
 flagellated organism in the rat's blood. Near the base of the 
 flagellum, Biitschli's protozoon presented a contractile vacuole, 
 but Lewis was unable to detect any such vacuole in the rat 
 hsematozoa. 
 
 In conclusion, Lewis observed that very probably these organ- 
 isms corresponded with the vermicules observed by Goss in the 
 
AMMAL MICKn-l'AKAH (;,,] 
 
 blood of a field mouse, and he aim ,,,,.,,ti,,iis that OhMHOU U.nd 
 minute " nematodes'' in the Hood of a Mark rat. 
 
 Wittich discovered in the blood ,,t tuunaten whip-like l^li,-, with 
 lively movements. They resembled frog, ^enn.w.oa, po*e> 
 a thick portion continued into a long lash-like thread. 
 considered them identical with the organi.M,,, dnerfbed 
 they also were observed in apparently healthy animal,. " K.H h ! 
 met with the same organisms. 
 
 Like Lewis, the author found that the Mood of th. 
 brown rat in England appeared to quiver with life, and that tfee 
 parasites were extremely difficult to examine until their inmen 
 was arrested for a moment or they became imprisoned in the serum 
 areas. After examining with various powers, from a ! d,\ 
 to a 2^5 o. i. of Powell and Lealand, the author came to th,- 
 following conclusion : That they are polymorphic, preKen 
 
 O 
 
 FIG. 240. A MONAD ix RAT'S BLOOD. The organism is n- presented t j*rtial 
 rest with its posterior filament impinging on a corpuscle, and Knowing the 
 undulating longitudinal membrane, the long flagellum, and the refractive 
 spherules in the granular protoplasm ( x 3000). 
 
 the most part slightly tapering bodies which terminate at one end in 
 a stiff, immotile, acutely-pointed flexible filament or spine-like prooeH, 
 and at the opposite end are provided with a long Hagellum, while. 
 longitudinally attached, a delicate undulating fin-like membrane can 
 be traced, which starts from the base of the pu>t.'rior filament, and 
 becomes directly continuous with the fla^ellnm (Fig. 
 
 With careful illumination the Udy i> t'"'nd to be di>tinrtly 
 granular, with one or more highly-refractive hpherule>. \\li.-n th-- 
 rapid movement is arrested the undulating m.-ml.r -\\n-\\\ 
 
 visible. The best opportunity . whi-n th- 
 
 organism comes to paitial re>t with it- >titV fil.un.-ni 
 corpuscle, as if to obtain a point (Tappiti, while : flag.-llum 
 
 in all directions (Fig. :M1, 1). At other time,, wh.-n the iinudtehiw 
 imj >inged with its posterior extremin 
 
 stiff filament is apparently entangled in '//////x. tin- movement* of 
 the organism give one the idea of it, n.d. -^..uring to ^^ ithelf 
 
602 APPENDICES. 
 
 but the author has not been able to persuade himself that they 
 " attack and disintegrate" the red blood -corpuscles. 
 
 In the active state the thicker portion, or body, appears to 
 twist and bend from side to side with great activity. The organism 
 can turn completely round with lightning rapidity, so that the 
 flagellum, at one moment lashing in one direction, is suddenly 
 observed working in the opposite direction. Then suddenly tne 
 organism makes progression, and it can be distinctly seen to move 
 in the direction of the flagellum, the flagellum threading its way 
 betiveen the corpuscles and drawing the rest of the orgcmism after 
 it. Currents set up by evaporation may undoubtedly here arid 
 there produce the appearance of the organism " wriggling along " 
 with its flagellum posterior ; but the author was convinced, after 
 
 FIG. 241. MONADS IN RAT'S BLOOD, x 1200. a, A monad threading its way among 
 the blood-corpuscles ; b, another with pendulum movement attached to a cor- 
 puscle ; c, angular forms ; d, encysted forms ; e and /, the same seen edgeways. 
 
 hours of patient observation, that in the normal mode of progression 
 the flagellum acts as a tractellum and not as a pulsellum. By 
 treating cover-glass preparations with osmic acid the appearances 
 obtained are very similar to those shown in Lewis's photographs, 
 so that there is no doubt, in spite of the descriptions not completely 
 according, that they are one and the same organism. There was a 
 great likeness to the organisms described by Mitrophanow, and to the 
 Surra parasite ; and when the author had stained the rat parasites, 
 the closest examination confirmed his belief that they were morpho- 
 logically identical with the stained parasites of Surra. 
 
 Cover-glasses with a thin layer of blood may be passed three 
 times through the flame of a Bunsen burner in the way commonly 
 employed for examining micro-organisms, and stained with an 
 
AMMAL MK KU-|'\ 
 
 aqueous solution of fuehsin. met h> 1- violet, or Bismarck- brown, 
 or with aurantia. nigrosin, un.l other anili,,,. dy.s. Th- foU 
 method will, however, be found I,,.M inftmtto:- I 
 prepared saturated solution of fuchsia or methy 1- violet in al. 
 alcohol, and put a drop with a pipette on the centre of the prepara- 
 tion; do not disturb the drop-form for a few moment ; th, , before 
 the alcohol has evaporated, wash off the excess . , s ,ll } 
 
 found that where the drop rested the organism* will he v.-ry d.-.-plv 
 stained, while in the surrounding area the colour will Vary ii, 
 intensity. By the effect of the different degrees ot 
 
 FIG. 242.M..NADS IN RAT'S BLOOU STAINKD WITH Mmm. YIOI.KI. 
 
 MEMBRANE UNDER DIKKKHKM A.-rK<T>, BLOOD-CORPUSCLES, HOME CBK- 
 XATEI> AND STAINED Discs (x 1200). 
 
 may be learnt (Fig. 242). In one organism the body and entire 
 membrane will be equally stained ; in another the margin of the 
 membrane only. In some the posterior still li lament is stained, 
 and at its base a darkly stained speck is \ery striking; and in 
 other cases, again, the posterior filament i> only taint ly tin;:*- 
 an unstained spot occurs near its base. 
 
 HJSMATOZOA OF I IMI. 
 
 In the year 1883 Mitrophanow pulli>li-d a paper in win 
 gave an account of organisms in the bl<*>d "t tin- mnd-tish and 
 carp. 
 
 In the blood of the mud-fish (''/,;//* /,,,,///*) the organisms at 
 the first glance looked like minute nematodes, but the appearance* 
 and changes which took place on further exa: showed 
 
 nothing in common with worms (Fig. 1'U). As a 1 p' : 
 solution had been add.-d to th.- blood und-r rxaii.i- occurred 
 
604 APPENDICES. 
 
 to Mitrophanow that they were possibly the cytozoa described by 
 Gaule ; but this idea was dismissed by the fact that they were 
 found in blood to which no salt solution was added. Their size 
 varied from 30 to 40 /A in length and 1 to 1| //, in width. At first 
 their rapid movements baffled examination, but as the rapidity 
 lessened there was the appearance of a curling movement in the 
 body portion and a swinging movement of the lash. The organism 
 moved in the direction of the lash, the anterior end of the body 
 being more pointed than the posterior, and gradually fining off into 
 the lash. When the body seemed to rest, the lash might be seen to 
 
 FIG. 243. ORGANISMS ix THE BLOOD OF MUD-FISH (Hcematomonas colitis), a, First 
 variety ; 6, second variety ; c, third variety, d, First variety in a state of 
 diminished activity, e, The same after treatment with osmic acid. (Mitro- 
 phanow.) 
 
 whip out in all directions. As the movement of the body gradually 
 diminished, it appeared to have a complicated screw form, the axis 
 of the screw corresponding to the body to which an undulating 
 membrane is fastened spirally. This could be distinguished when 
 the organism was dying, because the body in death contracted, and 
 the membrane then looked like a spiral addition. Thus the 
 organism consisted of a body, a spiral membrane, and a flagellum. 
 
 With higher magnification the organism appeared to consist of a 
 refractive, strongly contractile protoplasmic substance, which, when 
 death occurred, formed a shapeless mass. In the same blood two 
 other forms were observed : one without a membrane, but having 
 
ANIMAL MlCKo-l'AKAM | 
 
 ,,,1,,,,!,, ! . ,.., ,, 
 membra,, n,,, ,., , ..... ,, 
 
 plasm ,h seven, ,et,a,th, <,,,,,. .,,, ,,,,. ' , ...... 
 
 processes like pseudopodm. 
 
 In the carp (Fig. 2M ) the pa,,M,, h ,.,,,,,,,,, la , .,,, 
 poMMl an undulating membrane fastened al,,,,, ,|,,. ,.,,, , . 
 long body. When the body bent li M Uw* , ride .nd ,!,, . 
 the other a wave-like movement was ,,l,,,n.,l,l, .,, ,.. ,, 
 tins membrane. 
 
 These parasites were fo.m.l in all thr mml-H,!, ,x,,n,ned except 
 
 FIG. 244. ORI;AM>M> IN TMK lii.oor 
 
 , 6, c, Hmtwtowwtuis cara**-- . '. -. /. /. />, rli,-i ..p/;iiiMn> in tin- same blood 
 f.Mitn.ph.-ti. 
 
 one, and in greater immU'r> in tin- hot month.-. In the carp 
 
 they were only found occ;j>ion;illy. .Mitrojli;ino\\ le>rnl-.l ..th.-r 
 
 varieties, which he considfieil wi-rr j>oil.ly not c<>in|il-t. 
 
 )>ut developmental form-. ll- coii>idei--d that th-M- ..r^iui-iiiK were 
 
 i]it'u>oria between the r-ner.i ( '.i'C(.iii..na> and Tri 
 
 .ill-eat similarity to the Trichomona- d.-M-ril.i-d in the Li.-U-i kuhn'^ 
 
 glands of fowls and dnck> (KU-rth). 
 
 On account of their >|>vial hal.itat. Mitp.i, 
 IM-U ireiuis Haematomona-.d.'tininirtli j 
 of normal fish-blood, uorm-like, acti\vly mo\ i 
 indistinct diffeientiation of IMM|V paivnch;. fed at 
 
606 APPENDICES. 
 
 both ends, 30 to 40 //, long and 1 to 1 1 /x, wide. May possess in 
 front a flagellum, arid on one side an undulating membrane. 
 'Species : 
 
 Haematomonas cobitis. Body provided with a spiral membrane 
 and a flagellum at the fore-end. Parenchyma of body homogeneous. 
 Second variety, body and flagellum only. Movement undulatory, 
 body containing highly refractive spherules. Third variety, plasma - 
 like body, without membrane or flagellum ; quickly changes form 
 by sending out processes laterally, and contains two to four refractive 
 spherules. Blood of mud-fish. 
 
 Haematomonas carassii. Long bodies, with narrow membrane 
 attached along the whole length ; less actively motile. Several 
 forms also observed strikingly smaller than the above ; many disc- 
 shaped. Often seen attached to a red corpuscle, setting them in 
 motion by their movements. Blood of carp. 
 
 The morphological identity of the rat and Surra parasites has been 
 established by the author, and both ssem morphologically identical 
 with the organism of Mitrophanow. If we follow Mitrophanow, we 
 must obviously enlarge his genus of Hsematomonas. The author does 
 not agree with Mitrophanow in the advisability of adopting this 
 entirely new generic name. Mitrophanow suggested this new term 
 because of the special habitat, normal fish-blood, of the species he 
 discovered. But the characteristic features of these organisms are the 
 characteristic marks of the genus Trichomonas. It seems, therefore, 
 that they are embraced by the old genus Trichomonas, and that there 
 is no need to create a new one Haematomonas. The common habitat 
 of these species may be expressed by grouping them together in one 
 sub-genus Trichomonas sanguinis ; but the question arises whether 
 they are distinct species. If it were not for the different description 
 given by Mitrophanow of the organism in the mud-fish, the author 
 would be inclined to say that all these organisms belonged to one and 
 the same species, which might well be named Trichomonas sanguinis. 
 The monad in the rat and the Surra parasite are morphologically 
 identical with each other, and both, as far as one can judge from 
 the description, morphologically identical with the monad in the 
 blood of the carp. We have, however, seen that the organism in 
 Surra is believed to be pathogenic, and too much stress must not be 
 laid on morphological identity. There is strong evidence in favour 
 of believing in its pathogenic properties ; but at the same time it 
 must be borne in mind that the organism ha,s never been isolated 
 apart from the blood, and the disease then produced by its introduc- 
 tion into healthy animals. It is quite possible that the parasites in 
 
ANIMAL MICRO-PARASITES. 
 
 Surra are only associated with the disease, the impm-ei-Mied Mood 
 affording a suitable nidus for their development, while tin- con- 
 taminated water may be the common source of tin- I f 
 
 the disea.-e. On the other hand, the organism in the rat i- t<>und in 
 apparently perfectly healthy, well-nourUhed animal*. The author 
 suu-^sts that the parasites observed in the rat and liam>ier -hould 
 be named after Lewis, Trichomonas Lewisl; the organism in th- 
 mule, camel and horse after its discoverer, TricJumwnas Evan*i\ 
 and that the names Trickomonas cobitis and Trichvmonaa carasni 
 should be substituted for the names of the species described by 
 Mitrophanow. Thus we should have added provisionally to the 
 
 Genus TRICHOMONAS. 
 
 Sub-genus Trichomcmas sanguinis. Defi n i t ion : Elongated 
 tapering bodies, provided with a spiral (T. cobiti*), or 
 longitudinal (T. carassii, Lewiai, Evansi] ..... mbrsne, ter- 
 minating in a rigid filament and an anterior flagelhim. 
 Highly polymorphic. Habitat, the blood. 
 
 Species. Trickomows cobiti* (//.i,,.-if"numM < 
 
 Mitrophanow) Mud -li>h. 
 Tr'.chomonaa carassii (Hcematonwnas carateii 
 
 Mitrophanow) Carp. 
 Trickomona* Lewisi (Htrpetomoruu Lewi* 
 
 Kent) Rat, ham 
 Trichomonas Evansi(^ I''* 
 
 Steel) Horse, mule, rum-l ; (pathogenic ?). 
 
 H^MATOZOA OF THE FROG. 
 
 Tankester described an organism whirl. l. h'l 'H-verod n, 
 
 .-- 
 
 " ; 
 
608 APPENDICES. 
 
 One or more motionless filaments were occasionally observed attached 
 to these bodies. Gaule subsequently observed the same bodies, and 
 regarded them as resulting from the metamorphosis of the cells of 
 the frog's blood. Gaule's observations were refuted by Lankester in 
 1882, the parasitic nature insisted upon, and the organism named 
 Drepanidium ranarum. Lankester suggested that they were 
 probably the young stage of a sporozoon allied to Sarcocystis or to 
 Coccidium 
 
APPENDIX III. 
 
 PSOROSPERMS OR COCCIDIA. AMCEBA n.l 
 
 PSOROSPERMS OR COCCIDIA. 
 
 GREYISH-WHITE nodules may occasionally be found in the liver of a 
 rabbit, the result of a disease which may be mistaken for tttW 
 culosis. This disease often proves fatal, and may occur in an 
 epidemic form in rabbit warrens. The nodules have cheesy or 
 purulent contents, which are found, on microscopical examinat 
 to contain great quantities of Coccidium ovif&rme. 
 
 The coccidia pass from the intestine into the bile-ducte. The 
 walls of the bile-ducts become dilated and folded ; and irreg 
 cavities result from the partial or complete disappearance of the 
 dividing walls of the altered ducts. The folds are c-oin pose- 
 connective tissues lined with columnar epithelium, ;m<l tin- oxii.li.t, 
 in different stages of development, are found between the cells, and 
 free in the cavities of the nodules. 
 
 The individual coccidia are egg-shaped bodies. They possess a 
 thick smooth shell, with an opening, or ///"/"/<///<?, at one end, 
 protoplasmic contents which may completely fill the < i be 
 
 collected into a spherical mass. 
 
 After passing from the liver and intestine, these oval bodies 
 undergo a further development. According to Leuckart, who has 
 very fully described this parasite, the protoplasmic content* divide 
 into four masses, and from each is developed a C-shaped hyni 
 rod, the cavity of which is occupied by closely packed granules. 
 this condition they remain until they gain access to a f re>i 
 
 Coccidium oviforme has been found in the human liv.-r. :m<l also 
 in sheep, dogs, and cats. Similar, but not H.-nti.-al, bodies 
 mice, and also in fish and other cold-blooded animals. 
 
 M>'vscher'8 tubes are peculiar structures fc.uii'l in 
 sheep, deer, and mice. They consist of a firm envelope inclosing a 
 number of reniform or bean-shaped bodies. 
 
 808 - ; -' 
 
610 APPENDICES. 
 
 Pfeijfer's bodies. Pfeiffer has described certain appearances 
 which he attributes to coccidia, in epithelial cells in small- pox r 
 vaccinia, and other vesicular diseases. They are probably only 
 derived from the cell nucleus, and are not parasites. 
 
 Cancer bodies. In sections of malignant growths stained by 
 aniline dyes, certain bodies have been found and minutely described 
 and figured by various investigators, and a causal relation sug- 
 gested. Darier first described bodies like cysts, with spores, in 
 Paget's disease of the nipple. Wickham found similar structures 
 and figured them. Nils Sjobring described a cancer parasite, and 
 illustrated his researches with plates. Russell drew attention to 
 certain bodies in cancerous tumours, with a great affinity for 
 fuchsine. Soudakewitch, Podwyssozki, Sawtschenko, Buffer, and 
 Walker have, among others, contributed to the literature of the so- 
 called cancer parasites. These bodies appear in the form of ref ractile 
 spherical elements, which stain well with reagents, such as the 
 Ehrlich-Biondi stain. Sections are left in this stain for twenty-four 
 hours, washed in alcohol, cleared in xylol, and mounted in xylol 
 balsam. The spherical bodies have sometimes a radiate appearance. 
 These bodies have not been cultivated, and inoculation experiments 
 with cancerous tissue have been negative. The opinion is now very 
 generally held that these bodies are not parasites, but that changes 
 occur in the cells and nuclei, resulting in the formation of peculiar 
 structures, which have been brought to light by the use of aniline 
 dyes and complex staining methods. We are justified in concluding 
 that the cause of cancer is unknown. 
 
 Ballance and Shattock have made repeated attempts to cultivate 
 parasitic protozoa from malignant tumours, and they have extended 
 their researches to vaccinia and molluscum contagiosum, but with 
 negative results. Sand and water were used as the medium for 
 these experiments. Cultivations were made from nine scirrhous 
 carcinomata of the breast, five sarcomata from different sources, two 
 melanotic sarcomata from horses, and a sarcoma from a dog. In 
 every instance the result was negative. No traces of protozoic life 
 could be found, in spite of examinations at regular intervals, and 
 repeated for periods of many months. 
 
 AMOEBA COLI. 
 
 Losch, Grassi, Kartulis, and others have described an amoeba in 
 the intestines of patients suffering from dysentery. Losch adminis- 
 tered the fresh dejecta of a patient containing the amoebae to dogs, 
 
ANIMAL MICRO-PARASITES. Gil 
 
 and in one case a mucous mass was passed containing a nuu.U.r 
 of amoebae. Eighteen days afterwards the dog was killed, n, 
 mucous membrane of the intestine was reddened, iwofec 
 
 FIG. 245. AMCEBA COLI IN INTESTINAL M 
 
 ulcerated in three places. The mucus in the rectum ami in tin- 
 ulcers contained numerous amoebae. Cunningham, who hat- 
 the amoebae in choleraic and other cases, and in the intestine .f th. 
 cow and horse, does not attach any importance to their presence. 
 
APPENDIX IV. 
 
 APPARATUS, MATERIAL, AND REAGENTS EMPLOYED 
 IN A BACTERIOLOGICAL LABORATORY. 
 
 (A) HlSTOLOGICAL APPARATUS. 
 
 Microscope. For the investigation of micro-organisms a good 
 microscope with oil-immersion system and a condenser, such as 
 Abbe's, is essential. Such instruments are supplied by Zeiss, Leitz, 
 Reichert & Hartnack in Germany, and Powell & Lealand, Swift 
 <fc Baker in England. Zeiss supplies a micrometer eyepiece, with 
 directions for use. Some such arrangement is essential for the 
 measurement of bacteria. Other accessories to the microscope are : 
 
 A large bell-glass, for covering the microscope when not in use. 
 
 About a foot square of blackened plate-glass. 
 
 A white porcelain slab of the same size. 
 
 Glass bottles, with ground-glass stoppers, for alcoholic solutions of 
 aniline dyes, etc. 
 
 Glass bottles, with funnels, for aqueous solutions of the dyes, and 
 others provided with pipettes. 
 
 A small rod-stoppered bottle of cedar oil. This is recommended by 
 Zeiss in preference to other oils for his immersion lenses. 
 
 Set of small glass dishes or capsules and watch-glasses, for section- 
 staining, etc. 
 
 Stock of best glass slides, in packets of fifty. 
 
 Several boxes of round and square thin cover-glasses, in various sizes, 
 of the best quality. 
 
 Needle-holders, with a couple of platinum needles, and a packet of 
 ordinary sewing-needles. 
 
 Glass rods drawn out to a fine point ; useful for manipulating sections 
 when acids are employed. 
 
 Platinum or plated copper section-lifters. 
 
 One pair of small brass or spring-steel platinum-pointed forceps, for 
 holding cover-glasses. 
 
 One pair of brass tongs. 
 
 612 
 
APPARATUS, MATERIAL, AND RKAliKNTS. 
 
 
 Collapsible tubes, for containing Canada balsam ; very serviceable for 
 transport and general use. 
 
 Turn-table for sealing cover-glass preparations, with rings of cement. 
 
 Boxes for preparations, book-form. 
 
 Tickets and labels, various sizes. 
 
 Soft rags or old pocket-handkerchiefs, for removing cedar oil from 
 immersion lens, cleaning cover-glasses, etc. 
 
 Chamois leather for wiping lenses. 
 
 Warm Stages. In addition to those already described, Schafer 
 and Strieker have constructed warm stages for accurate observ; 
 Schafers apparatus consists of a vessel (/), filled with water 
 has been boiled to expel the air, and heated by means of a ga^ 
 at g. The warmed water ascends the indiarubber tube (c) to the 
 
 Fl(; M6. 
 
 WARM STAGE. 
 
 te, 
 
614 
 
 APPENDICES. 
 
 A more complicated apparatus, combining both a warm stage 
 and a gas chamber, is shown in Fig. 248. This consists of a rect- 
 angular piece of ebonite (E E) fixed to a brass plate which rests 
 on the stage of the microscope. On the upper surface of the ebonite 
 is" another brass plate (P), with an aperture (C) leading into a brass 
 
 FIG. 247. STKICKER'S WARM STAGE. 
 
 tube closed below by a piece of glass. To heat the apparatus the 
 copper wire B is placed on the tube , and its extremity heated by 
 the flame of the lamp. The nearer the lamp to the stage the higher 
 the temperature, which is indicated by the thermometer (t). To 
 
 C 
 
 n- 
 
 FIG. 248. STBICKER'S COMBINED GAS CHAMBER AND WARM STAGE. 
 
 employ it as a gas chamber the wire B is laid aside, and the gas 
 is conducted into the chamber by the tube a', and escapes by the 
 tube a. 
 
 Microtome. Schanze's is much in favour in Germany, but 
 Jung's of Heidelberg, though a somewhat cumbrous instrument, is 
 
APPARATUS, MATERIAL, ANI> KKAOENT8. 615 
 
 preferred by many workers. Smaller accessories which shmM be 
 within reach, are 
 
 A small can of sewing-machine oil. 
 
 A soft rag and chamois leather, for wiping the knives immediately 
 
 after use. 
 
 Stone and leather, for setting and sharpening the same. 
 Two or three camel's-hair brushes. 
 
 A freezing microtome is very useful : such as Swift 1 -, which is 
 used by the author ; and the method of embedding in oell< 
 combined with the ordinary process of freezing. 
 
 (B) REAGENTS AND MATERIAL EMPLOYKI. n an PROCESSES or 
 
 HARDENING, DECALCIFYING, EMBEDDING Kmxa AHI 
 
 OF TISSUES. 
 Alcohol, absolute. 
 Bergamot oil. 
 
 Celloidin. 
 
 Dissolved in equal parts of ether and alc< 
 
 Cork, or stock of ready-cut corks. 
 
 Ebner's solution. A mixture in the following proportions : 
 
 * 
 
 Hydrochloric acid . ^^ 
 
 Alcohol . 20 
 
 Distilled water . r> 
 Chloride of sodium 
 
 Formalin. 
 
 ........... ......... *.. v , 
 
 re-melted when required for i 
 
 Glycerine gelatine (Klebs). io 
 
 add 
 
 
 Gum. 
 
616 APPENDICES. 
 
 Kleinenberg's solution. 
 
 Saturated watery solution of picric acid . .100 
 
 Strong sulphuric acid . . 
 
 Filter, and add 
 
 Distilled water 300 
 
 Miiller's fluid. 
 
 Bichromate of potash ...... 2 
 
 Sulphate of sodium ...... 1 
 
 Distilled water 100 
 
 Osmic acid. 
 
 Distilled water ....... 100 
 
 Osmic acid ........ *5 
 
 Paper trays (or small glass capsules). 
 
 Paraffine. 
 
 Spermaceti. 
 
 Xylol. 
 
 (0) REAGENTS FOR EXAMINING AND STAINING MICROSCOPICAL 
 PREPARATIONS. 
 
 1. Acetic acid, strong. 
 
 2. Alcohol, absolute. 
 
 3. Alcohol, 60 per cent. 
 
 4. Alcohol, acidulated. 
 
 Alcohol ,100 
 
 Hydrochloric acid ...... 1 
 
 5. Alum Carmine (Grenadier). 
 
 Carmine ........ 1 
 
 Five per cent, solution of alum . . .100 
 
 Boil twenty minutes ; filter when cold. 
 
 6. Ammonia, strong. 
 
 7. Aniline. 
 
 8. Aniline water. 
 
 Distilled water .... . . .100 
 
 Aniline .... .... 5 
 
 Shake well, and filter emulsion. 
 
APPARATUS, MATERIAL, AND REAGENTS. I' 17 
 
 9. Bismarck-brown. 
 
 (a) Concentrated solution in equal parts of glycerine and water. 
 
 (b) Aqueous solution. 
 
 Bismarck-brown 
 
 Alcohol . .15 
 
 Distilled water 
 
 10. Borax-carmine (Grenacher). 
 
 Borax 
 
 Carmine . 
 
 Distilled water 
 
 To the dark purple solution add a 5 per cent, solution of acetic 
 acid until a red colour is produced; set aside twenty-four h.ur> ; 
 filter, and add a drop of carbolic acid. 
 
 11. Cedar oil. 
 
 12. Ehrlich-Biondi solution (Heidenhain). 
 
 Saturated aqueous solution of Orange. G. 
 
 Saturated aqueous solution of Rubin. S. 
 
 Methyl-green. 00. 50 
 
 To the mixtmv 
 
 All -1 "^ 
 
 Add water 
 
 13. Eosin. 
 
 (a) Saturated alcoholic solution. 
 
 (b) Aqueous solution. ^ 
 
 Distilled water. 
 Eosin 
 
 14. Ether. 
 
 15. Fuchsine. 
 
 (a) Saturated alcoholic solution. 
 
 (b) Aqueous solution. 
 
 Fuchsine . I - 
 
 Alcohol . g0 
 
 \VaU-r 
 
 16. Gentian- violet. 
 
 () &itur.-iu-d .-.Icoholic solution. 
 (6) Aqueous solution. 
 
 Gentian-viol JQQ 
 
 Distilled w;r 
 
<618 APPENDICES. 
 
 17. Gibbes' solution, for double staining. 
 Take of 
 
 Rosaniline hyclrochlorate ..... 2 
 
 Methylene-blue . . . . . . 1 
 
 Triturate in a glass mortar. 
 
 Dissolve aniline oil . . . . . . 3 
 
 In rectified spirit . . . . . .15 
 
 and add slowly to the above. 
 
 Lastly, slowly add distilled water . . .15 
 Keep in stoppered bottle. 
 
 18. Glycerine, pure. 
 
 19. Haematoxylin solution. 
 
 Hsematoxylin . . . . . . . 2 
 
 Alcohol . 100 
 
 Distilled water 100 
 
 Glycerine ....... 100 ' 
 
 Alum 2 
 
 20. Iodine solution. 
 
 Iodine, pure ....... 1 
 
 Iodide of potassium . . . . . 2 
 
 Distilled water ...... 50 
 
 21. Iodine solution (Gram). 
 
 Iodine . . . . . . . . 1 
 
 Iodide of potassium 2 
 
 Distilled water 300 
 
 22. Lithium- carmine solution (Orth). 
 
 Saturated solution of carbonate of lithium . 100 
 Carmine . . . . . . . . 2-5 
 
 23. Magenta solution (Gibbes). 
 
 Magenta . . . . . . . . 2 
 
 Aniline oil ....... 3 
 
 Alcohol (sp. gr. -830) . . . 20 
 
 Distilled water 20 
 
 24. Methylene-blue. 
 
 (a) Concentrated alcoholic solution. 
 
 (b) Aqueous solution. 
 
 Methylene-blue ... . . . . 2 
 
 Alcohol . . . . . . . .15 
 
 Water 85 
 
APPARATUS, MATERIAL, AM> KKAliKNTS. 
 
 {c) Koch's solution. 
 
 Concentrated alcoholic solution of methylene-blue 
 
 Ten per cent, potash solution . 
 
 Distilled water . . 200 
 
 (d) Loffler's solution. 
 
 Concentrated alcoholic solution of methylene-blue 30 
 Solution of potash, 1 to 10,000 
 
 25. Methyl- violet. 
 
 (a) Concentrated alcoholic solution. 
 
 {b) Aqueous solution. 
 
 Methyl-violet . 
 
 i AH 
 Distilled water 
 
 (c) Koch's solution. 
 Aniline water . 
 
 Alcoholic solution of methyl-violet 
 Absolute alcohol 
 
 26. Neelsen's solution. 
 
 Dissolve f uchsine . 
 
 In alcohol 
 
 Add a 5 per cent, watery solution of carbc 
 
 27. Nitric acid, pure. 
 
 28. Orseille (Wedl). 
 
 Dissolve pure ammonia-free orseille i ^ 
 
 Absolute alcohol 
 
 Acetic acid . ^Q 
 
 Distilled water 
 
 until a dark red liquid results. 
 
 29. Picric acid. 
 
 (a) Concentrated alcoholic solution. 
 
 (b) Saturated aqueous solution. 
 
 30. Picro-carmine (Ranvier). ^ 
 
 Carmine . ID 
 
 Distilled water 3 
 
 acid 
 
620 APPENDICES. 
 
 31. Picro-lithium- carmine (Orth). 
 
 To above-mentioned lithium- car mine solution 
 
 add saturated solution of picric acid . .2*3 
 
 32. Potash solution. 
 
 (a) 1 to 3 per cent. 
 
 (b) 10 
 
 (c) 33 
 
 33. Safranine. 
 
 (a) Concentrated alcoholic solution. 
 
 (b) Watery solution ..... 1 per cent. 
 
 34. Sulphuric acid, pure. 
 
 35. Salt solution 0'8 per cent. 
 
 36. Turpentine. 
 
 37. Vesuvin. 
 
 (a) Concentrated alcoholic solution. 
 
 (b) Watery solution. 
 
 Water, distilled. 
 
 Water, sterilised. 
 
 Distilled water can be kept for use in a wash bottle, or far 
 better in a siphon apparatus. Sterilised water is convenient in 
 plugged sterile test-tubes, which may be kept close at hand in a 
 beaker, or tumbler, with a pad of cotton wool at the bottom. The 
 numbered reagents can be conveniently arranged on shelves within 
 easy reach. Alcoholic solutions of the aniline dyes and other special 
 preparations should be kept in bottles with ground-glass stoppers, 
 Aqueous solutions of the dyes may be kept in bottles with fumie" 
 niters, and the solution filtered before use. To both aqueous and 
 alcoholic solutions a few drops of phenol, or a crystal of thymol 
 should be added as a preservative. For the rapid staining of cover 
 glass preparations, it is convenient also to have the most frequently 
 used stains (fuchsine, methyl- violet) in bottles provided with pipette 
 stoppers. 
 
 (D) REAGENTS FOR MOUNTING AND PRESERVING PREPARATIONS. 
 
 Acetate of potash. 
 
 Concentrated solution. 
 
 Asphalte lac. 
 
APPARATUS, MATERIAL, AND REAGENTS. 621 
 
 Canada balsam. 
 Dissolved in xylol. 
 
 Glycerine gum (Farrant's solution). 
 Glycerine. 
 Water. 
 
 Saturated solution of arsenious acid. 
 Equal parts ; mix, and add of picked gum arabic half a part. 
 
 Hollis' glue. 
 Zinc-white. 
 
 (E) DRAWING AND PHOTOGRAPHIC APPARATUS. 
 
 Camera Lucida. The camera lucida of Zeiss is an excellent 
 instrument, though many prefer the pattern made by Nachet of 
 Paris. Combined with the use of a micromillimeter objective, it 
 affords also a simple method for the measurement of bacteria. 
 
 For drawing microscopical appearances, and for illustrating 
 microscopical specimens with or without the use of a camera lucida, 
 the following materials should be within reach : 
 
 Pencils. 
 
 Etching pens. 
 
 Prepared Indian ink. 
 
 "Water-colour paints and brushes. 
 
 Ordinary and tinted drawing paper and other usual accessories. 
 
 Photo-micrographic Apparatus. Zeiss of Jena, Seibert & 
 Kraft of Wetzlar, Nachet of Paris, and Swift & Son of London, may 
 all be recommended for constructing an arrangement in which the 
 photographic camera is combined with the microscope. 
 
 The best models have been described fully in the chapter on 
 Photography of Bacteria. The accompanying figure (Fig. 249) 
 illustrates a model in which the microscope is used in the vertical 
 position. 
 
 For illumination either sunlight or artificial light may be em- 
 ployed. In the case of sunlight a heliostat is necessary to procure 
 the best results ; but as sunlight is not always available by day, and 
 it is also more convenient for many to work at night, it is better to 
 have recourse altogether to artificial light. Excellent results may 
 be obtained with an ordinary paraffine lamp, or with magnesium, 
 oxycalcium, or electric light. 
 
622 
 
 APPENDICES. 
 
 FIG. 249. VERTICAL MICRO-PHOTOGRAPHIC APPARATUS. 
 
 (F) STERILISATION APPARATUS. 
 
 Steam-steriliser. A cylindrical vessel of tin about half a metre 
 or more in height, jacketed with thick felt, and provided with a 
 conical cap or lid (Fig. 250). The lid is also covered with felt, has 
 handles on either side, and is perforated at the apex, to receive a 
 thermometer. Inside the vessel is an iron grating or diaphragm 
 about two-thirds the way down, which divides the interior into 
 two chambers the upper or . " steam-chamber," and the lower or 
 "water-chamber." A gauge outside marks the level of the water 
 in the lower chamber ; this should be kept about two-thirds full. 
 
APPARATUS, MATERIAL, AND REAGENTS. 
 
 623 
 
 The apparatus stands upon three legs, and is heated from below with 
 two or three Bunsen burners, or a Fletcher's 
 burner. It is employed for sterilisiiii: 
 nutrient media in tubes or flasks, for cooking 
 potatoes, or hastening the filtration of ag.ir- 
 agar. When the thermometer indicates 
 100 C. the lid is removed, and test-tul'<-> 
 are lowered in a wire basket by means of 
 a hook and string, and the lid quickly re- 
 placed. Potatoes or small flasks are lowered 
 into the cylinder in a tin receiver with a 
 perforated bottom, which rasts upon the 
 grating and admits of its contents being 
 exposed to the steam. A larger model is 
 shown in Fig. 33. 
 
