AGRICULTURE LIBRARY OF THE UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN no. 338 - 355 Return or rerww * Ubrary Material* The MMmum Fee for MCh Lost Book to $50.00. The person charging this material is responsible for its return to the library from which it was withdrawn on or before the Latest Date stamped below. Thefl. mutilation, and und+rtWng of book* arc reasons for discipli- nary action and may rasutt in dismissal from the University To ranaw cat Talaphorm Center, 333 8400 UNIVERSITY OF ILLINOIS UBRARY AT URBANA-CHAMPAK3N IHJ605 1999 LI61 0-1096 UNIVERSITY OF ILLINOIS Agricultural Experiment Station BULLETIN No. 343 STUDIES ON PORCINE INFECTIOUS ABORTION BY ROBERT GRAHAM, I. B. BOUGHTON, AND E. A. TUXXICLIFF URBAXA, ILLINOIS, MARCH, 1930 CONTENTS PAGE INTRODUCTION 179 REVIEW OF LITERATURE 183 Possible Relation of Swine Abortion to Undulant Fever 184 PORCINE ABORTION TRACEABLE TO DIFFERENT CAUSES 187 Infectious Type 187 Preventive Measures 188 SPREAD OF INFECTIOUS PORCINE ABORTION 189 Infected Animals May Farrow Normally 189 INFECTIVE CHARACTER OF BRUCELLA TRAUM 191 Antigenic Value of Live and Dead Cultures 193 Infected Boars Potential Spreaders 194 Relation of Brucella Bang and Brucella Traum 195 Normally Farrowing Sows as Carriers of Brucella Traum 199 Location of Brucella Traum in Infected Animals 201 Monthly Agglutination Tests of Vaccinated and Unvaccinated Pigs 215 SUMMARY AND CONCLUSIONS 235 LITERATURE CITED . . 239 Illustration on Cover. Each of the sows shown in the picture on the cover of this bulletin aborted one or more times and repeatedly gave positive agglutination tests of Bru- cella Traum. The boar of the herd, like the sows, proved infected. There are no clinical symptoms that enable the owner or veterinarian to diagnose this disease. While abor- tion often occurs in infected sows, many carry their litters for the full term. In breeding herds where the disease is sus- pected, the agglutination test is recommended. STUDIES ON PORCINE INFECTIOUS ABORTION' BT ROBEHT GRAHAM, I. B. BOUGHTON, AND E. A. TUNNICLIFF" Sporadic outbreaks of abortion in swine have occurred in Illinois for many years. The first important loss from this disease came to the attention of the Illinois Experiment Station in 1917, tho cultural and animal inoculation tests of the aborted fetuses failed to estab- lish conclusively the nature of the disease. In this outbreak a para- typhoid infection was encountered, which apparently was secondary to intestinal manifestations in the sows. Three years later (1920) abor- tions in sows were reported in several Illinois herds. Aborted ma- terials from one large herd at this time yielded evidence of a specific abortion infection which resembled the bovine abortion bacillus. Re- ports of aborting sows in other herds prompted a preliminary survey of the disease on different farms to determine the extent of the in- fectious type of the malady. A study of different sporadic outbreaks of swine abortion in Illi- nois suggested that the specific infection, tho an important factor, was not the exclusive cause of abortion in all herds. In small herds aborting sows of common breeding were generally fattened and sold, making it often impossible to obtain satisfactory material for in- vestigational work. Aborting sows in purebred herds, however, were not so promptly marketed. When valuable breeding sows aborted, an opportunity was presented to make bacteriologic studies of the aborted fetuses, fetal membranes, and vaginal discharges. At the same time samples of colostra and blood from aborting sows were tested for the presence of specific abortion agglutinins. Following the isolation of Brucella-like organisms from aborted materials in 1920 scarcely a farrowing season, during the past nine years, has passed without the infectious type of swine abortion coming "A popular discussion of the prevention and control of infectious abortion in swine is contained in Circular 271 of this Station, revised June, 1927. b RoBHT GRAHAM, Chief in Animal Pathology and Hygiene; I. B. BOUGHTON and E. A. TUNNICLJFF, formerly Associates. The bacteriologic findings and blood tests in different aborting herds as well as in breeding animals at time of slaughter have confirmed the occurrence of a specific type of abortion in swine and also abortions due to other factors. A herd that yielded positive (bacteriologic and serologic) evidence of abortion in 1920 was found to harbor the infection in 1929, suggesting the chronicity of the disease. In this herd abortion had subsided and had not been observed for five years until three sows aborted and one boar that suffered from orchitis proved infected at autopsy. 179 180 BULLETIN No. 343 [March, to the attention of the authors. In the spring of 1929 more than the usual number of sporadic outbreaks of swine abortion were reported by veterinarians and swine breeders. Upon examination, some of the important losses proved to be associated with the swine abortion organism. The recognition of the specific type of abortion in swine in 1920 prompted an experimental study of the disease in guinea pigs, gilts, and heifers. Guinea pigs inoculated subcutaneously developed lesions FIG. 1. PORCINE ABORTION IN GUINEA PIG A healthy guinea pig that was fed Brucella Traum from aborted guinea-pig fetuses. The causative organism was re- gained in pure cultures. involving the lymphatics, liver, and spleen that were more progressive than lesions induced by bovine strains of Brucella. Abortion in preg- nant guinea pigs invariably followed subcutaneous injection of cul- tures. Porcine strains encountered in different Illinois herds have proved consistently pathogenic for guinea pigs. This character is still regarded as an aid in distinguishing the porcine and bovine types of Brucella. All porcine strains isolated by direct culture as well as by guinea-pig inoculation have grown in open plates or tubes on plain nutrient agar without lowered oxygen tension or an increase of carbon dioxid in the atmosphere. Artificial exposure of gilts and sows by feeding, by subcutaneous and intravenous inoculation, and by intravaginal installation of cul- tures did not in all cases cause pregnant animals to abort, but without exception the agglutination titre became distinctly positive. The ex- perimental abortifacient character of the porcine strains, however, has been established by exposing healthy gilts, but all artificially exposed pregnant gilts in the Illinois experiments did not abort. 1930] STUDIES ON PORCINE INFECTIOUS ABORTION- A heifer injected intravenously with a culture of the porcine strain of the Brucella organism aborted and later died of septic metritis. Another heifer that was allowed to come in contact with positively reacting gilts and sows for a period of six weeks failed to reveal any evidence of abortion infection as judged by her breeding record and repeated agglutination tests. On the other hand, two of three heifers kept in the same lot with five positively reacting sows during the far- rowing period showed suspicious agglu- tination reactions three months later. In an effort to obtain information regarding the prevalence of the disease in swine, as judged thru the presence of agglutinins in the blood sera, 1,011 blood samples from old sows slaugh- tered at Chicago were tested in dilu- tions of 1 to 50 and 1 to 100. Of the group 5.6 percent were positive, while of 975 gilts 4.41 percent gave positive reactions. The percentage of positives in 1,034 barrows was 3.38. Evidence to support the contention that the porcine type of the disease may become established in cows was suggested in the results of injecting three heifers subcutaneously with por- cine cultures and later regaining the organism from the udder of one of the inoculated animals. The possibility of the porcine strain entering the udder thru natural channels is also suggested in the results of repeated examinations of the milk from eight naturally in- fected cows. One of the cultures iso- lated from the milk reveals some of the characteristics of the porcine type. The presence of specific Brucella agglutinins has been detected in human patients showing febrile symptoms, as well as in others with an indefinite or obscure history of the disease. While the evidence points to the pathogenic significance of the porcine Brucella in man, Fie. 2. BRUCELLA TRAUM The swine strain of Brucella grows luxuriantly on agar in air. Lowered oxygen tension is not advantageous to its growth. 182 BULLETIN No. 343 [March, there are as yet many unanswered questions regarding the swine and human infections. It is the purpose of this bulletin to report investigations conducted on the infectious type of swine abortion with particular reference to FIG. 3. RAPID AGGLUTINATION TEST The presence of abortion in swine can be recognized by testing the blood of each animal. Negative and positive results are shown in the above illus- tration. From left to right .02, .01, and .005 of the blood serum of the animal to be tested are mixed with .1 cc. of the porcine abortion organism. The anti- gen control without blood serum is at the extreme right. In the left-hand column is the identification of the blood samples, i.e., "1," "2," "3," and "4." Samples 1 and 2, showing clumping of bacterial suspension, with the antigen con- trol unchanged, are positive, or reactors, to the test; while 3 and 4 in the various serum dilutions are unagglutinated and therefore negative. The last sample, labeled "P. S.," is known positive serum. the location of the causative factor in the bodies of both naturally and artificially infected animals, and to record the preliminary results obtained from vaccines as a possible preventive measure. In view of the fact that wide differences of opinion exist with respect to the correct nomenclature for the Brucella organisms, the 1930] STUDIES ON POBCINE INFECTIOUS ABORTION 183 authors will follow the practice of the Illinois Undulant Fever Com- mittee 23 * until more evidence has been presented, using "Brucella Traum" for the porcine type and "Brucella Bang" for the bovine type. FIG. 4. STANDARD TUBE AGGLUTINATION TEST By the use of this test, as well as the rapid agglutination test, sows or boars harboring infectious abortion can be detected. The two cloudy tubes at the left are noninfected or healthy, while the eight tubes at the right show the mix- ture of blood serum and the bacterial suspension of the abortion organism agglutinated or clumped in varying degrees. Agglutination or clumping of the bacterial suspension by the serum of the animal indicates the presence of the infection in the animal. There is no other disease in swine that will produce the agglutinins for the abortion organism; therefore a positive result in the rapid or standard tube test is definite evidence that the animal is or has been infected. In the experiments discussed in this bulletin the standard test was used in the initial investigations and later supplemented by the rapid method. At the present time the rapid test is used in applying the agglutination test, supplemented in doubtful specimens by the standard method. REVIEW OF LITERATURE As early as 1914 Traum, 30 * of the Federal Bureau of Animal In- dustry, reported the isolation of the genus Brucella from the stomach contents, liver, and kidney of an aborted pig fetus. Many cows aborted in the herd from which Traum's swine specimens were re- ceived. The bacteriologic findings of Traura have been confirmed by several investigators, including Good and Smith 9 * in Kentucky (1917), Hagan 13 * (1917) of New York, Hayes and Traum 16 * (1920)* of Cali- fornia, Doyle and Spray 7 * (1920) of Indiana, Schlegel 25 * of Germany (1920), Connaway, Durant, and Newman'* of Missouri (1921). The latter investigators report that abortion in swine was experimentally produced by feeding abortion cultures isolated from cattle, but Hud- dleson 18 * (1921) of Michigan failed to infect swine, as judged by abortion, by feeding cow's milk which contained abortion organisms. Hadley and Beach 12 * (1922) in Wisconsin reported the occurrence of abortion in swine traceable to an organism of the Brucella genus 184 BULLETIN No. 343 [March, but biologically different from the bovine type. Artificial exposure of pregnant gilts by these authors showed that the average period of time from exposure to the act of abortion was 23.2 days. They found that young pigs were highly resistant, and that porcine abortion vaccines in a limited number of animals seemed to reduce the inci- dence of abortion. No evidence of infection carriers was found in their vaccinated animals. Traum, Schroeder and Cotton, and others recognized cultural and pathogenic characters in the porcine strains which differentiated them from the bovine strains. The carrier feature of the disease in swine was first studied by Hayes 15 * of California (1922). In his investi- gations the porcine strain was isolated from the udder of an artifici- ally infected sow three months after farrowing. The testicles of 17 artificially infected male pigs were examined with negative results. Weeter 32 * (1923), of the University of Chicago, in studying 435 sow blood samples collected on the killing floors of Chicago packing establishments found 9 percent positive to the agglutination test in a dilution of 1 to 100. Of 190 blood specimens from barrows, 5 or 2.6 percent were positive in a dilution of 1 to 200. From the nongravid uteri of 3 sows in 389 examined, the porcine strain was isolated. McAlpine and Slanetz 24 * (1927) of Connecticut studied the differ- ence between the porcine and bovine strains, and concluded thru meta- bolic studies that the porcine type is different from the bovine type, and that the strains isolated from man suffering from undulant fever display the characters of the porcine group. These authors also recog- nize that cows may become infected with the porcine variety should they come into close contact with infected swine. Such a conjecture suggests that cattle may be in more danger of contracting the porcine type than swine are of contracting the bovine type. In view of these observations the suggestion that the porcine type might possess ad- vantages in the form of a vaccine for cattle as immunizing agents seems unwarranted until more evidence is obtained regarding the patho- genic properties of the porcine type for cattle. Possible Relation of Swine Abortion to Undulant Fever A resume of the reported investigations on infectious porcine abor- tion over a period of several years suggests a rather widespread dis- tribution of a chronic infection which is recognized as a potentially dangerous disease to the swine industry, while more recent investiga- tions of Huddleson 19 * (1926), Carpenter and Merriam 4 * (1926), and Carpenter 2 * (1927), suggest the possible significance of the porcine type to public health. 1930] STUDIES ON POHCINE INFECTIOUS ABORTION 185 Evans first surmised the possible relation of the abortion organism of cattle to undulant fever in man, and classified the meletensis- abortus group under the genus Brucella, after Bruce, who discovered the cause of Malta (undulant) fever in man. The genus Brucella of Evans recognized the goat, cow, and swine strains. At the present time the consensus of opinion seems to be that the porcine type is a possible source of undulant fever in man in the middlewestern states. It has been shown by various investigators, such as Hayes and Traum, 16 * Cotton, 6 * Schroeder and Cotton, 26 * Hadley and Beach, 12 * Hayes, 15 * and Smith, 28 * that the porcine strains are more pathogenic for experimental animals than the bovine strains. Whether this is true in the case of Brucella types isolated from undu- lant fever patients remains to be definitely proved by future studies, but the evidence at this time tends to support this contention. In Illinois, Hull 22 * has observed that cases of undulant fever in man have occurred in localities that have the largest swine population. All the strains of Brucella obtained from human sources by Mo- Alpine and Slanetz 24 * of the Connecticut (Storrs) Station, belonged in the porcine group. These workers found that the biological charac- ters of the swine and human strains were more nearly related than those of the bovine and human. For instance, the human and porcine strains utilized from 4 percent to 18 percent glucose, increased the non-protein nitrogen, and produced