 Hot-air Steriliser. A cubical chest of 
 sheet iron with double walls, supported on 
 four legs ; it may also be suspended on the 
 wall of the laboratory, with a sheet of 
 FiG.250.-KocH's STEAM- asbestos intervening (Figs. 251 and 252). 
 
 STERILISER. It is heated with a rose gas-burner from 
 
 below, and the temperature of the interior in- 
 dicated by a thermometer inserted through * A J J | * J * 
 
 a hole in the roof ; in a second opening a 
 IM> regulator can be fixed. Test-tubes, 
 flasks, funnels, cotton wool, etc., may be 
 sterilised by exposure to a temperature 
 of 150 C. for an hour or more. 
 
 ?fii 
 
 fc 4 A 
 
 FIG. 251. HOT-AIR STERILI.-KK. 
 
 FIG. 252. SECTION OF HOT- 
 AIR STERILI^KK. 
 
624 
 
 APPENDICES. 
 
 (G) APPARATUS AND MATERIAL FOR PREPARING AND STORING 
 NUTRIENT GELATINE AND NUTRIENT AGAR-AGAR. 
 
 Water-bath. A water-bath on tripod stand is required for 
 
 boiling the ingredients of nutrient jellies and for general purposes. 
 
 The lid may be conveniently composed of a series of concentric 
 
 rings, so that the mouth of the vessel may be graduated to any 
 
 size required. 
 
 Test-tube Water-bath. This consists of a circular rack for 
 
 test-tubes within a water-bath. It is sometimes employed instead 
 
 of the steam cylinder for sterilising nutrient jelly in tubes, by 
 
 boiling for an hour for three successive days. 
 
 Hot-water Filter. A copper 
 funnel with double walls, the inter- 
 space between which is filled with 
 hot water. A glass funnel fits in- 
 side the copper cone, the stem of 
 the glass funnel passing through 
 and being tightly gripped by a per- 
 forated caoutchouc plug, which fits 
 in the opening at the apex of the 
 cone, The water in the cone is 
 heated by applying the flame of a 
 burner to a tabular prolongation 
 of the water- chamber. In a more 
 recent model, as represented in 
 Fig. 31, this prolongation is dis- 
 pensed with, and the temperature is 
 maintained by means of a circular 
 burner which acts at the same time 
 as a funnel ring. In Kohrbeck's 
 model the funnel of the filter is 
 connected with a flask, from which 
 
 the test-tubes can be easily filled with the liquid jelly (Fig. 253). 
 Glass Vessels. A number of glass vessels should be kept in 
 
 .stock according to requirements. 
 
 Bohemian hard glass flasks are employed in several sizes, for 
 
 boiling nutrient media. The conical forms are especially used in the 
 
 larger sizes for storing nutrient jelly. 
 
 Glass funnels, large and small, are necessary, not only in the 
 
 processes of preparing nutrient jelly, but for filtering solutions of 
 
 aniline dyes and for general purposes. 
 
 FIG. 253. HOT- WATER FILTERING 
 APPARATUS WITH KING BURNER. 
 
APPARATUS, MATERIAL, AND REAGFA 
 
 A liberal supply of test-tubes should always be kept in stock, as 
 they are not only employed for the tube-cultivations, but can be 
 conveniently used for storing bouillon, sterilised water, etc. 
 
 ( Ylindrical glasses graduated in cubic centimetres, 10 ccm., 100 
 ccm., 500 ccm., are required for measuring the liquid ingredients 
 of nutrient jelly, and also in preparing the various staining 
 solutions. 
 
 A large wide- mouthed glass jar, with a glass cover, is extremely 
 useful. It must be padded at the bottom with cotton wool for 
 containing a stock of tubes of sterilised nutrient jelly, and should 
 be placed within reach on the working table. 
 
 Balance and Weights A balance, with large pans and set of 
 gramme weights, is constantly required. 
 
 Cotton Wool. The best or " medicated " cotton wool should be 
 procured. 
 
 Gelatine. The gelatine for bacteriological purposes must be pf 
 the very best quality (gold label). 
 
 Agar-agar. This is also called Japanese Isinglass ; it consists 
 of the shrivelled filaments of certain Algae (Gracilaria lichenoides 
 and Gigartina speciosa). 
 
 Peptonum Siccum. 
 
 Table Salt. Prepared table salt can be obtained in tins or 
 packets. 
 
 Litmus Papers. Blue or red litmus paper in cheque-books, for 
 testing the gelatine mixture, etc. 
 
 Carbonate of Soda. A bottle, containing a saturated solution 
 of carbonate of soda, and provided with a pipette stopper, may be 
 kept, especially for use in the preparation of nutrient jelly. 
 
 Lactic Acid. 
 
 Filter Paper. For filtering gelatine, stout Swedish filter paper 
 of the best quality is recommended. 
 
 Flannel or Frieze. This is employed as a substitute for, or 
 combined with, filter paper in the preparation of nutrient agar- 
 agar. 
 
 (H) APPARATUS FOR EMPLOYMENT OF NUTRIENT JELLY IN TEST-TUBE 
 AND PLATE-CULTIVAT!' 
 
 Wire Cages. These cages or crates are used for containing 
 test-tubes. .-JM i.-lly wht-n they are to be sterilised in the hot-air 
 steriliser ; or for lowering tubes of nutrient jelly into the steam- 
 steriliser, etc. (Fig. 254). 
 
 40 
 
626 
 
 APPENDICES. 
 
 Test-tube Stands. The ordinary wooden pattern, or the 
 metallic folding stands, are called into use 
 for holding cultivations. Pegged racks are 
 also recommended for draining test -tubes 
 after washing. 
 
 Caoutchouc Caps. These are caps for 
 fitting over the cotton-wool plugs, and may 
 be used in different sizes for test-tubes and 
 stock- flasks. 
 
 Platinum Needles. A platinum 
 needle for inoculating nutrient media, ex- 
 amining cultivations, etc., consists of two or 
 three inches of platinum wire fixed to the 
 Several of these needles should be made with 
 platinum wire of various thicknesses. A piece of glass rod, about 
 seven inches long, is heated at the extreme point in the flame of 
 
 FIG. 254. WIRE CAGE 
 FOR TEST-TUBES. 
 
 end of a glass rod. 
 
 FIG. 255. PLATINUM NEEDLES; STRAIGHT, HOOKED, LOOPED. 
 
 a Bunsen. burner, and a piece of platinum wire, held near one 
 extremity with forceps, is then fused into the end of the rod. 
 Some needles should be perfectly straight, and kept especially for 
 
 FIG. 256. DAMP CHAMBER FOR PLATE-CULTIVATIONS. 
 
 inoculating test-tubes of nutrient jelly. For other purposes the 
 needles may be bent at the extremity into a small hook, and 
 others provided with a loop (Fig. 255). 
 
APPARATUS, MATERIAL, AND REAGENTS. 
 
 627 
 
 Tripod Le veiling -stand. A triangular wooden frame sup- 
 ported upon three screw-feet which enable it to be raised or lowered 
 to adjust the level. 
 
 FIG. 257. APPARATUS EMPLOYED FOB PLATE-CULTIVA i 
 
 Tripod Stand ; Glass Dish, filled with cold or iced water ; Sheet of Plate-glass ; 
 Spirit Level, and Glass Bell. 
 
 Large Glass Plate. A piece of plate-glass,, or a pane of 
 ordinary window-glass, about a foot square. 
 
 Spirit Level. 
 
 Glass Bells and Dishes. Shallow glass bells and dishes, for 
 making a dozen or more damp chambers 
 (Fig. 256), and for completing the apparatus 
 for pouring out liquefied nutrient jelly on 
 glass plates or slides (Fig. 257). 
 
 Iron Box. A box of sheet-iron 
 (Fig. 258), for containing glass plates during 
 their sterilisation in the hot-air steriliser, 
 and for storing them until required for use. 
 
 Glass Plates. Small panes of glass, 
 about six inches by four. Not' less than 
 three dozen are required for a dozen damp 
 chambers. 
 
 Glass Benches. These are necessary for arranging the glass- 
 plates or slides in tiers in the damp chambers (Fig. 256). MetaL 
 
 FIG. 258. Box FOB 
 GLASS PLATES. 
 
 FIG. 259. GLASS BENCHES FOB GLASS PLATES OB Si 
 
 shelves may be substituted for them, but the former are to be 
 preferred. They can be easily made, in any number required, by 
 
-628 
 
 APPENDICES. 
 
 cementing a little piece of plate-glass at either end of a 
 slip (Fig. 259). 
 
 Glass Rods. One dozen or more glass rods, twelve to eighteen 
 inches in length. They are employed for smoothly spreading out 
 the liquefied nutrient gelatine or agar-agar on the glass plates, etc. 
 
 Thermometers. Two or three centigrade thermometers. 
 
 (I) APPARATUS FOR PREPARATION OF POTATO-CULTIVATIONS. 
 
 Israel's Case. Sterilising instruments in the flame of a Bunsen 
 .burner is most destructive. It is better, therefore, to have a sheet- 
 iron case (Fig. 260) to 
 contain potato-knives, 
 scalpels and other in- 
 struments, and to ster- 
 ilise them by placing 
 the case in the hot-air 
 steriliser for an hour 
 at 150 C. The box 
 can be opened at the 
 
 FIG. 2GO. -ISRAEL'S CASE. side > and eacn instru- 
 
 ment withdrawn with 
 a pair of sterilised forceps when required for ase 
 
 Glass Dishes. Several shallow glass dishes are required for 
 preparing damp chambers for potato-cultivations (Fig. 261). The 
 upper, being the larger, fits 
 over the lower, and having 
 no handle, admits of these 
 damp chambers being placed, 
 if necessary, in the incubator 
 in tiers. The large size may 
 also be used in the same 
 way for plate-cultivations. 
 
 Potato Knives. A 
 common broad smooth-bladed knife set in a wooden handle is sold 
 for this purpose. 
 
 Scalpels. Half a dozen scalpels, preferably with metal handles, 
 may be kept especially for inoculating sterilised potatoes. 
 
 Brush. A common stout nail-brush, or small scrubbing-brush, 
 is essential for cleansing potatoes. 
 
 FIG. 2G1. DAMP CHAMBER FOR POTATO- 
 CULTIVATION. 
 
A1TAR.VITS, MATKKIAL, AND REAGENTS. 
 
 (J) APPARATUS FOR PREPARATION OP SOLIDIFIED STERILE 
 BLOOD-SERUM. 
 
 Glass Jar. A tall cylindrical glass jar, on foot, with a broad 
 ground stopper, for receiving blood. 
 
 Pipette. An ordinary or graduated pipette, for transferring the 
 serum from the jars to sterile test-tubes or glass capsules. 
 
 Koch's Serum Ster- 
 iliser. A cylindrical ra>r. 
 with double walls forming an 
 interspace to contain water, 
 closed with a lid, also double- 
 walled and provided with a 
 tubular prolongation of the 
 enclosed water-chamber (Fig. 
 262). The water in the 
 cylinder is heated from below, 
 and that in the lid by means 
 of the prolongation. 
 
 In the centre of the 
 cylinder is a column which 
 communicates with the water- 
 chamber of the cylinder, and 
 from it pass four partitions, 
 which serve to support the 
 test-tubes. 
 
 In the lid are three 
 
 openings, one of which communicates with the water-chamber in the 
 lid by which the latter is filled, and into which a thermometer is 
 
 then fixed. In the centre an 
 opening admits a thermo- 
 meter, which passes into the 
 central pipe of the cylinder ; 
 through a third opening a 
 
 FIG. 262. KOCH'S SERUM STERILISER. 
 
 thermometer passes to the 
 cavity of the cylinder. The 
 -r Under and cover are jacketed 
 with felt, and the apparatus 
 i> supported on iron legs. 
 
 Koch's Serum Inspis- 
 sator. A shallow tin case 
 with glass cover, both case and cover jacketed with felt (Fig. 263) 
 
 FIG. 2G3. SKI.I.M INM-ISSATOR. 
 
630 APPENDICES. 
 
 The case is double-walled, and the water contained in the interspace 
 is heated from below. It is supported on four legs, and the two 
 front ones move in grooves in the case, so that the latter can be 
 placed obliquely at the angle required and secured in position by 
 screw-clamps. It is employed for coagulating sterile liquid serum, 
 and for solidifying nutrient agar-agar so as to give them a sloping 
 surface. 
 
 Hueppe's Serum Inspissator. By the new process the serum 
 is obtained with every possible precaution, and solidified at once in 
 Hueppe's apparatus (Fig. 44). 
 
 Glass Capsules. Small capsules or hollowed-out cubes of 
 crystal glass are employed for cultivation on solid blood-serum, on 
 nutrient gelatine, and on agar-agar. They may be procured of 
 white and blackened glass, and are provided with glass slips as 
 covers. 
 
 (K) APPARATUS FOR STORING, AND FOR CULTIVATIONS IN, LIQUID 
 
 MEDIA. 
 
 Lister's Flasks. Lister devised a globe-shaped flask with two 
 necks a vertical and a lateral one. The lateral one is a bent spout, 
 tapering towards its constricted extremity. When the vessel is 
 restored to the erect position after pouring out some of its contents, 
 a drop of liquid remains behind in the end of the nozzle, and 
 prevents the regurgitation of air through the spout. A cap of 
 cotton wool is tied over the orifice, and the residue in the flask kept 
 for future use. The vertical neck of the flask is plugged with 
 sterilised cotton wool in the ordinary way (Fig. 60). 
 
 Sternberg's Bulbs. Sternberg advocates the use of a glass 
 bulb, provided with a slender neck drawn out to a fine point and 
 hermetically sealed (Fig. 62). 
 
 Aitken's Test-tube. This is an ingenious device for counter- 
 acting the danger of entrance of atmospheric germs on removal from 
 the ordinary test-tube of the cotton-wool plug. Each test-tube is 
 provided with a lateral arm tapering to a fine point, which is 
 hermetically sealed (Fig. 62). 
 
 Drop-culture Slides. About a dozen or more thick glass 
 slides with a circular excavation in the centre are required for 
 drop-cultures (Fig. 48). 
 
 Vaseline. A small pot of vaseline with a camel's-hair brush 
 should be reserved especially for use in the preparation of drop- 
 cultures. 
 
APPARATUS, MATERIAL, AND REAGENTS. 
 
 631 
 
 Bulbed Tubes. Glass vessels, such as test-tubes, flasks 
 and pipettes, which are used in dealing with liquid media, have 
 already been mentioned under other headings; but bulbed tubes, 
 Pasteur's bulbs, and various other forms are also required for special 
 experiments. 
 
 (L) APPARATUS FOR INCUBATION. 
 
 There are several forms of incubator, each of which has its 
 advocates. They are mostly rectangular chests, with glass walls 
 front and back, or in front 
 only. A cylindrical model 
 is preferred by some. Two 
 only will be described 
 here D'Arsonval's and 
 Babes'. The former admits 
 of very exact regulation of 
 temperature, and the latter 
 is a very practical form for 
 general use. 
 
 D'Arsonval's Incu- 
 bator. The " Etuve 
 D'Arsonval" (Fig. 264) is 
 a very efficient apparatus, 
 and is provided with a heat- 
 regulator, which enables 
 the temperature to be 
 maintained with a mini- 
 mum variation. It consists 
 of a cylindrical copper 
 vessel, with double walls, 
 enclosing a wide interspace 
 for containing a large 
 volume of water. The roof 
 of the water-chamber is 
 oblique, so that the wall 
 
 rises higher on one side than on the other. This admits of the inter- 
 space being completely filled with water. At the highest point is 
 an opening fitted with a perforated caoutchouc stopper, through 
 which a glass tube passes. The mouth of the cylinder itself is 
 horizontal, and is closed by a lid, which is also double-walled to 
 contain water. In the lid are four openings : one serves for filling its 
 
 FIG. 264. D'ARSONVAL'S INCUBATOR. 
 
632 APPENDICES. 
 
 water-chamber, and the others for thermometers and for regulating 
 the air supply in the cavity of the cylinder. The cylinder is con- 
 tinued below by a cone, also double- walled, and there is a perforated 
 grating at the line of junction of the cylinder and cone. The cone 
 terminates in a projecting tube provided with an adjustable 
 ventilator. The apparatus is fixed on three supports united to 
 one another below. One of them is utilised for adjusting the height 
 of the heating apparatus. Situated above this leg is the heat- 
 regulating apparatus (Fig. 265), attached to a circular, lipped 
 
 aperture in the outer wall of the 
 incubator. To the lip is fixed with 
 six screws the corresponding lip of 
 a brass box, with a tightly-stretched 
 diaphragm of indiarubber inter- 
 vening. Thus the diaphragm 
 separates the cavity of the box from 
 the water in the interspace of the 
 incubator. The cap of the box, 
 
 which screws on, is bored in the 
 FIG. 265. SCHLOSING'S MEMBRANE 
 
 REGULATOR. centre for the screw- pipe, by which 
 
 the gas is supplied. Another pipe 
 
 entering the box from below is connected with the gas-burners. 
 Around the end of the screw-pipe a collar loosely fits, and is pressed 
 against the diaphragm by means of a spiral wire spring. Close 
 to the mouth of the screw-pipe a small opening exists, so that the 
 gas supply to the burners is not entirely cut off even when the 
 diaphragm completely occludes the mouth of the screw-pipe. 
 
 To work the apparatus the tube and plug must be removed, and 
 the water-chamber filled completely with distilled or rain water at the 
 temperature required. The caoutchouc plug is replaced and the tube 
 pkced in position. Gas enters through d (Fig. 265), and passes through the 
 opening at its extremity into the chamber of the box. Thence it passes 
 through the vertical exit which is connected with the gas-burners. As 
 the temperature rises the water rises in the tube, and at the same time 
 exercises a pressure on every part of the walls of the incubator, and 
 hence on the diaphragm. In consequence of this, the diaphragm bulging 
 outwards approaches the end of the tube d, and gradually diminishes the 
 gas supply. As a result the temperature falls, the water contracts and 
 sinks in the tube, and the diaphragm receding from d, the gas supply 
 is again increased. By adjusting the position of the tube d to the 
 diaphragm, any required temperature within the limits of the working of 
 the apparatus can be regulated to the tenth of a degree provided (1) 
 that the gas supply is rendered independent of fluctuations of pressure 
 
APPARATUS, MATERIAL, AND REAGENTS. 
 
 by means of a gas-pressure regulator ; (2) that the height of the water 
 in the tube is controlled daily by the 
 withdrawal or addition of a few drops of 
 distilled water ; and (3) that the apparatus 
 is kept in a place with as even a tempera- 
 ture as possible, and sheltered from currents 
 of air. 
 
 The burners in Fig. 264 are protected 
 with mica cylinders similar to the burner 
 represented in Fig. 266. The flames of 
 these burners can be turned down to the 
 smallest length without danger of extinction, 
 and the temperature may be regulated very 
 satisfactorily without using the heat-regulator 
 just described, if the gas first passes through 
 a pressure-regulator (Fig. 269). To provide FlG - 266. GAS-BURNER 
 against the danger resulting from accidental 
 
 ... ., . TT !_ t i t UTIJKDWL 
 
 extinction of the gas, Koch has devised a 
 
 self-acting apparatus (Fig. 267), which, simultaneously with the extinction 
 
 of the flame of the burner, shuts off the supply of gas. 
 
 FIG. 267. KOCH'S SAFETY BURNER. 
 
 Babes' Incubator. The pattern used by Babes is a veiy 
 simple one, and may be recommended for economy and efficiency 
 (Fig. 268). 
 
 It consists of a double-walled chest with sides and roof jacketed 
 with felt. Water fills the interspace between the walls, and on 
 the roof are two apertures one for a ga>-n-Lriil;itor and the other 
 
634 
 
 APPENDICES. 
 
 for a thermometer. In front the chest is closed in by a sheet 
 
 of felt, a glass door, and a sliding 
 glass panel. The apparatus can be 
 suspended on the wall or supported 
 on legs, and is heated from below 
 by means of protected burners. 
 
 The gas should pass first through 
 a pressure-regulator, and then 
 through a thermo-regulator to the 
 burners. 
 
 Moitessier's Gas - pressure 
 Regulator. This apparatus is best 
 explained by reference to the dia- 
 gram (Fig. 269). In the bottom of 
 the cylinder (A) are the entrance (k) 
 and exit () gas-tubes. The tap (m) 
 regulates the size of the flame. The 
 cover (n n) roofs in the cylinder (A). 
 The bell (B) supports, by means of 
 e and /, the ball valve (cZ), which 
 lies in the cover (c c). The gas, 
 entering by k, passes through the 
 
 valve (cZ), and is thence conducted by the tube a 'to the tube I. The 
 
 bell (B) and the weighted dish (h) are screwed on to the connecting- 
 rod (g). To diminish as 
 
 much as possible the 
 
 friction of g in i, g 
 
 only touches i by three 
 
 projecting ridges. Section 
 
 of i and g is shown at s. 
 
 To put the apparatus in 
 
 use it is first levelled, then 
 
 h is screwed off, and the 
 
 cover (n n) removed. A 
 
 mixture of two parts of 
 
 pure acid-free glycerine to 
 
 one of distilled water is 
 
 poured into the . cylinder 
 
 until it flows out at q, 
 
 which is then closed, and FlG< 209. -MOITESSIER'S GAS-PRESSURE. 
 
 ,, REGULATOR. 
 
 the cover (n n) replaced. 
 
 The manometers are filled with coloured water, and k and I 
 
 FIG. 268. BABES' INCUBATOR. 
 
APPARATUS, MATERIAL, AND REAGENTS. 635 
 
 connected with the entrance and exit gas tubing respectively. The 
 pressure of the incoming gas raises the bell (B) ; and with it the valve 
 (d) is raised towards the opening at c c. The weight (A), which is 
 replaced on g, by its downward pressure counteracts this upward 
 pressure of the gas and opens the valve (c c). Thus the flame is 
 best regulated in the morning, when the pressure is at a minimum ; 
 then supposing an increase of pressure occurs, the weight of A is 
 overbalanced, B is raised, and with it d, and the gas supply pro- 
 portionately diminished by the gradual closing of the valved opening. 
 Reichert's Thenno-regulator. This regulator (Fig. 270) 
 consists of three parts a hollow J-piece, a stem and a bulb. 
 The T-piece fits like a stopper in the upper widened 
 portion of the stem. One arm of the J is open 
 and connected with the gas supply; the vertical 
 portion terminates in a small orifice, and is also 
 provided with a minute lateral opening. The stem is 
 provided with a lateral arm, and this arm, the stem, 
 and the bulb contain mercury. The regulator is 
 fixed in the roof of the incubator, so that the bulb 
 projects either into the interior of the incubator or 
 into the water-chamber. When the incubator reaches 
 the required temperature, the mercury is forced up 
 by means of the screw in the lateral arm, until it FIG. 270. 
 closes the orifice at the extremity of the vertical RKICHKRT'S 
 
 T^HKRMO- 
 
 portion of the T". The gas which passes through REGULATOR. 
 the lateral orifice is sufficient to maintain the 
 apparatus at the required temperature. If the temperature of the 
 incubator falls, the mercury contracts, and gas passing through the 
 terminal orifice of the T increases the flame of the burner, and 
 the temperature is restored. 
 
 Page's Thenno-regulator resembles the above, but instead 
 of the T-piece there are two pieces of glass-tubing. The outer 
 tubing envelops the upper part of the stem of the regulator, and 
 admits of being raised or lowered. The upper end of this tubing 
 is closed by a cork, which is perforated to admit the narrow glass- 
 tubing, which represents the vertical arm of the T> passing within 
 the stem of the regulator. This has a terminal and a lateral 
 opening, and is the means of entrance for the gas. This regulator 
 is adjusted by noting when the thermometer indicates the desired 
 temperature, and then pushing down the outer tube until the 
 terminal opening of the inner tube, which is carried down with 
 it, is obstructed by the mercury. 
 
636 
 
 APPENDICES. 
 
 Meyer's Thermo-regulator is represented in Fig. 271. No. I. 
 
 shows the construction of the regulator : its inner tube terminates 
 
 in an oblique opening, and is also provided with a minute lateral 
 
 perture, which prevents the complete shutting off of the gas supply. 
 
 FIG. 271. MEYER'S THERMO-REGULATOR. 
 
 No. II. illustrates the method of introducing the mercury by suction 
 through a filling tube, which is substituted for the inner tube of the 
 regulator. No. III. represents Frankel's modification of the same 
 instrument. 
 
 (M) INOCULATING AND DISSECTING INSTRUMENTS AND APPARATUS 
 IN COMMON USE. 
 
 Mouse-cages. As mice are the animals most frequently 
 employed for experimental purposes, mouse-cages have been 
 
APPARATUS, MATERIAL, AND REAli! VI -. 
 
 637 
 
 especially introduced, consisting simply of a cylindrical glass jar 
 with a weighted wire cover. 
 
 Dressing-case. A small surgu-ul divs>iiiLT-niM'. with it> usual 
 accessories forceps, knives, small, straight and curved scissors, 
 needles, silk, and so forth will serve for most purposes. 
 
 Pravaz' Syringe. Koch's modification of Pravaz' syringe 
 admits of sterilisation by exposure to 150 C. for a couple of hours. 
 
 Special Instruments and Material. Instruments required 
 for special operations, and the materials necessary for strict anti- 
 septic precautions, need not be detailed here.* 
 
 Dissecting-boards. Slabs of wood in various sizes, or gutta- 
 percha trays, provided with large-headed pins, are employed for 
 ordinary purposes. 
 
 Dissecting-case. A dissecting-case, fitted with scalpels, 
 scissors, hooks, etc., should be reserved entirely for post-mortem 
 examinations. 
 
 FIG. 272. SIPHON BOTTLE, WITH FLEXIBLE TUBE, GLASS NOZZLE, AND A 
 MOHR'S PINCHCOCK, 
 
 (X) GENERAL LABORATORY REQUISITES. 
 
 Siphon Apparatus. Two half-gallon or gallon glass bottles, 
 %vith siphons connected with long flexible tubes provided with 
 glass nozzles and pinchcocks (Fig. 272), should be employed for the 
 * Vide Cheyne, Antiseptic Surgery. 1882. 
 
638 
 
 APPENDICES. 
 
 following purposes : One is used to contain distilled water, with 
 the nozzle hanging clown conveniently within reach of the working 
 table; the other is to contain a solution of carbolic acid (1 in 20), 
 and may be placed so that the nozzle hangs close to the lavatory 
 sink or basin. The former replaces the use of the ordinary wash- 
 bottle, in washing off surplus stain from cover-glasses, etc., and the 
 latter is conveniently placed for disinfection of vessels and hands 
 after cleansing with water. They should be placed on the top of a 
 cupboard or on a high shelf. 
 
 Desiccator. The desiccator (Fig. 273) consists of a porcelain 
 
 pan containing concentrated 
 sulphuric acid and covered 
 over with a bell-glass receiver. 
 The sheet of plate-glass upon 
 which the pan rests is ground 
 upon its upper surface, and 
 the rim of the glass bell is 
 also ground and well greased. 
 In the centre of the pan is a 
 column supporting a circular 
 frame, which is covered with 
 wire gauze. Slices of potatoes, 
 upon which micro-organisms 
 have been cultivated, are 
 rapidly dried by the action of 
 sulphuric acid in confined air. 
 
 A detailed description of other kinds of apparatus commonly in 
 use in a research laboratory such as the various forms of apparatus 
 for filtering cultures in liquids, and the reagents necessary for 
 special chemical investigations must be sought for elsewhere. 
 Much information may be obtained about the most recent improve- 
 ments in bacteriological, chemical and physical apparatus by 
 reference to manufacturers' catalogues.* 
 
 * All bacteriological apparatus may be obtained from Berlin from Dr. 
 Muencke, 58, Louisen Strasse, or Dr. Hermann Eohrbeck, 24. Karlstrasse. 
 Dr. George Griibler, Leipzig, is recommended for special staining reagents. 
 In London, chemicals and bacteriological apparatus can be obtained from 
 Becker & Co., Hatton Wall, or from Baird & Tatlock, 14, Cross Street, Hatton 
 Garden, B.C. Mr. Baker, of High Holborn, W.C., is recommended as the 
 agent for microscopes and objectives by Continental makers, including 
 Zeiss' apochromatic objectives. 
 
 FIG. 273. DESICCATOK. 
 
APPENDIX V. 
 BIBLIOGRAPHY 
 
 CHAPTER I. 
 
 HISTORICAL INTRODUCTION. 
 
 Andry, De la Generation des Vers dans le Corps de THomme, 1701. Charlton 
 Bastian, Proc. Royal Soc. 1872. Bonnet, Considerations sur les Corps orga- 
 nises, 1768. Davaine, Corapt. Rend., T. Iviii. and lix. Gleichen. Dissertation 
 sur la Generation, 1778. Hill, Essays in Natural History and Philosophy. 
 Kircher, Ars magna lucis et umbrae, 1646. Koch, Beitrage zur Biologic der 
 Pflanzen, 1876. Lister, Pharmaceut. Journal and Transact., 1877. Miiller, 
 Animalia infusoria, 1786. Pasteur, Compt. Rend., 1859, 1880 ; Etude sur la 
 Maladie des Vers-a-soie, 1870. Plenciz, Opera Medico- Physica, 1762. Schroder 
 and Von Dusch, Ann. der Chem. und Pharm., vol. Ixxxix. Schultze, Poggen- 
 dorff's Ann., vol. xxxix. Schwann, PoggendorfFs Ann., vol. xli. Tyndall, Essays 
 on floating matter of the air, 1881. 
 
 CHAPTER II. 
 
 MORPHOLOGY AND PHYSIOLOGY OF BACTERIA. 
 
 MORPHOLOGY. 
 
 Baumgarten, Lehrbuch der Pathologischen Mykologie, 1890. Biedert. 
 Virch. Archiv, Bd. 100, 18S5. Billroth, Unters. iiber d. Veg. Form der Cocco- 
 bacteria septica, 1874. Brefeld, Botanische Untersuch. iiber Schimmelpilze* 
 Heft 1, 1*81. Cohn, Beitrage zur Biologie der Pflanzen, Bd. I., 1872, 1875. 
 Cornil and Babes, Les Bacteries, 1885. Dallinger and Drysdale, Monthly Micro- 
 scop. Journ., 1875. De Bary, Verg. Morph. und Itiolog. der Pilze, Mycetozoen, 
 und Bacterien, 1884 ; Vorlesungen iiber Bacterien, 1885. Dujardin, Histoire 
 Naturelle des Zoophytes, 1841. Ehrenberg, Die Infusionsthierchen als Volkom. 
 Organism, 1838. Fisch, Biolog. Centralbl., V., iHsf,. Fliigge, Handbuch 
 der Hygiene, 1883; Die Micro-organismen, 2nd Edition, 1886. Gram, 
 Fortsch der Med., II., No. 6. Grove, Synopsis of the Bacteria and Yeast 
 Fungi, 1884. Hallier, Die Pflanzlichen Parasiten, 1866. Hauser, Ueber 
 Faulniss Bacterien, 1H>5. Hueppe, Die Formen der Bakterien, issr, Lankester, 
 Quart. Journ. Microscop. Science, 1*73. Leunis, Synopsis d. Pflanzenkunde : 
 
 639 
 
640 APPENDICES. 
 
 Hanover, 1877. Lister, Quart. Journ. Microscop. Science, 1873. Lutz, 
 Fortschr. d. Med., 1881. Marpmann, Die Spaltpilze, 1884. Miller, Deut. Med. 
 Woch., 1884. Nageli, Die Niederen Pilze, 1877. Neelsen, Biol. Centralbl., III., 
 No. 18, 1883. Sachs, Text-book of Botany, 1882. Van Tieghem, Compt. 
 Rend., 1879 ; Traite de Botanique, 1883. Zopf, Die Spaltpilze, 1885. 
 
 GENERAL BIOLOGY. 
 
 Arloing, Archiv de Physiol., 1886. Bordoni-TJffreduzzi, Fortschr. d. Med., 
 1886. Cheyne, Brit. Med. Journ., 1886. (John and Mendelsohn, Beitr. z. 
 Biol. d. Pflanzen, Bd. III., Heft 1. Cortes, Compt. Rend., T. 99, p. 385, 1884. 
 Downes, Proc. Roy. Soc., 1886. Duclaux, Compt. Rend., 1885. Engelmann, 
 Arch. f. d. Ges. Physiologic, Bd. 26, 1881 ; Botan. Zeitg., 1882. Fodor, 
 Archiv f. Hygiene, 1886. Hauser, Archiv f. Exper. Patholog. u. Pharmacologie, 
 1886. Hofmann, Allgem. Med. Centralbl., S. 605. Liborius, Zeitschrift f. 
 Hygiene, 1886. Nageli, Die Niederen Pilze : Miinchen, 1877 ; Unters. iiber 
 Niedere Pilze : Miinchen, 1882. Nencki, Virchow's Archiv, 1879 ; Beitrage zur 
 Biol. der Spaltpilze : Leipzig, 1879 ; Ber. d. Deutschen Chem. Gesellsch., S. 
 2605, 1884. Begnard, Compt. Rend., T. 98, p. 744, .1884. Tumas, St. Petersb. 
 Med. Wochenschr., 1879. Wyssokowitsch, Zeitschrift f. Hygiene, 1886. 
 
 CHROMOGENIC BACTERIA. 
 
 Babes, Biolog. Centralbl., Bd. 2, 1882. Chabert and Fromage, D'une Altera- 
 tion du Lait de Vache, designee sons le Norn du Lait Bleu, 1880. Charrin, 
 Communication faite a la Societe Anatomique, 1884. Cohn and Miflet, Cohn's 
 Beitrage zur Biol. d. Pflanzen, Bd. III., Heft 1. 1879. Eberth, Centralbl. f. d. 
 Med. Wissensch., 1863. Ehrenberg, Micr. Prodigiosus, Verhandl. d. Berl. 
 Acad., 1839. Fordos, Compt. Rend, de 1'Acad. de So., 1860. Frank, Cohn's 
 Beitr. z. Biol. d. Pflanzen, Bd. I., Heft 3, 1875. Gessard, De la Pyocyanine 
 et de son Microbe, 1882 ; Ann. de 1'Institut Pasteur, T. iv. Gielen, Mag. f. 
 Ges. d. Thierheilkunde, 1852. Girard, Unters. uber Blauen Eiter ; Chirurg. 
 Centralbl., II., 1875; Revue des Sc., T. 5, 1877. Hermstadt, Ueber die 
 Blaue und Rothe Milch., 1833. Hugues, Echo Veterinaire, 1884. Klein, 
 Quart. Journ. of Micr. Sc., Vol. 15, 1875. Lankester, Quart. Journ. of Micr. Sc., 
 Vol. 13, 1873, 1876. Liicke, Arch. f. Klin. Chir., 1862. Mosler, Virchow's 
 Archiv, Bd. 43, 1868. Neelsen, Cohn's Beitr. z. Biologic d. Pflanzen, Bd. III., 
 Heft 2, 1880. Schrb'ter, Cohn. Beitr. z. Biol. der Pflanzen, Bd. I., Heft. 2, 
 1872. Steinhoff, Neue Ann. d. Mecklenb. Landw. Ges., 1838. Wernich, Cohn's 
 Beitrage zur Biol. d. Pflanzen, Bel. III., Heft 1, 1879. Van Tieghem, Bull, de 
 la Soc. Bot. de France, 1880. 
 
 ZYMOGENIC BACTERIA AND FERMENTATION. 
 