very little free ammonia, while those of bovine origin used very little if any glucose, decreased the non-protein nitrogen, and produced large amounts of free ammonia. They also noted that porcine and human strains were inhibited or unaffected by carbon dioxid, while the bovine strains required car- bon dioxid for initial growth. Huddleson, 20 * in studying the characters of the Brucella genus, found that the porcine group was inhibited by methyl violet in a dilution of 1 to 100,000, by basic fuchsin in a dilution of 50,000, but not by thionin in a dilution of 50,000. The bovine group was inhibited by thionin in a dilution of 1 to 50,000, while the caprine group was not inhibited by any of these dilutions of dyes. He also found that the Brucella organisms could be divided into groups according to their hydrogen sulfid production. The porcine group produced a consider- able amount of gas over a period of four days, the bovine group a considerable amount for two days, while the caprine group failed to produce any detectable amount of gas. In addition to the biological characteristics of the porcine and human strains of Brucella, McAlpine and Slanetz 24 * call attention to 186 BULLETIN No. 343 [March, the fact that the "history and distribution of various human cases in the United States lends some support" to the theory that the porcine type is the causative factor in the spread of the abortus infection in man. Their data show that the undulant fever incidence is much higher in states where the swine industry is large. This does not necessarily mean that the majority of cases are contracted di- rectly from swine, however, for it has been proved by Hadley and Beach, 12 * Schroeder and Cotton, 27 * and Carpenter and King, 3 * that porcine strains may readily infect cattle and become established in the mammary glands, where they may be eliminated with the milk. In his differentiation of the species of the genus Brucella, Huddle- son 20 * finds a lower percentage of porcine strains than McAlpine and Slanetz, 24 * yet his results point toward the porcine type as an impor- tant etiological agent in undulant fever. Out of 96 strains isolated from cows he found that 8 belonged to the porcine type ; of 75 strains isolated from man, 15 were porcine, while all the strains isolated from swine were of the porcine type. All of Huddleson's strains which w r ere isolated from cases of undulant fever in Michigan resembled the bo- vine abortus species, and those sent from cases outside of Michigan, with two exceptions, resembled the porcine species. Huddleson tested the virulence of the Brucella organisms by feeding monkeys and found that the porcine type was more pathogenic than the bovine re- gardless of the source of isolation. Further evidence in behalf of the possible relation of the porcine type to undulant fever in man has recently been presented by Hardy 14 * (Iowa, 1925). He reports the isolation of Brucella-like organisms from 43 cases of undulant fever of which 29 were of the porcine type. Smith 28 * likewise places Brucella types isolated from human sources in the porcine group. His investigations are sum- marized as follows: "We must, therefore, be prepared to look for the source of human infections in the bacillus of swine abortion, pro- vided the caprine type is not in evidence." Smith 29 * also holds the porcine type "largely responsible either directly in the handling of swine or raw pork or indirectly when the swine bacillus is introduced into the cow's udder in one of several ways." Evans, 8 * Blake and Oard, 1 * Viviani, 31 * and other investigators cite cases which support the contention that man may, by contact with infected animals or as a result of wound infection, suffer from the porcine type of infection. 1930] STUDIES ON PORCINE INFECTIOUS ABORTION 187 PORCINE ABORTION TRACEABLE TO DIFFERENT CAUSES In aborting herds coming to the attention of the authors the clinical, bacteriologic, and serologic findings suggest that abortion in swine, aside from the infectious type, may occur as a sequel to other diseases. Sows suffering from cholera, pneumonia, bronchitis, or en- teritis may abort or give birth to weak, unthrifty litters. Such herds upon examination invariably prove negative to the agglutination test for the infectious type of porcine abortion caused by Brucella Traum. Abortion in sows or gilts traceable to other diseases often causes weakness and emaciation of the animals, while the death rate in such herds suggests the severity of a disease independent of abortion infec- tion. In sick herds, apparently healthy sows that farrow normally sometimes fail to supply milk in quantities sufficient to nurse their litters. Five severe outbreaks of abortion in swine, the causes of which were not determined, have been observed by the authors. The history of the herds, as well as the examination of aborted materials and of blood samples from aborting sows, failed to indicate the character of the malady. The general health of the aborting animals appeared normal. Likewise the ration seemed properly balanced and whole- some, tho the possibility of a transitory dietary disturbance as the result of toxic or harmful substances in the feed was recognized as a possible cause. In one herd, where abortions occurred from unestab- lished causes (1928), it appeared that a sleet storm might have been a predisposing factor. The pregnant sows which aborted in rapid suc- cession had been a few days before in a lot that was covered with a thin layer of icy sleet. The slippery footing might have been re- sponsible for injuries which terminated in abortion. Swine abortions coming to the attention of the authors, even tho due to different causes, have appeared suddenly and often have oc- curred in rapid succession, but they have displayed little or no tendency to recur in a serious form on the same farm, even when aborting animals were rebred. One infected herd under observation has harbored the disease for nine years, tho few of the sows have aborted since the initial outbreak. Infectious Type The specific infectious type of abortion in swine, as judged by symptoms and gross pathologic lesions in the placenta, is not dis- tinguishable from abortions due to other causes. Often the first symp- tom noticed is the expelled fetus, tho a discharge may precede abor- 188 BULLETIN No. 343 [March, tion. The discharge continues from one to four weeks following abor- tion. A large number of aborting animals in a herd suggests the presence of the infectious type of the disease, yet herds harboring the infectious type may suffer but a mild loss following the initial out- break. Causes of abortion other than a specific infection may appar- ently be responsible for severe losses. Abortions in infected herds have occurred and disappeared rather abruptly and in subsequent breeding seasons aborting sow r s that were rebred frequently farrowed normal litters. The abortion rate may be higher in gilts than in old sows, tho pregnant animals of all ages FIG. 5. AN ABORTION INFECTED Sow THAT GAVE BIRTH TO A NORMAL LITTER OF PIGS have aborted. In one lot of forty gilts practically all aborted from the infectious type of the disease (1926), while a large number of mature sows on the same farm farrowed normally. The following year none of the sows on that farm aborted. In this respect the course of the disease differs quite markedly from the disease in cattle, which displays a tendency to recur year after year. There can be little doubt, however, that the infectious agent is harbored indefinitely in aborting sows or in the male reproductive organs. Preventive Measures Prompt isolation and segregation of aborting sows is recommended as a control measure, yet it must be recognized that in initial out- breaks many animals may be infected before the owner is aware of the disease. Sanitary measures are effective if applied before the infection spreads thruout the herd. As in cows, infected sows may develop a tolerance to the abortion infection and carry their young 1930] STUDIES ox PORCINE INFECTIOUS ABORTION 189 for the full term. Since the infectious type of abortion may be present in a herd without manifesting itself in definite symptoms, it is