 Be"champ, Compt. Rend., T. 60, p. 445, 1865 ; T. 93, 1881. Boutroux, Compt. 
 Rend., T. 86, 1878. Brefeld, Landwirthsch. Jahresber., Bd. 3, 1874 ; Bd. 4, 
 1IS75; Bd. 5, 1876. Cienkowski, Die Gallertbildungen rl. Zuckerriibensaftes, 
 1878. Colin, Bull, de 1'Acad. de Med., 1875. Dubrunfaut, Compt. Rend., 
 T. 73, 1871. Duclaux, Theses presentees a la Faculte de Paris, 1865. 
 Dumas, Compt. Rend., T. 75. Nr. 6, 1872 ; Ann. de Chim. et de Phys., 1874. 
 Eriksson, Unters. aus. d. Botan.' ; Institut in Tubingen, Heft 1, 1881. Feltz 
 and Bitter, Journ. de 1'Anat. et Phys., 1874. Fermi, Centralb. f. Bact., 
 1891. Fitz, Berichte d. Chem. Ges., Bd. 6, S. 48, 1875; Bd. 10, p. 216, 1878 ; 
 
BIBLIOGRAPHY. 641 
 
 Bel. 11, pp. 42 and 498, and Bd. 12, p. 474, 1879; Bd. 13, p. 1309, U 
 Bd. 15, p. 857/1882; Bd. 16, p. 841. L888; IJd. 17. ,>. HSK, issj. Fleck, 
 Ber. d. Chem. Centralst. : Dresden, 1*7*;. Frankland, < 'suitor Lectures, 
 Gessard, De la Pyocyanine et de son Microbe. Ouiard, These ! P:iris. 
 Hallier, (Jjihrungserscheinungen, 1867. Hansen, I'litcrsuch. aus. d. 1'rax. .1. -r 
 (iahrungsindustrie, 1890, 1892. Harz, Grundziige der alkoholischen - 
 rungslehre, 1877. Hiller, C'entralbl. f. d. M-d. Wiss.. S. :,::. 1^71. Hofmann, 
 Arr/tl. Yen-in /u Wien. Mai 1873; Allgem. Med. Centralbl., S. f,o:,, 1873. 
 Hoppe-Seyler, Medic.-Chem. Untersuchungen, Heft 4, 1871. Hueppe, Mittli. 
 a. d. Ges. Amt, Bd. ii., 1884; Deut. Mi-d. W,oh., 1884. Jacksch, Zeitschr. f. 
 I'liysiol. Chemie, Bd. 5, 1881. Jorgensen, Microorg. of Ferment Trans., 1893. 
 Karsten, Cheraismus der Pflanzenzelle, 1869. Kern, Bull. <le la Soc. Imp. des 
 Xaturalistesde Moskau, Xo. 3, 1881. Krannlials, Deut. Arch. f. Klin. M.-.I.. 
 Bd. 35, 1884. Ladureau, Compt. end., T. 99, p. 877, 1884. Le"pine and Eoux, 
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 Akad. d. Wiss.. 1861 ; 5 Nov. 1869 ; Ueber Gahrung, Quelle der Muskel- 
 kraft und Ernahrung ; Leipzig u. Heidelberg, 1870. Lister, The Pharmac. 
 Journ. and Transact., 1877. Mayer, Lehrbuch der Gahrungschemie ; 2 Aufl., 
 1876. Monoyer, These de Strassburg, 1862. Mttller, Journ. f. Prakt. Chem., 
 IM;O. Musculus, Ber. d. Chem. Ges., S. 124, 1874; Compt. Rend., T. 78, 1874. 
 Nageli, Theorie der Gahrung : Munchen, 1879. Pasteur, Annal. de Chim. et 
 de Phys., III. Ser., T. 58. 1860 ; Compt. Rend., 1860, 1861, 1863, 1864, 1871, 
 l^Ti' ; Bull, de la Soc. Chim., 1861; Ann. de Chim. et de Phys., T. 64, 1862; 
 Etudes sur le Vin, 1866 ; Bull, de 1'Acad. de Med., Xo. 27, 1876 ; Etudes sur 
 la Biere, 1876. Pasteur and Jonbert, Compt. Rend., T. 83, 1876. Popoff, Botan. 
 Jahresber., 1875. Prazmowski, Untersuchungen Uber die Entwicklungs- 
 geschichte und Fermentwirkung einiger Bakterien, 1880. Bichet, Compt. 
 Rend.. T. 8H, 1879. Scheibler, Zeitschr. f. Riibenzuckerindustrie, 1874, 
 Schtitzenberger, Die Gahrungserscheinungen, 1874. Sheridan Lea, Journ. 
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 des Sc., Ser. 7, T. 7, 1867. Tyndall, Compt. Rend., T. .>. l>iil : Kssays on 
 the Floating Matter of the Air, 1881. Van Tieghem, Compt. Rend., 1864, 
 1874, 1879, 1880, 1884. 
 
 PHOTOGENIC BACTBBIA. 
 
 Beyerinck, Archiv Xeerland, XXIII. Fischer, Zeitschr. f. Hygiene, 1887; 
 Centralbl. f. Bakteriolog., 1888. Forster, Centralbl. f. Bakteriolog., 1887. 
 Girard, Compt, Rend., 1890. Oirard and Billet, Compt. Rend.. 1889. Katz, 
 < V-ntralbl. f. Bakteriol., 1891. Lehmann, Centralbl. f. Bact., ls<9. Ludwig, 
 Centralbl. f. Bact.. 1887. 
 
 CHAPTER III. 
 
 EFFECT OF ANTISEPTICS AND DISINFECTANTS ON BACTERIA. 
 
 Arloing, Cornevin and Thomas, Ly<m Med., 1883. Blyth, Proc. Roy. Soc., 
 1885. Buchholz, U-ber das Verhalten von Bakterien zu einigen Antiseptics, 
 1 -7'; : Arch. f. Exp. Pathol., P,d. 7, 1^77. Chairy, Compt. Rend., 1 88 1. Chamber- 
 land and Eoux, Compt. Rend., 1883. Chauveau, Compt. Rend.. lss;{, 1884. 
 Cheyne, Antiseptic Surgery, 1**'2. Colin, Compt. Ki-nd.. T. '.''.', 1884, De la 
 
642 APPENDICES. 
 
 Croix, Arch. f. Exp. Pathol.. Bd. 13, 1881. Dujardin-Beaumetz, Bull, de 1'Acad.. 
 de Med. de Paris, 1884. Eidam, Cohn's Beitr. zur Biol., Bd. I., Heft 3, 1875. 
 Fischer, Berl. Klin. Woch., 1882. Fischer and Proskauer, Mitth. a. d. Kaiserl. 
 Ges. Amt, Bd. II., 1884. Frank, Ueber Desinfection von Abtrittsgruben, 1885. 
 Frisch, Sitzungsber. d. Wiener Akad., Bd. 75 u. 80, 1877. Gartner and Plagge, 
 Deut. Med. Woch., 1885. Haberkorn, Das Verhalten von Harnbakterien gegen 
 einige Antiseptica ; Dissert. Dorpat., 1879. Handford, Brit, Med. Journ., 1885. 
 Heydenreich, Compt. Rend., T. 98, 1884. Hoffmann, Experimentelle Unter- 
 suchungen liber die Wirkung der Ameisensaure Diss. Greifswald, 1884. 
 Hueppe, Mittheilg. a. d. Kaiserl. Ges. Amt, Bd. I., S. 341, 1881 ; Deut. Militar- 
 arztl. Zeitschr., 1882. Koch, Cohn's Beitr. zur Biol. der Pflanzen, Bd. II., 
 Heft 2, 1876 ; Mitth. a. d. Ges. Amt, Bd. I., S. 234, 1881. Koch and Gaffky, 
 Arbeit, a. d. K. Gesundh. Amt, 1885. Koch, Gaffky and Lbffler, Mitth. a. 
 d. Kaiserl. Ges. Amt, Bd. I., S. 322, 1881. Koch and Wolff hiigel, Mitth. a. d. 
 Kaiserl. Ges. Amt, Bd. I., S. 301, 1881. Kbnig, Chirurg. Centralbl., 1885. 
 Laillier, Ann. d'Hygiene, 1883. Larrive, L'Eau Oxygenee : These de Paris, 
 
 1883. Lassar, Deut. Med. Woch., 1880. Lebedeff, Arch, de Physiol. Norm, et 
 Pathol., 1882. Maly and Emich, Sitzungsber. d. Kais. Akad. d/Wiss. zu Wien. 
 Jan. 1883. Marie-Davy, Eevue d'Hygiene, 1884. Merke, Virchow's Archiv,. 
 Bd. 81, 1880. Meyer, Ueb. d. Milchsaureferment u. sein Verhalten gegen 
 Antiseptica, 1880. Mignet, Annuaire de 1'Observatoire de Montsouris, 1884- 
 Miojiel, Semaine Medicale, 1883. Mbrscheli, Deut, Med. Woch., 1880. Nageli, 
 Die Niederen Pilze, 1877. Pasteur, Ann. d'Hyg., 1880 ; La Vaccination Char- 
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 CHAPTER IV. 
 
 CHEMICAL PRODUCTS OF BACTERIA. 
 
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BIBLIOGRAPHY. 643 
 
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 Rend., T. 92, 1881. Tappeiner, Med. Centralbl., 1885. Vandevelde, Arch, de 
 Biol. par van Beneden, 1884. Willgerodt, Ueber Ptomaine, 1884. Ziilxer and 
 Sonnenschein, Berl. Klin. Woch., 1869. 
 
 CHAPTER V. 
 
 IMMUNITY. 
 
 Arloing, Cornevin and Thomas, Du Charbon Bactrien; Pathogenic et 
 Inoculations Preventives, 1883. Behring, Deutsche Med. Woch.. 1890. 
 Behring and Kitasato, Deutsche Med. Woch., 1890. Blazekovic, Oesterr 
 Monatschr. f. Thierheilk., 1884. Bouchard, Compt. Rend., 1889. Bouley, 
 L'Inoculation Preventive de la Fievre Jaune: Compt. Rend., T. 100, 1885. 
 Brieger and Frankel, Berl. Klin. Woch., 1890. Biichner, Eine neue Theorie 
 iiber Erzielung v. Immunitat gegen Infectiouskrankheiten, 1883. Chamberland, 
 Le Charbon et la \ 7 accination Charbonneuse d'apres les Travaux RScents de 
 M. Pasteur, 1883. Chamberland and Roux, Compt, Rend., T. %, Ni. ]:,, 1883. 
 Chauveau, Compt. Rend., T. 89, 1879 ; T. 96, Nr. 'J ; Nr. lo : Nr. 11, 1883; Gaz. 
 Hebdom. de Med. et de Chir., 22, 1884. Felte, Compt. Rend.. T. '.'.. i 
 1884. Frank, Jahresber. d. K. Thierarzneischule in Munchen, is*:;. Gamaleia, 
 La Semaine Med., 1890. Grawitz, Die Theorie der Schutziinpfunu' : Virrhow's. 
 Arch.. Bd. 48, 1881. Hankin, Brit. Med. Journ., 1889 and 1S'.M ; i' n ,<-. Roy. 
 Soc., 1890; Centralb. f. Bacteriolog.. IM. IX., Lancet, 1891. Hess, S.-h\\-<-iz. 
 Arch. f. Thierheilk., Bd., 27, 1885. Kitasato, Zcitsdir. f. IK-i,.,,,., i:d. X. Koch, 
 Ueber die Milzbrandimpfung, 1882. Koch, Gaffky and Ldffler, Miith. a. d. Ges. 
 Amt, Bd. U., 1884, Loffler, Mitth. a. d. Ges. Amt, Bd. I., issi . Martin, Reports 
 
644 APPENDICES. 
 
 Med. Dept. Loc. Govt. Board, 1890-91 ; Brit. Med. Journal, 1891. Masse, Des 
 Inoculations Preventives dans les Maladies Virnlentes, 1883. Metschnikoff, 
 Virchow's Archiv XCVI. and XCVIL, Ann. de 1'Institut Pasteur, 1887, 1889, 
 
 1890, 1891, 1895. Nuttall, Zeitschr. f. Hygiene, 1888. Oemler, Arch. f. Wiss. 
 u. Pract. Thierheilk., 1876, 1881. Ogata, Centralb. f. Bacteriolog. , 1891. Ollive, 
 Compt. Rend., T. 89, 1879. Pasteur, Bull, de 1'Acad. de Med. and Gaz. 
 Med. de Paris, Nr. 18, 1880; Compt. Kend., 1883; La Vaccination Charbon- 
 neuse, 1883; Revue Scientifique, 1883; Bull, de 1'Acad. de Med., 1883. 
 Perroncito, Atti R. Ace. d. Lincei., 1883. Piitz, Vortrage f. Thierarzte, 
 Ser. 7, Heft 1, 1884. Boux, Ann. de 1'lnstitut Pasteur, 1888. Roszahegyi, 
 Pester Med.-Chir. Presse, 1882. Salmon and Smith, Centralb. f. Bacteriolog., 
 1887. Semmer, Virchow's Arch., Bd. 83, 1881. Semmer and Krajewski, 
 Centralbl. f. d. Med. Wiss., 1880. Strebel, Schweiz. Arch. f. Thierheilk., 1885. 
 Tizzoni and Cattani, Centralb. f. Bacteriolog., 1891. Toussaint, Bull, de 
 1'Acad. de Med. ; and Compt. Rend., 1880, 1881 ; Gazette Medicale de Paris, 
 Nr. 32, 1881. Wooldridge, Proc. Roy. Soc., 1887 ; Archiv f. Anat. and Phys., 
 1888. 
 
 CHAPTER VI.. 
 
 ANTITOXINS AND SERUM-THERAPY. 
 
 Be*clere, Chambon and Menard, Ann. de 1'Institut Pasteur, 1896. Behring 
 and Kitasato, Deutsche Med. Woch., 1890 ; Trans. Internat. Cong. f. Hygiene, 
 
 1891. Behring and Wernicke, Zeitschrift f. Hygiene, 1892. Buchner, Munich 
 Med. Woch., 1891. Calmette, Ann. de 1'lnstitut Pasteur, 1895. Emmerich and 
 Mastbaum, Archiv f. Hygiene, 1891. Fedoroff, Zeitschr. f. Hygiene, 1893. 
 Gromakowsky, Ann. de 1'Institut Pasteur, 1895. Heubner, Trans. Internat. 
 Med. Congress, 1894. Hewlett, Practitioner, 1895. Kossel, Zeitschrift f. 
 Hygiene, 1894. Marchoux, Ann, de 1'Institut Pasteur, 1895. Marmorek, Ann. 
 de 1'Institut Pasteur, 1895, 1896. Ogata, Centralb. f. Bakteriolog., 1891. 
 Report, Med. Sup. Metropolitan Asylums Board, 1896. Roux, Trans. Internat. 
 Congress of Hygiene, 1894 ; Ann. de 1'Institut Pasteur, 1894. Roux, Martin, 
 Chaillou, Ann. de 1'Instit. Pasteur, 1894. Tizzoni and Cattani, Centralb. f. 
 Bakteriolog., 1891. Welch, Trans. Assoc. Amer. Physicians, 1895. Wladmoroff, 
 Zeitschr. f. Hygiene, 1893. 
 
 CHAPTERS VII., VIII., IX., X. 
 
 VII. THE BACTERIOLOGICAL MICROSCOPE. VIII. MICROSCOPICAL 
 EXAMINATION OF BACTERIA. IX. PREPARATION OF NUTRIENT 
 MEDIA AND METHODS OF CULTIVATION. X. EXPERIMENTS 
 UPON THE LIVING ANIMAL. 
 
 Almquist, Hygeia, XLV. ; Stockholm, 1883. Banti, Manuale di Tecnica 
 Batteriologica, 1885. Baumgarten, Zeitschr. f. Wissensch. Mikr., 1884. Behrens, 
 Hilfsbuch zur Ausfiihrung Mikroskop. Untersuch., 1883. Bizzozero and Firket, 
 Manuel de Microscopic Clinique, 1885. Blanchard, Rev. Inter. ScL, III., 1879. 
 Bordoni Uffreduzzi, Microparasitici, 1885. Brefeld, Bot. Untersuch. iiber 
 Schimmelpilze, Bd. IV., 1881; Botan. Unters. liber Hefenpilze, Bd. V., 1883 ; 
 Verhandl. d. Physik. Med. Ges. in Wiirzburg, 1875. Buchner, In Nageli's 
 
BIBLICH.KAl'HY. 645 
 
 linn-such. uber Nie;lere Pilze: Munich. ls*2: Aer/tl. Intelligenzbl., No. 33, 
 11. Carpenter, The Microscope aii'l its Krv,-l:itious (iith Edition), 1-^1. 
 Cohn, Beitriige zur Biologic der Pflanzen, Bd. I., Heft it, 1^7.1, }^~<\. Cornil 
 and Babes, Les Baet.'-ries. issi 1 ,. Crookshank, Manuel Pratique .li- l'.;n-t'ri- 
 logie, traduit par Bergeaud; avcc I Phtimiir.ngraphii-s. lss;. Dolley, Tech- 
 nology of Bacteria Investigation. INS."). Duclaux, Ferments et Maladies, 1881. 
 Ehrlich, Deut. Mod. Woch., No. 19. ls>2 : Zeits.-hr. f. Klin. Med.: Bd. I. 1880. 
 Bd. II., Heft 3. 1881. Eisenberg, Bakteriologische Diagnostik, 1886. Esmarch, 
 Zeitschrift f. Hygiene. ISM',. Fehleisen, I'eber Ncnc Method, der t'ntersucli. 
 11. Cultur Pathogen. Bakteriun ; Physik. Med. (res. /.u Wiirzburg, 1882. Fliigge, 
 Handbuch der Hygiene und der Gewerbe Krankheiten, 1883. Friedlander, 
 Microscopische Technik (1st Edition), 1881. Gibbes, Practical Histology and 
 Pathology, 1885. Gram, Fortsch. d. Med., II., No. 6, 1H*4. Kauser, Ueber 
 Faulnissbacterien; mit 15 Tafeln in Licbtdnick, 188~>. Hazlewood, American 
 Monthly Microscop. Journ., isss. Hiiber and Breker, Die Path. Histolog. und 
 Bacteriologischen Untersuch. Methoden, 1880. Hueppe, P.aktcriologische 
 Apparate : Deut. Med. W6ch., 188ti ; Die Methoden der Bakterien F'r><-liung. 
 !>M; ; translated by Biggs, 188(5. Johne, Ueber die Kochschen Keinculturen, 
 1885. Klebs, Archiv f. Exp. Pathol.. Bd. L, 1873. Klein, Micro-organisms and 
 Disease, 1886. Koch, Biol. Klin. Woch., No. 15, 18x2 ; Mitth. a. d. Kais. Ges. 
 Amt, Bd. I., 1881.; Bd. II., 18*4 ; Untersuchungen Uber Wundinfections 
 Krankheiten, 1878; Beitrage z. Biol. d. Pflanzen, Bd. II., Heft 3, 1877. Lee, 
 The Micro tourist's Yade Mecum. lss ; >. Magnin and Sternberg, Bacteria, 1884. 
 Malley, I'hotomicrogra})!!}-, ixs.i. Orth, Path. Anat. Diagnostik, 1884. Pasteur, 
 Etudes sur la Biere, 1867. Perty, Zur Kenntniss Kleinster Lebensform, 
 l- s .'.2. Plant, Fiirbungs Methode z. Nachweis. der Micro-organismen, 1885; 
 Salomonsen, Bot. Zeit. ; No. 39, 1879 ; No. 28, 1880. Schafer, Course of Practical 
 Histology, ls77. Woodhead and Hare, Pathological Mycology, 1885. 
 
 CHAPTER XI. 
 
 EXAMINATION OF AIR, SOIL AND WATER. 
 
 Angus Smith, Rep. to the Loc. Gov. Board, 1884 ; Sanitary Kecord, 1883. 
 Becker, Reichsmedicinalkalender, 18s.l. Beumer, Deut. Med. Woch., 1886. 
 Bischof, Journ. Hoc. Chem. Industry, iss;. Buchner, Vortrage im Aerztl. 
 Verein zu Miinchen, Issi. Chamberland, ('..mpt. Rend., T. 99, p. 247, 1886. 
 Cramer, Die Wasserversorgung von Zurich. ls*.~,. Crookshank, Notes from a 
 Bact. Labin-., Lancet, 1>*.~>. Cunningham, Micr. Exam, of the Air: Calcutta, 
 1-71. Fodor, Hygienische Unters. Uber Luft, Boden u. Wasser., 1882. 
 C. Frankel, Zeitschr. f. Hygiene. !>-;. Frankland, P. and G., Proc. Roy. Soc., 
 L885y 1886 j .Microorganisms in Water. ls;4. Gunning, Arch. f. Hyg., 3, 1883. 
 Heraeus, Zeitschr. f. Hygiene, ISM;. Hesse, Deut. Med. Wo, -I,.. Nr. 51, 1884; 
 L'. 1884; Mitth. a. d. Ges. Amt, Bd. II., 1884; Ueber Was^rliltralion : D.-ut. 
 Med. Woc-h.. 1^85; Zeitschr. f. Hygiene, 1886. Klebs and Tommasi-Crudeli, 
 Archiv f. Exper. Path.. Bd. 11. 1*79. Koch, Mitth. a. d. (Jes. Amt, Bd. I.. 
 l^^l. Laurent, Journal de Pharmacie et de Chimie, 1885. Lemaire, Compt. 
 Rend., T. r,7. L863. Letzerich, ?:xp. Unters. ill), die Aetiologie des Typhus 
 mit bes. Berucksichtigung der Trink. u. Gebrauch>wiier. lss:j. Maddoz, 
 Month. Microscop. Journal. 1870. Meade Bolton, Zeitschr. f. Hygiene. L886. 
 Miflet, Cohn's Beitr. z. Biol. d. Ptlan/.en, Bd. III.. 1>79. Miquel, Annuaire 
 
646 APPENDICES. 
 
 de 1'Observat. de Montsouris, 1877, 1882 ; Compt. Rend., T. 86, 1878 ; Bull, 
 de la Soc. China., 1878; Ann. d'Hygiene, 1879; Les Organismes Vivants 
 de 1'Atmosphere, 1883. Miquel and Freudenreich, La Semaine Medicale, 1884. 
 Moreauand Plantymausion, La Semaine Medicale, 1884. Nageli, Unters. liber 
 Niedere Pilze., 1882. Olivier, Les Germes de 1'Air, These, Rev. Scientif.. 
 1883. Pasteur, Ann. de Chim. et de Phys., T. 64, 1862 ; Compt. Rend., T. 50. 
 1860; T. 52, 1861; T. 56, 1863; T. 85, 1877. Pfeiffer, Zeitschr. f. Hygiene. 
 1886. Pouchet, Compt. Rend., T. 47, 1858. Schrakamp, Archiv f. Hygiene. 
 Bd. II., 1884. Sehlen, Fortschr. d. Med., Bd. II., S. 585, 1885. Smart, Germs. 
 Dust and Disease, 1883. Soyka, Sitz.-Ber. der. K. Bayr. Akad. d. Wiss. : Math. 
 Physik. Classe, 1881 ; Vortrage im Aerztl., Vereiri in Munchen, 1881; D. 
 Vierteljsch. f. Oeff. Ges., Bd. 14, 1882; Prager Med. Woch., 1885; Fortschr. 
 d. Med., 1885. Tissandier, Compt. Rend., T. 7H, 1874. Torelli, La Malaria 
 in Italia, 1883. Tyndall, Brit. Med. Journ., 1877 ; Essays on the Floating- 
 Matter of the Air, 1881 ; Med. Tim. and Gaz., 1870. Wernich, Cohn's Beitrage 
 z. Biol. d. Pflanzen, Bd. III., S. 105, 1879. Wolffhugel and Riedel, Arbeit. 
 a. d. K. Ges. Amt, 1886. Wollny, Viert. f. Oeff. Ges., S. 705, 1883. Zander, 
 Centralbl. f. Allg. Ges., 1883. 
 
 CHAPTER XII. 
 
 PHOTOGRAPHY OF BACTERIA. 
 
 Crookshank, Photography of Bacteria, 1887. Frankel and Pfeiffer, Mikro- 
 photog. Atlas, 1889. Giinther, Photogram. Path. Mikroorg., 1887. Itzerott 
 and Niemann, Atlas der Microphotograph, 1895. Koch, Cohn's Beitrage zur 
 Biol. der Pflanz., 1877 ; Mitth. a. d. K. Gesundheitsamte, Bd. I., 1881. Neuhaus, 
 Centralbl. f . Bakteriolog., Bd. IV. Sternberg, Photomicrographs and How to 
 Make them, 1884. Woodward, Rep. to Surg. Gen. U. S. Army, 1870. 
 
 CHAPTER XIII. 
 
 SUPPURATION. PY^MIA. SEPTIOEMIA. ERYSIPELAS. GONORRHOEA. 
 
 OPHTHALMIA. 
 
 Ainstie, Lancet, 1870. Arloing, Recherches sur les Septicemies, 1884. 
 Babes, Compt. Rend., 1883. Balfour, Edinb. Med. Journ., 1877. Barthold, 
 Pyaemisch-Metast. Dissert. Berlin, 1875. Bastian, Brit. Med. Jonrn., 1878. 
 Be"champ, Compt. Rend., 1881; Trans. Internat. Med. Cong. London, 1881. 
 Beck, Rep. LOG. Govt. Board, 1880. Birch-Hirschfeld, Untersuchungen iiber 
 Pyamie, 1873. Braidwood and Vacher, Brit. Med. Journ., 1882. Burdon- 
 Sanderson, Trans. Path. Soc., 1872; Brit. Med. Journ., 1875. Crookshank, 
 Trans. Internat. Congr. of Hygiene, 1892. Lowdeswell, Quart. Journ. Micr. 
 Sc. London, Vol. 18, 1878 ; Proc. Roy. Soc. London, Vol. 34, 1883. Dreschfeld, 
 Brit. Med. Journ., 1883. Drysdale, Pyrexia, 1880. Garre, Fortschritte d. 
 Med., 165, 1885. Heiberg, Die Puerperalen u. Pyamischen Processe, 1873. 
 Hoffa, Fortsch. d. Med., 1885. Horsley, Rep. Med. Officer Loc. Govt. Board, 
 1881. Klemperer, Zeitschr. f. Klin. .Med., 1885. Koch, Wundinfectionskrank- 
 heiten: Leipzig, 1878; Mittheil. d. Kais. Ges. Amts, Bd. I., 1881. Lister, 
 Lancet, 1867; Med. Times and Gazette, 1877; Quart. Journ. Microscop. Science, 
 
BIBLIOGRAPHY. 647 
 
 1887. Oertel, Zur Aetiologie der Infectionskrankheiten, 1881. Ogston, Urit. 
 M. .1. Journ., Vol. I., 1881 ; Journ. of Anat. and Phys., Vol. 17, 1882. Passet, 
 Ueber Mikroorganismen der Eitrigen Zellgewebsentziindung des Menschen ; 
 Fortschritte d. Med., Nr. 2, 1885, Bd. 3, 1885. Perret, De la Septicemie : 
 Paris, 1880. Rindfleisch, Lehrb. der Pathol. Gewebelehre : 1 AutL S. 204, 
 1866. Eosenbach, Mikrooganismen bei den Wundinfectionskrankheiten des 
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 zu Breslau, 1871. Watson-Cheyne, Trans. Path. Soc., xxxv., 1884. 
 
 OSTEOMYELITIS. 
 
 Becker, Deut. Med. Woch., and Berl. Klin. Woch., 1883. Collmann, Bak- 
 terien im Or^anistnus eines an einer Verletzung am Oberschenkel verstorbenen 
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 Compt. Rendus, T. 99, 1884. Rosenbach, Centralbl. f. Chirurgie, 1884. 
 
 Schiiller, Centralbl. f. Chirurgie, Nr. 12, 1876. 
 
 ENDOCABDITIS. 
 
 Birch-Hirschfeld and Gerber, Archiv d. Heilkunde, 1876. Bramwell, 
 Diseases of the Heart, 1H84. Bristowe, Brit. Med. Journ., 1884. Coupland, 
 Brit. Med. Journ., 1885. Gibbes, Brit. Med. Journ., 1884. Goodhart, Trans. 
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 Bd. I., 1881. Koester, Virchow's Arch., Bd. 72, 1875. Kundrat, Sitz.-Ber. 
 d. Kais. Acad. d. Wissensch. zu Wien, 1883. Leyden, Zeitschr. f. Klin. Med., 
 1881. Nocard, Recueil de Med. Vet., 1885. Oberbeck, Casuistische Beitrage 
 zur Lehre von der Endocarditis Ulcerosa : Inaug.-Diss., 1881. Orth, J^ Ver- 
 sammlung Deutscher Naturf. zu Strasburg, 1885. Osier, Brit. Med. Journ., 1885 ; 
 Trans. Int. Med. Congress, 1881. Eibbert, Fortsch. d. Med., 1886. Eosenbach, 
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 Weichselbaum, Wien. Med. Woch., 1885. Weigert, Virchow's Arch., Bd. 84, 
 1881. Wilks, Brit. Med. Journ., 1882. Wyssokowitsch, Centralbl. f. d. Med. 
 Wissensch., Nr. 33, 1885. 
 
 EKYSIPELAS. 
 
 Baader, Schweiz. Naturf. Gesellsch., 1875. Denuce, Etude sur la I'athogenie 
 et 1'Anatomie Pathologique de I'Erysipele, 13K ; >. Dupeyrat, Recherches 
 Cliniques et Experimentales sur la Pathogenic de 1'Erysipele, 1881. Fehleisen, 
 Wiirzburger Phys. Med. Ges., 1881 ; Deut. Zeitschr. f. Chin, Bd. 16, 1882; Die 
 Aetiologie des Erysipsls. : Berlin, 1883. Hiiter, Med. Centralbl., Nr. 34, 1868. 
 
648 . APPENDICES. 
 
 Janicke and Neisser, Centralbl. f. Chir., Nr. 25, 1884. Klebs, Archiv f. Exper. 
 Fathol. u. Pharmacol., Bd. 4, 1875. Lukomsky, Virchow's Archiv, Bd. 60, 1874. 
 Nepven, Des Bacteries dans 1'Erysipele, 1885. Orth, Archiv f. Exper. Pathol. 
 u. Pharmacol., Bd. I., 1873. Kaynaud, Union Med., 1873, Kecklinghausen and 
 Lankowski, Virchow's Arch., Bd. 60, 1874. Rheiner, Virchow's Arch., Bd. 100 r 
 Heft 2, 1884. Tillmanns, Verhandl. d. Deutsch. Ges. f. Chirurgie, 1878; 
 Archiv f. Klin. Chirurgie, Bd. 23, 1879. Trosier, Bull. Soc. Anat. de Paris, 
 1875. Wolff, Virchow's Arch., Bd. 81, 1880. 
 
 PUEEPEEAL FEVER. 
 
 Aufrecht, Naturforsch. Versamml., 1881. Doleris, La Fievre Puerperale et 
 les Organismes Infect., 1886. Heiberg, Die Puerperalen und Pyamischen 
 Processe, 1873. Karewski, Zeitschr. f. Geburtsh. u. Gynakologie, Bd. 7, 1881. 
 Laffter, Bresl. Aerztl. Ztg., 1879. Mayrhofer, Monatsschr. f.- Geburtsk. u. 
 Frauenkrankheiten, Bd. 25, 1865. Orth, Virchow's Arch., Bd. 58, 1873. 
 Pasteur, Bull, de 1'Acad. de Med., T. 9, 1880. Recklinghausen and Lankowski, 
 Virchow's Arch., Bd. 60, 1873. Waldeyer, Arch. f. Gynakologie, iii., 1872. 
 
 GONOEBHCEA. 
 
 Arning, Viertelj. f. Dermatol. u. Syph., S. 371, 1883. Aufrecht, Patho- 
 logische Mittheilungen, 1881 ; Centralbl. f. d. Med. Wiss., Nr. 16, 1883. 
 Bockhart, Sitzungsbericht d. Phys. Med. Ges. zu Wurzburg, 1882; Viertelj. f. 
 Dermatol. und Syph., 1883. Bokai, Allgem. Med. Centralzeitung, Nr. 74, 
 1880. Bokai and Finkelstein, Prager Med. Chir. Presse, 1880. Biicker, 
 Ueber Polyarthritis Gonorrhoica. Diss., 1880. Bumm, Der Mikroorganismus 
 der Gonorrhoischen Schleimhauterkrankungen, 1885. Campona, Italia Medica, 
 1883. Chameron, Progres Medical, 43, 1884. Eschbaum, Deut. Med. Woch., 
 S. 187, 1883. Frankel, Deut. Med. Woch., Nr. 2, 1883 ; S. 22, 1885. Haab, 
 Der Mikrokokkus der Blennorrhoea Neonator, 1881. Hirschberg and Krause, 
 Centralbl. f. Pract. Augenheilk., 1881. Kammerer, Centralbl. f. Chirurgie, 
 Nr. 4, 1884. Krause, Die Mikrokokken der Blenorrhcea Neonator, 1882. 
 Kroner, Naturforschervers. in Magdeburg, Arch. f. Gyn., xxv., S. 109, 1884. 
 Leistikow, Charite-Annalen, 7 Jahrg., S. 750, 1882. Lundstrom, Studier ofver 
 Gonokokkus : Diss. Helsingfors, 1885. Martin, Kech. sur les Inflamm. Metast. 
 a la Suite de la Gonorrhee, 1882. Neisser, Centralbl. f. d. Med. Wiss., 
 Nr. 28, 1879; Deutsche Med. Woch., 1882. Newberry, Maryland Med. Journ.,. 
 1883. Petrone, Bivista Clin., No. 2. Eeter, Centralbl. f. d. Med. Wiss., 1879. 
 Sanger, Naturforschervers. in Magdeburg; Ibid., S. 126, 1884. Schrotter and 
 Winkler, Centralbl. f. Bact, Bd. ix. Smirnoff, Vrach.. 1886. Steinschneider, 
 Verhandl. der Deutsch. Dermat. Gesellsch., 1889; Berl. Klin. Woch., 1890. 
 Sternberg, Med. News, Vol. 45, Nr. 16, 1884. Weiss, Le Microbe du Pus 
 Blennorrhagique, 1889. Welander, Gaz. Med. de Paris, 1884. 
 
 OPHTHALMIC DISEASES. 
 
 Balogh, Med. Centralbl., xiv., 1879. Bock, Virchow's Arch., Bd., 91, 1883. 
 Cornil and Berlioz, Compt. Rend, de 1'Acad. d. So., 1883. Deutschmann, 
 v. Graefe's Archiv, Bd. xxxi., 1885, Gifford, Archiv f. Augenheilkunde, 1886. 
 Soldzieher, Centralblatt f. Prakt. Augenheilk., 1884. Kahler, Prager Zeitschr. 
 1 Prakt. Heilk., Bd. I., 1882. Klein, Centralbl. f. d. Med. Wiss., Nr. 8, 1884, 
 
BIBLIOGRAPHY. 649 
 
 Kroner, \Vrh. d. Xaturforscher-Yers. Magdeburg, ISM. Kuschbert, Dcutsrh. 
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 lecture : Lancet, 1886. 
 
 CHAPTER XIV. 
 
 ANTHRAX. 
 