sug- gested that newly purchased bred sows or gilts be held in quaran- tine until after farrowing and then tested for abortion. Males should be tested before being employed for breeding purposes. SPREAD OF INFECTIOUS PORCINE ABORTION The infectious type of swine abortion is generally introduced into a herd thru the purchase of infected animals. Bred gilts and sows FIG. 6. SHOTES ix AX ABORTION HERD THAT PROVED POSITIVE TO THE SERUM AGGLUTIXATIOX TEST The dams of these pigs were infected naturally with porcine infectious abortion. may be infected and show no symptom;?. The diagnosis of infectious abortion in swine by the agglutination test may not be accurate if the sows have been or are being fed milk from abortion-infected cattle. Independent of positive agglutination tests in swine receiving milk from abortion-infected cattle, the organism has been isolated from aborted materials in five herds in Illinois since 1920. These results confirm the existence of the infectious type of abortion as reported by Traura, 80 * Good and Smith 8 * of Kentucky; Hayes and Traum 16 * of California; Doyle and Spray 7 * of Indiana; Schlegel 25 * of Germany; Connaway, Durant, and Newman 5 * of Missouri; Hadley and Beach 12 * of Wisconsin; Hayes 15 * of California; and Weeter 82 * of Illinois. Infected Animals May Farrow Normally In one group of aborting sows that suffered from the infectious type of the disease it was observed that a majority of the infected sows subsequently farrowed healthy litters and showed no clinical evidence of the disease notwithstanding a positive agglutination test. 190 BULLETIN No. 343 [March, The possibility of animals breeding normally and yet being carriers of the disease, first suggested by Hayes, 15 * is supported not only by FIG. 7. BARROWS IN AN* ABORTION INFECTED HERD These animals (eight months old) gave a positive serum aggluti- nation test at the time they were marketed. field observations of the authors, but also by the results of bacterio- logic examinations of the colostrum and vaginal discharge of experi- mentally infected gilts that farrowed normal litters. The number of sows that actually abort would appear therefore to be an inaccurate index to the extent of the disease in a herd. The FIG. 8. ABORTED FETUSES This sow (2012) suffered from porcine infectious abortion. From the fetuses pure cultures of Brucella Traum were obtained. 1930] STUDIES ON PORCINE INFECTIOUS ABORTION 191 chronic character of the disease, unaccompanied by abortion, has for many years provided an unsuspected opportunity for infection to spread from herd to herd thru apparently normal tho infected animals. INFECTIVE CHARACTER OF BRUCELLA TRAUM The first investigational studies of infectious abortion in swine at the Illinois Station dealt with the infective character of Brucella Traum isolated from aborted pig fetuses. Cultures of Brucella Traum 2400 tooo 1600 1200 800 4OO MARCH APR. MAY JUNC JULY I9C2 G. itpr. OCT. Nov. DEC. JAN. Fee. MARCH APR. MAY 1923 FIG. 9. AGGLUTINATION REACTION OF GILT AFTER RECEIVING INTRAVENOUS INJECTION OF BRUCELLA TRAUM Healthy Gilt 2061, at the age of 7 months, was treated with Brucella Traum injected intravenously on March 24, 1922. This gilt farrowed 6 healthy pigs on October 10, 1922, and 8 healthy pigs on April 2, 1923. At farrowing time Brucella Traum was not demonstrated in colostrum or fetal membranes. Agglutinations March? Neg. March 24 Neg. March 31 1-10000 April 7 -2700 April 14 -1000 April 21 -200 April 28 -50 May 5 -200 May 12 -200 May 19 -200 May 26 1-666 June 2 1-666 June 9 1-1000 June 16 1-1000 June 23 1-1000 June 30 1-500 J922 July 7 1-200 1922 Oct. 27 -200 -100 -500 -100 Meg. Meg. Meg. -100 -50 -50 -100 -20 -50 -20 Meg. July 15 1-100 Nov. 3 July 22 Neg. Nov. 10 July 28 1-50 Nov. 17 Aug. 4 Aug. 11 ....Neg. 1-400 Nov. 24 Dec. 1 Aug. 18. . . . 1-200 Dec. 8 Dec. 15 Dec. 22 Aug. 25. . . . . .1-20 Sept. 1 Neg. Sept. 8 1-50 Dec. 29 Sept. 15 ..1-50 ises Jan. 5 Jan. 12 Sept. 22 Sept. 29 Oct. 6 ....Neg. . . . . Neg. 1-50 Jan. 19 Oct. 13 Oct. 20 1-20 ....Neg. Jan. 26 Feb. 2 ] W23 Feb. 9 Neg. Feb. 16 1-1CO Feb. 25 Neg. March 2 Neg. March 9 1-50 March 16 Neg. March 23 Neg. March 30 Neg. April 6 Neg. April 13 Neg. April 20 Neg. April 27 1-20 Nlay 4 1-20 May 11 1-50 May 18 Neg. May 25 Neg. isolated from the internal organs of aborted fetuses and fetal mem- branes proved capable of producing abortion when fed or injected intravenously into pregnant gilts. While all experimental gilts ex- posed by feeding or intravenous injection gave a positive agglutina- tion reaction, some farrowed normal litters. Sows infected via the 192 BULLETIN No. 343 [March, CH te*. MAY JUNE. JULY AUG. ^EPT. OCT. Nov. DEC. JAN. FES. fVecu Ape. HAY Q79 fQ7T aoo 1922 FIG. 10. AGGLUTINATION REACTION OF GILT AFTER BEING FED BRUCELLA TRAUM Healthy Gilt 2060 at the age of 7 months was fed Brucella Traum on March 25, 1922. On October 13, 1922, this gilt farrowed 1 dead and 8 healthy pigs, and on April 7, 1923, 5 healthy pigs. At farrowing time Brucella Traum was not demonstrated in colostrum or fetal membranes. Agglutinations 1988 1922 1922 1922 1 923 March 7. . Neg. June 9. . .1-2500 Sept. 1.. .1-50 Dec. 1 1-50 Feb. 25. . .Neg. March 24 . Neg. June 16. . 1-1250 Sept. 8. . .1-50 Dec. 8 1-50 March 2.. .Neg. March 31 .1-100 June 23. .1-1250 Sept. 15. . Neg. Dec. 15. . . .1-50 March 9.. .1-20 April 7. . .1-100 June 30. . 1-1666 Sept. 22. .Neg. Dec. 22. . ..Neg. March 16. .Neg. April 14. .1-500 July 7... .1-200 Sept. 29. .Neg. Dec. 29. . . . 1-100 March 23. .Nets. April 21. .1-100 July 15. . .1-50 Oct. 6... .1-500 192$ March 30. .Neg. April 28 . .1-200 July 22.. .Neg. Oct. 13.. .1-1000 Jan. 5 . . .1-50 April 6 . . .Neg. May 5. . . .1-200 July 28. . .1-100 Oct. 20.. . 1-1250 Jan. 12. . . . 1-100 April 13. . .Neg. May 12.. .1-200 Aug. 4. . . Hemol. Oct. 27.. .1-400 Jan. 19.. ..Neg. April 20. . .Neg. May 19. . .1-200 Aug. 11. .1-500 Nov. 3 . . .1-100 Jan. 26.. ..1-20 April 27.. .1-20 May 26.. .1-500 Aug. 18. .1-666 Nov. 10. .1-400 Feb. 2. . . ..Neg. May 4. . . .1-20 June 2 .... 1-1000 Aug. 25. .Neg. Nov. 17. .1-100 Feb. 9. . . . .1-20 May 11. . .1-20 Nov. 24. .1-400 Feb. 16. . ..1-20 May 18. . .Neg. May 25. . .1-50 , Y . . ^ ...3 _1 ,3 -_2 _ Z A 6 .4 MAY JUNE JULY Au&. JEPT. OCT. Nov. DEC. JAN. FE&M*RCH AR&. MAY 1922 1923 FIG. 11. AGGLUTINATION REACTION OF GILT AFTER RECEIVING INTRA VAGINAL INJECTION OF BRUCELLA TRAUM Health3' Gilt 2059 at the age of 7 months was injected intravaginally with Brucella Traum on June 15, 1922, and bred fifteen minutes later. On October 7, 1922, this gilt farrowed 2 dead and 6 live pigs, and on May 20, 1923, 14 live pigs. At farrowing time Brucella Traum was not demonstrated in colostrum or fetal membranes. Agglutinations 1922 March 7 Neg. April 7 Neg. May 5 Neg. June 2 Neg. June 16 Neg. June 23 1-50 June 30 1-500 July 7 1-1000 July 15 1-200 July 22 1-500 July 28 Neg. Aug. 4 1-20 Aug. 11 1-400 1922 Aug. 18 1-500 Aug. 25 Neg. Sept. 1. Neg. Sept. 8 Neg. Sept. 15 Neg. Sept. 22 Neg. Sept. 29 Neg. Oct. 6 1-100 Oct. 13 1-50 Oct. 20 1-400 Oct. 27 1-200 Nov. 3 Neg. Nov. 10 1-100 Nov. 17 1-100 1AM Nov. 24 1-100 Dec. 1 1-500 Dec. 8 1-500 Dec. 15 1-200 Dec. 22 Neg. Dec. 29 1-100 1923 Jan. 5 1-100 Jan. 12 1-50 Jan. 19 Neg. Jan. 26 1-50 Feb. 2 Neg. Feb. 9 1-50 Feb. 16 1-50 1923 Feb. 25 Neg. March 2 1-20 March 9 1-100 March 16 1-50 March 23 1-20 March 30 Neg. April 6 1-50 April 13 1-50 April 20 Neg. April 27 Neg. May 4 1-20 May 11 1-20 May 18 1-50 May 25 Neg. 1980} STUDIES ON PORCINE INFECTIOUS ABORTION 193 vagina at the time of breeding likewise reacted to the agglutination test, while cows and horses inoculated with Brucella Traum gave positive agglutination reactions. 3000 - 2000 1000 1200 800 400 MMBX APR. HAY 1922 UNL Jtcr AUG. 3o>r. OCT. Nov. DEC. JAM. FL&.rt*KH >Vx. HAY JUNE JULY AUG. 1U3 FIG. 12. AGGLUTINATION REACTION OF BOAR AFTER BEING FED BRUCELLA TRAUM Healthy Boar 2058 at the age of 7 months was infected by being fed live culture Brucella Traum on March 24, 1922. When the animal was killed on August 23, 1923, Brucella Traum was isolated from the bulbo-urethral gland and seminal vesicles. Agglutinations 1913 April 13 Neg. April 20 Neg. April 27 Neg. May 4 Neg. May 11 1-100 May 18 Neg. May 25 1-100 June 2 1-20 June 9 Neg. June 16 Neg. June 23. Neg. June 30 Neg. July 7 Neg. July 14 Neg. July 21 Neg. July 28 Neg. Aug. 4 Neg. Aug. 18 1-50 19tt March? 