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 Yirchow's Arch., Bd. 91, 1**3. Chauveau, Compt. Rend., T. 90 and 91, 1880; 
 T. !2, isM : T. '.'4. 1*82; T. 96, 1883. Chelchowsky, Der Thierarzt. lss4. Colin, 
 Bull. Acad. de Med.: Paris. T. 2, ls73; T. 7, 1878; T. 8, 1-7'.' : T. 9, 1880; 
 T. 10, 1881. Crookshank, Rep. Agric. Dept., 1888. Davaine, Compt. Rend.: 
 Paris, T. 77. 1*73 : T. 57 and 59, 1863 ; T. 60, 1865 ; T. 61, 1866, 1877 ; Rec. de 
 Med. Yet., T. 4, 1*77. Dowdeswell, Rep. Med. Off. Local Gov. Board, 1883. Esser 
 and Schiitz, Mitth. a K. Preuss Amtl. Yet. Sanitatsbericht, 1882. Ewart, 
 Quart. Journ. of Microsc. Sc., 1878. Feltz, Compt. Rend., T. 95, 1882. Fodor, 
 Dent. Med. \Yoch., 1886. Fokker, Centralbl. f. d. Mad. \\">sensch., Bd. is. 
 1880; Centralbl. f. d. Med. Wiss., 1881; Virchow's Archiv. Bd. 88, 1882. 
 Frank, Zeitschr. f. Hygiene, 1886. Friedrich, Zur Aetiologie des Milzbrands., 
 !"">. Greenfield, Quart. Journ. Mi<T. S<-. Lmuion, 1*71 ; Proc. Roy. Soc. 
 London, Isso. Hoffa, Die Natur des Milzbrandgiftes : Wiesbaden, 1886. 
 Huber, Deut. Med. Woch., Bd. 7, 1881. Johne, Ber. ii. d. Veter. Wesen. i. K. 
 Sachsen, iss*;. Kitt, Sitzungsb. d. Ges. f. Morphol. u. Physiol. zu Miinchen, 
 1885. Klein, Rep. of the Medical Officer of the Local Govt. Board, 1881 ; Quart. 
 Journ. Micr. Sc., 18*:'.. Koch, Beitrage zur Biologic der Pflanzen. P,d. II.. 
 Heft 2, 1876; Wundinfectionskrankheiten, 1878; Mitth. aus d. Ges. Amt. 
 Bd. L, 1881 ; Milzbrand und Rauschbrand : Stuttgart, lss;. Martin, Proc. 
 
 1890; Rep. Med. Off. Local Govt. Board, 1890-JM : Brit. Med. Journal. 
 1'.U. Oemler, Archiv f. Wiss. u. Pract. Thierheilk., Bd. 4, 1878, 1879, 
 Osol, Experiment. Untersuch. U. das Anthraxgift : Inaug. Diss. Dorpat, 1885. 
 Pasteur, Bull. Acad. de Med., 1877, 1879, 1880 ; Compt. Rend.. 1 1 877 ; 
 
 T. 90 and 91, 1880: T. t'2. 1881 : T. :*:>, 1882. Pollender, Yk-rteljahr>chr. 
 f. Ger. Med., Bd. B, l^.V,. Prazmowski, A-ad. d. Wissensch. in Krakau, 1884 ; 
 
<650 APPENDICES. 
 
 Biol. Centralbl., Bd. 4, 1884. Rodet, Contribution a 1'Etude Experimentelle 
 du Carbon Bacteridien, 1881 ; Compt. Rend., T. 94, 1882. Roloff, Archiv f. 
 Wissensch. u. Pract. Thierheilk., Bd. 9, 1883 ; Der Milzbrand : Berlin, 1883. 
 ;Schmidt, Deut., Zeitschr. f. Thiermed. u. Vergl. Pathol., 1879. Schrakamp, 
 Archiv f. Hygiene, Bd. 2, 1884. Semmer, Centralbl. f. d. Med. Wissensch., 
 Bd. 18, 1880, 1884 ; Der Milzbrand und das Milzbrandcontagium, 1882. 
 Sternberg, Am. Monthly Micr. Journ., 1881. Szpilman, Zeitschr. f. Physiol. 
 Chemie : Strassburg, Bd. 4, 1880. Toepper, Die Neueren Erfahrungen iiber 
 d. Aetiologie d. Milzbrands., 1883. Toussaint, Compt. Kend., T. 85, 1877, 
 1878, 1880 ; Eecherches Experimentales sur la Maladie Charbonneuse, 1879. 
 Wachenheim, Etude Experimentelle sur la Septicite et la Virulence du Sang 
 Charbonneux, 1880. 
 
 CHAPTER XY. 
 
 QUARTER-EVIL. MALIGNANT OEDEMA. RAG-PICKER'S SEPTICAEMIA 
 OF GUINEA-PIGS. SEPTIOEMIA OF MICE. 
 
 QUAKTEE-EVIL. 
 
 Arloing, Cornevin and Thomas, Compt. Rend., 1880 ; Bull, de 1'Acad. de 
 Med., and Revue de Med., 1881 ; Du Charbon Bacterien, Charbon Symptoma- 
 tique, etc., 1883 ; Revue de Med., 1884 ; Chabert's Disease : Transl. by Dawson 
 Williams in Micro-parasites and Disease (New Syd. Soc.), 1886. Babes, 
 Journ. de 1' Anatomic, 1884. Bellinger and Feser, Wochenschr. f. Thierheil- 
 kunde, 1878. Ehlers, Unters. lib. d. Rauschbrandpilz : Inaug. Diss. Rostock, 
 1884. Hess, Bericht iiber die entschadigten Rauschbrand u. Milzbrandfalle 
 im Canton Bern, 1886. Kitt, Jahresber. der K. Thierarzneisch. in Miinchen, 
 1884. Neelsen and Ehlers, Ber. d. Naturforsch. Ges. zu Rostock, 1884. 
 .Strebel, Schweiz. Archiv f. Thierheilk., 1886. 
 
 MALIGNANT (EDEMA. 
 
 Brieger and Ehrlich, Berl. Klin. Wochenschr., N. 44, 1882. Chauveau and 
 Arloing, Archiv Vet., 1884 ; Bull. Acad. de Med., 1884. Davaine, Bull, de 
 1'Acad. de Med., 1862. Gaffky, Mitth. as. d. K. Ges. Ami., 1881. Hesse, W. 
 and R., Deut. Med. Woch., 1885. Kitasato and Weyl, Zeitschr. f. Hygiene, 
 Bd. VIII. Kitt, Jahresber. der K. Thierarzneischule in Miinchen, 1884. Koch, 
 Mitth. aus dem Ges. Amt, I., S. 54, 1881. Lebedeff, Arch, de Phys. Norm, et 
 Path., 1882. Lustig, Jahresber. d. K. Thierarzneischule zu Hannover, 1884. 
 Pasteur, Bull, de 1'Acad. de Med., 1877, 1881. Roger, Compt. Rend. Soc. de Biol., 
 1889. Eoux and Chamberland, Ann. de 1'Institut Pasteur, 1887. Trifaud, Rev. 
 de Chirurg, T. iii. Van Cott, Centralb. f. Bact., 1891. Verneuil, La Semaine 
 Med., 1890. 
 
 RAG-PICKER'S SEPTIC^MIA. SEPTICAEMIA OF GUINEA-PIGS. SEPTIOEMIA 
 
 OF MICE. 
 
 Bordoni-Uffreduzzi, Zeitsch. f. Hygiene, 1888. Klein, Centralbl. f. Bac- 
 teriolog., 1890. Koch, Wundinfectionskrankeit, Trans. New Syd. Soc., 1880. 
 Paltauf, Wiener Klin. Woch., 1888. 
 
BIBLIOGRAPHY. 651 
 
 CHAPTER XVI. 
 
 ILEMORRHAGIC SEPTICAEMIA. 
 
 si:rnc.MiA OP BUFFALOES. SEPTIC PLEUBO-I-NKI MOMA OF CALVIN 
 SWINE FEVEB. SEPTIC^MIA OF DEEB. SEPTICAEMIA OF RABBITS. 
 FOWL CHOLEBA. FOWL ENTEBITIS. DUCK CHOLEBA. GBOUSE DISEASE. 
 
 Babes, Compt. Rend, de 1'Acad. d. Sc., 1883; Arch, de Physiol., 1884. 
 BarthelSmy, Compt. Rend., T. 96, No. 18, 1883. Bunzl-Federn, Centralbl. f. 
 Bacteriolog., 1891. Camera, Centralbl. f. Bacteriolog., 1891. Cornil, Ar< h. <! 
 Physiol., Bd. 10, 1882. Condi and Chantemesse, Le Bulletin Mu-1., 1887. 
 Cornil and Toupet, Bull, de la Soc. Nat. d' Acclimation., 1888. Davaine, 
 Bull, de 1'Acad. de Med., 1879. Eberth and Schiminelbusch, Virr 
 Archiv, 1889 ; Fortschr. d. Med., Bd. VI. Gaffky, Mitt, aus dem K. G. Amte, 
 1881. Gamaleia, Centralb. f. Bacteriolog., 1888. Hueppe, Berl. Klin. Woch., 
 1886. loannes and Me"gnin, Journ. d'Acclimatation, 1877. Kitt, Allg. Dent. 
 Gefliigelzeitung, 1885. Klein, Rep. Med. Off. Local Govt. Board, 1877-7- : 
 Fortschr. der Med., 1888 ; Centralb. f. Bakteriolog., 1889. Koch, Aetiologie 
 der Wundinfections R., 1878. Oreste and Armani, Atti de R. Instit. d'Incorrag. 
 Alle Sci. Nat., Econ e Tech., 1887. Pasteur, Compt. Rend., T. 90, 1886. 
 Perroncito, Arch. f. Wiss. u. Prakt. Thierheilk., 1879. Petri, Centralbl. f. d. 
 Med. Wiss., 1885. Kietsch and Jobert, Compt, Rend., 1888. Salmon, Reports 
 Bureau of Animal Industry, 1886, 1887, 1888. Salmon and Smith, Amer. 
 Monthly Med. Journ., 1881. Schiitz, Archiv f. Wiss. und Prakt. Thierheilk., 1888. 
 Semmer, Deut. Zeitschr. f. Theirmed. u. Vergl. Path., 1878. Smith, Journ. f. 
 Comp. Med. and Surg., 1887; Zeitschr. f. Hygiene, 1891. Smith and Veranus 
 Moore, Rep. Bureau of Animal Industry, 1895. Ziirn, Die Krankheiten des 
 Hausgefliigels, 1882. 
 
 CHAPTER XVII. 
 
 PNEUMONIA. INFECTIOUS PLEURO-PNEUMOMA OF CATTLE. 
 INFLUENZA. 
 
 PNEUMONIA. 
 
 Afanassiew, Compt. Rend. Soc. de BioL Paris, T. 5, 1884. De Blasi, Rivista 
 Internaz. di. Med. e Cbir., 1885.- Bruvlant and Verriers, BulL de 1'Acad. 
 Beige, 1880. Dreschfeld, Fortschr. d. Med., Bd. 3, 389, 1885. Poa and 
 Bordoni-TJffreduzzi, Deut. Med. Woch., 1886. Frankel, Verhandl. d. Congr. 
 f. Innere Med., Fortschr. d. Med. Nov., 1884; Deut. Mod. Woch., 1886; 
 Zeitschr. f. Klin. Med., Bd. x. and xi., 1886. Friedlander, Virchow's Arch., 
 Bd. 87, 1882; Fortschr. d. Med., Bd. I., 1883; Bd. II., 1884; Bd. 3, 92, 1885. 
 Friedlander and Frobenius, Berl. Klin. Woch., 1883. Germain-8e*e, Compt. 
 Rend. Acad. de Sc. Paris, 1884; Des Maladies SpScifiques du Poumon, 1885. 
 <Hles, Brit. Med. J., Vol. II., 1883. Oriffini and Cambria, Centralbl. f. d. Med. 
 Wi->., 1883. Jaccoud, La France Medicale, 1*86. Jurgensen, Berl. Klin. 
 Woch., Bd. 22, 1884. Klein, Centralbl. f. d. Med. Wissensch., 1*X4. Koranyi 
 and Babes, Tester Med. Chir. Presse, 1884. Kflhn, Deutsch. Arch. f. Klin. 
 Med., 1878 ; Berl. Klin. Woch., Nr. 38, 1881. Lancereaux and Besancon, 
 
652 APPENDICES. 
 
 Archiv Gen. de M6d., 1886. Maguire, Brit. Med. Journ., Vol. II., 1884. 
 Manfredi, Fortsch. d. Med., 1886. Matray, Wien. Med. Presse, Nr. 23, 1883. 
 Mendelssohn, Zeitschr. f. Klin. Med., Bd. 7, 1884. Nauwerck, Beitr. zur 
 Pathol. Anat, von Ziegler, 1884. Neumann, Berl. Klin. Woch., 1885. Paw- 
 lowsky, Berl. Klin. Woch., 1885. Peterlein, Bericht ii. d. Vet.-Wesen. i. K. 
 Sachsen, 1885. Pipping, Fortsch. d. Med., No's. 10 and 14, 1886. Platanow, 
 Mitth. a. d. Wiirzburg. Med. Klinik, 1885 ; Ueber die Diagnostische 
 Bedeutung d. Pneumoniekokken : Inaug.-Diss. Wiirzburg, 1884. Ribbert, 
 Deut. Med. Woch., Nr. 9, 1885. Salvioli and Zaslein, Centralbl. f. d. Med. 
 Wissensch., 1883. Arch, pour les Sc. Med., T. 8., 1884. Schou, Fortschr. der Med., 
 Bd. 3, Nr. 15, 1886. Sternberg, Amer. Journ. Med. Sciences, 1885 ; Journ. 
 Roy. Micr. Soc., 1886.' Talamon, Progr. Medic.. 1883. Thost, Deut. Med. 
 Woch., 1886. Weichselbaum, Wien. Med. Woch., 1886. Ziehl, Centralbl. f. d. 
 Med. Wiss., 1883; Centralbl. f. d. Med. Wiss., 1884. 
 
 CEEEBBO-SPINAL MENINGITIS. 
 
 Bonome, Centralbl. f. Bact. u. Parasitolog., IV. Foa, Zeitschr. f. Hygiene, 
 IV. Leichtenstern, Deut. Med. Woch., Nr. 23 u. 31, 1885. Ley den, Centralbl. 
 f. Klin. Med., Nr. 10, 1883. Weichselbaum, Wien. Klin. Woch., 1888. 
 
 PLEUKO-PNEUMONIA. 
 
 Arloing, Compt. Rend, cix., 1889. Bruce and Loir, Ann. de 1'Institut 
 Pasteur, 1891. Bruylants and Verriers, Bull, de 1'Acad. Belg., 1880. Cornil 
 and Babes, Arch, de Physiol. Norm, et Path., T. 2, 1883. Lustig, Centralb. f. 
 die Med. Wiss., 1885. Mayrwieser, Woch. f. Thierheilk. u. Viehzucht, 19, 1884. 
 Pasteur, Recueil de Med. Vet., 1882. Poels, Fortsch. d. Med., 1886. Poels 
 and Nolen, Centralbl. f. d. Med. Wiss., Nr. 9, 1884; Fortsch. d. Med., 1886. 
 Putz, Thier. Med. Vortrage, Bd. 1, 1889. Report on Pleuro-pneumonia and 
 Tuberculosis, 1888. Schiitz and Steffen, Archiv f . Wiss. und Prakt. Thierheilk.. 
 Bd. xv. Sussdorff, Deutsche Zeitschr. f. Thiermed. u. Vergl. Pathol., 1879. 
 
 INFLUENZA. 
 
 Babes, Centralbl. f. Bacteriolog., 1890; Deutsche Med. Woch., 1892. Bein, 
 Zeitschr. f . Hygiene, 1890. Bouchard, La Semaine Med., 1890. Canon, Deutsche 
 Med. Woch., 1892. Fischel, Prager Med. Woch., 1890; Zeitschr. f. Heilkunde, 
 1891. Jolles, Wiener Med. Blatt,, 1890. Kirchner, Centralbl. f. Bacteriolog., 
 1890 ; Zeitschr. f. Hygiene, 1890. Kitasato, Deutsche Med. Woch., 1892. Klein, 
 Brit, Med. Journ., 1892. Klebs, Centralbl. f. Bakteriolog., 1890 ; Deutsche Med. 
 Woch., 1890. Pfeiffer, Deutsche Med. Woch., 1892. Prudden, New York Med. 
 Rec., 1890. 
 
 CHAPTER XVIII. 
 
 ORIENTAL PLAGUE. RELAPSING FEVER. TYPHUS FEVER. YELLOW 
 
 FEVER. 
 
 ORIENTAL PLAGUE. 
 
 Aoyama, Mitth. ii. d. Pest. Epidemie im Jahre 1894; in Hong Kong, 1895. 
 Yersin, Ann. de 1'Institut Pasteur, 1894. Yersin, Calmette and Borrel, Ann. 
 de 1'Institut Pasteur, 1895. 
 
BIBLIOGRAPHY. 653 
 
 Albrecht, St. Petersb. Med. W. . h .. i*7'.. Carter, Lancet. l*7'.i; Trans. 
 Intermit. Med. Congress, issi. Cohn, J)rut. Med. Wodi.. 1*711. Engel, 
 Klin. Woch., 187H. Gunther, Fortschr. d. Med., ls*f,. Guttmann, Vir< 
 Arch., 1*80. Heydenreich, St. Petersb. Med. Woch., ls7; : l>,-r Para>ir 
 falltyphus, 1877. Jaksch, Wien. Med. Woch., .Juli. isyo. Koch, Pent. Med. 
 Woch., 1879. Laptschinsky, Centralbl. f. d. M.-d. Wi>s.. P.d. li!. 1*7:,. Manas- 
 sem, St. Peter.sb. Med. Woch., No. 18, 1876. Metchnikoff, Vin-ho\\'s Archiv. 
 1*77. Moczutowsky, Deutsches Archiv fiir Klin. Med., Bd. xxiv.. 1-7',. 
 Muhlhauser, Virchow's Arch., Bd. 97, 1880. Obermeier, Med. Centralbl., 1 1 : 
 Berl. Med. Ges. ; Berl. Klin. Wochenschr., 1873. Soudakewitch, Ann. de 
 Tlnstitut Pasteur, 1891. Weigert, Deut. Med. Woch., 1876. 
 
 VKLI.OW FI:VER. 
 
 Babes, Compt. Rend., 17 Sept., 1883. Bouley, Compt. Rend., T. 100, p. 1276, 
 1885. Carmona y Valle, Lecons sur TEtiol. et la Prophylax. de la F' 
 Jaune, 18H5. Cerecedo, El Siglo Medico, 1886. Domingos Freire, Rechcnhe- 
 sur la Cause de la Fievre Jaune, 1884 ; La Fievre Jaune et ses Inoculations 
 Preventives, 1896. Domingos Freire and Eebourgeon, Compt. Rend., 1 
 p. 804 ? 1884. 
 
 CHAPTER XIX. 
 
 SCARLET FEVER AND MEASLES. 
 
 SCARLET FEVER. 
 
 Coze and Feltz, Les Maladies Infectieuses, 1872. Crooke, Lancet, 1 
 Fortsch. d. Med., 1885. Crookshank, Report Agric. Dept., 1887. Hahn, Berl. 
 Klin. Woch., No. 38, 1882. Heubner and Bahrdt, Berl. Klin. Woch., Nr. 44, 
 1884. Klein, Nature, xxxiv., 1886 ; Report of the Medical Officer of the 
 Privy Council, 1887, 1888, 1889. Laure, Lyon Medical, 1886. McKendrick, 
 Brit. Med. Journal, 1872. Pohl-Pincus, Centralbl. f. d. Med. Wiss., No. 36, 
 1883. Both, Miinch. Aerztl. Intelligenzbl., 1883. 
 
 M KASLES. 
 
 Cornil and Babes, Archiv de Phys., 1883. Keating, Phil. Med. Times, 1882, 
 
 CHAPTER XX. 
 
 >M ALL-POX. CATTLE FLAG IK. 
 SMALL-POX. 
 
 Chauveau, Compt. Rend., IH>. Cohn, Virchow's Archiv, Bd. 55, 1872. 
 Copeman, Brit. Med. Journ., 1896; The Practiti..n-r, 1 *'.;. Cornil and Babes, 
 Med. des Hop.. 1883. Crookshank, History and Path, of Vaccination, 1 *-'.. 
 Haccius, Variola-vaccine, 1 S( .L'. Hamerink, Ueberdie sog. Vac. u. Variola, 1 ^ J. 
 Ischamer, Aerztl. Verein. Steiermark. issu. Klebs, Arch. f. Exp. Path. u. 
 Pharmakol., Bd. 1". 1*7-1. Klein, ll.-p. Me<l. Off. Loc. Govt. Board, 
 
654 APPENDICES. 
 
 Luginbuhl, Verhdl. d, Phys. Med. Ges. in Wurzburg, 1873. Marchand, Revue- 
 Mycologique, 1882. Pfeiffer, Die Protozoen als Krankheitserreger, 1890 ; 
 Behandl. und Prophylax. der Blattern, 1893. Pissin, Berl. Klin. Woch., 1874. 
 Plaut, Das Organis. Contagium der Schafpocken, 1883. Pohl-Pincus, Vaccina- 
 tion, 1882. Quist, St. Petersburg Med. Woch., Nr. 46, 1883. Beports, Royal 
 Vaccination Commission, 1888-96. Buffer and Plimmer, Brit. Med. Journ., 
 1894. Weigert, Ueber Bakterien in der Pockenhaut, 1871 ; Anat. Beitr. z. 
 Lehre v. d. Pocken, 1874. Wolf, Berl. Klin. Woch., Nr. 4, 1883. Ziilzer, Berl. 
 Klin. Woch., 1872. 
 
 CATTLE PLAGUE. 
 
 Crookshank, History and Pathology of Vaccination, 1889. Beport of the 
 Cattle Plague Commission, 1865. Beport on Indian Cattle Plague, 1871. 
 Semmer and Archangelski, Ueber das Rinderpestcontagium und dessen Miti- 
 gation ; Centralbl. f.'d. Med. Wiss., 1883. Simpson, Brit. Med. Journal, 1896. 
 Smart, Reports on Cattle Plague, Edinburgh, 1865. 
 
 CHAPTER XXI. 
 SHEEP-POX. FOOT AND MOUTH DISEASE. 
 
 SHEEP-POX. 
 Crookshank, History and Path, of Vaccination, 1889. 
 
 FOOT AND MOUTH DISEASE. 
 
 Klein, Report Med. Off. Local Govt. Board, 1885. Schottelius, Centralb. f. 
 Bakteriolog., 1892. 
 
 CHAPTER XXII. 
 
 HORSE-POX. COW-POX. 
 
 HORSE-POX. 
 Crookshank, History and Pathology of Vaccination, 1889. 
 
 COW-POX. 
 
 Bucknill, Prov. Med. Journ., 1895. Crookshank, Brit. Med. Journ., 1888;: 
 History and Pathology of Vaccination, 1889 (Vol. II. contains reprints of the 
 works of Jenner, Pearson, Woodville, Loy, Bogers, Birch, Bousquet, Estlin, 
 Ceely, Badcock, Auzias-Turenne, Dubreuilh, Layet). Beports of the Royal 
 Vaccination Commission. 
 
 CHAPTER XXIII. 
 
 DIPHTHERIA. 
 
 Abbot, Bull. Johns Hopkins Univ., 1891, 1893 ; Journ. of Path, and Bact., 
 1893. Babes, Virchow's Archi-v. 1890. Behring, Deutsche Med. Woch. 
 1890. Behring and Kitasato, Deutsche Med. Woch., 1890. Birch-Hirschfeld, 
 Archiv fur Heilk., 1872. Brieger and Frankel, Berl. Klin. Woch., 1890. 
 
BIBLIOGRAPHY. 656 
 
 Buhl, Zeitschr. fur Biol., 1867. Cornil, Arch, dp 1'hysiol., 1881. Eberth, Med. 
 Centralbl., XI., Nr. s. 187:5. Emmerich, Compt. Rendus et M6moirt>s du V. 
 Congres Internat. d'Hygiene, 1884 ; Deut, Med. Woch., 1884. Everett, Med. 
 ;md Surg. Reporter, 1881. Forster, Wien. M-d. \V...-h., issi. Francotte, La 
 Diphtheric, 1883. Freidberger, Deut. Zeitschr. fiir Tliiermed.u. Vergl. 1'athol., 
 1879. Fiirbringer, Virch. Arch., Bd. 91, 1883. Gerhardt and Klebs, Verhandl. 
 d. Congresses f. Inn. Med., 1882, 1883. Heubner, Die Experimentelle Diph- 
 theric, 1883. Hueter and Tommasi, Centralbl. f. d. Med. \\i^.. 1868, Klebs, 
 Arch. f. Exp. L'athol., Bd. 4, 187.">. Klein, Report Med. Dept. Local Govt. 
 Board, 1889. Letzerich, Berl. Klin. Woch., xi., 1874 ; Virch. Arch., Bd. 55 r 
 1872; Bd. 68, 1876. Loffler, Mittheil. a. d. Kais. Ges. Amt, Bd. II., 1884 ; 
 Microparasites and Disease (New Syd. Soc.), 1886 ; Centralbl. f. Bacteriolog. r 
 1887, 1890 ; Deutsche Med. Woch.,* 1890. Lumner, Aerztl. Int. Bl., No. 31, 
 1881. Martin, Rep. Med. Off. Loc. Gov. Board, 1890. Neumayer, Neue 
 Thesen zu Diphtheritisfrage, 1880. Nieati, Compt. Rend., T. 88, 1879. Oertel, 
 Deut. Arch. f. Klin. Med., Bd. 8, 1871 ; Zur Aetiologie der Infectionskrank- 
 heiten, 1881. Park, New York Med. Record, 1892, 1893. Prudden, Amer. 
 Journal of Med. Sci., 1889 ; New York Med. Rec., 1891. Eivolta, Giornale de 
 Anat. Fisiol. e Patol. delli Anim., 1884. Roux and Yersin, Ann. de 1'Institut 
 Pasteur, 1888, 1889, 1890. Salisbury, Gaillard's Med. Journ. : New York, 1882. 
 Talamon, Bull, de la Soc. Anat. de Paris, T. 56, 1881. Welch, Amer. Journal 
 of the Med. Sci., 1894. Welch and Abbott, Bull. Johns Hopkins Univ., 1891. 
 Wood and Formad, Bull. Nat. Board of Health, Wash, and Med. Times and 
 Gazette, 1882. Zahn, Beitrage zur Pathol. u. Histol. der Diphtheric, 1878. 
 Zarniko, Centralbl. f. Bact, 1889. 
 
 CHAPTER XXIV. 
 
 TYPHOID FEVER. 
 
 Almquist, Typho'idfeberus-Bakterie, 1882. Beumer and Peiper, Zeitschr. f. 
 Hygiene, 1886. Birch-Hirschfeld, Zeitschr. f. Epidemiologie, I., 1874. Boens,. 
 Acad. Roy. de Med. de Belgique : Bull. 3 Ser., T. 17, 1883. Brautlecht, 
 Virchow's Arch., Bd. 84, 1881. Coats, Brit. Med. Journ., lss2. Crooke, Brit. 
 Med. Journ., 1882. Eberth, Arch. f. Pathol. Anat., Bd. 81, 1880; Volkmann, 
 Klin. Vortrage, 1883. Eppinger, Beitr. zur Pathol. Anatomic aus d. Patholog, 
 Institut. Prag., 1880. Feltz, Compt. Rend., T. 85, 1877. Fischel, Beitr. zur 
 Pathol. Anat. aus d. Pathol.- Anat. Inst. Prag., 1H80. Fraenkel and Simmonds, 
 Die Aetiologische Bedeutung des Typhus-Bacillus, 1886. Gaffky, Mitth. a. d. 
 Ges. Amt, Bd. II., 1884. Klebs, Arch. f. Exper. Pathol. u. Pharmakol., 1880. 
 Klein, Med. Centralbl., xii., 1874. Letzerich, Yin-how's Archiv, Bd. 68,1876;. 
 Archiv f. Exper. Pathol., Bd. 9, 1878 ; Bd. 10, 1881 ; Experimentelle Unter- 
 suchungen iiber die Aetiologie des Typhus Abdominalis : Leipzig, 1H83. Luca- 
 tello, Bollet. d. R. Accad. Med. di Geneva, l*si;. Maraghano, Centralbl. f. d. 
 Med. Wiss., Bd. 15, 1SS2. Meisels, Wien. Med. \Vic:h.. 1-ssr,. Meyer, Unters. 
 iiber den Bacillus des Abdominaltyphus : Inaug. Diss., 1881 . Michael, Fortsch. 
 d. Med., 1885. Neuhauss, Berl. Klin. \Vu-h.. Pfeiffer, Deut. Mi-d. 
 
 Woch.. Nr. i".'. 1 **.",. Rappin, (.'out rib. ft 1'Ktude des Bact. de la Bouche, 
 a 1'Etat Normal et dans la Fievre Typho'ide, 1HS1. Seitz, Bakteriolog. Studien 
 /.. Typhusiitiologie, 1886. Sirotinin, Zeitsch. f. Hygiene, 1886. Tayon, Compt. 
 Rend., T. 99, p. .'531, 1*84. Tizzoni, Studi. di Pat. Sperim. sulla Gen. d. Tifo. : 
 Milan,' 1*H>. Wernich, Zi-itsrhr. f. Klin. Med., Bd. 6. 
 
656 APPENDICES. 
 
 CHAPTER XXV. 
 
 SWINE-TYPHOID. 
 
 Klein, Eep. of the Mecl. Offic. of the Privy Council, 1877-78 ; Virchow's 
 Archiv, 1884. M'Fadyean, Journ. of Comp. Path, and Therapeutics, 1895. 
 Heport of a Conference on Swine-fever : Board of Agriculture, 1896. Rietsch, 
 Jobert and Martinaud, Compt. Bend., T. 106. Selander, Centr. f. Bakt. u. 
 Parasitenk., 1888 ; Ann de llnstitut Pasteur, 1890. Smith and Veranus Moore, 
 Bull, of Bureau of Animal Industry U.S., 1894. Welch and Clement, Trans. 
 Internat. Vet. Congress of Amer., 1893. 
 
 CHAPTER XXVI. 
 
 SWINE-MEASLES. DISTEMPER IN DOGS. EPIDEMIC DISEASE OF 
 FERRETS. EPIDEMIC DISEASE OF MICE. 
 
 SWINE-MEASLES. 
 
 Cornil and Babes, Arch, de Physiol., 1883. . Detmers, Science, 1881. 
 Eggeling, Fortschr. d. Med., 1883. Lb'ffler, Arbeiten aus dem Kaiserl. Gesund- 
 heits Amt, Bd. I., 1885. Lydtin and Schottelius, Der Kothlauf der Schweine : 
 Wiesbaden, 1885. Pasteur, Compt. Rend., T. 95, 1882. Pasteur and Thuillier, 
 Bull, de 1'Acad. de Med. de Paris : Compt. Rend., T. 97, 1883. Salmon, 
 Report Depart. Agricul. Washington, 1881, 1884. Schiitz, Ueber den Rothlauf 
 der Schweine und die Impfung desselben, 1885. 
 
 CHAPTER XXVII. 
 
 ASIATIC CHOLERA. CHOLERA NOSTRAS. CHOLERAIC DIARRHCEA 
 FROM MEAT-POISONING. DYSENTERY. CHOLERAIC DIARRHCEA 
 OF FOWLS. 
 
 CHOLEEA. 
 
 Ali-Cohen, Fortschr. der Med., 1887, 1888. Almquist, Zeitschr. f. Hygiene, 
 1887. Babes, Virchow's Archiv, 1885. Bianchi, Lancet., 1884. Bitter, Archiv f. 
 Hygiene, 1886. Bochefontaine, Exper. pour servir t\ 1'Etude des Phenomenes 
 determines chez 1'Homme par 1'Ingestion Stomacale du Liquide Diarrheique 
 -du Cholera: Compt. Rend., 1884. Brieger, Deutsche Med. Woch., 1887; Berl. 
 Klin. Woch., 1887; Virchow's Archiv, 1887. Brunetti, Fatti Considerazioni 
 Conclusioni sul Colera, 1885. Biichner, Archiv f. Hygiene, 1885; Miinchn. Aerztl. 
 Intelligenzbl., S. 549, 1884. Biichner and Emmerich, Miinch. Med. Woch., 1885. 
 Bujwid, Zeitschr. f. Hygiene, Centralbl. f. Bacteriolog., 1888. Cameron, Brit. 
 Med. Journ., 1884. Cattani, Deut. Med. Woch. 1886. Carter, Lancet, 1884. 
 Cheyne, Brit. Med. Journ., 1885. Crookshank, Lancet, 1885. Cunningham, 
 Scientific Memoirs of Med. Off. of Indian Army, 1885, 1891. Deneke, Deut. 
 Med. Woch., Nr. 3, 1885. Doyen, Soc. de Biol. de Paris, 1884. Drasche, 
 Allg. Wien. Med. Zeit., 1885. Dunham, Zeitsch. f. Hygiene, 1887. Emmerich, 
 Deut. Med. Woch., Nr. 50, 1884. Ermengem, Deut. Med. Woch.. Nr. 29, 1885 ; 
 
P.II'.LKHJKAIMIV. 657 
 
 Recherches sur k- Micmln- <lu Cholrra Asiati.pi.-, INS.". Ferran, Coinpt. Rend., 
 T. K>(. p. !.V.i. 1^:>. Finkler and Prior, Naturforschenersainmliu 
 burg. 1SS4; Deut. Med. Woch., Nr. 86, 1884; Kr-an/.im-sheftr zum Centralbl. 
 f. Allg. Gesundbeitepflege, Bd. I., ll.-ft :, n. r,. [885, Fltigge, D.-ut. 
 Woch., Nr. 2. Iss:,. Gaffky, Arb.. aus <1. K. C.'Mmdh.-iisainte, ]ss;. Gamaleia, 
 Compt. Rend., lsvv : Compt. Rend. Soc. de Biol.,._r . issji. Oibier and van 
 Ermengem, Compt. Rend., T. 101, isx:,. Haffkine, Brit. Med. .Journ., 
 Hericourt, Compt. Rend., 1885. Hueppe, Berl. Klin. Woeh.. iss?; Deut><-h.- 
 Mcd. Woch., 1887; Compt. Rend., ]>^'.i; rra<_r. Mel. Wo<-h., ixxn. Hunter, 
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 Kitasato, Zeitschrift f. Hygiene. l^ v . l^'.i. Klebs, IVber Cuolera Asiatica, 
 l^s."). Klein, Brit. Med. Journ. and Proc. Roy. 8oc. London. N >. :N, : 
 Bacteria of Asiatic Cholera, 1889. Klein and Gibbes, An Inquiry into the 
 Etiology of Asiatic Cholera: Bluebook, issr,. Koch, h.-ut. .Mr 1. W.c'.i.. l 
 Etiolog-y of Cholera : Barlin Cholera Conference : Translated by Laycock, 
 in Microparasites and Disease (New 8yd. Soc.), lss.i. Lewis, M.-d. Tinn-- 
 and Gazette, 1884. Lustig, Centralbl. f. die Mi-d. \Viss., 1S^7: Xeitsjhrift f. 
 Hygiene, iss?. Macnamara, P.rit. Med. Journ., 1-ssj. Miller, Dent. 
 \V< ,rh.. Nr. t, is.v.-). Neuhana, Centralbl. f. Bacteriolog., 1889. Hicati and 
 Eietsch, Arch, de Physiol.. INS:, : i;t-\ue dc Medecin, T. :>, 1885; Revue 
 d'Hygiene, 1885. Petri, Centralbl. f. Bacteriolog., 1889. Pettenkofer, Lmcet, 
 l^^f.. Pfeiffer, Corresp.-Bl. des Allgem. Aerztl. Ver. inThiiringen, Nr. it, 1 -^ i : 
 Deut. Med. Woch., 1886-88. Pfuhl, Zeitschr. f. Hy-i, , : , . l-^i>. Boy, Brown 
 and Sherrington, Proc. Roy. Soc., Vol. xli. Salkowski, Virchow'.s Arc iiv, 1^-7. 
 Schottelius, Deut. Med. Woch., Nr. 14, 18^. Strauss, Eoux, Thuillier and 
 Nocard, Compt. Rend. Soc. de Biol., T. 4, 1*83. Tizzoni and Cattani, 
 Centralb. f. die Med. Wiw., 1886, 1887. Vincenzi, iK-utM-h.- Med. Woch., 1887, 
 Wassiljew, Zeitschr. f. Hygiene. 1*>7. Weisser, Zeitsshrift f. Hygiene, 1 ";. 
 Weisser and Frank, Zeitschr. f. Hygiene, ls>C. Zislein, Deutsche M -I. 
 Zeitung, 18X7-"^. 
 