1 S*eg. March 24 ] S~eg. March 31 ] April 7 *eg. -20 April 14 -100 April 21 -50 \\,r:\ _>> ,..1 -20 MayS -50 May 12 1 3 12 19 26 2 9 16 2530.6 14 Zl Zff .4 II IS 23,, 1 APR. MAY 132Z JUNE JULY . AUG. ,, -JEPT. 15 2Z 29 6 13 20 2T 3 N OCT. ov. FIG. 15. AGGLUTINATION REACTION OF Sow TO REPEATED SUBCUTANEOUS INJECTIONS OF BRUCELLA TRAUM Sow 2080 farrowed 2 healthy pigs on April 16, 1922. She was bred on May 12, 1922, and rebred May 31, 1922. Treatments: March 13, 1922, fed fetuses. April 21, % of one 48-hour agar slant of Brucella Traum killed by heating for 15 minutes at 65 C. injected. April 28, 1 48-hour agar slant heated 15 minutes at 65 C. injected. On the following dates the number of 48-hour agar slants, live culture, injected were as follows: May 5, 1; May 12, 1^; May 19, 2; May 26, 3; June 2, 4; June 9, 5; June 16, 6; June 23, 7; June 30, 8; July 6, 9; July 14, 10; July 21, 11; July 28, 12; Aug. 4, 13; Aug. 11, 14. Agglutinations April 28 1-100 May 5 1-500 May 12 1-500 May 19 1-1250 May 26 1-2500 June 2 1-666 June 9 1-500 June 16 1-1666 June 23 1-1250 June 30 1-500 July 6 1-50000 July 14 1-20000 July 21 1-20000 July 28 1-20000 Aug. 4 1-10000 Aug. 11 1-666 Aug. 18 1-666 Aug. 25 1-666 Sept. 1 Neg. Sept. 8 Neg. Sept. 15 Neg. 1922 Sept. 22 Neg. Sept. 29 Neg. Oct6 1-50 Oct. 13 Neg. Oct. 20 1-500 Oct. 27 Nov. 3 1-500 fed infected milk each day, beginning one month after breeding and continuing until farrowing time. Two gallons of a composite sample of milk taken from ten abortion-infected cows were fed each day. By guinea-pig inoculation the milk was shown to be infected. Abor- 1990} STUDIES ON PORCINE INFECTIOUS ABORTION 197 tion did not occur in any of the seven gilts receiving infected cows' milk, while animal inoculation and bacteriologic examination of the fetal membranes and colostra at farrowing time proved negative to Brucella Traum. Similar negative results were encountered in gilts which were fed milk from infected cows that had been injected subcutaneously with porcine abortion strains. -SCO . oL__ , /8 R 30, 6 73 21 28.4 II 75 Z3.1 8 13 22 29.6 13 20 27. .3 JUNE JUUY Au&. JEPT. OCT. Nov. on FIG. 16. AGGLUTINATION REACTIONS OF Two GILTS FOLLOWING SUBCUTANEOUS INJECTIONS OF LIVE AND KILLED CULTURES OF BRUCELLA TRAUM Healthy Gilts 2035 and 2079, at the age of one year, were given subcu- taneous injections of Brucella Traum. Reaction of Gilt 2035 is shown by the continuous line; of Gilt 2079, by the broken line. Treatments Gilt tOSS, I agar slant 19S8 /ire culture June 16 66-day growth June 23 73-day growth June 30 75-day growth July 6 48-day growth July 13 93-day growth July 21 108-day growth July IS 130-day growth 19tt June 16. June 23. June 30. July 6. . July 13. July 21. July 28. G. tOSS G. t079 Agglutinations Neg. 1-20 1-20 1-20 1-200 .1-100 .1-100 Neg. 1-100 1-500 l-.VK) 1-200 1-100 i-aoo Aug. 4.. .1-500 Aug. ll..l-66 Aug. 18. . 1-1000 Aug. 25. . 1-1000 Sept. l...Neg. Sept. 8... Sept. 15 G. tOSS G. K>79 Neg. l-.VX) 1-500 1-100 Neg. 1-20 Neg. Gilt K>79, 1 ajar slant. dead culture 108-day growth 71-day growth 107-day growth 34-day growth 131-day growth 127-day growth 134-day growth 19tt G. 90S6 Sept. 22. Sept. 29. Oct. 6... Oct. 13.. Oct. 20.. Oct. 27.. Nov. 3. . 1-500 1-Jixl 1-500 .1-20 1-500 G.t079 Neg. Neg. Neg. 1-50 Neg. ('From September 1 to October 6 no weekly readings were made for Gilt 2035.) Since subcutaneous and intravenous injections of pregnant gilts with virulent saline suspensions of Brucella Traum, as well as the feeding of porcine abortion cultures, have not consistently produced abortion, negative results in swine receiving Brucella Bang in milk may not be conclusive, and it would seem probable that the under- lying facts regarding the intercommunicability of Bang abortion dis- ease of cattle to swine can be determined only by more extensive studies. It is apparent that Bang's disease in cattle is not usually com- municated to swine, yet until more evidence is available it is not ad- visable to ignore the possible danger of spreading infectious abortion from cattle to swine and vice versa. 198 BULLETIN No. 343 [March, 3000 2400 2000 1600 1200 too 400 O . n n J2 19 26 2 ITAY J 9 16 23 30. 7 W 2J ttJ.4 11 J8 Z5.J 8 13 ONE JULY AUG. OEPT. FIG. 17. AGGLUTINATION REACTION OF FILLY AFTER RECEIVING INTRAVENOUS INJECTION OF BRUCELLA TRAUM Treatments: May 5, 1922, 1 48-hour live culture was injected. On the following dates agar slants, in the numbers indicated, were injected. May 12, 1; May 19, 1^; May 26, 2; June 2, 3; June 9, 4; June 16, 5; June 23, 6; June 30, 7; July 7, 8; July 14, 9; July 21, 10; July 28, 11. 1922 May 5 1-20 May 12 1-500 May 19 1-1000 May 26 1-2500 June 2 1-1666 Agglutinations June 9 1-2500 June 16 1-2500 June 23 1-1666 June 30 1-5000 July 7 1-5000 1922 July 14 1-2000 July 21 1-2000 July 28 1-5000 Aug. 4 1-5000 Aug. 11 1-1000 1922 Aug. 18 1-1000 Aug. 25 1-1250 Sept. 1 1-200 Sept. 8 1-500 Sept. 15 1-666 isas JuLv AD&. FIG. 18. AGGLUTINATION REACTIONS (WEEKLY) OF TWENTY-TWO MALE PIGS, JUNE 2 TO AUGUST 4, 1923 Pigs 2949 to 2070, from 45 to 93 days old, with one exception, were each fed 1 agar slant of old culture Brucella Traum on June 1, 1923. Average Agglutinations 19S3 June 2 NCR. June 8 1-2 1923 June 16 1-10 June 23 1-22 1913 June 30 Neg. July 7 1-5 1923 July 14 1-3.1 July 21 1-3.3 1923 July 28 1-3.5 Aug. 4 1-3.5 1930] STUDIES ON PORCINE INFECTIOUS ABORTION 199 Normally Farrowing Sows as Carriers of Brucella Traum In a group of 17 aborting sows, abortions occurred as early as three weeks after breeding, tho the majority took place between the fourth and twelfth weeks of pregnancy. Ten of the aborting sows farrowed normal litters in the first pregnancy following abortion. Four of the sows aborted two consecutive times. One sow, after aborting in the spring of 1920, farrowed normally in the fall and aborted during the next period of pregnancy in the spring of 1921. Two of the origi- nal aborting sows repeatedly failed to conceive and were regarded as FIG. 19. PURERBEO CHESTER WHITE Sow WHICH ABORTED MARCH, 1920 On September 10, 1920, this sow farrowed 6 healthy and 4 weak and dead pigs. Abortion bacilli were present in the internal organs of the dead pigs. nonbreeders. The result of monthly agglutination tests on aborting animals (Table 1) indicated that some of these sows probably re- mained actively infected and were potential spreaders of the disease for many months. This suspicion was further supported by the breed- ing records of four sows (of the experimental herd) that aborted the second time and one sow that farrowed normally in the fall of 1920 and aborted in the spring of 1921. The presence of Brucella Traum in the fetal membranes and colostra of normally farrowing sows that were experimentally fed or injected intravenously with the organism prompted an examination of the fetuses and fetal membranes of 23 normally farrowing sows in an infected herd.' One fetal membrane was too badly contaminated "All fetuses and fetal membranes were delivered immediately to the labora- tory. The fetuses were washed in tap water and the skin was dissected back over the thorax and abdomen, care being taken not to open the thoracic and abdominal cavities. The fetuses thus exposed were flamed thoroly. The thora- cic and abdominal walls were removed with sterile instruments in order to ex- pose the heart and stomach. The parts exposed were again flamed. The heart 200 BULLETIN No. 343 [March, c I li o B J2 S & ! , . o . . . . ^ ;;;... ea os "Si "o U3 . 'I ^ 3*3 c_^ g ^^-fi S d-ci^^' e i < T J'"''^ weak and dead) 9-14 (12 healthy) 9-19 (3 healthy) 9-7 (7 healthy) 9-12 (5 healthy) 12-14 (5 healthy, 1 immature) * | o 1 B M B Oj I \ 1 + ++++ 1 + 1 1 1 ++ 1 O TJ o 1 + 1 I+++ +++++ 1 + 1 rcine a p O 1 + + |,, ++ ++ 1 ++++ 1 O c & CO ++ ++ 1 + 1 + l+l 1+1+ 1 at 37.5 1 1 1 ++ ++I I++ ++ 1 ++++ 1 E o .= .| "3 1-9 + + +1 1 1 ++ + 1 1 1 ++ 1 1 T3 1 o B 1-5 ++ ++ 1+ 1 + ++ 1 1 + 1 + + u E I + + ++++++ ++ 1 + 1 ++ 1 a i B a! i + + ++++++ ++++ 1 ++ 1 tj o 1 ++ ++ 1 + 1 + I++I I++ 1 o B al S S", is i II mill 4 *o o o *1 -- 1 II 3Ju -H cq 0) N oo oo d-i rt NO IN O | * *" -"- -H -,-, ~ - H - H "C 0. . . "3 g A |Q cSocijQo S2SS2S2 S < O SE ^^ X X X ^^ .> 8 1 - S S < 3 .2 - a rn fill O O O o CO CO ^ . . co . i Sft w d Q ro Q'"' al* 55 A A A *M O Be ?| fc| ; gl sj| ; &| ifl 5 fi 00 M OiflQJI CQ OD O X 5^ S M ^c| e S ^ S 1 c o I 2*** o = ^ o I?* .f 1 J ^ 8 M 'S S m 1 y - t- "" ~ T: "x ^ examil 3 O a o " 5 3 A M H V ^ fl> ^H^ .^ yj y; tf QJ tj ^"S " "^*S ec C S "3 flu i i s! i i il l! Jt || i| i| : i| fcg : ?> > : 55* Z-S ^d 55* 55-os 55"S .s s ! W 00 r 3 00 rt s g. g. "Si S ^ T" "O o .2 oS u .- o BI 3 ts ^1 Si 1 j ^s fe l& il il Is.l i^ ija ^ | i a 0+ osS. a o, "l"e' s o ^ "^"^ ^*~f" S S " 33o2O xco StK OJ"3Z^ wS O"e5o w K 02 S o a lU -s- -a- -zU fe " -1 ^ ~ __ ^ OS ^ a a^ g"^ "3 a"^ g^"" o: oj * Q Q fe Q Q fa E fa E 1 N I T TT T TT TTT co coco co coco cococo t t^OO 0>O *4 it M 55 8^^ N^" M CS > ~' C*"~' IN M^' IN OJ / 00] STUDIES ON PORCINE INFECTIOUS 203 ^1 Iii Us 5s Us i || M 1 i 11 i 11 II 5 i A 88 8 ?* ?* P - -.3 ' - Z 1 "Sft O M O M i Q ft 3 L a X 5 3* "' ^ 00 *e S a A i3| s o f-SJ Cli- 2 2 cc* 2 2 s 2 s 5* ^ ;l 56 I 22 6 x* a* d^ c? " 2 JL ._ c i icteic u 2^ 2^ 2 'a / - x - x ~ 20 / - / - S ll IE % 3jj i ci^ J;J=: Ji J-^J: a 3 ft T "C S s <- T C T T - C T Q 0* S 5 ^? m - Si S SSS "/. CC w O **4 GC QC CQ CC QQ QC ~ : * I "= =g s 1= "8 3 a II 51 1 3 1 . f|| I Si Sil of < xii in ina c 3 O i =? S II JSl^ f ^i~= Bi 1 * =:=&-= ?= = S" i . a g t> isg $ 1 & . = aSft'l* ftSftcgft ! &"2.5^'t a - XT * M,* ^ "cs ' QQ ^*-M^ tco - cc X cc = M a co A ib cc i u I 1 31 u : || || |l : || : |l |l || 1 "z + a E If s i oi m r i 3 - _ ^ Jl 1 i 35 -Si ^-a"i -Si's 2= ^ Ss Jag S w -= =-a 5'- Jl c-c*!- 4 "^* S >>! "-S5& sf^? 3 99 fl^S t* 1 "^ P*"5 t-S J=_ T3 J? _^> S3 5-i 1 "tl | "jCS 1| |^ IftJ ^ ' g> 3 c-3 Q g'fg +5 e-S 3 e-S 5 .2-f'l a^ ftt 1 1+ + 1-3 *+ H^S Ox SOu XO t?x C"oO O OC 3 Material C tt' -v tt-^ &"^ C & C tt*^ ex'"- A-^ il II 1 1 1 ll 1 If 1 1 1 i 1 tf 'tf s r. M to. |ji opopoeae A Z Z Z Z Z Z Z Z Z Z 6 s ? s- s~ s^ s s 88-8- 2 Z 8 8 8"^ 8^ S~' 8 8 s S^* 8 ^ s^ 204 BULLETIN No. 343 [March, if? frl? &i- co ~co frl? M g CO c ^-fl-5 x i " 10 3 12 "M ^l . r *~* 5 0.0 SI i* M C i, 03 i* M C i ffi 0).- 55 55* ^s 55-5 55 -S 5 1 s?l 1 m frj fr! U5 O o fc| *1 -3 pa Jz;* 55* ^ fc S c 1 Ha .5 CO t CO p^ g Qa o I S _ CD o 00 00 IN M a 3a o ffi 06 fcS^B o O O o o fill- * * * * 55 55 CO ca CO ^ c CO O3 ffi . 2 >n "J ^ CN H E d* d>. NiK K fe IN "I flj W cJ ^* J ^* J W OS 1 " 1 m c , K || 1 0> J | M_> ^ > 1 Q) rj 1 O> ^?> 1 0) 55^ 55* 55 S^a *Z "*^ 55? 55? 55 IS 55-g 55 "S 55 "S h a c "'c' e , o S 1 |J V C o V a V "il V "a 1 0& f I 5 5 n w c w 8 i as oil I 1 al H s V B ^ >, X !>, _ e 1 "S c B Guinea-p Autopsy 1-31 Spleen slight nlarged, follicles rery prominent t-3 Splenic follicl lightly enlarged 1-3 Spleen rather mall. Follicles lightly visible 2 '3 "S-o J| -3 No gross lesio '2 -2 1-10 Spleen slight nlarged. Follicle >rominent. Some lecrotic areas in iver 1-10 Spleen slight nlarged. Follicles ather prominent [-10 Spleen slight nlarged. Follicles ather prominent 1-10 Spleen slightl nlarged. Follicles ather prominent o 1 1 00 = ^ *" ^ ** *" * > n _: COTS it" ' M c i> <^3 553 55 "S 55 -S 2 ,.! >> 8 . "S m 1 i "f II Direct culti G + tetrads, 1 and in fine col Small G + sta lococcus Staphylococcu* albus Staphylococcu, albus G+ rod sprc G rods G + spreader Staphylococcu, aureus Staphylococcu, albus ^ So S -l & I'S+I iilas G+ rods Staphylococcui citrius | "f BEJ 'S t* !1 oi Fine spreadin growth T 0) g 8 s S CO 3 c a c c a M M C 'C g g g g a~ & 3 g ^ j2 Js (^ _4S -fi 1 r 11 W 5 ^ 3s s g 3s il "* "S g fog S fo g fog fog fog fog fog Q Q fog *8 oo o _ CO "5 CO co CO ?? 83 co CO CO i CO i co CO 1 CO CO CO l CO c^ *C CO co 1C "5 CO 00 00 oo o IN N "c8' o \ IN CN CN IN CN CN N ?. gS K- Xl4 '4- 2^J) M i it -2"s _ i 2'i Xs ->.i 1 r - fei s If ^ s 4 u S it 00 o 'b fgj 2 - o i i u tjo> tb * , i e o o o Z {|J -z z 2: Z O * 2 o a p . J. - . - 11 t- ** ** OD * ^Jj *^ ^ < 6 m i II if| ?| S?| S.| g ?| a _ 6 ._ ^ S .0 ii 0-5 a ^L^: e ,2^ ^ g 8 * * 1 11 +1-J i oo oo 02^ O SO^s oo oo B Q 1 s J J 1 K| I H Q a 2 ! Guinea- 1 |i || ^ 1 ;! ] < 9 8 8 a o o "S-o !" -5 S | < eiJ? 58 S5 2 M Z 5 T.C > 00 i C 5 c 1 | 03 -0 i "S ?-B S 2- .g c 2 I -~ *S ; *3'?"S'~^e > " * c c > O < 35 S QQ fisXC--E^.S;^cjw'ctic^ oo 'A oo OQ oo 3 a 888 e>i i s 1 II 1 il III? I s f f Tf 3 ii S S I !!!!!!! H d 352 3 3! *^ *c *o ^^ "5 *^<^ " -^ *" .-/. 2222 c T i i ~ i^S /. 3C CC 00 OD 00 TTTTTTX^T '/. /. '/."/. 'A I 1 i I siii iJJjll" 1 "a j . s -~ a a a o o 115 " srtu 2 2 2 22 ^ Sfi I-. J-JSHO.' L r ?* ^r-''" cte cvccceci6c 111 11111111111 K o o ] oooooooooo 22> ^2222222222 210 BULLETIN No. 343 [March, E 3 g ^.^.5:.::.::.::.^ QJ M a a a a a a J5 M M M M M M l^^zz^S!: *> 1 EE EEEE 33 3 3 3 3 o. g CO o M PH 1 S3 3333 1_ I- U (~ (_ Q ^ g z ( J 3 ^ c E ^ 3 3 3 5 1 lllilii . S i < 1 h O M a (* Q s O I o co oo x oo oo x E W "3 h < O z o H EESE EE 3 3 3 3 C 3 3 a a a a.o a a < 1 S|feS^S| T >s Illllln i IO c c QJ o>Z 3) a) a j fri te *i fe ^ PQ < H Je M x QCD*O*O tN.co 'S "^* 6 fc r^ jjCM-^i-t^p CO CO CO CO COCO Source and history Date castrated to obtvo eo "O o g | 1 1 1 | | |J ||| 1 | | | 1 '/. O O O oo g C O d OO QJ C O (3 ( c ( ^ > t^ CO 00 r I s * W r* r* 00 CO 00 r 6 55 IK sim^isSSlfeliliii! 88 ^!!^ s| ^ggg^^^g I 1 1 1 1 1 1 1 1 1 1 1 1 1 1 Identification Date examined 1923 *"* 7 l rj?77 c i < 57'T5' : i lc i <<: i l 2 " 6 iiiHHiiiniu 1930] STUDIES ON PORCINE INFECTIOUS ABORTION 211 pigs. One testicle yielded Brucella Traum in cultures. Other guinea pigs injected with the different saline suspensions of testicular tissue yielded negative results. Twenty-one days following exposure, 21 pairs of testicles from the five groups were cultured and injected into guinea pigs. Brucella Traum was found in one testicle on direct cul- ture, altho guinea pigs injected with the testicular emulsion yielded negative results. Thirty-eight days following exposure, 21 pairs of testicles from the five groups were cultured and injected into guinea JUS- FIG. 20. MULTIPLE ABSCESS OF TESTICLE AND EPIDIDYMUS OF BOAR CAUSED BY BRUCELLA TRAUM Tho the animal gave a positive blood test and re- peatedly failed to impregnate sows, there was no visible enlargement of either testicle. Chronically enlarged testicles in male hogs are frequently traceable to infec- tion with the abortion organism. Such infected ani- mals are potentially dangerous in the spread of the disease. pigs. Direct cultures in all cases proved negative, but Brucella Traum was isolated from the spleen of four guinea pigs inoculated with testi- cular tissue. The positive specimens came from the infected pigs in the first four groups. Forty-five days following exposure, 43 pairs of testicles from the five groups of exposed pigs were examined for Brucella Traum. The testicles from one pig in Group 3 gave positive cultures, and the injection of a testicular tissue of one pig in Group 1 into a guinea pig yielded positive cultures from the spleen. The agglutination test of the blood of the pigs made 30, 60, and 90 days after castration showed that some animals continued to aggluti- nate Brucella Traum for a period of 90 days following castration. The results of examining testes of artificially infected pigs suggest that the testes may harbor Brucella Traum for 14 to 45 days after exposure even tho no gross lesions are manifested. Examination of Lymph Glands and Spleen. Seven pigs weighing 15 to 60 pounds infected naturally as well as by fed cultures were killed after intervals of 30 to 80 days for examination of the lymphatic 212 BULLETIN No. 343 [March, glands and spleen for Brucella Traum. Two of the seven pigs yielded cultures from the body lymph nodes, six from the visceral lymph glands, and one from the spleen. One pig proved positive in the body visceral lymph glands as well as the spleen, while five of the pigs yielded positive cultures via guinea pigs from visceral lymph glands only. One pig proved positive in body lymph glands only (Table 4). Examination of Epididymi. The positive findings in testes, lymph glands, and spleen of young pigs artificially infected with Brucella Traum suggested the possible localization of the organism in the ac- cessory