 GHAPTEU XXVni. 
 TUBERCULOSIS. 
 
 Albrecht, Arch. f. Kinderheilk., Bd. .".. Il. Andrew, Lancet, 
 Arloing, Compt. Rend.. 1>-I. Aufrecht, Centralbl. fiir d. Med. Wi-.. 1882, 1883. 
 Babes, Compt. Rendu-. 1883 : Centralbl. f. d. Mel. Wissensch.. 1883. Babes 
 and Cornil, Journ. de TAnat. et de la PlivMol. Norm et 1'atlml., l^sj. Balogh, 
 Wien. Med. Woch., !**_>. Baumgarten, J'.eil. Klin. Wo h.. l',d. 17. ]s7 ( .t : 
 Centralbl. f. d. Med. Wiss.. P.d. I 1 .*. 1 1 : J82 ; Md. i'l. 1883 : I! 1. L'2, 
 
 1884; Deut. Med. Woch., Bd. 8, ISM' : Zeitschr. f. Klin. M.-d., J5d. ;. 
 I886b Biedert, Vir.-h<.\v's Archiv. Hd. '.'>. 1884 Klin. W,,ch . 1886. Black, 
 
 Lancer. L88& Bock, Vin-hnws Archiv. Bd. 1M. 1^::. Bollinger, (Vntralbl. f. 
 ,1. M, !!. -2\. S. C,(M). 1--:;: Miim-h. At-rztl. :. Intclligenzbl.. Nr. ir, 
 
 1883. Bouley, La Nature Vinmte de 1 Contagion, ContagiosiW d 1 Tnber- 
 culose: Paris. lss4. Brouilly, Rev. de Chir.. T. :;.. 1883. Celli and Guarneri, 
 Arch, pom 1883. Cheyne, Brit. M.-d. Journ.. V,,l. I.. ; 
 
 Practitioner. Vol. XXX., 1-vt. Chiari, Wi,-n. Mr 1. I'r . 1888. Cochet, 
 
 Compt. Rend. Su<-. de Bi.l. : Pari>, I. 5, 1888. Cohnheim, r.-l)-rtra-_l, .rk.-it 
 der Tuberculose: Berlin, Is77. Cornil and Leloir, Arch, de Physiol. Norm, et 
 
 42 
 
658 APPENDICES. 
 
 Pathol., 1884. Cramer, Sitzungsber. d. Phys. Med. Soc. zu Erlangcn, 1883. 
 Creighton, Trans. Path. Soc., 1882 ; Brit. Med. Journ., 1885 ; Lancet, 1885. 
 Crookshank, Rep. Agric. Dept,, 1888 ; Proc. Phys. Soc., 1890 ; Trans. Path. Soc., 
 1891. Damscli, Deut. Med. Woch., Nr. 17, 1883. Dejerine, Rev. de Med. : Paris, 
 T. 4, 1884. Demme, Jahresber. d. Jennerschen Kinderspitals : Bern, 1883. Dett- 
 weiler, Berl. Klin. Woch.. Bd. 21, 1880. Dieulafoy and Krishaber, Arch, de 
 Physiol. Norm. etPath., T. I., 1883. Doutrelepont, Vierteljahrschr. f. Dermato- 
 logie u. Syphilis, 1884 ; Deut. Med. Woch., Nr. 7, 1885. Ehrlich, Dent. Med. 
 Woch., 1882, 1883. Ermengem, Ann. de la Soc. Beige de Microscopie, 1882. 
 Ewart, Lancet, 1882. Formad, The Bacillus Tuberculosis : The Philad. 
 Medical Times, 1882. Frantzel and Palmers, Berl. Klin. Woch., 1882, 1883. 
 Fiitterer, Virch. Arch., Bd. 100, Heft 2, 1885. Gaffky, Mitth. a. d. Kaiserl. 
 Gesundheitsamte, Bd. II., 1884. Giacomi, Fortschr. d. Med., Bd. I., S. 145, 
 1883. Gibbes, Lancet, 1883. Goldenblum, Vrach., Nos. I. and XL, 1886. 
 Green, Brit. Med Journ., 1883 ; Lancet, 1887. Harries and Campbell, Lupus: 
 London, 1886. Harris, St. Barthol. Hosp. Reports, 1885. Heron, Lancet, 
 
 1883. Hiller, Deut. Med. Woch., Bd. 8, 1882. Johne, Die Geschichte der 
 Tuberculose, 1883 ; Ber. iib. d. Veterinarwesen im Kb'nigr : Sachsen, 1883 ; 
 Fortschr. d. Med., Bd. 3, 198, 1885. Karg, Centralbl. f. Chir., 1885. Kirstein, 
 Deut. Med. Woch., 1886. Klebs, Virchow's Arch., Bd. 44, 1868 ; Arch. f. Exp. 
 Pathol. u. PharmakoL, Bd. I., 1873 ; Bd. 17, 1883. Koch, Die Aetiologie der 
 Tuberculose : Berl. Klin. Woch., 1882 ; Deut. Med. Woch., Nr. 10, 1883 ; 
 Mittheilungen aus dem Kais. Ges. Amt, Bd. II., 1884. Kundrat, Wien. Med. 
 Presse, 1883. Kiissner, Deut. Med. Woch., Nr. 36, 1883. Landouzy and 
 Martin, Rev. de Med., T. 3, 1883. Leube, Sitzungsber. der Phys.-Med. Soc. zu 
 Erlangen, 1883. Levinsky, Deut. Med. Woch., Bd. 9., 1883. Leyden, Zeitschr. 
 f. Klin. Med., VIII., 1884. Lichtheim, Fortschr. d. Med., Bd. I., 1883. Lustig, 
 Wien. Med. Woch., Nr. 48, 1884. Lydtin, Badische Thierarztl. Mittheil, 1883. 
 Malassez and Vignal, Compt. Rend., T. 97, 1883 ; Coinpt. Rend., T. 99. p. 200. 
 
 1884. Marchand, Deut. Med. Woch., Nr. 15, 1883. Max-Bender, Deut. Med. 
 Woch., 1886. Meisels, Wien. Med. Woch., 1883. Middeldorpf, Fortsch. d. 
 Med., 1886. Miiller, Centralbl. f. Chir., 3, 1884, 1886. Nauwerck, Deut. 
 Med. Woch., 1883. Obrzut, Deut, Med. Woch., Nr. 12. 1885. Pfeiffer, 
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 Tuberculose des Menschen zu der Thiere, 1883. Raymond, Arch. Gen. de 
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 Wien., 1883. Sticker, Centralbl. f. Klin. Med., 1885. Strassmann, Virchow's 
 Archiv, Bd. 96, 1884. Sutton, Trans. Path. Soc. London, Vol. XXXV., 1884. 
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 1885. Veraguth, Arch. f. Exp. Path. u. PharmakoL, Bd. 16, 1883. Vignal, 
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BIBLIOGRAPHY. 
 
 CHAPTER XXIX. 
 
 LEPROSY. SYPHILIS. RHINOSCLEROMA. TRACHOMA. 
 
 LEPBOSY. 
 
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(i(>0 APPENDICES. 
 
 CHAPTER XXX. 
 
 \( T1NOMYCOSIS AND MADURA DISK AH . 
 
 ACTINOMYCOSIS. 
 
 Acland, Brit. Mod. Journ. and Trans. Path. s,,,-.. issr, : :i nd Alllmfs Systom oi 
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 1883, Kansome, Brit. Mod. Journ.. 1S5M>. Report of tho Hoard of Livo-Stork 
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 -lahrl... S. 177, L888, 
 
 MADIKA PISKASB. 
 
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 CIIA1TK1I XXXI. 
 GLANDERS. 
 
 Babes, Aoad. do. Mod.. L888, Baumgarten. Contrail*!, f. Haoteriolog., 1888. 
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wm.iMi,:.\niY. 
 
 rilAlTKi: \\.\ll. 
 \M 8, i;\!:<i>. LOUPUCG-ILL, 
 
 l'i: i LHUB, 
 
 Carle and Rattone, Studio s^'rim.-ntalr siill' Kii.-l 
 
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 Vaillard, Ann. do ITnstitut 1'asirur. ls;::. Vogel, IVut. Mrd. \V... 
 I88i 
 
 K AIIIIIS. 
 
 Babes. L.-s Baotfoies, 1886, Bauer, Mun<-h. lied, Wbob., I^M*.. Bert, 
 
 r.-mpt. lU-nd.. !>>L>. Colin, Hull. Aoa<l. d- .M.-.I. Paris, T. in, Issl. DoUrit, 
 . dr rai-is. T. :? : Tribune Mrd. 1'aii^. T. U. INS]. Dowdeiwell, 
 .lourn. liny. Micro. S,x-. and 1 Fol. Ao*d l vv l. 
 
 Frisch, \Vit-n. I Gibier. Oompt. lirud.. is^:{. Kerr, Urn. 
 
 Mil. .lourn.. Issr,. Pasteur, Cunpt. IN-nd.. l^^l. l^l. l^^t; : Ann. <! M.-d. 
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 Klein, .Tourn. of Koyal A.irrio. 8oo., l x< .'". MTadyean, .lour: 
 1896, and .lourn. of Comp. Path, and Bacteriology, 1 S 
 
 CHAPTER XXX II I. 
 
 FOOT-ROT. 
 
 Brown, Journ. Itoyal Agi 99t, Nott, .lourn. Koyal 
 
 1890. 
 
 rilAlTKU XXXIV. 
 
 FOUL-BROOD. INFK(Tlors II>KA>K OP BEES l\ ITALY. 1'KIIKIM-:. 
 FLACHKRIK. IMK< i'lOlS PISKASK < >F CATF.RIMLLAKS. 
 
 B^champ, ('omitt. Kend., ls;7. Cheshire and Cheyne, .lourn. of Hoy. M 
 
 y. Cowan, .lourn. Koyal Apr: - '-. Forbes, Hull. Illin. - 
 
 La'n. of Nat. Hist., INSI',. Klamann, lUi-nenwirth-. -Ijal't. < 'entralbl., i 
 Pasteur, KtmU-s des mala- -ie. 187O. 
 
 OHAFTBB \\.\\ . 
 
 ( -L.\ II K ATlnN AND I>KS( |{1 1' I'K t.N F - 
 
 Acostaand Rossi, Centndbl. : Adametz, hi.- l',akt. ,! r Nut/. 
 
 u. Trinkwii-.T. 1888 : Landwirth-rhaft. -Jarhb.. ISM. Afanassiew, Bl 
 burg M-d. \V. !:.. 1-^7. Ali-Cohen, <Vntr. f. J!a.-t., i:d. V'.. l--.. Almquist, 
 
662 APPENDICES. 
 
 Zeitschr. f. Hygiene. Bd. X., 1891. Alvarez, Compt. Bend., T. cv., 1887. 
 Arloing, Compt. Kend., cvi. and cvii. Arthur, Proc. Acad. Nat. Sci. Phil., 1836. 
 Babes, Bakt. Untersuch. ii. Sept. Proz. des Kindesalters, 1889 ; Virch. Archiv, 
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P.lP.LIOCihAPHY. (Hi.) 
 
 APPENDIC I 
 
 \ 
 
 H/KMATO/OA. 
 
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SUPPLEMENTARY APJ'KNDIX. 
 
 EXTRACTS FROM THE FINAL REPORT OF THE ROYAL 
 VACCINATION COMMISSION. 
 
 THE filial Report of the Royal Commission ito inquire into the 
 subject of Vaccination was published on September 18th, 1896.* 
 The author desires to gratefully acknowledge the permission granted 
 him, by the Controller of Her Majesty's I Stationery Office, to make 
 extracts bearing more especially on the history and pathology of 
 protective inoculation and the prevention of small-pox. The reader 
 is recommended by the author to study the whole of the report. 
 
 History of 
 
 The early history of small-pox, like that of many similar diseases, is 
 obscure, is subject to much debate, and, save perhaps on one point, is of 
 antiquarian interest only. 
 
 The records of the eighteenth century show that the disease was very 
 prevalent in western Europe during the whole of that century. The 
 records of the seventeenth century also show that small-pox was a very 
 common disease during that century : this is especially the case as regards 
 the latter half of the century. The statistics which exist with respect to 
 Geneva, and various scattered statements, further show that small-pox 
 was a well-known disease in the sixteenth century ; but, except for the 
 records which are said to exist of severe epidemics in Iceland taking 
 place as early as 1241, as we go further back the evidence as to the 
 existence of the disease becomes less and less clear, and indeed debate- 
 able, depending as it does largely on the interpretation of incidental 
 
 * The report may be obtained either directly, or through any bookseller, 
 from Eyre & Spottiswoode, East Harding Street, Fleet Street, E.C., and 32, 
 Abingdon Street, Westminster, S.W. ; or John Menzies & Co., 12, Hanover 
 Street, Edinburgh, and 90, \\Yst Nile Street, Glasgow; or Hodges, Figgis & 
 Co., Limited, 104, Grafton Street. Dublin. 
 
 f The headings to the extracts from the Kt-jx-rt <>f tin- Cinmi>H"ii un mine. 
 E. M. 0. 
 
f)68 SUPPLEMENTARY APPENDIX. 
 
 statements in various medical and other writings. There seems, how- 
 ever, to be adequate proof of the prevalence of small- pox in the East, 
 in Asia Minor and other countries, even in the earlier centuries of the 
 Christian era. 
 
 A view very generally taken teaches that small-pox, introduced from 
 the East, began to be common in western Europe during the fifteenth 
 century, though perhaps existing still earlier ; that it increased during 
 the sixteenth and seventeenth centuries, especially the latter ; and that it 
 was very prevalent during the eighteenth century. 
 
 In dealing with the eighteenth century it must be borne in mind that 
 during the second half of the century the natural conduct of small-pox, 
 as we shall see later on, was modified by the practice of inoculation that 
 is, by the artificial giving of the disease by the introduction of the virus 
 through a wound in the skin. 
 
 Our knowledge of the history of small-pox in western Europe during 
 the seventeenth and eighteenth centuries is very largely based on the 
 official records known as the '' London Bills of Mortality." Official 
 records bearing on small-pox are furnished by Geneva, going back as 
 far as the sixteenth century, by Sweden, going back to the year 1749, 
 and by some other places. Data are also furnished, especially for the 
 latter part of the eighteenth century, by parish records in various parts 
 of Great Britain reaching over a variable number of years, as well as by 
 scattered statements in various works. 
 
 These Bills of Mortality form by far the most complete source of our 
 knowledge of small-pox in England in past times ; but it must be borne 
 in mind that in respect to any contagious disease like small-pox the 
 conditions of London were peculiar. The population was to a marked 
 extent a moving one ; a large number of persons were continually 
 entering London or leaving it, were passing to and from it, from and to 
 the provinces of England and other countries. Of these persons, some, 
 coming from infected districts, brought into London fresh sources of 
 contagion ; others again, coming from districts free from small-pox, and 
 never having had the disease, brought into London fresh material to serve 
 as food for the disease. Further, London presented in an exaggerated 
 degree the two features of a great city which have a great influence on 
 the progress and characters of a contagious disease like small-pox. The 
 crowding both of the dwelling-places and the thoroughfares, as well as 
 the movement continually going on, multiplied the opportunities for the 
 spread of disease, and the accompanying insanitary conditions, as well as 
 the greater inducement to irregular living, tended to increase the severity 
 of the disease when taken, and to heighten the mortality from it. The 
 history of small-pox in London must not be taken as representative of 
 the history of small-pox in England generally. 
 
 Inoculation of ^ntnU-^ox. 
 
 The practice of inoculation for the small-pox that is, the artificial 
 introduction of the virus into the system by the insertion of fluid from 
 a variolous pustule into wounds of the skin made for the purpose 
 began definitely in England towards the end of the first quarter of the 
 
REPORT OF THE ROYAL YA I.VV1 I. >N COMMISSION. C69 
 
 eighteenth century. Attention was directed to the matter by letters 
 from Timoni of Athens (dated 1713) and Pylarini. published in th, 
 21) th volume of the 7 '/<;/< -.<;,/, ; t w '/>///*// ;//* (1716), and especially 
 by a letter from Lady Mary Wortley Montagu in 1717. Though 
 there aie indications that in Great Britain and Ireland, as in other 
 countries, some sort of inoculation -had occasionally been practised at 
 a much earlier date, the first clearly recorded case in England is that 
 of the daughter of Lady Mary Wortley Montagu (whose son had some 
 time before been inoculated at Constantinople), inoculated by M;iitland, 
 iu London, in April 1721. Other cases soon followed in England, and 
 about the same time the practice was also introduced in other count rit-> 
 of western Europe, and into the United States of America, namely, at 
 Boston. 
 
 It was found that the attacks induced by inoculation were as a rule 
 milder, and very much less fatal, than the attacks of the u natural ' 
 disease, the fever and constitutional disturbance being less and of shorter 
 duration, and the eruptive pustules much fewer : the number of these 
 varied, being commonly a dozen or two, j-omiti nes only two or three, 
 sometimes a hundred or more. In som >. cases there was no eruption 
 at all, the effect being limited to constitutional disturbances and to 
 changes in the wounds of inoculation themselves ; it was maintained that 
 in such cases the disease had really been taken, and immunity against 
 a subsequent attack secured, as in case* of natural small-pox or of 
 inoculated small-pox manifesting itself in an eruption of pustules. 
 
 In England the practice of inoculation at its introduction, though 
 much lauded and strongly urged by some, was bitterly opposed by others. 
 Moreover, the initial enthusiasm in favour of it soon declined, so that 
 in the years 1730-40 very little inoculation seems to have been practised. 
 About 1740, however, a revival appears to have taken place: in 174> 
 an Inoculation and Small-pox Hospital was started in London ; and 
 during the whole of the latter half of the eighteenth century the practice 
 may be said to have been very general. It was especially so during the 
 last quarter of the century, the increase being at least largely due 
 to the -'improved methods" of inoculation introduced by one Sutton 
 iu 17G3, and known as " the Suttonian method." 
 
 Since an inoculated person was infectious, each inoculation was 
 a source of danger to those, not protected by a previous attack, who 
 came into the company of, or even near, the inoculated person during 
 the attack ; and this danger was increased by the fact that the mild 
 character of the inoculated disease permitted, in many cases at least, 
 the patient to move about among his fellows. Moreover, as Haygarth, 
 himself a zealous advocate of inoculation in a systematic regulated 
 manner, points out, the beneficial results of inoculation had robbed 
 the disease of its terrors to so great an extent that the rich and powerful 
 no longer made the efforts which they formerly did to prevent its 
 entrance into, or its spread in, their neighbourhood, and thus favoured 
 its spread among the unprotected poor ; so that inoculation " though 
 eminently useful to the rich appeared to be injurious to the poor." 
 Adding, therefore, together the cases of inoculated small-pox, and the 
 
670 SUPPLEMENTARY APPENDIX. 
 
 cases of natural small-pox of which the inoculated cases were in one way 
 or other the cause, it seems probable that inoculation did tend to increase 
 the prevalence of small-pox ; but there are no recorded data to show that 
 this really was the case, and this supposed influence may have been 
 counterbalanced by other influences. 
 
 The evidence as to the influence which inoculation had on the 
 mortality from small-pox is in many respects conflicting. Haygarth, 
 though he admits that in other parts of the kingdom the practice may 
 have saved many lives, was persuaded that in his own part of England 
 and Wales the deaths by the small-pox had been augmented by it ; and 
 he points out that in London, Geneva, and other " towns in different 
 situations and circumstances, the mortality from this distemper has 
 increased since the introduction of inoculation." Several writers in the 
 latter part of the last, and the early part of the present century, held 
 a similar view. Other writers, again, opposed this view. 
 
 Tradition of the Dairy-folk. 
 
 There was at the close of the eighteenth century, if not earlier, in 
 districts where cow-pox had appeared, a belief among the dairy-folk that 
 those who had taken the cow-pox never took the small-pox ; and indeed 
 one Jesty, a Dorsetshire farmer, had in 1774, in the case of his wife and 
 sons, purposely introduced the matter of cow-pox into the human subject 
 with the view of protecting from small-pox. 
 
 Cow-pox. 
 
 Vaccinia or cow-pox is a disease affecting milch cows, and marked by 
 an eruption on the udder and teats. The disease can be communicated 
 from the cow to man. Dairymen and maids engaged in milking cows 
 affected with cow-pox are apt to have sores of a special kind on their hands 
 or elsewhere, the development of the sores being frequently accompanied 
 by febrile symptoms. There can be no doubt that, in a certain number 
 of cases at all events, such sores are the local manifestations of cow-pox : 
 the virus from the eruption on the cow being introduced into some scratch 
 or other imperfection in the skin of the milker and there producing its 
 local effects, accompanied more or less by general symptoms. 
 
 Inoculation of Cow-pox. 
 
 The practice, however, of inoculating with the matter of cow-pox, or 
 vaccination as it was subsequently called, may be considered as dating 
 from the publication of the u Inquiry into the Causes and Effects of the 
 Variolae Vaccinse " of Edward Jenner, published in the summer of the 
 year 1798. The practice rapidly spread, and prevailed widely in this 
 country and other parts of western Europe during the first quarter of the 
 present century. It was, beyond all question, so adopted in the genuine 
 belief that it afforded protection against small-pox. 
 
 In the treatise to which reference has been made Jenner records in the 
 first place a number (19) of cases in which a person who had accidentally 
 taken cow-pox from the cow had never had small-pox, and appeared 
 
REPORT OF THE ROYAL WACONA11O9 COMMISSION, (171 
 
 incapable of taking that disease ; the insusceptibility being shown on the 
 one hand by the failure to contract the disease after ample exposure to 
 contagion, such as nursing and attending to or even sleeping with persons 
 suffering from small-pox, and on the other hand by the fact that when 
 the person in question was inoculated with the matter of small-pox in the 
 manner then usual (the matter being tested as to its efficiency on 
 susceptible persons) the inoculation failed to excite small-pox. In the 
 course of the inoculation practice it had been observed that when the 
 operation was performed upon a person who had already had small-pox, 
 either naturally or by inoculation, the wound of inoculation, instead of 
 developing, as it did when the operation was successful in a person who 
 had not had the small-pox, into a vesicle and so into a pustule with the 
 variolous characters (the development being accompanied by febrile 
 symptoms and, save in exceptional cases, by the appearance of a smaller 
 or greater number of variolous pustules on parts of the skin other than 
 the seat of inoculation), presented as a rule nothing more than some slight 
 inflammation, dying away in a few days without any other symptom, or even 
 healed at once without any symptoms at all, local or general : and in the 
 exceptional cases in which further changes took place in the wound, these 
 were not accompanied or followed by an eruption of pustules or even by 
 the febrile and other general symptoms of small- pox. Accordingly, in 
 cases of small-pox inoculation where it was doubtful whether the disease 
 had been communicated, it had become not an uncommon practice to 
 repeat the operation, in order to judge by the effects produced whether 
 the earlier inoculation had or had not produced the disease ; and the 
 practice, thus originating in connexion with small-pox inoculation, had 
 come to be spoken of as the " variolous test." 
 
 In his treatise Jenner distinguishes between what he calls true cow-pox 
 and other eruptions which he speaks of as spurious, and which he regarded 
 as not affording protection against small-pox, although he gives no details 
 to show that the cases quoted by him as affording protection were cases 
 of his true cow-pox. He also developed the view that matter derived from 
 horses suffering from the disease known as the grease is capable of giving 
 rise to cow-pox in the cow, and indeed is the real origin of the true 
 disease. It may be added that Jenner also expressed the opinion that the 
 protection thus afforded by cow-pox was permanent in character. 
 
 Jenner further recorded in the same treatise how he had in IT'.'f, 
 inoculated a healthy boy of eight years of age in the arm with cow-pox 
 matter taken from a sore on the hand of a dairymaid who had been 
 infected with the disease by milking cows suffering from cow-pox. He 
 describes the appearances subsequently presented by the wounds, and 
 states that, six weeks afterwards, the results of inoculating the l>oy with 
 variolous matter were those commonly seen to follow the inoculation of 
 persons who had previously had the cow-pox or the small-pox : that is 
 to say, the " variolous test" showed the boy to be insusceptible to small- 
 pox. Some months afterwards the boy was again inoculated, but no 
 sensible effect was produced on the constitution. Jenner then relates that 
 subsequently, in the spring of 1798, he inoculated a child, and obtained a 
 similar result with matter taken directly from the nipple of a cow infected 
 
672 SUPPLEMENTARY APPENDIX. 
 
 with cow-pox ; from the pustule on the arm of this child he inoculated 
 another, and from this again several, and from one of these latter a fourth 
 in Succession, and then a fifth. To three of these the " variolous test" 
 was applied, and it is stated with the same results. 
 
 Woodville's Lymph . 
 
 The experiences of Jenner did not stand alone. His results and 
 views attracted great attention, and in the early part of the year 1790 
 Woodville and Pearson, who were physicians to the Small-pox Hospital 
 in London, commenced making experiments with vaccine matter with 
 a view to ascertain whether it afforded protection against small-pox. 
 They arrived, like Jenner, at the conclusion that it did. 
 
 In January 1799 Woodville, having found cow-pox to be present in 
 a " dairy " at Gray's Inn Lane, inoculated seven persons at the Small- 
 pox Hospital with matter from one of the cows at the "dairy," and 
 other persons with matter from sores on a dairymaid employed at 
 the same place who had become infected from the cows. From these 
 cases he inoculated in succession others at the Hospital, eventually to 
 the number of many hundreds, and thus established the stock of what 
 has been spoken of as " Woodville's lymph." Pearson also at the same 
 time occupied himself with the question of inoculation with the 
 cow-pox, writing a pamphlet about it. Woodville and he distributed 
 to many persons in this country and abroad quantities of the lymph 
 from the Hospital ; and this was the beginning of the more general 
 practice of vaccination, for Jenner's stock of lymph, the results of 
 which he had described in his treatise, had come to an end. 
 
 Although Woodville's "Hospital lymph" appears to have been 
 widely distributed by himself and by Pearson, and thus to have been 
 the source of the lymph used in vaiious places in the early days of 
 vaccination, it was not the only source, even in those days. Pearson 
 also obtained lymph from cow-pox at a dairy in the Marylebone Road, 
 and used this " in certain situations," which may be presumed to 
 include places elsewhere than in the Hospital. He also speaks of 
 having obtained lymph from the cow from a third source. Jenner 
 again, who received and used some of Woodville's Hospital lymph, also 
 obtained lymph from some other sources : for instance, from a cow 
 at a Mr. Clark's farm in Kentish Town. Further, Woodville in 1800 
 speaks of his having at various times procured the vaccine virus as 
 produced in different cows, which when used at the Hospital produced 
 the same effects as the Gray's Inn Lane lymph. We are not justified 
 in assuming that an account of every new source of lymph was 
 published ; and there may have been others, it is impossible to say 
 how many, than those just mentioned. In any case Woodville's 
 Hospital lymph was not the only lymph used in those early days ; not 
 only, however, was it largely used (indeed, we have no evidence of so 
 widespread a use of lymph derived from any other source), but the 
 use of it marks the definite beginning of the practice of vaccination ; 
 and the history of it demands special notice. 
 
REPORT OF THE ROYAL VACCINATION COMMISSION. 673 
 
 Of the cases recorded by Woodville in his Reports, the larger 
 number (about three-fifths) presented an important, and, as compared with 
 Jenner's cases, a new feature, in that, in addition to the changes taking 
 place at the seat of inoculation and constituting what Woodville called 
 the "cow-pox tumour," which may here be spoken of as the "vaccine 
 vesicle," an eruption over the body of a greater or less number of 
 pustules was observed. These eruptive pustules occurred in the very 
 first cases : of the seven cases inoculated from the cow, four, and of 
 the five inoculated from the dairymaid, four, had such pustules ; and 
 their appearance is recorded again and again in the series, down to the 
 case which appears last but one in the tabular statement forming part 
 of the Reports. 
 
 Moreover, an eruption of pustules is described in certain of the cases 
 of which accounts were published at about the same time by Pearson 
 and many others. In some of these cases the lymph used was supplied 
 from the Small-pox Hospital by Woodville or Pearson. 
 
 It must be admitted that these pustules were pustules of small-pox,, 
 and that, therefore, Woodville's cases, which did so much to establish 
 the practice of vaccination, were not cases simply of cow-pox but of 
 cow-pox mixed, so to speak, with small-pox. It has indeed been 
 maintained that Woodville's cases were not cases of cow-pox at all 
 that small-pox was inadvertently introduced into the very first cases ; 
 that the history of the whole series is the history of a series of small-pox 
 cases putting on special characters, and that therefore the lymph used 
 and distributed by Woodville and Pearson was in reality not cow-pox 
 lymph but small-pox lymph. A review of all the evidence available 
 leads to no other conclusion than that, however much in Woodville's, 
 Pearson's and other cases cow-pox was mixed up with small-pox, the 
 lymph used and distributed by Woodville and Pearson and called by 
 them cow-pox lymph (excluding of course all the cases, of which there 
 were not a few, in which matter was taken not from the local <; cow-pox 
 tumour" at the seat of inoculation, but from one of the eruptive 
 pustules) was veritable cow-pox lymph having the true characters of 
 cow-pox lymph only. 
 
 It of course follows that the cases, both in Woodville's practice and 
 in that of others, in which the inoculation of cow-pox matter was 
 accompanied by an eruption of pustules, due to small-pox being present 
 as well as cow-pox, when appealed to as showing immunity against 
 small-pox (by the test either of exposure to contagion or of inoculation), 
 furnished false evidence as to that immunity being due to cow-pox ; 
 it might have been due to the accompanying small-pox. So far then as 
 the adoption of vaccination was assisted by\cases of this description, it 
 may be held to have rested on erroneous data. 
 
 77"- J}i-ri;,t, nf >//,///-, 
 
 One effect of the introduction of vaccination was a very great decrease 
 in the practice of inoculation, which had become very prevalent during the 
 later part of the previous century. And the view has been put forward 
 
 43 
 
674 SUPPLEMENTARY APPENDIX. 
 
 that, the prevalence of inoculation having greatly increased the amount 
 of small-pox, the diminution of small-pox in question was the result of 
 the decrease of inoculation. \ 
 
 The question how far the behaviour of small-pox in the eighteenth 
 century and earlier was influenced by sanitary conditions, is one rendered 
 difficult by the lack of exact information. We may distinguish between 
 overcrowding as one insanitary condition and all other insanitary condi- 
 tions, such as lack of cleanliness and the like. A priori we should ex- 
 pect that a dense population, especially one of great internal movement, 
 and one in continual interchange with surrounding populations, by 
 offering greater facilities for the conveyance of contagion, would lead to 
 a greater amount of small-pox. London was a conspicuous instance of 
 the above, and the apparent greater prevalence of small-pox in London 
 than in the provinces may be attributed to these causes : but it would 
 appear that the increase was felt as indeed would, a priori, seem pro- 
 bablerather in the constant presence of small-pox to a considerable 
 amount at all times than in the mortality of the epidemics when these 
 occurred. And the same seems also to be shown to a less extent in other 
 large cities, such as Liverpool. But in this matter of dense and moving 
 populations the eighteenth century did not differ markedly from the 
 arly part of the nineteenth. We might a priori expect the other 
 acknowledged imperfect sanitary conditions of the eighteenth century to 
 increase the fatality of, and so to a corresponding extent the mortality 
 from, small-pox ; but there is no exact evidence to confirm this supposi- 
 tion. If on the contrary we recognise that in the course of the eighteenth 
 century the general mortality, the relative number of deaths from all 
 causes, went on decreasing, and attribute, as has been done, this decrease 
 to improved sanitary conditions, no like decrease of small-pox took place. 
 Again, the places which were deemed the most salubrious appear to have 
 been visited by epidemics of small-pox as severe as those which fell on 
 unhealthy places. Thus the epidemic in Chester in 1774 was undoubtedly 
 a severe one, and yet Haygarth writes, " The healthiness of Chester," as 
 shown by statistics, " must appear so very extraordinary as to be almost 
 incredible." And in general both the incidence of, and mortality from, 
 small-pox seem to have been far less affected by sanitary conditions than 
 might a priori have been expected. 
 
 It may be urged against the view that the decline of small- pox was 
 due to improved sanitary conditions, in the first place, that, admitting 
 the introduction of sanitary improvements, no evidence is forthcoming 
 to show that during the first quarter of the nineteenth century these 
 improvements differentiated that quarter from the last quarter, or half, 
 of the preceding century in any way at all comparable to the extent 
 of the differentiation in respect to small-pox. In the second place, 
 admitting a priori that crowded dwellings tend to increase the liability to 
 contagion, and so the prevalence of the disease, while other insanitary 
 conditions tend in addition to increase the fatality among those attacked, 
 so that insanitary conditions as a whole must tend to increase the 
 mortality from small-pox, no evidence is forthcoming which distinctly 
 shows that the dependence of the prevalence of, or the mortality from, 
 
REPORT OF THE ROYAL VACCINATION COMMISSION. 675 
 
 small-pox, on the lack of sanitary conditions, was a feature of the history 
 of small-pox during the eighteenth century. 
 
 Upon the whole, then, we think that the marked decline of small-pox 
 mortality in the first quarter of the present century affords substantial 
 evidence in favour of the protective influence of vaccination. 
 
 Age Incidence of 
 
 A study of the age incidence of small-pox mortality is very instructive. 
 In connexion with this point it is necessary to bear in mind that experi- 
 ence has led to the conclusion that, whatever be the protective effect of 
 vaccination, it is not absolutely permanent ; the most convinced advocates 
 of the practice admit that after the lapse of nine or ten years from the 
 date of the operation its protective effect against an attack of small-pox 
 rapidly diminishes, and that it is only during this period that its power in 
 that respect is very great ; though it is maintained that, so far as regards 
 its power to modify the character of the disease and render it less fatal, 
 its effect remains in full force for a longer period, and never altogether 
 ceases. The experience upon which this view is founded is derived almost 
 exclusively from the case of infantile vaccination. It has been supposed 
 by some that the transitory character of the protection results from 
 changes connected with the growth from infancy to adult years. Whether 
 this be so or not, we have no means of determining. 
 
 No doubt, when Jenner drew the attention of the public to the value 
 of vaccination, he believed that a single successful inoculation of vaccine 
 matter secured absolute immunity for the future from an attack of small- 
 pox. It is certain that in this he was mistaken. It may well be doubted 
 whether the anticipation was a reasonable one. No such immunity is 
 secured by an attack of small-pox, though there are few who would 
 maintain the proposition that it is without protective influence against 
 another attack. ^1 priori there would seem to be no sound ground for 
 expecting that vaccinia would afford more potent protection than small- 
 pox itself. The extent of the protection afforded (assuming that there is 
 some protective influence) could only be determined by experience. It soon 
 became apparent that Jenner had, in the first instance, overrated the 
 effect of vaccination. That he should thus have overestimated it 
 is not to be wondered at, when the tendency to be unduly sanguine, 
 which besets the discoverer of any new prophylactic, and, indeed, every 
 discoverer, is borne in mind. 
 
 We think, taking it all together, that the evidence bearing upon 
 the question whether the vaccinated are less liable to be attacked by 
 small-pox than the unvaccinated, points to two conclusions : first, that 
 there is, taking all ages together, less liability to attack among the 
 vaccinated than among the unvaccinated ; and next, that the advantage in 
 this respect enjoyed by vaccinated children under ten years of age is 
 greatly in excess of that enjoyed at a more advanced period of life. 
 
 In considering whether vaccination has been the principal cause of the 
 decline, we must inquire whether the other causes suggested by those who 
 deny the efficacy of vaccination will satisfactorily account for it. 
 
676 SUPPLEMENTARY APPENDIX. 
 
 Effect of Sanitation. 
 
 It is said that the decline has, in the main, been due to changes in the- 
 general conditions of life in the different parts of the United Kingdom, 
 apart from the spread of the practice of vaccination, amongst other 
 things, to improvement of sanitary conditions. 
 