reproductive organs. Fifteen male pigs farrowed in March and April, 1923, were fed Brucella Traum on June 1. Fourteen to 16 days later these animals were castrated and the epididymi were cultured and injected into guinea pigs. All epididymi proved negative with the exception of 2969 (Table 5). At autopsy one of the guinea pigs (937) injected with this specimen, showed lesions of abortion in the spleen. Agglutination test of the blood serum of this pig was positive, while Brucella Traum was isolated from the spleen. Altho no gross lesions were found in the epididymi examined, the positive serologic and bacteriologic findings suggested that the epidi- dymi of young pigs, as well as testes, body and visceral lymph glands, and spleen, may temporarily harbor Brucella Traum. Presence of Brucella Traum in Bulbo-Urethral Glands and Semi- nal Vesicles of an Actively Breeding Boar. 10 * On March 24, 1922, a grade Duroc-Jersey boar (2058) seven months old was fed one agar slant of a freshly isolated porcine strain of Brucella Traum. Two consecutive agglutination tests, with an interval of 17 days preceding the feeding of the abortion organism, indicated that the animal was probably free from abortion infection. Two weeks after the feeding, agglutinins appeared in the blood serum of the boar, and four months after feeding, agglutination was complete in a dilution of .0002. The agglutination tests were continued weekly from the date of exposure until August 18, 1923 (Fig. 12) . The titre declined to negative in September, 1922, and following mild fluctuations suggestive of oc- cult infection, returned to negative in April, May, June, July, and August, 1923. The decline in the agglutinating titre of the blood serum of the boar indicated the probability of recovery notwithstanding the fact that certain evidence pointed to the service of this boar as a pos- sible factor in the transmission of abortion infection to Sow 2063. The source of the infection in one sow was suggested by positive re- action to the agglutination test following breeding on neutral ground at the time the boar (2058) was reacting strongly to the agglutination test. Other sows bred to the same boar (2058) were exposed to abortion infection thru other channels, and information could not be obtained thru the breeding records to suggest clearly the active role of the boar 1930] STUDIES ON PORCINE INFECTIOUS ABORTION 213 in transmitting the disease at the time of breeding. In order to secure evidence which might throw light on the possible relation of the boar to the transmission of abortion infection in Sow 2063, the boar was slaughtered and the genito-urinary organs were examined pathologically and bacteriologically for the presence of Brucella Traum. No gross pathologic lesions of infectious abortion could be detected. Saline suspensions of testicular tissue, epididymi, bulbo- FIG. 21. LIVER OF GUINEA PIG SHOWING LESIONS OF BRUCELLA TRAUM INFECTION Guinea pigs injected with cultures of Brucella Traum or tissues containing this organism show marked necrotic lesions in the liver, spleen, and lymph glands. urethral glands, prostate glands, seminal vesicles, synovial joint fluid, and synovial fluid of the tendon sheaths were each injected subcu- taneously into two healthy guinea pigs. The guinea pigs were placed in separate cages and were killed 21 days later. The blood serum from each guinea pig in diagnostic dilutions was mixed with abortion antigen and incubated at 37 C. After three weeks the inoculated guinea pigs were autopsied and examined for lesions of abortion. Cultures from the spleen and the liver were also made on plain agar. The sera of guinea pigs which had received the saline tissue emul- sion of bulbo-urethral glands and seminal vesicles reacted positively to the agglutination test, and the spleens of these pigs showed lesions typical of abortion. Pure cultures of Brucella Traum were isolated from the splenic tissues of these guinea pigs. The saline emulsions of testes, epididymi, prostate glands, synovial joint fluid, and synovial fluid of tendon sheaths, when injected into guinea pigs, gave negative bacteriologic, pathologic, and serologic findings. The presence of Brucella Traum in the accessory organs (bulbo-urethral glands and 214 BULLETIN No. 343 [March, seminal vesicles) of an actively breeding male suggests the part which an infected male might play in the spread of abortion infection among swine even tho reacting irregularly or negatively to the agglu- tination test. The boar (2058) in question gave a negative weekly reaction from September 1 to September 29, and from March 23 to May 4. and again from June 9 to August 11 (Fig. 12). FIG. 22. LESIONS OF BRUCELLA BANG IN SPLEEN OF GUINEA PIG The above spleen was removed from a guinea pig six weeks after injection with 2 cc. of a suspension of Brucella Traum grown on an agar slant. Examination of Uteri and Ovaries. Six sows were slaughtered and the ovaries and uteri examined for Brucella Traum in September. 1923. Four of these animals had been artificially infected by the sub- cutaneous injection of cultures of the organism; two of the four had been given a second injection of the virus approximately eleven months after the first exposure. The other two had contracted the infection thru association with infected animals. The reproductive organs of each animal were removed at the time of slaughter and brought to the laboratory. Three of the uteri were gravid. The surface of the uterus was washed with tap water and then flamed before opening with sterile instruments. The stomach contents of the fetuses and the amniotic fluid were injected subcu- taneously into guinea pigs. Saline emulsions of the uterine mucosa from the three nongravid uteri and the ovaries of four of the sows were macerated in sterile saline solution and injected into guinea pigs. The uterine mucosa of sows Duroc-Jersey 7, Hampshire 3, and Hamp- shire 13, upon injection into guinea pigs, gave positive serologic or bacteriologic evidence of Brucella Traum, while the ovarian tissues of sows Hampshire 13, Duroc-Jersey 66. and Hampshire 3 also proved 1930] STUDIES ON PORCINE INFECTIOUS ABORTION 215 positive. The location of the abortion virus in the nongravid uterine mucosa and the ovaries of a pregnant sow may be regarded as some- what at variance with the location of the virus of abortion disease in cattle (Table 6). These findings confirm the observations of Weeter 82 * on the presence of the organism in the nongravid uterus. In November of the same year four artificially infected sows of another group were slaughtered, and their ovaries and uteri were ex- amined for Brucella Traum. Three of these sows had been infected at least twenty months prior to the examination, two by the subcu- taneous injection of cultures and one by intravaginal injection. One sow had been infected by association, as judged by the serum agglu- tination test. The same technic of examining the uterine mucosa and ovaries by animal inoculation as in the first group was carried out. The mucosa of gravid and nongravid uteri of sows Duroc-Jersey 73 and Duroc-Jersey 22, respectively, were found to harbor Brucella Traum (Table 7). Five artificially infected sows, 9907, 9908, 9909, 9910, and 9911, were slaughtered in December, 1923, and the uteri and ovaries were examined for Brucella Traum. One sow had been fed virulent porcine abortion cultures nineteen months prior to the time of slaughter and on two other occasions had been given an intravenous injection of the same cultures. Two sows that contracted the disease naturally also received an intrauterine injection of Brucella Traum. Of the remaining sows one had been infected by an intravenous injection and the other by a subcutaneous injection. Later both were injected with cultures into the uterus just before breeding. The uterine mucosa and ovarian tissue of these sows proved negative to Brucella Traum (Table 8). Monthly Agglutination Tests of Vaccinated and Unvaccinated Pigs In an experiment designed to test the carrier feature of Brucella Traum in young pigs, monthly agglutination tests were made on one group of pigs of which eleven were vaccinated and eleven remained unvaccinated. The abortion