 It is beyond doubt that an infectious disease like small-pox is, other 
 things being equal, more likely to spread in towns than in country districts, 
 and more likely to spread in crowded town districts than in others not so 
 densely populated ; so that we should expect a lessened proportion of 
 overcrowded dwellings, by diminishing the opportunities for contagion, 
 to check the prevalence of the disease and consequently to render its 
 mortality less. 
 
 Effect of Isolation. 
 
 It has been maintained that the decline in small-pox mortality is 
 largely due to more frequent and systematic attempts to isolate those 
 suffering from small-pox. We think an answer to this contention is to be 
 found in the fact that it is only in quite recent years that there has been 
 any systematic practice of isolating small-pox patients, and that it has 
 been confined even then to a very limited number of localities. The 
 fact to which we are about to call attention in greater detail than 
 hitherto, that the decline in the deaths from small-pox is found almost 
 exclusively among those of tender years, appears also to militate against 
 the contention. The risk of contagion is not confined to children. 
 Adults also are subject to it. If a better system of isolation had been 
 a main cause of the reduced mortality, we should have expected to see it 
 operate in the case of adults as well as of children. At the same time 
 we are far from thinking, as will appear when we come to deal with 
 that subject, that the efforts at isolation which have characterised recent 
 years have been without a beneficial effect on small-pox mortality. 
 
 Sanitary Legislation. 
 
 We have already pointed out that on d priori grounds it is reasonable 
 to think that improved sanitary conditions would tend to diminish 
 the fatality of, and so to a corresponding extent the mortality from, 
 small-pox. And there can be no doubt that the period with which 
 we are dealing has been characterised by an improvement of this 
 description. There has been better drainage, a supply of purer water, 
 and in other respects more wholesome conditions have prevailed. 
 
 It may be useful at this point to furnish a brief summary of the 
 principal Sanitary Acts which have been passed relating to the different 
 parts of the United Kingdom. 
 
 In 1848 was passed the first great and comprehensive measure which 
 may be called the groundwork of our sanitary legislation as regards 
 England. The Public Health Act of 1848 was, however, principally 
 designed for towns and populous places in England and Wales, not 
 including the Metropolis, which- was dealt with in Acts passed in the 
 same year. The powers of local government supplied by the Act were 
 generally an extension of those before given by sundry local Acts to 
 
REPORT OF THE ROYAL VACCINATION COMMISSION. 677 
 
 -Commissioners of Sewers in the Metropolis, and to authorities in a few 
 large towns. Many provisions corresponding to sections in the Towns 
 Improvement Clauses Act of 1847 are found in the Public Health Act, 
 and communities were thus enabled to obtain by a simple process powers 
 which they could not previously obtain except by a local Act incorporating 
 sections of the Towns Improvement Clauses Act. 
 
 In 1848 was also passed the Nuisances Removal and Diseases Preven- 
 tion Act of that year, in substitution for a similar Act of 1846 which was 
 about to expire ; and in 1849 this Act of 1848 was amended. The 
 provisions of all these three Acts extended to England, Scotland and 
 Ireland. In 1855 a comprehensive Nuisance Removal Act was, as regards 
 England, substituted for the Acts passed in 1848 and 1849 ; and in the 
 following year there was similar legislation for Scotland. In 1860 the 
 English Act was amended ; and in 1806, by the Sanitary Act of that year 
 (to which we shall again refer), the provisions of the English Acts of 1855 
 and 1860, as then amended, were applied to Ireland. 
 
 In 1855, by the Metropolitan Local Management Act of that year, 
 provision was made for the appointment of a medical officer of health 
 and an inspector of nuisances by every vestry and district board in 
 the Metropolis. This provision did not extend to the City of London, 
 where, in 1848, a medical officer of health had been appointed under 
 power given by a local Act. 
 
 In 1858, the Local Government Act of that year, to be construed with 
 the Public Health Act of 1848 as one Act, was passed, and took effect in 
 all places where that Act was in force at the time of its passing ; and, 
 as regards England, these two Acts together constituted until 1872 the 
 principal sanitary legislation on the statute-book. 
 
 There followed, however, within the next ten years many public Acts 
 having sanitary objects, some applying to all, and some to particular, parts 
 of the United Kingdom, besides numerous other Acts of local application. 
 We need only now specially refer to one of these public statutes the 
 Sanitary Act of 1866, which was probably the most important, and applied, 
 in part at least, to England, Scotland and Ireland. This Act, amongst 
 other things, extended the powers of local authorities for the disposal of 
 sewage, and, in amending the English Nuisances Removal Acts of 1855 
 and 1860, added to the definitions of nuisances, especially as regards 
 crowded houses and workshops, and to the duties and powers of local 
 authorities for their abatement, especially in the way of providing means 
 for disinfection and places for ihe reception of dead bodies. 
 
 In 1867 the Public Health (Scotland) Act was passed a comprehensive 
 measure which consolidated into one Act, with certain amendments, the 
 whole statute law relating to the public health in Scotland. 
 
 In 1872 a complete distribution of England into sanitary districts took 
 place, and some further amendments were made in the sanitary laws. In 
 1875 these laws were consolidated in the Act of that year. In 1891 
 a Sanitary Act was passed relating to the Metropolis. 
 
 In 1874 an Act was passed for Ireland, containing substantially the 
 same provisions as those which had been enacted in the case of England 
 in 1872. 
 
678 SUPPLEMENTARY APPENDIX. 
 
 Value of Vaccination. 
 
 We have not disregarded the arguments adduced for the purpose of 
 showing that a belief in vaccination is unsupported by a just view of the 
 facts. We have endeavoured to give full weight to them. Having done so r 
 it has appeared to us impossible to resist the conclusion that vaccination 
 has a protective effect in relation to small-pox. 
 
 We think : 
 
 1. That it diminishes the liability to be attacked by the disease. 
 
 2. That it modifies the character of the disease, and renders it 
 
 () less fatal, and (5) of a milder or less severe type. 
 
 3. That the protection it affords against attacks of the disease is 
 
 greatest during the years immediately succeeding the operation 
 of vaccination. It is impossible to fix with precision the 
 length of this period of highest protection. Though not in 
 all cases the same, if a period is to be fixed, it might, we think, 
 fairly be said to cover in general a period of nine or ten years. 
 
 4. That after the lapse of the period of highest protective potency, 
 
 the efficacy of vaccination to protect against attack rapidly 
 diminishes, but that it is still considerable in the next quin- 
 quennium, and possibly never altogether ceases. 
 
 5. That its power to modify the character of the disease is also 
 
 greatest in the period in which its power to protect from 
 attacks is greatest ; but that its power thus to modify the 
 disease does not diminish as rapidly as its protective influence 
 against attacks, and its efficacy during the later periods of life 
 to modify the disease is still very considerable. 
 
 6. That re -vaccination restores the protection which lapse of time 
 
 has diminished ; but the evidence shows that this protection 
 again diminishes, and that, to ensure the highest degree of 
 protection which vaccination can give, the operation should be 
 at intervals repeated. 
 
 7. That the beneficial effects of vaccination are most experienced 
 
 by those in whose case it has been most thorough. We think 
 it may fairly be concluded that where the vaccine matter is 
 inserted in three or four places, it is more effectual than 
 when introduced into one or two places only and that if the 
 vaccination marks are of an area of half a square inch, they 
 indicate a better state of protection than if their area be at all 
 considerably below this. 
 
 Question of Specific Protection or of Antagonism. 
 
 When an attack of disease secures immunity or protection against 
 another attack of disease, the two attacks are, as a rule, attacks of the 
 same disease. Some pathologists have, it is true, of late years been led to 
 suppose that one disease may confer some degree of immunity or protec- 
 tion against another different disease ; but instances of this are few, and, 
 moreover, cannot be regarded as thoroughly established. The ordinary 
 instances of immunity are so clearly those in which the attack, natural or 
 
REPORT OF THE ROYAL VACCINATION COMMISSION. 679 
 
 artificial, which confers the immunity is of the same disease as that towards 
 which immunity is conferred, that identity of disease has been considered 
 as essential to the conferring of immunity. And it has been argued that 
 it is a /-;>,} improbable that cow-pox should confer immunity from small- 
 pox, seeing that the two are different diseases. Such a purely theoretical 
 argument can have little weight against positive evidence of vaccination 
 having actually conferred immunity. If this be definitely proved to be the 
 fact, proof is thereby at the same time afforded that the theory is unsound, 
 either because a particular disease may confer immunity against a different 
 disease, or because small-pox and cow-pox are not different diseases. For 
 the practical object with which alone we are concerned, it is not material 
 that we should reach any conclusion upon the question what is the real 
 source of error in the theory alluded to, supposing it to be erroneous ? 
 We shall content ourselves, therefore, with a very brief notice of the 
 subject. 
 
 It appears to us that we may dismiss for practical purposes the 
 theoretical questions which were discussed before us so fully. If the fact 
 be established that the introduction of vaccine matter and the consequent 
 vaccinia produce some effect upon the human body which renders it less 
 susceptible to small-pox, or which modifies that disease when the small-pox 
 virus enters the system, it will not be a strange or unwonted experience 
 that we should be unable to explain how this comes about. Science 
 has not yet succeeded in freeing therapeutics or kindred subjects from 
 obscurity, or in solving all the problems which they present. The precise 
 mnilii* njH-1-innJi by which a previous attack of a disease furnishes security 
 against a subsequent attack by the same disease has not yet been elucidated. 
 There can be no cause for astonishment, then, if we are unable to trace 
 the steps by which vaccination exerts a protective influence, supposing 
 the fact that it does so be established, nor is it essential that we should 
 succeed in tracing them. Our inability to accomplish this does not seem 
 to us to be the slightest reason for regarding with doubt the conclusions 
 to which the facts lead us. 
 
 Professor Crookshank, than whom no one has more strongly insisted on 
 the theoretical arguments against the protective influence of vaccination in 
 relation to small-pox, gives it as his opinion that vaccination creates a 
 transient antagonism to that disease. We understand his view to be that 
 an attack of disease can only afford protection against the same disease, 
 and that small-pox and cow-pox are not the same but different diseases. 
 We gather, however, that, in his opinion, so long as the state of antagonism 
 lasts, the person in whose system it exists is less likely to suffer from 
 small-pox than he would be if the state of antagonism were wanting. 
 This seems to us to amount in effect to the same thing as saying that 
 during that period vaccination has conferred some protection. Whether 
 the effect be to create antagonism or to confer protection, and whatever 
 difference there be between the influx ,,/>< r<i/i</i in the one case and in the 
 other, we know equally little about it. If a condition of transient 
 antagonism to small-pox is induced by vaccination, theoretical considera- 
 tions will not afford a guide of the slightest value to the conclusions how 
 long this transient antagonism will last, or how soon it will pass away. 
 
680 SUPPLEMENTARY APPENDIX. 
 
 Experience, and experience alone, can answer that question. A priori we 
 -do not see that there is any better reason for supposing that it would last 
 for two or three years than that its duration would extend to ten years. 
 
 Cow-pox and Smallpox not Convertible. 
 
 Jenner himself, in his first paper, advanced the view that the cow-pox 
 and small-pox were identical with each other ; and since his time 
 numerous observers have attempted to prove the identity of the two 
 diseases experimentally namely, by giving rise to cow-pox in the cow 
 through the inoculation of small-pox matter, or by the introduction of 
 contagion in other ways. It may at once be stated that while cow-pox 
 is readily transferred from the cow to man and back again from man to 
 the cow, the disease in man being identical with that in the cow, small- 
 pox cannot be transferred from man to the cow so as to give rise to a 
 disease in the latter identical in its features with the small-pox of man. 
 Nor can cow-pox be so transferred to man as to give rise in him to 
 small-pox. The two diseases are not in this sense convertible. 
 
 Small-pox Vaccine. 
 
 In most cases the attempt to transfer small-pox from man to the 
 cow has had simply a negative result ; no obvious effect of any kind has 
 been observed. This has been the case in the attempts to introduce the 
 contagion through absorption by the respiratory or digestive organs, and 
 in most of the attempts to introduce the contagion by inoculation. In 
 certain instances these latter attempts have produced results which may 
 be briefly described in three categories. (We may pass over the isolated 
 experience of Thiele, who in 1838 asserted that by keeping small-pox 
 matter sealed between glass plates for ten days before using it, and 
 by diluting it with milk when using it for inoculation, the matter thus 
 treated through ten removes through the human body the cow not 
 intervening at all was converted into something which gave results 
 identical with those of ordinary vaccine matter. We are not aware of 
 any attempt to corroborate this experiment.) 
 
 The first category includes the experiments in which the inoculation 
 of small-pox matter into the udder, or adjoining parts, of the cow gave 
 rise at or near the seat of inoculation to a vesicle, either identical in 
 visible characters with the ordinary vaccine vesicle produced by inocula- 
 tion with the matter of cow-pox, or to a vesicle the features of which, 
 while not corresponding wholly with those of a perfect vaccine vesicle, 
 so closely resembled them as to justify the vesicle being called a vaccine 
 vesicle. Further, the matter from a vesicle which at the first inoculation 
 had not the characters of a perfect vaccine vesicle, when carried through 
 a second or third remove in the cow, fully acquired those characters, and 
 when transferred to man gave results indistinguishable from the ordinary 
 vaccine vesicle. Indeed, lymph of such an origin has come into general 
 use for vaccination purposes. Of the experiments, the best known or 
 most quoted are those of Thiele (1838), Ceely (1840), Badcock (between 
 1840 and 1860), Voigt (1881), Haccius and Eternod (1890), King (1891), 
 
REPORT OF THE ROYAL VACCINATION 00111088109. 681 
 
 Simpson (1892), and Hime (1892) ; but there are several others. The 
 details of the experiment are very scanty in the cases of Thiele and 
 Badcock, but more full in the others, especially, perhaps, those of Ceely 
 and Haccius. 
 
 In the second category may be placed the experiments of Klein and 
 Copeman. Klein, who had in 1*79 obtained in 31 trials what then 
 appeared simply negative results, found in a renewed research in 1 *.'_' 
 that the result of the first inoculation in the cow of small-pox matter 
 was not a distinct vesicle but merely a thickening and redness of the 
 wound. Lymph pressed from the thickened wound produced, when 
 inoculated into a second cow, a like result, but rather more mark. < I : 
 the thickening and reddening still further increased with a third and 
 a fourth cow. Lymph squeezed from the wounds of the fourth cow 
 produced in a child typical vaccine, and crusts from the child produced 
 typical vaccine in a cow. Copeman obtained somewhat similar result* : 
 the appearances increasing in three removes and approaching those of 
 typical vaccine, but not reaching them. 
 
 The third category consists of the results obtained in an elaborate 
 inquiry conducted by a Commission of the Society of Medical Sciences at 
 Lyons, with Chauveau at its head. Those results, reported in 1865, were 
 briefly as follows : 
 
 Inoculation of the cow with small-pox matter in any one of the 30 
 animals used did not give rise to a vaccine vesicle. Nevertheless a 
 definite result was obtained ; in the form, however, not of a vesicle, but 
 of a thickening and inflammation of the wound ; when a puncture was 
 employed this became a papule. Lymph squeezed from such a papule 
 and inserted into a second animal gave rise to a like papule : and this, 
 again, might be used for a third animal, but often failed ; and the effect 
 could in no case be carried on through more than three or four removes. 
 
 When the inoculation was repeated on an animal in which a previous 
 inoculation had produced such a papule, no distinct papule was formed ; 
 and, moreover, lymph squeezed from the seat of inoculation produced no 
 effect at all when used for the subsequent inoculation of another animal. 
 This shows that the development of the papule was the result of the 
 specific action of the virus. The same is shown by the fact that no such 
 papule was produced when the small-pox matter was inserted into an 
 animal which had previously had cow-pox naturally or artificially, as well 
 as by the fact that when an attempt was made to vaccinate, with vaccine 
 matter of proved efficacy, an animal on which a papule had been so 
 developed by inoculation with small-pox, the vaccination failed, though 
 the animal had never had natural cow-pox or had never been vaccinated. 
 The specific nature of the lymph of the papule is further shown by the 
 fact that such lymph when used on the human subject gave rise to veri- 
 table small-pox. It has been urged that in this case the virus producing 
 the effect was simply the old virus used in the inoculation, producing 
 the papule and still clinging to the wound. This is disproved by the 
 experience that lymph //<//* a />"// f> ,,f tin Iso gave rise in 
 
 the human subject to veritable small-pox. 
 
 Thus Chauveau and bis Commission found that small-pox implanted 
 
682 SUPPLEMENTARY APPENDIX. 
 
 in the cow gave rise to a specific effect which was not cow-pox but was 
 of the nature of small-pox, though its manifestations in the cow were 
 different from those of small-pox in man. They also obtained similar 
 results in attempting to transfer small-pox to the horse. 
 
 It must be admitted that the results finally obtained in some of the 
 successful cases were indistinguishable from those of vaccination ; the 
 characters of the local vesicle, the absence of eruptive pustules and of 
 contagiousness, show that the lymph thus apparently originating from 
 small-pox in the hands of Ceely, Badcock, and others, was vaccine lymph. 
 It has been urged that a vaccine vesicle making its appearance in the 
 wound of inoculation with small-pox was due to the accidental intro- 
 duction of cow-pox matter into the wound ; the small-pox matter in 
 the wound produced no effect, and the cow-pox matter its usual effect. 
 Several considerations support this view. The cow is peculiarly suscep- 
 tible to cow-pox. In some cases (Ceely, Voigt), the animal was 
 vaccinated as well as inoculated with small-pox : thus, in Ceely 's first 
 case, the animal was inoculated with small-pox on one side of the body, 
 and a few days after vaccinated on the other side. In many cases the 
 experiments were conducted in an animal vaccine establishment, the 
 stalls, the operating tables, and the assistants being those used or engaged 
 in vaccination. It is true that in some cases at least special precautions, 
 sterilisation of instruments and the like, were taken to avoid the accidental 
 introduction of cow-pox ; but in observations of this kind the difficulties 
 of avoiding all such sources of error are notorious. Still the successful 
 cases are now so numerous that it is difficult to resist the conclusion that 
 the same accident could not have occurred in all, and that a transformation 
 of small-pox into cow-pox that is to say, into the artificially inoculated 
 cow-pox which we call vaccine * really took place. 
 
 Accepting this view provisionally, it may be remarked that in most 
 cases the transformation was sudden and complete ; the small-pox virus, 
 under the influence of the tissues of the cow, became immediately 
 converted into vaccine virus, and this produced a typical vaccine vesicle. 
 In some cases (ex. gr., that of Hime) the transformed virus produced its 
 effect not in the wound of inoculation, or not chiefly so, but at some little 
 distance from it. In some cases the characters of the vesicle first formed, 
 though sufficiently distinct to justify the vesicle being called a vaccine 
 vesicle, were not those of a perfect vaccine vesicle, but the lymph from 
 such a vesicle, at least after one or two removes, gave rise to most typical 
 vaccine vesicles. 
 
 In Klein's experiments the transformation was gradual. In his fourth 
 cow, the virus was as yet not typical vaccine, since it did not produce a 
 typical vesicle ; yet it was so far already vaccine that, transferred to the 
 child, it produced typical vaccine (unless we suppose some accidental 
 introduction of vaccine to have taken place). That the vesicle on the 
 child was vaccine, and not small-pox unaccompanied by eruptive pustules, 
 was shown not only by its characters but also by the fact that lymph from 
 it produced typical vaccine in the cow. 
 
 In Chauveau's experiments no transformation at all took place. 
 * The italics are mine. E.M.C. 
 
REPORT OF THE ROYAL VACCINATION COMMISSION. 683 
 
 As has been urged in another place, there are no adequate reasons 
 leading us to believe that in the human subject the small-pox virus and 
 the cow-pox virus can so act on each other as to form a hybrid disease. 
 But this does not preclude the view that, accepting the conclusion that 
 the body of the cow has the power to convert small-pox into KMXOM, the 
 virus may exist for a while in a phase in which, while ceasing to be 
 typical small-pox, it has not yet fully acquired the characters of vaccine, 
 and we may regard Klein's results as illustrating this. In some of the 
 experiments for instance, those of Ceely and Voigt the results obtained 
 with the lymph of the vesicle produced by the inoculation of small-pox 
 give rise to the suspicion that the lymph had small-pox qualities, as seen, 
 for example, in the case of Ceely's assistant, Taylor ; but the facts cannot 
 be said to be more than suspicious they are not decisive. Moreover, 
 admitting that the vesicle itself in such cases was the result of the trans- 
 formed virus, some not transformed old virus might still remain dormant 
 in the wound, and might be present in the lymph of the vesicle, mixed 
 with the transformed and generating virus ; this old virus might have 
 happened to be in excess on the point of the lancet which wounded 
 Taylor. 
 
 Small-pox Vaccine Cow-pox Vaccine Horsebox Vaccine Cattl*- 
 j>l<i(jni' Vaccine Sheep-j>u.,- V<n-cn . 
 
 Taking all the various facts into consideration, we seem led to the 
 provisional conclusion that under certain conditions the tissues of the 
 cow are able to transform small-pox into ni<-chn\ that these conditions 
 may be such as to lead to the transformation being sudden and complete, 
 that under certain other conditions the transformation may be gradual 
 and incomplete, and that under certain other conditions (and these seem 
 most commonly to obtain) the transformation into vaccine does not take 
 place at all. But what the above conditions are has not as yet been 
 clearly made out. It has been suggested that one condition favourable 
 to the transformation is extreme youth of the subject : to effect the 
 change the animal used should be a calf of not more than three or four 
 months old : but this is not definitely proved. 
 
 l"ntil these favourable conditions have been clearly recognised, so that, 
 the conditions being fulfilled, the transformation will always be secured, 
 the conclusion cannot be regarded as indisputable. Moreover, it must 
 be borne in mind that effects more or less closely resembling a vaccine 
 vesicle have been at various times obtained by various observers through 
 inoculating man or the cow or another animal with material other 
 than that obtained from the pustules of the small-pox of man. Much 
 discussion has taken place concerning the " grease " of the horse, which 
 Jenner believed to be the origin of the cow-po* of the cow. Without 
 entering into any discussion of the matter, it may be said that investiga- 
 tion has shown that horses do suffer from a malady which, transferred to 
 the cow, gives rise to a vaccine identical apparently with that produced 
 by the inoculation of the natural cow-pox. Hence this malady is spoken 
 of as the " horse-pox," and some cases at least of so-called " grease " 
 appear to t>e cases of this horse-pox. But it is at least not proved that 
 
684 SUPPLEMENTARY APPENDIX. 
 
 all the cases of " grease " which by inoculation were found to give rise to 
 vaccine vesicles in man were cases of true horse-pox. And this at least 
 must be said, that no investigations as complete and varied as those which 
 have been carried out with regard to the development of vaccine vesicles 
 through the inoculation of small-pox matter, have been carried out with 
 regard to the alleged development of vaccine vesicles by the inoculation 
 of other material, such as the matter from the eruptions of the sheep-pox, 
 the cattle plague, and the like. Nor have there been like extended in- 
 quiries as to the production of vesicles resembling those of vaccine by the 
 inoculation of small-pox matter into animals other than the cow or the 
 horse ; such results as have been obtained by observers are conflicting. 
 There is still room for much inquiry ; meanwhile it may be said that, in 
 any case, the evidence in favour of a possible transformation of small-pox 
 into vaccine is sufficiently strong to remove the force of the theoretical 
 objection to the power of vaccination to secure immunity towards small- 
 pox, on the ground that the two diseases are absolutely distinct. 
 
 Risks of Vaccination. 
 
 It must not be forgotten that the introduction into the system of even 
 a mild virus, however carefully performed, is necessarily attended by the 
 production of local inflammation and of febrile illness. If these results 
 did not in some measure follow, the practice would probably fail in its 
 protective influence. As a rule, the inflammation and illness are of a 
 trifling character .; in exceptional cases, however, they may exhibit more 
 severity, and, as certain facts submitted to us in evidence have shown, 
 there are cases, though these are rare, where a general eruption may 
 follow vaccination. 
 
 In order to determine how far the risk of erysipelas is a necessary 
 incident of vaccination, what is the extent of that risk, and how it may 
 best be avoided, it is necessary to consider the various circumstances 
 which may occasion erysipelas and allied diseases in the case of vaccinated 
 children. It is established that lymph contains organisms, and may 
 contain those which under certain circumstances would be productive of 
 erysipelas. It is therefore possible that some contagious material (the 
 specific virus of erysipelas, for instance,) may be conveyed at the time of 
 vaccination, owing either to its presence in the lymph employed, or to its 
 being conveyed by the vaccinator himself, or by those with whom the 
 child comes in contact at the time of vaccination. We believe that the 
 cases in which the virus is conveyed at the time of vaccination are rare. 
 It has, however, in some instances been clearly established, the immediate 
 occurrence of erysipelas in several co-vaccinees making it practically 
 certain that some virus was conveyed at the time of the operation. In 
 some instances where this has been the case, and there is every reason for 
 believing that the contagion was conveyed through the medium of the 
 lymph, it is nevertheless in evidence that the vaccinifer did not display 
 anything more than a slightly inflamed arm. The scrupulous avoidance 
 of inflamed arms in vaccinifers will do much to reduce the risk of 
 conveying erysipelas in the act of vaccination (a risk which, as we have 
 
REPORT OF THE ROYAL VACCINATION < MM MISSION. 685 
 
 seen, has been proved to be a very slight one), but it is possible it would 
 not wholly remove it. 
 
 We have dwelt upon features presented by the cases of erysipelas and 
 various forms of septic disease which have followed vaccination, be.- 
 they suggest precautions which may be adopted to lessen, if not to 
 prevent, such evils in the future. If, for example, vaccination were 
 performed at the patient's home instead of at a public vaccination \ 
 the chance of disease being contracted at the time of vaccination would 
 be to some extent diminished : and the same may be said of the inspection 
 of the vaccinated person which takes place eight days after the operation. 
 On these points we shall have some remarks and recommendations to 
 make at a later stage of our report. 
 
 A study of the cases which have been made the subject of careful 
 examination and report points to the conclusion that an exercise of 
 greater care would largely diminish the risk, already small, of erysipelas- 
 contagion and blood-poisoning. 
 
 Although it may be confidently hoped that by additional care on the 
 part both of vaccinators and parents, the number of inflamed arms and 
 of cases of erysipelas may be reduced to very few, yet it is not to be 
 expected that such occurrences will be wholly prevented. A vaccination 
 wound is, like one from any other cause, so long as it exists, a source of 
 some risk. 
 
 The use of calf-lymph, though it may be supposed to be more free 
 from the risk of conveying erysipelas, does not appear to prevent inflamed 
 arms. Some witnesses have indeed supposed that it is attended with 
 more risk of inflammation than the employment of that taken from the 
 human subject. This opinion has not, however, been corroborated by 
 some of those of widest experience. 
 
 Nothing has produced so deep an impression hostile to vaccination as 
 the apprehension that syphilis may be communicated by it. It was at 
 one time doubted whether syphilis could result, and it was even confi- 
 dently asserted that it could not. The fact that this was possible had 
 been fully established, and was generally acknowledged by the medical 
 profession, before we commenced our inquiries. 
 
 The very close resemblance in certain very rare cases of the results 
 of vaccination, whether with calf-lymph or humanised lymph, to those 
 attributed to syphilis (a resemblance so close that it has caused in a few 
 cases a difference of opinion whether the disease was syphilis or vaccinia) 
 has led to the expression by Dr. Creighton of the opinion that there is 
 some essential relationship between the two diseases. This, however, 
 is a point of speculative, almost it might be said of transcendental 
 pathology, upon which for practical purposes it is useless to enter. It 
 must be sufficient to remark that, whatever may be the relationship 
 alluded to, it exists, if it exists at all, equally between small-pox and 
 syphilis as between vaccination and syphilis. For all practical purposes 
 variola and vaccinia are both wholly distinct from syphilis, and their 
 differences are, with the rarest exceptions, easily recognised. They are 
 alike in being attended by affections of the skin and mucous membranes, 
 and exceptionally by disease of the bones, eyes, and other p.-irt> : but in all 
 
686 SUPPLEMENTARY APPENDIX. 
 
 these it is a question of resemblance and not of identity with which we 
 have to deal. 
 
 Only a few items of the evidence produced before us appear to require 
 special notice : among these, the most prominent is what has been known 
 as the " Leeds case," upon which we have heard the evidence of Mr. 
 Ward, Mr. Littlewood and Dr. Barrs. The witnesses named regarded it as 
 a case of syphilis, conveyed by vaccination, but all of them admitted that 
 the course of events was most unusual. We have carefully investigated 
 this case, and notwithstanding the opinion formed by the witnesses, there 
 appears good reason to doubt whether it was one of syphilis. The case was 
 made the subject of careful inquiry by Dr. Barlow on our behalf, who 
 shared the doubt we have expressed. The view taken by the medical 
 inspector of the Local Government Board who in the first instance in- 
 vestigated the case was that it was a case of hereditary syphilis. It seems 
 certain, however, that the parents of the child whose death was in question 
 were not in any way affected with syphilis. The vaccinifer also appeared 
 to be free from any taint of that disease, and its family history confirmed 
 this view. The co-vaccinees from the same lymph also exhibited no trace 
 of syphilis. These facts of themselves make out a strong case against 
 that having been the nature of the disease. Coupled with the fact that 
 it could not have been communicated by the vaccinator himself, they seem 
 to render it practically impossible that syphilis was the cause of death. 
 If the symptoms exhibited had in all respects corresponded with those 
 which are known to characterise syphilis, the proper inference might have 
 been that there was some error in ascertaining the facts of the case. But 
 it is beyond question that the course of events was very different in some 
 respects from that experienced in undoubted cases of syphilis, and we 
 think the true conclusion is that it was not a case of that disease. It may 
 probably be classed with a few others as examples of gangrene and blood- 
 poisoning, the direct result of vaccination, which are not to be explained 
 by supposing the introduction of any syphilitic or other poison. Fortu- 
 nately, such cases are extremely rare so much so that the witnesses 
 concerned knew of no case precisely parallel. 
 
 The evidence offered to us would lead to the belief that, whilst with 
 ordinary care the risk of communication of syphilis in the practice of arm- 
 to-arm vaccination can for the most part be avoided, no degree of caution 
 can confer an absolute security. The rejection as vaccinifers of young 
 infants, say below four months of age (in whom congenital syphilis may 
 be as yet undeclared), and of adults (in whom the disease may possibly 
 have been recently acquired) are precautions which would probably shut 
 out almost the whole of the risk. The outbreaks of syphilis in connection 
 with vaccination which have been mentioned to the Commission (all of 
 which had been previously published) have occurred chiefly in arm-to-arm 
 vaccination amongst soldiers, or from the use as vaccinifers of young 
 infants the offspring of parents whose history was not known to the 
 vaccinator. It must, however, be admitted that neither the examination 
 of the vaccinifer if taken alone, and without a knowledge also of the 
 parents, nor the most scrupulous avoidance of any visible admixture of 
 blood with the lymph, are in themselves, however valuable, sufficient 
 
REPORT OF THE ROYAL \A<< INATIoN OOMMI8SH (1*7 
 
 al.x.lutcly to exclude risk. The evidence given by Dr. Husband, of the 
 Vaccine Institution of Edinburgh, established the fact that all lymph, 
 however pellucid, does really contain blood cells. Absolute freedom from 
 risk of syphilis can be had only when calf- lymph is used ; though where 
 the antecedents of the vaccinifer are fully ascertained, and due care is 
 used, the risk may for practical purposes be regarded as absent. 
 
 It i> obvious that the employment of calf-lymph only would wholly 
 exclude the risks as regards both syphilis and leprosy. Respecting the 
 latter disease, however, there appears to be reason to doubt whether any 
 risk exists, and at any rate it does not concern the British population. 
 Even in leprosy districts the employment of English human lymph would 
 be, so far as leprosy is concerned, as safe as that from the calf. 
 
 There can be no doubt that vaccination ought to be postponed when 
 erysipelas, scarlet fever, measles, or chicken-pox are prevalent in the 
 neighbourhood of the child's residence, or, if the child is not to be 
 vaccinated at home, either there or near the place of vaccination. Here 
 again there would be a gain if the home was more often the place of 
 vaccination. 
 
 It would, in our opinion, be an advantage if the postponement of 
 vaccination were expressly permitted, not only on account of the state of 
 the child, but of -its surroundings and any other conditions rendering the 
 operation at the time undesirable. If more discretion in this respect 
 were possessed and exercised, we think untoward results would become 
 even rarer than they are. 
 
 We are quite alive to the objections which may be urged against a 
 prolongation of the period within which vaccination must be performed. 
 It will naturally be said that a number of children, who otherwise would 
 be protected against small-pox, would be left without that protection, 
 and would thus be liable to suffer from the disease themselves, and be a 
 source of danger to others. It must be remembered, however, that so 
 long as children cannot walk, the risk of their contracting contagion is 
 less than if they were able to move freely about and mix with other 
 people, and that, for the same reason, the risk of their communicating 
 contagion to others is less. We cannot trace in the statistics relating to 
 Scotland any grounds for believing that the later compulsory vaccination 
 age which prevails in that country as compared with England has affected, 
 to any substantial extent, the general small-pox mortality of Scotland, 
 though no doubt it may have led to some deaths among children under 
 six months of age which otherwise would not have taken place. 
 
 We have already shown how satisfactory a position Germany has 
 occupied in relation to small-pox since the year 1874. The age of com- 
 pulsion in that country is the end of the next calendar year after birth. 
 It is true that re- vaccination has been there made compulsory as well as 
 primary vaccination : but we think the experience of Germany is not 
 without its bearing on the question we are now considering. Wherever 
 the line is drawn, whether at three months or six months, it will always 
 leave a class of unvaccinated persons. The age to be fixed is a question 
 of policy into which many considerations must enter. If an extension 
 of the age within which vaccination was required rendered its untoward 
 
688 SUPPLEMENTARY APPENDIX. 
 
 incidents fewer in number, and diminished hostility to the operation, it 
 may be that on the whole it would promote the cause of vaccination, and 
 secure, as its result, that the number of vaccinated persons would be 
 greater than at present. 
 
 Means, other than Vaccination, for diminishing the Prevalence of Small-pox ; 
 and how far such means could be relied on in place of Vaccination. 
 
 Another question upon which we are asked to report is, what means, 
 other than vaccination, can be used for diminishing the prevalence of 
 small-pox ; and how far such means could be relied on in place of 
 vaccination. 
 
 The means, other than the inoculation of small-pox or cow-pox, which 
 have been referred to by witnesses as being capable of diminishing the 
 prevalence of small-pox, are such means as have been employed against 
 infectious diseases generally ; they may be summarised as (1) Measures 
 directed against infection, e.g., prompt notification, isolation of the 
 infected, disinfection, etc. ; (2) Measures calculated to promote the public 
 health, the prevention of overcrowding in dwellings or on areas, 
 cleanliness, the removal of definite insanitary conditions, etc. 
 
 The principle underlying the practice of isolation with its accompany- 
 ing machinery is obviously the very opposite of that which recommended 
 the practice of inoculation ; it aims at exclusion of the disease, whereas 
 inoculation aimed at universal acceptance by artificially ; ' sowing " or 
 " buying " the disease. Except in regard to the plague, our knowledge 
 and practice of measures of isolation and quarantine against epidemics is 
 of relatively recent growth. As the result of increased knowledge of the 
 mode of propagation of infectious diseases, of greater sanitary activity,, 
 and under the stimulus of legislation, organised effort, more or less 
 thorough, is now, in this as in other countries, directed against the spread 
 of dangerous infectious diseases. Side by side with a vaccination system, 
 means of isolation, etc., have been successfully employed to check the 
 spread of small-pox. They have also been sometimes so employed in 
 recent years in places where vaccination has fallen into disuse. 
 
 It will be well to commence with a brief statement of the growth of 
 our knowledge on the subject of isolation as a means of dealing with 
 infectious or contagious diseases. We have already adverted to the fact 
 that small-pox is highly contagious, and that contagion from those 
 suffering from it is the means by which the disease is propagated. 
 
 Although reference to infection appears in some of the Arabian writers^ 
 the contagiousness of small-pox attracted little attention in this country 
 and in western Europe until the eighteenth century. Sydenham (1624-89) r 
 though he refers to the contagiousness of srnall-pox, did not dwell upon 
 the matter, and did not regard it as so important an element in the spread 
 of the disease as some peculiar constitution of the atmosphere to which 
 he attributed epidemics. Boerhaave was the first, at the commencement 
 of the eighteenth century, distinctly to formulate the now generally 
 accepted doctrine that small-pox arises only from contagion. 
 
 In 1720, Mead drew up an elaborate system of notification, isolation,, 
 disinfection, etc., in view of a threatened invasion of the plague ; but no 
 
REPORT OF THE ROYAL VACCINATION COMMISSION 689 
 
 atu-mpt to deal with small-pox in a similar fashion appears to have l-,-n 
 made until the last quarter of the eighteenth century. This was in all 
 probability largely due to the adoption of inoculation as the recognised 
 defence against small-pox, and the acceptance of Sydenham's doctrine 
 of epidemic causation may have exercised an influence in the same 
 direction. 
 
 Xo writer appears to have suggested methods of isolation, disinfection, 
 etc., against small-pox prior to 1 763. In that year Rast of Lyons publ i 
 his " Reflections on Inoculation and Small-pox, and upon the means which 
 might be taken to deliver Europe from that malady.'' He maintained 
 ( 1 ) That small-pox was not a necessary and inevitable malady ; (2) That it 
 arose only from contagion : (3) That it resembled plague in most of its 
 features. His conclusion was expressed in these terms : u I say, that to 
 "deliver Europe from small-pox we must act upon principles directly 
 opposed to inoculation : far from multiplying the contagion, we must 
 keep it away by taking the same precautions and employing the same 
 " measures against that malady as have proved so successful against leprosy 
 and the plague." 
 
 The earliest account of the practical employment of such means is 
 from Rhode Island, U.S.A. Haygarth, on the authority of Drs. Moffat and 
 Waterhouse, states that for many years prior to 177* small-pox had been 
 successfully prevented from becoming epidemic there by regulations for 
 isolation of the infected on a neighbouring small island specially used for 
 that purpose, and for quarantining infected vessels, destruction of infected 
 clothing, etc. Moreover, inoculation was discouraged at Rhode Island, 
 and those who wished to be inoculated had to go to some place away from 
 the Island, and were not to return until there was no danger of their 
 infecting others. 
 
 A passage in Dimsdale's work on Inoculation, published in 1781, shows 
 that in some towns of England pest-houses were beginning to be used for 
 small-pox. In 1784 Haygarth, of Chester, published his ki Inquiry how to 
 prevent the Small-pox," and in 1793 '' A Sketch of a Plan to exterminate 
 the Small-pox from Great Britain." 
 
 The great epidemic of small- pox at Chester in 1774, to which allusion 
 has already been made, was the occasion of Haygarth's first attempts at 
 organised dealing with epidemics of small-pox with a view to preven- 
 tion. In his " Inquiry '' he combated Sydenham's doctrine that epidemics 
 are due to some occult condition of the atmosphere, and argued that 
 small-pox was always spread by infection only. He further maintained 
 that the variolous poison could be carried as an infection for a little 
 distance only through the air. and " consequently that the small-pox may be 
 ' prevented by keeping persons liable to the distemper from approaching 
 within the infectious distance of the variolous poison till it can be 
 "destroyed." These views led him, upon the return of an epidemic in 
 1777. to propose a plan for the prevention of the natural small-pox. an<l 
 in 1778 a society was formed to carry out the plan in Chester. The plan 
 consisted on the one hand of a general inoculation at people's homes at 
 stated intervals, on the ground that the inoculated small-pox was far less 
 fatal or injurious than the natural small-pox, and on the other hand of 
 
 H 
 
690 SUPPLEMENTARY APPENDIX. 
 
 11 Rules of Prevention " based on Haygarth's views of infection. In the 
 report of the Society, called shortly "The Small-pox Society," dated 
 September 1782, it is stated that in the four and a half years of its 
 existence two general inoculations had been held, and that the deaths 
 from small-pox had been greatly lessened. Great difficulties, however, 
 were met with. u A large proportion of the inhabitants " refused inocula- 
 tion, and a large proportion also, " being fearless, or rather desirous, that 
 their children should be infected with the natural small-pox," refused to 
 obey the Rules of Prevention. Hence, though the same report states 
 that the example of Chester had been followed by Liverpool, where 
 " a general inoculation was successfully executed in the autumn of 1781 
 and another in the spring of 1782," and in Leeds, where a general 
 inoculation was held in 1781 and another proposed in 1782, with such 
 success that the Royal College of Physicians in Edinburgh appointed a 
 committee to inquire into " the modes of conducting the general inocula- 
 tions of the poor " thus adopted in these places, the plan met with such 
 difficulties that it was ultimately abandoned. It will be observed that a 
 general inoculation was an essential part of the plan proposed and 
 carried out in 1778-82 ; but, writing in 1784, Haygarth looked forward 
 to being able ultimately to dispense with inoculation, and in the preface 
 to his later edition, published in 1 793, he states more definitely that the 
 adoption of his Rules of Prevention without any general inoculation 
 might exterminate small-pox in some country other than Great Britain. 
 It must be remembered, however, that Haygarth entertained the opinion 
 that the infection of small-pox could not be carried through the air above 
 about half a yard, and that no one could be infected by the clothes of a 
 person visiting a small-pox patient provided that he kept beyond this 
 distance from the patient. It is obvious that if this had been established 
 the control of the disease by isolation would be a much simpler matter 
 than it really is. 
 
 In the Medico-Chirurgical Review for 1796 there appeared an account 
 of a work by Dr. Faust, of Leipsic, entitled " An Essay on the Duty of 
 ; 'Man to separate persons infected with the Small-pox from those in 
 " Health, thereby to effect the extirpation of that disease equally from 
 "' the towns and countries of Europe," in which it was argued that the 
 first person ill in a place is the only source from which all the rest, 
 perhaps hundreds and thousands, become affected, and that if he were 
 put immediately into a situation where he 'could not injure by contact 
 those who had not had the disorder, the spread of the disease would be 
 prevented. 
 
 In the same Review for 1799 appeared an account of establishments 
 for the extirpation of small-pox. The failure of inoculation to attain the 
 desired end is referred to, and legislation is urged to facilitate isolation. 
 It is further stated that in 179G the Prussian College of Physicians re- 
 ported favourably to the King on the project, and that at Halberstadt it 
 had been resolved to establish a house for the purpose. At Cote d'Or in 
 France a similar plan had been tried with success. 
 
 In 1798 Jenner's " Inquiry " was published, and in the early years of this 
 century inoculation began to be discouraged ; for a while the prospects of 
 
RKPOKT OK THE ROYAL VACCI5ATIOB OOMMffiSKMf, 
 
 annihilating small-pox by vaccination appear to have superseded, in the 
 minds of many, the plans of Haygarth and others. Some vaccinators, 
 however, like Willan and Ring, still looked to methods of <iuar;uitine and 
 to national and municipal regulations promoting isolation to exterminate 
 the small-pox. 
 
 It is worthy of notice, too, that Haygarth himself, in a letter quoted 
 by Dr. Cappe of York in a communication to the LH<\,I M.il,,-nl ,id 
 PJiytiwI Jnumnl (vol. iv., p. 42H), dated October 13th, ISIHI. remark.-d. 
 " An introduction of the vaccine still more than of the variolous inocula- 
 tion would effectually promote the great object of my publications." 
 
 Prior to the year 1866 there was no provision made by law for enabling 
 ^unitary authorities to establish hospitals for infectious diseases, and thus 
 to promote the isolation of such cases. The only institutions of that 
 description then existing were the result of private effort. So far as 
 regards small-pox there was, practically speaking, no provision for its 
 treatment by means of isolation. 
 
 The Sanitary Act of 1866 empowered, though it did not compel, 
 local authorities throughout England and Wales, Scotland and Ireland, 
 to provide or to join in providing isolation hospitals for the use of the 
 inhabitants of their districts. There was further legislation on the 
 subject by the Public Health Act, 1875 ; the Public Health (London) Act, 
 1891 : the Public Health (Scotland) Act, 1867 : and the Public Health 
 (Ireland) Act, 1878, into the details of which it is not necessary to enter. 
 The most recent Act relating to the matter is the Isolation Hospitals Act 
 of 1*!>3, which applies to the small towns and rural districts of England 
 and Wales. 
 
 in 
 
 Leicester suffered severely from small-pox in 1*72. ;U6 deaths having 
 been registered as caused by it. Two deaths from that disease occurred 
 in 1873, but no other until 1877, when there were six, and one in the 
 following year. The next year in which small-pox deaths were registered 
 was 1881. There were two in that year, and five and three in the follow- 
 ing years. Xo other death took place until 1892 and 1893, in which 
 years the fatal cases numbered '1 \ . 
 
 Prior to 1875 the vaccination laws were well observed in Leicester. 
 In that year the number of children born who were unaccounted foi 
 only some 4 per cent. Since then there has been, as we have seen, a 
 marked and progressive decline in the number of vaccinations, especially 
 since 1883, until at the present time 80 per cent, of the children born 
 remain unvaccinated. 
 
 The borough hospital for infectious diseases was erected in 1*71-1* 
 outride the town : though within the last few .years houses have been 
 built in proximity to it. It appears to have been with Dr. Crane, the 
 Medical Officer of Health in 1875, that the quarantining the inmat 
 an infected house, in addition to isolating the patient, originated. II I 
 successor, Dr. John- blished it in 1*77 as a regular system. He 
 
 was aided in this, after 1*7'.'. by the notification of infection 
 then rendered compulsory by a private Act which Leicester, anticipating 
 
092 SUPPLEMENTARY APPENDIX. 
 
 most other towns, obtained in that year. Dr. Johnston reported that up 
 to 1884 the spread of small-pox from imported cases had been arrested in 
 20 instances by the means he adopted. 
 
 His successor, Dr. Tomkins, though, like his predecessors, regretting 
 the increasing disuse of vaccination, bore testimony in his annual reports 
 to the efficacy of the measures adopted in Leicester, and expressed his 
 opinion that had such a system been in force at Sheffield in 1887 it would 
 not have suffered in the way it did. 
 
 In 1892 small-pox became prevalent in different parts of England, 
 especially in Lancashire and Yorkshire. Many of the large provincial 
 towns suffered, and Leicester amongst them. There were, in 1892-3, 357 
 cases of small-pox in Leicester, of whom 21, or 5*8, died ; 193 households 
 were invaded, containing 1234 persons. The first importation was by a 
 tramp, whose disease, passing unrecognised, caused infection at a common 
 lodging-house and at the workhouse. Eleven other importations of the 
 disease by tramps occurred in the course of 1892-3. 
 
 Leicester suffered less than many of the other large towns which have 
 been invaded by small-pox during recent years, both in the number of 
 cases and in the number of deaths. In connection with this, however, 
 a point to which we have already called attention must be borne in mind. 
 The disease was remarkably slight there in its fatality, even as regards 
 those who, by reason of their age, could not be affected by the change of 
 practice in relation to vaccination. Dr. Priestley, the Medical Officer of 
 Health, claims, in his report to the Sanitary Committee for 1893, that 
 it was by reason of the energetic methods adopted that the disease had 
 been prevented running riot through the town. His claim may be well 
 founded. At all events, the experience of Leicester affords cogent 
 evidence that the vigilant and prompt application of isolation, etc., even 
 with the defects which were brought to light during the recent epidemic, 
 is a most powerful agent in limiting the spread of small-pox. It is true 
 that the system and appliances which appeared adequate for some years 
 failed to prevent a serious outbreak of small-pox in 1892-3. We think 
 its value was none the less real. 
 
 Stampiiig-out System in London. 
 
 In the Report of the Royal Commission of 1881, already alluded to, 
 suggestions were made with regard to notification and isolation which 
 have since been largely carried into effect. As we have said, it was con- 
 sidered proved that the existing small-pox hospitals had caused a spread 
 of the disease in their neighbourhood. We cannot but think that this 
 may in some measure account for the greatly increased mortality from 
 small-pox in London during the 1871-72 epidemic as compared with the 
 rest of the country. It is true that the statistics relating to England and 
 Wales outside the Metropolis include those of other large towns where the 
 same evil was present ; but it probably did not exist there in so aggravated 
 a form, and the effect may be neutralised by the statistics relating to 
 smaller towns and rural districts with which they are combined. This 
 idea has been suggested to us, as the result of the inquiry, how it has come 
 about that whilst the Metropolis, in the decennium 1867-76, and again 
 
REPORT OF THK KnYAl, VAO INATh'N OOMMI8SIOH, 
 
 down to 1885, compared so unfavourably with the rest of the country, the 
 condition has since that date become so entirely changed ? We think it 
 is impossible to attribute this change to vaccination. There is no n 
 to suppose that the position of the Metropolis in respect to vaccination 
 has, since the year 1885, become superior to the rest of England anl 
 Wales : rather the other way, as the decrease in infantile vaccination has 
 been greater during the last few years than in the rest of England and 
 Wales. The change, therefore, must be due to some other cause. 
 
 The hospitals which, in the opinion of the Commissioners, were 
 propagating the disease in their neighbourhood, were in operation 
 down to July 1882, when their Report was made. In 1877 and 1*78. and 
 again in 1881. small-pox was epidemic in London to a considerable extent. 
 
 We have stated in detail in paragraph 471 * the steps which were taken 
 by the Metropolitan Asylums Board in consequence of the recommenda- 
 tions of the Royal Commission. It will be seen that the intra-urban 
 hospitals still continued in use. and that complaints were made in 1 884 that 
 they were spreading small-pox in their vicinity, although the number in 
 each of them was not allowed to exceed 50. In October 1884 this number 
 was reduced to 25. It was not, however, until 1885 that the system now 
 in operation was inaugurated, and all cases of small-pox were treated in 
 hospital ships. It is impossible not to be struck with the fact that it is 
 since the year 1885 that the Metropolis has presented so satisfactory an 
 aspect as regards small -pox mortality. The facts to which we have been 
 calling attention certainly seem to point to the conclusion that this has 
 been due to a system of isolation, well organised and administered, the 
 beneficial effect of which is no longer neutralised by a spread of the 
 disease from the hospitals in which the isolation is carried out. 
 
 Upon the whole, we think the experience of London affords cogent 
 evidence of the value of a sound system of isolation in checking the 
 spread of small-pox. 
 
 The experience of isolation systems in Australia is interesting and 
 worthy of special notice, because whilst in this country the quarantining 
 of persons who have come in immediate contact with those suffering from 
 small-pox has only been possible with the consent of the persons whom it 
 was proposed to subject to quarantine, in Australia their removal to a 
 place of isolation has been made compulsory. 
 
 Australia, by virtue of its geographical position, and the consequent 
 separation by long sea voyage from infected ports, enjoyed for a long 
 time a sort of natural isolation. Thus, Hirsch, in his *' Historical and 
 Geographical Pathology," vol. i.. pp. i:.:i-l ( 1>*1 ,. n -mark* : 
 
 ' The continent of Australia up to 1 838 had enjoyed an absolute 
 immunity from small-pox : towards the end of that year the disease 
 41 appeared at Sydney, having been imported probably from China : it 
 lasted, however, only a short time, and remained absent from the 
 continent until l*t*. In that year it was introduced into Melbourne \>y 
 " a ship, and again it spread only to a slight extent, and quickly died cut. 
 
 * Final 
 
694 SUPPLEMENTARY APPENDIX. 
 
 " By a rigorous inspection of ships on their arrival, it has been found 
 " possible to prevent subsequent importations, a notable instance of pre- 
 " vention having occurred in 1872. Tasmania has hitherto quite escaped 
 " the disease ; so also has New Zealand, where an importation of it in 
 " 1872 was prevented by strictly isolating a vessel that had arrived with 
 ' ; small-pox on board." 
 
 In New South Wales, Dr. MacLaurin, who has been President of the 
 Board of Health since 1889, informed us that the Government act on the 
 assumption that small-pox is an exotic disease, and that every case must 
 have come from outside the colony, and it is therefore dealt with under 
 a quarantine Act of William IV., originally instituted for dealing with 
 cholera. By an Act passed in 1882, notification of small -pox was made 
 compulsory on medical men and householders under heavy penalties. At 
 Sydney notification of small-pox is followed up by the compulsory 
 removal of the patient and all persons who have been in the house with 
 the patient to the quarantine station at North Head. This station is 670 
 acres in extent, and situated on the peninsula at the mouth of Sydney 
 Harbour, and is seven miles from the Health Office, with which there is 
 telephonic and telegraphic communication. The persons are conveyed to 
 the station by a steamboat comfortably fitted expressly for the purpose, 
 and no difficulty has been experienced in effecting their removal. It was, 
 in Dr. MacLaurin's opinion, by carrying out this practice of isolation and 
 quarantine that " the epidemic of 1881-82 was suppressed," and small-pox 
 " has 'never become epidemic since this plan has been adopted." The 
 persons who have been in the house with the patient are detained 21 days 
 in quarantine from the date of the last possible contagion. Should a case 
 of small-pox arise among them, those who had been in contact with such 
 infected person would be detained for a further period of 21 days, and so 
 on. To facilitate this, the exposed persons are distributed in separate 
 groups within the station. They are allowed to receive letters or parcels, 
 etc., and a telegraph operator is employed, " whose special business it is 
 " to work the telegraph at their request." " Reasonable compensation is 
 " given by the Government for loss ; " and there are heavy penalties under 
 the original Act whereby the quarantine is secured. The station is, accord- 
 ing to Dr. MacLaurin, " a pleasant place to stay in, and everything is done 
 " that can be done to make the people comfortable : they have nothing 
 "whatever to do, and are free from all care, and they can spend the day 
 " pleasantly enough ; but they do not like it." No one, however, raises 
 any objection to the Sydney system : " the people are all very sensible 
 about it." In all Australian towns the same system is carried out as 
 strictly, with the result that there was not a case of small-pox in Australia 
 on February 5th, 1890 ; and Dr. MacLaurin is of opinion that the risk of 
 dying of small-pox in Australia is smaller than in any other part of the 
 world. As regards vaccination : In New South Wales it is very little 
 practised ; there is no compulsory Act ; and though medical opinion is in 
 favour of it, an opinion shared by Dr. MacLaurin, it is not likely that a 
 compulsory Vaccination Act could be passed or would be tolerated. The 
 proportion of young persons in New South Wales who are not vaccinated 
 is accordingly very large ; probably much more than half of those under 
 
REPORT OF Till-: KnYAL \ AC( INATK >.\ COMMISSION, li!).' 
 
 ten years of age are unvaccinated. Although Dr. MacLaurin 
 vaccination and respects it highly, he is satisfied that th, system of isola- 
 tion as supervised by him is perfectly successful. As President of the 
 Board of Health he considered it his business to produce extinction of the 
 disease ; he does not consider vaccination a sufficiently absolute protect i..n 
 for such purpose ; and he is " fully of opinion that the only way in which 
 
 you can bring to an end an outbreak of small-pox, that is to say, bring 
 
 it under control, and not leave it to work itself out, is by notification 
 " and isolation. Of course, in any small community, if you let the di 
 
 " in it will work itself out in time, because all the susceptible people will 
 have had it ; but the only way in which you can absolutely control an 
 li epidemic of small-pox is by a system of notification and isolation." 
 
 Small-pox has never been epidemic in Western Australia. < )nly one 
 case has occurred within the last 31 years, and that was an imported one ; 
 quarantine was carried out, and no infection occurred ; the immunity from 
 the disease is mainly at least due to isolation. Before 187 ( J vaccination 
 was not generally practised a great majority of those born in the colony 
 were unvaccinated : in that year a compulsory Vaccination Act was passed 
 in consequence of Sir H. Ord and Dr. Waylen's representations, and in 
 consequence of reports of small-pox in other colonies, and not on account 
 of the existence of small- pox in Western Australia. 
 
 In Tasmania there was a compulsory vaccination law, but it was found 
 to be inoperative because no one was appointed to conduct the prosecu- 
 tions. and it has now fallen into desuetude. The same system of isolation 
 and quarantine is exercised as in the other Australian colonies. Small-] ><>\ 
 was for the first time introduced into Tasmania in 1887, and although 
 preparations for isolation were inadequate, the disease was soon stamped 
 out. Communication between Launceston and Melbourne was temporarily 
 suspended, and to this precaution the non-invasion of Victoria was 
 attributed. The particulars of this, the first introduction of small-pox 
 into Tasmania during the history of that colony, are to be found in a 
 report to the Central Board of Health by Mr. A. Man It, dated Novem- 
 ber 17tb, 1887. The origin of the outbreak is not clear, but it 
 presumed to have been imported, probably by a ship from China, into 
 Launceston. The earliest case reported to the Local Board of Health 
 was on September 23rd, though it appears that earlier cases had p.-. 
 unnoticed, or had been notified as measles. Thirty-three cases in all 
 occurred, every one of which was traced to direct infection from th< 
 case. By September 27th a temporary hospital had been erected, and 
 thither patients and suspects to the number of 72 were removed. Th 
 case appeared on October 1 .'5th . < Hher persons who had been to the infected 
 houses were isolated in their houses and watched. Only four of th- 
 persons quarantined at the station were attacked. The clothing was 
 burnt, and very thorough disinfection of tin- infected houses was carried 
 out, and the dead were interred in a special cemet 
 
 were communicated with, and quarantine, at first unduly rigid and ti 
 wards relaxed, was practised against ships procet diip_ r from Tasmania. 
 Although vaccination had been nominally compulsory in Tasmania, it 
 estimated that two-fifth* <f the population were unvaccinated. 
 
696 SUPPLEMENTARY APPENDIX. 
 
 Suggested Stamping-out System tit the United Kingdom. 
 
 We have no difficulty in answering the question, what means other 
 than vaccination can be used for diminishing the prevalence of small-pox ? 
 We think that a complete system of notification of the disease, accom- 
 panied by an immediate hospital isolation of the persons attacked, together 
 with a careful supervision, or, if possible, isolation for sixteen days of 
 those who had been in immediate contact with them, could not but be 
 of very high value in diminishing the prevalence of small-pox. It would 
 be necessary, however, to bear constantly in mind, as two conditions of 
 success : first, that no considerable number of small-pox patients should 
 ever be kept together in a hospital situate in a populous neighbourhood : 
 and secondly, that the ambulance arrangement should be organised with 
 scrupulous care. If these conditions were not fulfilled, the effect might 
 be to neutralise or even do more than counteract the benefits otherwise 
 flowing from a scheme of isolation. 
 
 When we turn to the other branch of the inquiry, how far such means 
 could be relied on in the place of vaccination, we find ourselves -involved 
 in questions of a much more complicated nature. We have little or no 
 experience to fall back upon. The experiment has never been tried. 
 The nearest approach to a trial of it has probably been in Australia. But 
 even in the parts of that country to which we have alluded the population 
 has not been entirely unvaccinated, though there has been a large 
 unvaccinated class amongst it. Moreover, in applying the experience of 
 Australia to this country, two things must be borne in mind. In the first 
 place small-pox has only appeared from time to time, introduced from 
 without at one or other of the ports of the country, and the several 
 colonies of which Australia is composed are of great territorial extent, 
 with few large centres of population. In this country small-pox is 
 always present in some part of it. There has not been a single year 
 without several deaths from the disease. Large centres of population 
 are numerous, and the intercourse between them constant. In the several 
 colonies of Australia the number of ports is not great, the vessels which 
 enter them are comparatively speaking not numerous, and the ports from 
 which they arrive are many days' voyage distant ; and there are careful 
 arrangements for quarantining vessels to exclude disease. The shipping 
 which enters English ports is of vast quantity, and passengers are brought 
 in large numbers from the continent of Europe not only daily, but it may 
 almost be said hourly ; the voyage, too, is but brief. The other matter to 
 be remembered is, that part of the Australian system is the compulsory 
 removal to quarantine for 21 days of those who have been in the house 
 with the patient, in addition to the transfer of the patient himself to a 
 hospital. There can be no doubt that such a system, if completely carried 
 out, would be of the highest efficacy. But it is obvious that in this 
 country the practical difficulties of working such a scheme in the large 
 towns would be really insuperable, to say nothing of the difficulty of 
 procuring legislative sanction for it. 
 
KKPURT OF THE ROYAL \ A( ( I N A 1 K -N n EMISSION. 
 
 ,,f /.</,/,/ 
 
 \\\ ran see nothing, then, to warrant the conclusion that in this country 
 vaccination might safely be abandoned, and replaced b\ m of 
 
 isolation. If such a change were made in our method of dealing with 
 small-pox, and that which had been substituted for vaccination proved 
 ineffectual to prevent the spread of the disease (it is not suggested that it 
 could diminish its severity in those attacked), it is impossible to contem- 
 plate the consequences without dismay. 
 
 To avoid misunderstanding, it may be well to repeat that we are very 
 far from underrating the value of a system of isolation. We have already 
 Iwelt upon its importance. But what it can accomplish a^ an auxiliary 
 t<> vaccination is one thing : whether it can be relied on in its stead is quite 
 another thing. 
 
 Even admitting fully the protective effect of vaccination, it does not, 
 in our opinion, diminish the importance of measures of isolation or dis- 
 pense with their necessity. We think that steps should be taken to secure 
 a more general provision for the isolation of small-pox patients than 
 exists at present. We have already called attention to the fact that 
 mischievous results are likely to follow the use as a small-pox hospital of 
 a building situate in a populous place. We think that wherever it is 
 placed it should have sufficient space around it to enable the sanitary 
 authority to add rapidly to the accommodation by the erection of tem- 
 porary buildings. 
 
 Compulsory I'mcixinn <>f I*<ilntinn Hospitals, 
 
 Sanitary authorities are now sometimes reluctant to provide isolation 
 hospital-. We think that, on a petition by a prescribed number of the 
 ratepayers in a sanitary district, the Local Government Board, if satisfied 
 that the hospital accommodation ought to be provided, should have power 
 to make an Order for such provision. 
 
 VV- think that notification of small-pox should everywhere be compul- 
 and, whenever the disease showed a tendency to become epidemic, 
 a notice should be served by the sanitary authority upon all persons in 
 the neighbourhood who would be likely to come within the reach of con- 
 tagion, urging them to submit to vaccination or re-vaccination, as the case 
 might be, if they had not been recently successfully vaccinated or re- 
 vaccinated : and attention should be called to the facilities afforded for 
 their doing so. Attention should also be called to the importance of 
 avoiding contact with persons suffering from the disease, or coming into 
 proximity to them, and of avoiding contact with any person or thing 
 which may have become infected. It is important to notice that, even 
 where vaccination has l>een neglected, there is great readiness to submit 
 to it in the presence of a threatened epidemic : a large number of vacci- 
 nations are then obtained willingly and without opposition. Whenever 
 
698 SUPPLEMENTARY APPENDIX. 
 
 a sanitary authority has received notification of a case of small-pox r 
 we think the fact should be at once communicated to the vaccination 
 authority of the district in which the case of the disease has occurred. 
 
 Regulations an to Tramps, Inmates of Lodging-houses, etc. 
 
 Our attention has been drawn to the circumstance that outbreaks of 
 small-pox have not unfrequently had their origin in the introduction 
 of the disease to common lodging-houses by tramps wandering from place 
 to place. In view of this we make the following recommendations : 
 
 (i.) That common shelters which are not now subject to the law 
 relating to common lodging-houses should be made subject 
 to such law. 
 
 (ii.) That there should be power to the local authority to require 
 medical examination of all persons entering common lodging- 
 houses and casual wards to see if they are suffering from 
 small-pox, and to offer a reward for prompt information of 
 the presence of the disease. 
 
 (iii.) That the local authority should have power to order the keeper 
 of a common lodging-house in which there has been small- 
 pox to refuse fresh admissions for such time as may be 
 required by the authority. 
 
 (iv.) That the local authority should be empowered to require the 
 temporary closing of any common lodging-house in which 
 small-pox has occurred. 
 
 (v.) That the local authority should have power to offer free 
 lodgings to any inmate of a common lodging-house or casual 
 ward who may reasonably be suspected of being liable to 
 convey small-pox. 
 
 (vi.) That the sanitary authority should give notice to all adjoining 
 sanitary authorities of the occurrence of small-pox in 
 common lodging-houses or casual wards. 
 
 (vii.) That where the disease occurs, the Public Vaccinator or the 
 Medical Officer of Health should attend and vaccinate the 
 inmates of such lodging-houses or wards, except such as 
 should be unwilling to submit themselves to the operation. 
 
 Relaxation of the Vaccination Law. 
 
 After careful consideration and much study of the subject, we have 
 arrived at the conclusion that it would conduce to increased vaccination 
 if a scheme could be devised which would preclude the attempt (so often 
 a vain one) to compel those who are honestly opposed to the practice to 
 submit their children to vaccination, and, at the same time, leave the law 
 to operate, as at present, to prevent children remaining unvaccinated 
 owing to the neglect or indifference of the parent. When we speak of 
 an honest opposition to the practice, we intend to confine our remarks to 
 cases in which the objection is to the operation itself, and to exclude 
 
HKI'OKT OF THK KoYAL VACCINATION COMMISSION, 699 
 
 cases in which the objection arises merely from an indisposition to incur 
 the trouble involved. We do not think such a scheme impossible. 
 
 It must of course be a necessary condition of a scheme of this 
 description that it should be such as would prevent an objection to the 
 practice being alleged merely as an excuse to save the trouble connected 
 with the vaccination of the child. We may give the followin. 
 examples of the methods which might l>e adopted. It might be provil-<l 
 that if a parent attended before the local authority and satisfied them 
 that he entertained such an objection, no proceedings should be taken 
 against him. Or, again, a statutory declaration to that effect before any 
 one now authorised to take such declaration, or some other specified 
 official or officials, might be made a bar to proceedings. We do not think 
 it would l>e any real gain to parents who had no conviction that the 
 vaccination of their children was calculated to do mischief, to take either 
 of these steps rather than submit them to the operation. 
 
 It is in England that the point we have been recently discussing is of 
 most practical importance, but if our suggestion were adopted the change 
 should, of course, be made in all parts of the United Kingdom. 
 
 (Signed) HERSCHELL. 
 
 JAMES PAGET. 
 
 CHARLES DALRYMPLE. 
 
 W. (il'YER HUNTER 
 
 EDWIN H. GALSWORTHY 
 
 JOHN S. DUGDALE. 
 
 M. FOSTER. 
 
 JONATHAN HTT< HINSON. 
 
 FREDERICK MEADOWS WHITE. 
 
 SAM. WHITBREAD. 
 
 JOHN A. BRIGHT. 
 
 BIJKI I.\< i:. 
 August 1 Secretary. 
 
 The undersigned do not find themselves able to go so far in recom- 
 mending relaxation of the law as is implied. We think that in all cases- 
 in which a parent or guardian refuses to allow vaccination, the person ><> 
 refusing should be summoned before a magistrate, as at present, and that 
 the only change made should be to permit the magistrate to accept a 
 sworn deposition of conscientious objection. ;m<l to abstain from the 
 infliction of a fine. 
 
 \\ V are also of opinion that, in spite of the difficulties as set forth in 
 paragraph :3:'.* a second varcination at the age of twelve ought to 
 compulsory. 
 
 w. <,! YI:I; m xn:i; 
 
 .JONATHAN 
 
 Of thf Fiiwl K-p -it. 
 
700 SUPPLEMENTARY APPENDIX. 
 
 We the undersigned desire to express our dissent from the proposal to 
 retain in any form compulsory vaccination. 
 
 We cordially concur in the recommendation that conscientious ob- 
 jection to vaccination should be respected. The objection that mere 
 negligence or unwillingness on the part of parents to take trouble might 
 keep many children from being vaccinated would be largely, if not wholly, 
 removed by the adoption of the Scotch system of offering vaccination at 
 the home of the child, and by providing for medical treatment of any 
 untoward results which may arise. 
 
 We therefore think that the modified form of compulsion recommended 
 by our colleagues is unnecessary, and that in practice it could not be 
 carried out. 
 
 The hostility which compulsion has evoked in the past toward the 
 practice of vaccination is fully acknowledged in the Report. In our 
 opinion the retention of compulsion in any form will in the future cause 
 irritation and hostility of the same kind. 
 
 The right of the parent on grounds of conscience to refuse vaccination 
 for his child being conceded, and the offer of vaccination under improved 
 conditions being made at the home of the child, it would in our opinion be 
 best to leave the parent free to accept or reject this offer. 
 
 SAM. WHITBREAD. 
 JOHN A. BRIGHT. 
 W. J. COLLINS. 
 J. ALLANSON PICTON. 
 
 Note. Dr. Collins and Mr. Picton */V/w the abuce note of reservation, 
 though they have not signed the Report. A statement of their grounds of 
 dissentfrom the Report will be found in the form of an Appendix (05 pages) 
 to the Report. They make the following recommendations : 
 
 In accordance with the sub-head No. 2 of the reference to the 
 Commission, we would suggest the following as the means other than 
 vaccination which should be employed for protection of a community 
 from small-pox : 
 
 1. Prompt notification of any illness suspected to be small-pox. 
 
 Improved instruction in the diagnosis of small-pox. 
 
 2. A hospital, suitably isolated, of adequate accommodation, in per- 
 
 manent readiness, and capable of extension if required. No 
 other disease to be treated at the same time in the same 
 place. 
 
 3. A vigilant sanitary staff ready to deal promptly with first cases, 
 
 and if necessary to make a house-to-house inspection. The 
 medical officer of health to receive such remuneration as to 
 render him independent of private practice. 
 
 4. Prompt removal to hospital by special ambulance of all cases 
 
 which cannot be properly isolated at home. Telephonic com- 
 munication between Health Office and hospital. 
 
REPORT OF THE BOTAL \ \( < |.\.\TI< .\ COMMISSION, 701 
 
 5. Destruction of infected clothing and bedding, and thorough dis- 
 infection of room or house immediately after removal of the 
 patient. 
 
 0. Daily observation (including, where possible, taking the tempera- 
 ture and inspection for rash) of all persons who have l>een in 
 close contact with the patient during his illness ; such super- 
 vision to be carried out either in quarantine stations (away 
 from the hospital) or at their own homes. 
 
 7. Closure of schools on the occasion of the occurrence of small-pox 
 among the scholars or teachers. 
 
 *. Hospitals and quarantine stations to be comfortable and attractive, 
 and so administered as to secure the confidence of the public. 
 Hospital treatment to be free to all classes, and compensation 
 to be paid to those detained or otherwise inconvenienced in the 
 public interest, at the public expense. 
 
 ',). Tramps entering casual wards to be medically inspected, their 
 clothing to be disinfected, and bath provided. The measures 
 for detection and isolation of small-pox in common lodging- 
 houses suggested in section 507 of the Report to be carried 
 out. 
 
 10. International notification of the presence of small-i>ox, and 
 
 special vigilance at seaports in communication with infected 
 places, after the plan adopted in the case of cholera. 
 
 1 1 . Attention to general sanitation prevention of overcrowding. 
 
 abundant water supply, and frequent removal of refuse. 
 
 They conclude as follows : 
 
 We believe the methods of isolation of the infected, disinfection, and 
 the observance of strict cleanliness, are both more successful and more 
 legitimate methods for the State to encourage. They have the advantage 
 of applying the preventive only where it is required : and they do not 
 necessitate an operation upon the person of every healthy individual. 
 
 We therefore recommend that the law be amended by the repeal of 
 the compulsory clauses of the Vaccination Acts. But in consideration 
 of the prevalent belief in the value of vaccination as a prophylactic for 
 an indefinite period, we suggest that in other respects the law should be 
 left as it is. subject, however, to such modifications as are recommended 
 for the diminution of attendant risks. The precedent established in the 
 case of the abolition of compulsory church rates might be followed with 
 advantage. In that case all machinery for laying and collecting the 
 was left intact though the power of enforcement was taken away. The 
 effect of our recommendation, if adopted, would be that vaccination 
 would continue to be provided as at present for those who desire to avail 
 themselves of it, but efforts to secure vaccination would be limited to 
 moral influence in a word, the whole country would be in the position of 
 those unions in which the guardians have abandoned compulsion. 
 
 The grounds on which we object to the enforcement of vaccination 
 by penalties necessarily lead us to object to any nu -tlio<! of indirect 
 
702 SUPPLEMENTARY APPENDIX. 
 
 .compulsion. We regard as both inexpedient and unjust exclusion from 
 any branch of the public service because of the refusal to submit to 
 vaccination or re- vaccination. The injustice is perhaps most severely felt 
 in the case of candidates for employment as pupil-teachers in public 
 elementary schools. There are now districts in which, owing to the 
 general opposition to vaccination, scarcely a girl or boy can be found who 
 is legally eligible, and candidates have to be brought in at great incon- 
 venience from surrounding districts. The existence of an exceptional 
 <;ase or cases in which such rejected candidates have at some time after- 
 wards taken small-pox is in our view no justification for the continuation 
 of this grievance. Statistics furnished to the Commission prove that 
 large numbers of vaccinated or re-vaccinated persons have taken the 
 disease ; and we are not aware of any evidence to show that vaccinated 
 pupil-teachers have any special immunity. If our recommendations were 
 carried out, the danger of contagion would be greatly diminished in 
 schools, as elsewhere. 
 
 On the whole, then, while there is much in the report of our colleagues 
 from which we dissent, and we have accordingly abstained with reluctance 
 from adding our signatures to theirs, we are at one with them in holding 
 that it is unwise to attempt to enforce vaccination on those who regard it 
 as useless and dangerous. We. however, go further, and agree with our 
 colleagues, Mr. Whitbread and Mr. Bright, that it would be simpler and 
 more logical to abolish compulsory vaccination altogether. 
 
INDEX 
 
 
INDEX. 
 
 Abba's condenser, 74 
 Aberration, chromatic, 67 
 
 spherical, 67 
 Abscess, 173 
 
 Achorion Schonleinii, 584 
 Actinomycosis, 413 447 
 
 bovis, 434 
 
 cattle to cattle, 444 
 
 cultivation of, 436 
 
 hominis, 431 
 
 in cattle, 429 
 
 in man, 426 
 
 transmission of, from man to 
 
 lower animals, 443 
 Agar-agar, 625 
 Air, compressed. 24 
 
 examination of, 140 
 
 Hesse's method, 141 
 
 Koch's 141 
 
 Petri's 142 
 
 Pouchet's .. 143 
 
 - Sedgwick's 143 
 Aitken's tubes, 129, 630 
 Alexines, 57 
 Alum carmine, 616 
 Amoeba coli. 610 
 Anaerobes, classified, 494 
 
 cultivation <>f, 130133 
 
 Aniline water. ''.17 
 Animals Order, 1>7-. l.V, 
 Anthrax. 12. 1*3. 11)1 21> 
 
 -bacillus of. 1U l'J7 
 
 in horses. 1' (I 7 
 
 in swine. L 
 
 origin and spread of, 198 
 
 preventive inoculation, 209 
 
 measures in, 1 
 
 Anthrax, stamping out system, 210 
 Antiseptics, 30 
 Antitoxins, 56, 57 
 
 diphtheria, 5862 
 
 septic infectious, 63 
 
 tetanus, 62 
 
 typhoid, 64 
 Arthrospores, 19 
 Artificial fluids as media, 120 
 Ascococcus, 14 
 
 - Billrothii, 498 
 
 citreus, 498 
 Ascomycetes, 584 
 Asiatic cholera, 360 
 Aspergillus albus, 588 
 
 clavatus, 588 
 
 flavescens, 588 
 
 flavus, 586 
 
 fumigatus, 586 
 glaucus, 586 
 
 nidulans, 588 
 
 niger, 587 
 
 ochraceus, 588 
 
 repens, 586 
 
 subfuscus, 588 
 
 B 
 
 Babes' incubator, 633 
 Bacillus, 13, 488 
 
 acidiformans, 498 
 
 acidi lactici, 498 
 
 aerogenes 
 : capsulatus. 
 
 aerophilu- 
 
 albns, i 
 anaerobiescen- 
 
 put, 
 
 
706 
 
 INDEX. 
 
 Bacillus allantoides, 499 
 - allii, 499 
 
 alvei, 470 
 
 amylobacter, 502 
 
 amylozyma, 500 
 
 anaerobicus liquefaciens, 500 
 
 anthracis, 192 
 
 aquatilis, 500 
 
 fluorescens, 500 
 
 graveolens, 500 
 
 sulcatus, SCO 
 arborescens, 500 
 
 argenteo-liquefaciens, 501 
 
 phosphorescens, (J01 
 
 aurantiacus, 501 
 
 aureus, 501 
 
 beroliniensis indicus, 502 
 brassicae, 502 
 
 brevis, 502 
 
 brunneus, 502 
 
 buccalis fortuitus, 502 
 
 maxiinus, 502 
 
 minutus, 502 
 
 butyricus, 502, 503 
 
 cadaveris, 503 
 
 canalis capsulatus, 504 
 
 parvus, 504 
 
 candicans, 504 
 
 capsulatus, 504 
 
 mucosus, 504 
 
 - suis, 504 
 
 carabiformis, 504 
 carnicolor, 504 
 
 carotarum, 505 
 cavicida, 505 
 
 chromo-aromaticus, 505 
 Havaniensis, 505 
 
 circulans, 505 
 
 citreus cadaveris, 505 
 
 cloacae, 505 
 
 cceruleus, 505 
 
 : coli communis, 344 
 si mills, 506 
 
 constrictus, 506 
 
 coprogenes f cetidus, 506 
 
 parvus, 506 
 
 crassus aromaticus, 506 
 
 sputigenus, 506 
 
 cuniculicida, 228 
 
 cuticularis, 506 
 
 albus, 506 
 cyaneo-fuscus, 507 
 
 Bacillus cyaneo-phosphorescens, 507 
 
 cyanogenus, 507, 508 
 
 cystiformis, 508 
 
 delicatulus, 508 
 
 dentalis viridans, 508 
 
 dentriticus, 508 
 
 devorans, 508 
 
 diffusus, 508 
 
 diphtherias, 332336 
 
 columbarum, 336 
 
 vitulorum, 509 
 dysodes, 509 
 
 endocarditidis capsulatus, 509' 
 griseus, 509 
 
 enteritidis, 372 
 
 epidermidis, 509 
 
 erysipelatis suis, 356 
 
 erythrosporus, 509 
 
 figurans, 510, 511 
 
 filiformis, 511 
 
 Havaniensis, 511 
 
 flavescens, 511 
 flavocoriaceus, 511 
 
 fluorescens aureus, 511 
 
 liquefaciens, 511 
 
 longus, 512 
 
 minutissimus, 512 
 
 nivalis, 512 
 
 non-liquefaciens, 512 
 
 putidus, 512 
 
 tenuis, 512 
 
 foetidus, 513 
 ozaenae, 513 
 
 fulvus, 513 
 
 fuscus, 513 
 
 limbatus, 513 
 
 gallinarum, 513 
 
 gasoformans, 513 
 
 glaucus, 513 
 
 gliscrogenus, 513 
 
 gracilis, 514 
 
 granulosus, 514 
 
 graveolens, 514 
 guttatus, 514 
 
 hsemorrhagic septica3mia, 231 
 
 halophilus, 514 
 
 Hansenii, 514 
 
 Havaniensis, 505 
 
 liquefaciens, 514 
 
 helvolus, 515 
 
 heminecrobiophilus. 515 
 
 hepaticus fortuitus, 515 
 
INDEX. 
 
 707 
 
 Bacillus Hessii, 515 
 
 hyacinth! septicus, ol .1 
 
 - hyalinus, 515 
 
 hydrophilus fuscus, 515 
 
 ianthinus, 516 
 
 implexus, 516 
 
 in acne contagiosa in horses, 516 
 in cancer, 516 
 
 incanus, 518 
 
 in cholera in ducks, 516 
 
 in choleraic diarrhoea, 516 
 
 indicus, 518 
 indigogenus, 519 
 
 in diphtheritic disease of calves, 
 
 516 
 - in erythema nodosum, 517 
 
 in infectious disease of bees, 471 
 
 inflatus, 519 
 
 in fowl enteritis, 230 
 
 in gangrene, 517 
 
 in grouse disease, 517 
 
 in hog cholera, 517 
 
 in infantile diarrhoea, 517 
 
 in intestinal diphtheria in rabbits, 
 
 517 
 
 in jequirity infusion, 517 
 
 in measles, 517 
 
 in noma, 517 
 
 in potato rot, 517 
 
 in purpura haemorrhagica, 517 
 
 in putrid bronchitis, 518 
 
 in "red-cod," 51 8 
 
 in rhinoscleroma, 518 
 
 in saliva, 518 
 
 in swine fever, 346-352 
 
 . erysipelas, 354 
 
 measles, 354 
 
 inunctus, 519 
 
 invisibilis, 518 
 
 in whooping cough, "> 1 ^ 
 
 iridescens, 519 
 
 iris, 519 
 
 lactericeus, 520 
 
 lactis aerogenes, 519 
 
 albus, 520 
 
 erythrogenes, 520 
 
 pituitosi, 520 
 
 leporis lethalis, 520 
 
 leprze, 407 
 
 leptosporus, 520 
 
 limbatus acidi lactici, 520 
 
 limosus, 520 
 
 Bacillus liodermos. .v_>i 
 
 liquefaciens. :,i'l 
 
 communis, 521 
 
 magnus, 521 
 
 parvus. ."iL'l 
 
 liquidus, :.L' 1 
 
 - litoralis, 521 
 
 - lividus, 524 
 luteus, 524 
 
 - maidis, 52 I 
 mallei, 452 
 
 megatherium, r>L' I 
 
 rnembranaceus amethystinus, 525 
 
 meningitidis purulentse, 525 
 
 meseutericus fuse IK. .",!'."> 
 
 ruber, ,V_T, 
 
 vulgatus, 526 
 
 multipediculus, .VJii 
 
 muscoides, 522 
 
 mycoides, 522 
 
 roseus, 522 
 
 neapolitanus, 522 
 necrophorus, :,>:', 
 
 nitrificans, 523 
 nodosus parvus, 523 
 
 nubilus, f>L':; 
 
 ochraceus, 523 
 
 oedematis aerobicus, B 
 
 maligni, 220 
 
 of Belfanti and Pascarola, 523 
 
 of Colomiatti, 526 
 
 - of Fulles, 526 
 
 of Guillebeau, 526 
 
 of Letzerich. ."iL'; 
 
 of Martinez, 526 
 
 - of Nocard. r>2; 
 
 of Okada, 52(5 
 
 of quarter-evil, 217 
 
 of Rot! 
 
 of Sattler. 526 
 
 of ScbaftYi 
 of Sclieurlci 
 
 !,.:. HN 
 
 of septicaemia <>f biifTaloes, L 
 
 of guinea-pigs, 2L' I 
 
 - of mio . 
 
 of swine plagu< 
 
 of Tommasoli. .VJ7 
 
 of Utpadel. 
 
 of Winograd-'r.v. v_'7 
 
 ophthalmia I 1 .'" 
 
 ovatus minutissimus, 
 
708 
 
 INDEX. 
 
 Bacillus oxytocus perniciosus, 527 
 - pestifer, 528 
 
 phosphorescens gelidus, 528 
 
 indicus, 528 
 
 indigenus, 528 
 
 plicatus, 528 
 
 pneumonias crouposse, 233 
 
 pneumosepticus, 528 
 
 polypiformis, 528 
 
 prodigiosus, 528 
 
 proteus fluorescens, 529 
 
 pseudo-diphtheriticus, 529 
 
 tuberculosis, 529 
 
 pulpae pyogenes, 529 
 
 punctatus, 529 
 putrificus coli, 529 
 
 pyocyaneus, 529 
 
 pyogenes fcetidus, 530 
 soli, 530 
 
 radiatus, 530 
 
 aquatilis, 530 
 
 ramosus, 531 
 
 reticularis, 531 
 
 rhinoscleroma, 411 
 
 rosaceus metalloides, 531 
 rubefaciens, 531 
 
 rubellus, 531 
 
 ruber, 531 
 
 rubescens. 532 
 rubidus, 532 
 
 sanguinis typhi, 532 
 
 saprogenes, 532 
 
 scissus, 532 
 
 septicaemias hasmorrhagicae, 231 
 septicus, 532 
 
 acuminatus, 533 
 
 agrigenus, 533 
 
 keratomolaciae, 533 
 
 ulceris gangrasnosi, 533 
 
 vesicae. 533 
 
 sessilis, 533 
 
 smaragdino-phosphorescens, 533 
 
 smaragdinus foetidus, 534 
 solidus, 534 
 
 spiniferus, 534 
 
 spinosus, 534 
 
 stolonatus, 534 
 stoloniferus, 534 
 
 striatus albus, 534 
 
 flavus, 534 
 
 subflavus, 535 
 
 subtilis, 535 
 
 Bacillus subtilis similans, 536 
 
 sulfureus, 536 
 
 superficialis, 537 
 
 syphilis, 410 
 
 tenuis sputigenus, 537 
 
 termo, 537 
 
 tetani, 457 
 
 thelassophilus, 537 
 
 thermophilus, 537 
 
 tremelloides, 537 
 
 tuberculosis, 378 
 gallinarum, 402 
 
 tumescens, 537 
 
 typhi abdominalis, 342 346 
 murium, 359 
 
 ubiquitus, 537 
 
 - ulna, 538 
 vacuolosis, 538 
 
 varicosus conjunctivas, 538 
 
 venenosus, 538 
 . brevis, 538 
 
 invisibilis, 538 
 
 liquefaciens, 539 
 
 ventriculi, 539 
 
 vermicularis, 539 
 
 vermiculosus, 539 
 
 violaceus, 539 
 
 laurentius, 539 
 
 virescens, 539 
 
 viridis pallescens, 539 
 
 viscosus, 540 
 
 Zurnianus, 540 
 
 Bacteriacese, 480 
 
 Bacteria, chemical action on, 25 
 - products of, 39 
 
 composition of, 11 
 
 distribution of, 29 
 
 form of, 13 
 
 growth of, 23, 25 
 
 in cattle plague, 296 
 
 in diphtheria of pigeons, 336 
 
 in distemper, 358 
 
 in foot and mouth disease, 300 
 
 in horse-pox, 312 
 
 - in liquids, 83 
 
 in louping-ill, 463 
 
 in measles, 283 
 
 in pus, 176 
 
 in rabies, 460 
 
 in scarlet fever, 262 
 
 in sheep-pox, 298 
 
 in small-pox, 285 
 
INDEX. 
 
 Bacteria in vaccine lymph, 324326 
 
 in yellow fever, 2H 
 
 microscopic examination of, 83 
 nitrifying, i'7 
 
 pathogen! 
 
 saprogenic, 26 
 
 stained, >." 
 
 unstained, 84 
 
 Bacterium aerogenes, 540 
 
 brunneum, ."!' 
 
 fusiforme, 5 In 
 
 gingivae pyogenes, 540 
 hyacinth!, 5 In 
 
 hydrosulfureum ponticum, 540 
 
 litoreum, 540 
 
 luteum, 540 
 
 merismopedioides, 541 
 navicula, 541 
 
 photometricum, 541 
 
 synxanthum, 541 
 termo. 541 
 
 tholoeideum. 51 I 
 
 urese, 541 
 
 violaceum, 541 
 - Zopfii, 542 
 
 Beggiatoa alba, 5 1 2 
 
 roseo-perticina, 512 
 
 Bibliography, 639 
 
 Bilious fever, 183 
 
 Biondi's stain, 6 1 5 
 
 Bismarck-brown, 617 
 
 Blepharadenitis. 1M 
 
 Blood-serum, 113. 114, 119 
 
 Borax carmine, 617 
 
 Botrytis Bas>i;ui 
 
 Bread paste, 118 
 
 Broncho-pneumonia, 183 
 
 Broth, 118 
 
 Bunge's method of staining flagella, 
 
 Camera lucida, 621 
 Cancer bodies, ',l<> 
 Caoutchouc caps. 
 Caterpillars, disease of, 
 Cattle plague, 293- 2 
 
 tuberculosis of. 
 
 Cerebro-spinal menin.tr it K 1 - I 
 Chemical action on L. 
 
 disinfectair 
 
 products of bacteria, 
 
 Chemiotaxi-. 51 
 Chi.inyphr CUI-KM 
 Cholera 
 
 bacteria <.f. :;:i 
 
 comma bacilli of, :;r,l :;r,7 
 
 diarrhoea from meat-poisoning. 
 
 871 
 
 diarrhoea in fowls, : 
 nostra>. 
 
 protective inoculation. 
 
 I'toina'iii'--. 1 1 
 
 spirillum of, 361 
 Chromogenic bacteria. L'."< 
 Cladothriceae, -1><> 
 Cladothrix, 47'J 
 
 dichotoina, 51.; 
 
 Forsteri, 5 ! 1 
 
 intricata, 544 
 
 invulneral.ilis. 5H 
 Classification, 47.-.. 1-7. 1-2,481 
 Clostridium butyricum, 544 
 
 -fcetidum. 515 
 Coccaceze. I-M 
 Cocci, 485 
 Coccidia. tin'.i 
 Comma bacilli. 361- 
 Cover-glass preparatio: 
 Cow-pox, 274282, 312324, 326 
 
 Crenot'nrix Kiihniana. 5J5 
 
 D 
 
 Dacryory>tK 1M 
 Damp chan 
 
 D' Arson val\s in<:ubat..r. ;:!! 
 I>avaincV s-ptic:umia. 
 J'r Ha: KfcioO, 1-1 
 
 Decalcifying, 9.",, ;15 
 
 Defensive j 
 
 Deneke's comma bacillus. 
 
 l>iarrhoea, cholfrai--. :;71 
 Diphti 182 
 
 .11, 5s 62 
 
 in mil; 
 
 in'i'- 
 
 LJ'ifH.-: . 88 
 
 1'ti.in;.! 
 
 DIploCO ci;>. 1 | 
 
 albicans amp ii~. 517 
 
 tarlissimn- 
 
710 
 
 INDEX. 
 
 Diplococcus citreus conglomeratus. 
 547 
 
 liquefaciens, 547 
 coryzae, 547 
 
 flavus liquefaciens tardus, 547 
 
 fluorescens foetidus, 547 
 
 intercellularis meningitidis. 548 
 
 - luteus, 548 
 
 - pneumoniae, 233238 
 
 - of horses, 548 
 
 roseus, 548 
 
 subflavus, 548 
 
 Distemper in dogs, 358 
 Drop-cultures, 120 
 Duck cholera, 230 
 Dysentery, 373 
 
 B 
 
 Ebner's solution, 615 
 Egyptian ophthalmia, 190 
 Ehrlich's staining method, 89 
 Electricity, 24, 127 
 Embedding, 93, 615 
 Empusa muscae. 581 
 
 radicans, 582 
 
 Endospores, 18 
 Enteric, 41, 340 
 Enzymes, 48 
 Eosin, 89, 617 
 
 Epidemic disease of deer and boars, 
 227 
 of ferrets, 358 
 
 - of mice, 358 
 Erysipelas, 185189 
 Esmarch's roll-cultures, 113 
 Experiments on animals, 134, 394 
 
 F 
 
 Farrant's solution, 621 
 Favus, 584 
 
 Filter, hot water, 624 
 Flacherie, 472 
 Flagella, 17, 90 
 Fliigge's classification, 481 
 Foot rot, 464 
 Form of bacteria, 13 
 Foul-brood, 469 
 Fowl cholera, 228 
 enteritis, 230 
 
 scab, 586 
 
 Friedlander's bacillus, 233 
 Fuchsine, 617 
 
 G 
 
 Gangrene. 182 
 
 Gas chamber, 127 
 
 Gases produced by bacteria, 24 
 
 Gentian violet stain, 617 
 
 Giant cells, 376 
 
 Gibbes' solution, 618 
 
 Glanders, 451 
 
 . bacillus of, 452 
 
 mallein in, 454 
 
 ptomaine, 48 
 
 Glycerine agar, 104 
 
 Glycerine gelatine, 615 
 
 Gonococcus of Neisser, 189 
 
 Gonorrhoea, 189 
 
 Gram's method, 88, 89, 97, 618 
 
 Grease, 303 
 
 Grouse disease, 230 
 
 H 
 
 Haematococcus bovis, 548 
 Hasmatomonas carassii, 605 
 
 cobitis, 603 
 Hasmatoxylin, 618 
 Hgematozoa, 589, 593, 599, 603 
 Hardening, 93, 615 
 Helicobacterium aerogenes, 549 
 Herpetomonas Lewisi, 599 
 Hessert's stain, 92 
 Historical introduction, 1 
 Horse-pox, 303, 312 
 Hot air and steam, 36, 623 
 Hot water filter, 624 
 Hueppe's classification, 482 
 
 inspissator, 630 
 
 Hydrophobia, 459 
 Hyphomycetes, 581 
 Hypodermii, 581 
 
 Illumination, 75 
 
 Immersion system, 69 
 
 Immunity, 50 57 
 
 Impression preparations, 92 
 
 Incubators, 631 
 
 Infectious pleuro-pneumonia, 239, 247 
 
 Influenza, 247249 
 
 Inspissators, 629 
 
 Involution forms, 15 
 
 Iodine, 618 
 
 Isolation of micro-organisms, 139 
 
 Israel's case, 628 
 
INDEX. 
 
 711 
 
 Klein's micrococcus of pneumonia, 238 
 Kleinenberg's solution, 616 
 Koch's comma bacillus, 3t>l 
 
 postulates, 9 
 
 serum steriliser. \V2\i 
 
 steam steriliser, t^i- 
 
 Leprosy, 406 
 
 bacillus of. H'7 
 
 stamping out system, 409 
 
 I^eptothrix, 480 
 
 buccalis, 549 
 
 gigantea, 549 
 
 Leuconostoc, 549 
 Leukaemia, 184 
 
 Light, effect of, on bacteria, 24 
 Listers flasks, 128, 630 
 Lithium carmine solution, 61 H 
 Loffler's solution, 619 
 
 stain, 88, 90 
 Louping ill, 462 
 
 bacillus of, 463 
 
 Lutesch's stain, 91 
 
 M 
 
 Madura disease, 447 
 Magenta solution, 618 
 Malaria, 589 
 Malignant oedema, 220 
 Mallein, 454 
 
 Measles, bacteria in. 1-.!, L^:< 
 Media, 99 
 Merismopedia, 14 
 Methyl violet, 619 
 Methylene blue, 618, r>l!> 
 Micrococcus acidi lactici, 550 
 
 liquefacien-. ",< 
 
 aerogenes, 5."- 1 
 
 agilis, 551 
 
 citreus, 551 
 albus liquefaciens. 551 
 
 amylivorus, .">.">! 
 
 aquatilis. 551 
 
 invisibilis, .V>1 
 
 auranti-au>, .".."> 1 
 
 botyogenus. ~>~>- 
 
 candi 
 
 candidus. 
 
 ca: 
 
 cerasinus siccus. 
 
 Micrococcus cereus albus, 1 T - 
 
 llavus. 1> 
 
 cinnabareus, 552 
 
 citreus, 552 
 
 concentri'-u-. 
 
 cremoides, 553 
 
 crepusculum, 553 
 
 - cumulatus tenuis, 553 
 
 endocarditidis rugatu-. 
 
 fervidosus, 553 
 
 - Finlayensis. 668 
 
 - flavus desidiMis, ">:>:* 
 
 liquefaciens, .V, 1 
 
 tardigradus, 554 
 
 fcetidus. ."..", i 
 
 Freudenreichi, .V> I 
 
 fuscus, 554 
 
 gingivae pyogenes, 554 
 
 gonorrhoeae, 190 
 
 Havaniensis, 5.V> 
 
 in abscess in rabbits. .V)i; 
 
 in Biskra button, ?>'>'> 
 
 in gangrenous mastitis in sheep, 
 
 555 
 
 in infectious pleuro-pneumonia, 
 
 555 
 
 in influenza, 5."i.~i 
 
 in pemphigus, 555-6 
 
 - in pneumonia. !':;;. !>:;>. .V.; 
 
 - in py^mia in rabbit-. 
 
 -- in septicaemia in rabbi ts. 
 
 in syphilis, 557 
 
 in trachoma, lint 
 
 lactis viscosu-. 
 
 luteus, 557 
 
 melitensis (Malta fever). B 
 ochroleucus, :>~> 7 
 
 plumosu>. 
 
 pneumonia} crouposae, '2M 
 pyogenes tenuis, 557 
 
 rosaceus, 558 
 
 rosettaceus, 558 
 
 salivarus septicus. 
 
 stellatus, 588 
 
 tetragenus. 558 
 
 mobilis vvntricnli. ."."- 
 
 subl' 
 
 versitil,-. 
 
 ureae liquefaciens. 559 
 
 vereicolor. 
 
 violaceu- 
 viticulosus, 
 
712 
 
 INDEX. 
 
 Micrometer, 80 
 Microscope, 65, 612 
 Microsporon furfur, 586 
 Microtomes, 94, 614 
 Miescher's tubes, 609 
 Mildew, 581 
 Milk, 120 
 
 scarlatina, 265, 282 
 
 tubercular, 291 
 
 Miquel's bulbs, 129 
 Moist chambers and cells, 121 
 Moitessier's gas regulator, 634 
 Monads, 601 
 Monas Okenii, 560 
 Vinosa, 560 
 
 Warmingii, 560 
 
 Mouse favus, 586 
 Movements of bacteria, 15, 24 
 Mucors, 583 
 Miiller's fluid, 616 
 Myconostoc gregarium, 560 
 Myco-protein, 11 
 
 Nature of soil for bacteria, 23 
 
 Neelsen's solution, 89, 98, 619 
 
 Nicols' stain, 92 
 
 Nitromonas of Winogradsky, 560 
 
 Nutrient agar, 103 
 
 Nutrient gelatine, 100 
 
 O 
 
 Oidium albicans, 586 
 
 lactis, 584 
 
 - Tuckeri, 584 
 Oil immersion, 70 
 Orseille, 619 
 
 Parietti's method, 148 
 Pasteur's apparatus, 130 
 Pathogenic bacteria, 27 
 Pebrine, 471 
 Pediococcus acidi lactici, 560 
 
 cerevisias, 560 
 
 Penicillium glaucum. 588 
 Peronospora infestans, 582 
 Petri's method exam, of air, 142 
 Pfeiffer bodies, 610 
 Phagocytosis, 27, 55 
 Photogenic bacteria, 25 
 Photography of bacteria, 150, 168 
 
 Photo-micrographic apparatus, 621 
 
 Phycomycetes, 582 
 
 Phylaxins, 57 
 
 Picric acid, 619 
 
 Picro-carmine, 619 
 
 Picro-lithixim carmine, 620 
 
 Pilobolus, 583 
 
 Plague, bacillus of, 250, 252 
 
 Plate cultivations, 106 
 
 Platinum needles, 83, 616 
 
 Pleuro-pneumonia, 239 
 
 Pneumo bacillus liquefaciens bo vis, 
 
 242, 560 
 Pneumonia, 233238 
 
 ptomaines, 48 
 
 Potash solution, 620 
 Potato medium, 115 117 
 Pouchet's aeroscope, 143 
 Preservation of preparations, 93 
 Protective inoculation, 49 
 
 in anthrax, 209 
 
 in cholera, 369 
 in rabies, 460 
 
 in sheep-pox, 298 
 
 in small-pox, 285, 293 
 
 i in swine erysipelas, 336 
 
 Proteus capsulatus septicus, 560 
 
 hominis capsulatus, 224 
 
 in gangrene of the lung, 560 
 
 microsepticus, 560 
 mirabilis, 561 
 
 septicus, 561 
 
 sulfureus, 561 
 
 vulgaris, 561 
 
 Zenkeri, 562 
 
 Pseudo-diphtheritic bacillus, 335 
 Pseudo-diplococcus pneumonias, 562 
 Psorosperms, 609 
 
 Ptomaines, 39 
 Pus, bacteria in, 176 
 Putrefactive bacteria, 26, 28 
 Pyaemia, 175 
 
 Quarter-evil, 217 
 
 Q 
 
 K 
 
 Babies, 459462 
 Kag-pickers' septicaemia, 224 
 Refraction, 65 
 Relapsing fever, 257 
 Reproduction, 18 
 
INDEX. 
 
 Respiration of bacteria, 22 
 Rhabdomonas rosea, 562 
 Rhinoscleroma. 1 1 1 
 Ringworm, 585 
 
 S 
 Saccharomyces acidi lactici, 580 
 
 albicans, E 
 
 anomalus, 579 
 
 apiculatus, 578 
 
 aquifolii, 580 
 
 cerevisiae, 577 
 
 conglomeratus, 578 
 
 ellipsoideus. ."J77 
 
 exiguus, 578 
 
 glutinus, 579 
 
 Hansenii, 580 
 
 ilicis, 580 
 
 Jorgensenii, 578 
 
 Ludwigii, 580 
 
 Marxianus, 580 
 
 membranaefaciens, 580 
 
 minor, 580 
 
 niger, 580 
 
 ! pastorianus, 578 
 
 pyriformis, 579 
 
 rosaceus, 580 
 
 sphaericus, 579 
 
 Safranine, 620 
 iSapraimia, 175 
 Saprogenic bacteria, 26 
 Saprolegin;, 
 
 8aprophytic bacteria, 27 
 Sarcinae, 487 
 Sarcinaalb; 
 
 aurantiaca, 562 
 
 Candida, 563 
 flava, 563 
 
 byalina. 563 
 intestinal!-. 
 
 - litoralis, 563 
 
 lutea, 563 
 
 mobilis, 563 
 
 pulmonum, 563 
 
 Reitenbachii. ."",;} 
 
 - rose;. 
 urina?. 
 
 ventriculi, 564 
 
 Scarlet fever. 'jr. I 
 
 Schizomycett ~, 177 
 
 Schlosing's membrane regulator, 632 
 
 Sclavo's stain, '.! 
 
 Septicaemia, 175 
 
 of calves, L'L'7 
 
 of Davainc, L'L' I 
 
 of guinea-pigs, UL' 1 
 
 of mice, ! 
 of rabbits, 228 
 
 of rag-pickers, L'L' I 
 Serum therapy, 56 
 Sheep-pox, i".i7 
 Small-pox, 182, 284-293, 326 
 Smut, 581 
 
 Soil, examination of, 144 
 
 Sozins, 57 
 
 Sphaerotilus natans, 564 
 
 Spirilla, I:'.', 
 
 Spirillum amyliferum, 564 
 
 anserum, 564 
 
 attenuatum, 564 
 
 aureum, 564 
 
 cholerae Asiaticae, 361 
 
 choleroides, 565 
 concentricum, 565 
 
 dentium, 565 
 
 - Finkler and Prior, 258 
 flavescens, 565 
 
 flavum, 565 
 Giinther, 566 
 
 leucomelaneuin, 565 
 
 linguae, 565 
 
 marinum, 565 
 
 Metchnikovi, 373 
 
 Miller, 566 
 
 nasale, 566 
 
 Neisser, 566 
 
 Obermeieri, L>.> 
 
 plicatile, 466 
 
 Renon, 566 
 rosaceum, 566 
 
 Rosenbergii. 566 
 
 rubrum, 569 
 rufum, 566 
 
 rugula, 567 
 sanguineum. 
 
 saprophiles, 567 
 
 serpen- 
 
 Smith, 566 
 
 sputigenum, 568 
 
 tenue, 568 
 
 - tyrogenum, 568 
 undula, 568 
 
 Volut: 
 
714 
 
 INDEX. 
 
 Spirillum Weibel, 566* 
 Spirochoetas, 596 
 Spiromonas Cohnii, 569 
 
 volubilis, 569 
 Spontaneous generation, 3 
 Spores, 18 
 Staining spores, 90 
 Staphylococcus, 14 
 
 cereus albus, 178, 324 
 - flavus, 178 
 
 pyogenes albus, 178 
 
 aureus, 176, 324 
 
 citreus, 178 
 
 pyosepticus, 569 
 
 salivarius pyogenes, 569 
 
 viridis flavescens, 569 
 Sternberg's bulbs, 128, 630 
 Streptococcus, 14 
 
 acidi lactici, 569 
 
 albus, 569 
 
 bombycis, 472 
 
 brevis, 569 
 
 cadaveris, 570 
 
 coli gracilis, 570 
 
 conglomerates, 570 
 
 flavus desidens, 570 
 
 giganteus urethras, 570 
 
 Havaniensis, 570 
 
 in contagious mammitis in cows, 
 
 570 
 
 in progressive necrosis in mice 
 
 571 
 - in strangles, 571 
 
 liquefaciens, 571 
 
 mirabilis, 571 
 
 of Bonome, 571 
 
 of erysipelas, 185, 189 
 of Manneberg, 571 
 
 perniciosus psittacorum, 572 
 pyogenes, 178-184 
 
 radiatus, 572 
 septicus, 572 
 
 liquefaciens, 572 
 
 vermiformis, 572 
 Streptothrix, 496 
 
 actinomycotica, 431 
 albus, 572 
 
 asteroides, 573 
 
 aurantiaca, 573 
 
 carnea, 573 
 
 chromogena, 573 
 farcinica, 573 
 
 Streptothrix Forsteri, 573 
 
 Hoffmanni, 573 
 
 liquefaciens, 573 
 
 Madurse, 449 
 musculorum suis, 573 
 
 odorifera, 573 
 
 violacea, 573 
 
 Substage condenser, 74 
 Suppuration, 48 
 Surgical fever, 182 
 Surra, 593 
 
 Swine erysipelas, 354 
 fever, 46, 227 
 
 measles, 354 
 
 Syphilis, 183, 410 
 
 Tarichium megaspermum, 582 
 Temperature for bacteria, 24 
 Test-tube cultivations, 105 
 Tetanus, 457 
 
 ptomaine, 41 
 
 Thermo-regulators, 635 
 Tilletia caries, 581 
 Toxalbumins, 39 
 Trachoma, 412 
 Trenkmann's stain, 91 
 Trichomonas, 607 
 Tricophyton tonsurans, 585 
 Tripod levelling apparatus, 627 
 Tuberculosis, 43, 375 
 
 - and meat, 397 
 
 and public health, 391 
 antitoxin, 64 
 
 - bacillus of, 378 
 
 - giant cells in, 376 
 
 - in animals, 389, 394, 399, 400 
 
 - in man, 387 
 Typhoid, 340 
 Typhus, 259 
 
 U 
 
 Ulcerative endocarditis, 183 
 Jndulina ranarum, 607 
 Uredo, 581 
 
 Urine as a medium, 120 
 Urobacillus Duclauxi, 573 
 Freudenreichi, 574 
 
 - Maddoxi, 574 
 
 Pasteuri, 574 
 
 Schutzenbergi, 574 
 
INDEX. 715 
 
 Urocystis occulta, 581 Weigert's stain, 
 
 fstilago carbo, ">M t gelatine, 104 
 
 V X 
 
 Van Ermengen's stain, 91 Xylol. <JH'> 
 
 Vegetable infusions as media, 120 
 
 \Vsuvin. 620 Y 
 
 Vibrio rugula, 574 j Yeast>. ',:: 
 
 ! YeUow fever, 182, 259 
 W 
 
 Warmstage, 123. 1 'J \ 
 
 Water bath, r.i! ! Zooglea, 13 
 
 examination of, 145 Xoj.fs classification, 480 
 
 Z 
 

 
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