vaccine was given subcutaneously. The vaccinated pigs showed a positive agglutinating titre about one month following treatment. A similar reaction was observed in the unvac- cinated group two to four months later. The character of the reactions in the two groups were comparable, but the vaccinated pigs reacted to the agglutination test more promptly. They showed an average maxi- mum of agglutination reaction six months after treatment, with a secondary elevation of the agglutination curve two months following the initial rise. Three months previous to farrowing, the agglutination reaction was negative in the vaccinated pigs (Fig. 23). Unvaccinated pigs allowed to associate with the vaccinated pigs apparently con- tracted the disease. Each gave a positive agglutination reaction three 216 BULLETIN No. 343 [March, o o 5 I h O see s s s 3 -O 9 eJ TSTJ -3 -O = .3 11 11 "3t> 4)" OU (. b t.S "o1 " "* S M M M *i 00 c _o 2o 22 oo oo oo o2 22 PQZ Z<5 ZZ ZZ Z0 (Q PQ 1 i s i S gg 5$ ^S 1 .2.2 * i llTi.rf oo 2 2 o* as go o o H 1 ^ p p MM MM MM -p O O S| *o < G MO. ZZ ZZ ZZ i i ' t. c i'' 3 o> s s B B > - C '5 -" o o o gag ^*"**M^ 1 b O s D 1 ill} S 1 ii. M a I So? S^S 4>tt) aid; a; 0) a; *^ ^ S 55 ^J c ^-2^2-2^- c= c ^S^oj^'* 3 y}^> co i cc b / x, xx ZZ x / _^- ^ /. y QQ 3 >, o < H a kiSS oo oo oo oo oo oo oo , 1 1 >> r s il c *^ ^ ^ "2 j 1 a o ) : ill ! Ill H il 111 ill!| |||!j n li &1 5l II; llil lljii 51 II o s^ s ;: 35 s^ 1 g w ?: * IN M OOlN OOC- W ON 3 i a 1 ^ Is i ! s oS ol 3 -PS -r 3 -r 3 d 3 .S i'3 s - ^- - i- E E g E 56 - - u MV MV 030; ~ i C3 D MOJ 03 2"^ SZ BJ| Ej| C-3 g 1 ^ >; d ifl 53 5 3 C 3 U3 O 3 o 3 o a. Z Z 55 . | 1 g '3-2 "3-S 3 ~3 O 3 O 3 c; c B -^ ^ -- ^ N >o) B B B BE 1 "5 *H '5 "5 "i '> O< .O _C d _O B bU O>P* WIN j> O _g _cj P _g _O -^ V O U 1 i 3-3,0 S^^ -2 1 5 !s 3 1 i* 1 i^ 1 >> >> 1 >> 3 ffl S^ J S d J a -3 -s S TJ flS^goS^B * "cj "cj ^ "o 1 213 J5il 2S 1 1 2! 1 a Q NM NCO M C<5 O V - 0*0' 1<2 "S "'B ~* "Q "o "o "Q 1930} STUDIES ON PORCINE INFECTIOUS ABORTION 217 to 9 - - r- 4"t = = s B * a - = 'E = am .-g 11 cS KCQ 552 11 1 ,5 g o 33 SS ^Ji II mi O& r^T^T 0& >& fig - fi ra "5 8 8 I 1 O O =.5 R , N . e 3 o t -3 ml s i| ? i i i i 8 * . C - . Hi s i| 8 ?? 9 ?? = - * fi S 2 *- i 218 BULLETIN No. 343 [March, ~s o3 1*1 i 1 i 4?*~ ^ 2 Bacterio- logic ex- amination "S ~ "C I* OS e! OS I* It Is Is " . 3O -e* 53 ea! S 3 C O OOOO " ~^ D 8 W 9 H 8 M U t- t- u o w g C 3 -: = C = a; M a M S: 0. 2gSg ggo oooo +-s O 2: aq P5HWH PQE-iZ Z 55 fc OS I ? 8 Aggluti- nation guinea- pig sera 0) OQJ'O QJO4) VVOJ'flVnS .^ .i .: o . .: .^; .S; .^ .^ o .> o _> s as 3 is sss Silili 3 !!- 3 !! 3? ojoto ** ojC'Qj a?ojGJOoJO g-S * .SS-o IS 1 ^ o a^ Bj ft S SSaooa)2 >p aj i _ t vujcmiuvv g o e a c a ca cca- ccccecc g-sfeo o -3. o> _ oo oo-s ooooooo ti^ z^g^z ^^^ ^^^^^^^ 1 O Mi -4 31 S PS PS PS |o |o o |o 6 fc L-5t^ >O-H *! 1OO31N t^^TjtCOQOOO SIN -HTJI COO -COO T^iOiCOiO Oi-H iCi-H "3W Ob-CO CO ^i i-i US i-( 1 4> 13 73 T3 'P ^>_g 'P M 'P T37JS g 3 ""S g s g d '? 3 "S - C 'C - 'E M^"SS g> M g g g ' > ,|3g| 03 03 0^0 Artificial infection Brucella Traum 2J"l'o -3*0 a ^"3 S3 25 u i'o s" 5 rt< S llll f8 | c'| iijl U 5S g S'18 1111 18'P ^-2 III IH" 8l rt -2 ill 111! 11! l 11! 1111 11! l g 13? Is!! !il HI I!! lil i! in III S?33 1|I lil 1>-I 5S1 |.|I i-i 13S fill 52i ji !! !|^2 'li ^S! -!si Hi! SI 381 SIS 1111 1s 111 |||S ggg ||| g|g ll^ 8g |1| *fg h-lrH O^S rHJ2 M'B 8 100^5 J^r^O rt JS >5'SS UJOJ3 o "S Q NOCO (NCO CO 4O00 40 "5'0 INININ ON d i-ciN CMNIN N M W >-H IN COO300 ujob US COOO COO500 U5CO US ^ CO ^ CO ^_* Oi ^ O> OS ^ O5 ^ Ol " Oi Oi ^ Oi ^ a US^ US^ US^ u,^ ,,Q US^ US^ STUDIES ON PORCINE INFECTIOUS ABORTION 219 r r XX gg gg gg :g ~~ 33 33 -r 22 22 22 :2 S 33 22 gg gg 33 *3T ^ 22 22 ~ -- -= JSJS -CJ3 k ;* ?? MM MM UK O O 22 :2 II o e 22 If 1? U II o e o e 22 22 33 ** 22 3 3 3 3 eg SB SfSf 22 22 1! 22 33 -3'3 I! 11 22 22 Ee |1 c-^. : ; C B 22 ee o* 22 TT ^ aT I! II II TT II II 2$! i-i| hi] Ills isli is "5'* SI"" p " I r is O 3 2 03 2 jj| -ii if g-B IJJIJ Ill 2 -g .- i- y. u y, fc. r. 5 31 11 st - ~. M *^ M ** Erf"' il ii il ii CUJ CUS B > - v s c ) et> 3 - a ill Illl JlSI ^1 41 ill * ' 8* iIII II!! *l II 11 ll e , t ifi M !!U i=l ^| *s! lltl is | is is | II!! ill III 111 t* I 333 III 333 33 3 & $ U 4. ^^ -3 ,0? 5^ a*^ S* 5 fe 5 Q *ffl 220 BULLETIN No. 343 [March, months following exposure, but the dilution was relatively low. The agglutination curve was similar to that displayed by vaccinated pigs. The development of agglutinins was prompt in vaccinated pigs, while unvaccinated exposed pigs showed similar reactions 90 days following 10 19ZZ I 26 IAN. 1923 26 APR. FIG. 23. AGGLUTINATION REACTIONS (MONTHLY) OF ELEVEN VACCINATED AND ELEVEN UNVACCINATED PIGS, OCTOBER 26, 1922, TO MAY 26, 1923 Pigs PCM, DJ-34, PC-7, DJ-6, DJ-32, DJ-7, DJ-11, DJ-3, PC-23, PC-11, and DJ-4 were infected subcutaneously with Brucella Traum at the time of weaning, May 24, 1922. Pigs DJ-66, DJ-65, DJ-63, DJ-92, PC-41, PC-66, DJ-22, DJ-15, DJ-95, PC-32, and DJ-9 were associated with infected pigs. The vaccinated and unvaccinated pigs were kept in the same lot. Reaction of the vaccinated pigs is shown by the continuous line; of the unvaccinated pigs, by the broken line. Average Agglutinations Vacci- Unvac- 1988 noted cinated Oct. 26 1-77 1-63 Nov. 26 1-113 1-90 Dec. 26 1-49 1-77 1983 Jan. 26 1-9 Neg. Vacci- 1983 noted Feb. 26 1-24 March 26 1-12 April 26 May 26 1-1.8 1-4 Untac- cinated 1-114 1-19 contact with the vaccinated group. As in other instances, the titre in young pigs following exposure was not high, and it gradually receded and often disappeared. In the unvaccinated pigs 9.9 per- cent aborted, while of the vaccinated group showing the early aggluti- nation reaction all farrowed normally (Fig. 23). Brucella Traum in Vaccinated Pigs. In an effort to determine the relation of a vaccine prepared from Brucella Traum following 1930} STUDIES ON POBCINE INFECTIOUS ABORTION 221 subcutaneous injection of gilts, to the carrier feature of the disease, three groups of young pigs following weaning in the infected herd were placed at the disposal of the Animal Pathology division each year from 1921 to 1924. The vaccine consisted of a porcine culture isolated from an aborted pig fetus, grown in nutrient agar and sus- pended in sterile saline. For three consecutive years approximately one-half of the female pigs (Duroc-Jersey, Poland China, and Hamp- shire) at two to three months of age were treated with abortion vac- cine. Untreated female pigs selected from the same litters as the vac- cinated pigs were kept in the same lots under similar conditions. In November and December the gilts were bred to boars of their re- spective breeds. At the time the gilts farrowed in March and April the fetal membranes and aborted pigs were examined for Brucella Traum, while the blood and milk of the vaccinated gilts at farrowing time were subjected to the agglutination test. 11 * A recheck of the blood of the gilts by the agglutination test was made one to two months after farrowing. The technic used in examining these speci- mens was the same as that used in the examination of fetuses and fetal membranes from normally farrowing sows described on page 199. Examination of Fetal Membranes of Vaccinated and Unvaccinated Gilts, 1921-22. In 1921, ten female pigs were injected with Brucella Traum live vaccine at weaning time. Eleven unvaccinated female pigs were placed with the vaccinated pigs. In November and Decem- ber the gilts were bred and at the time of farrowing, March and April, 1922, fetal membranes and aborted fetuses were examined for Brucella Traum. None of the vaccinated gilts aborted. Two of the 88 pigs in ten litters, or 2.27 per cent, were born dead. Nine fetal membranes from the vaccinated group (1921-22) upon direct culture proved nega- tive. The negative bacteriologic findings in direct cultures were con- firmed by guinea-pig inoculation, while the presence of agglutinins in the blood of the inoculated guinea pigs could not be demonstrated three weeks later at the time of autopsy. Blood samples obtained from three gilts at the time of farrowing or within two months after farrowing showed a low titre for Brucella Traum. Bacteriologic evi- dence that the vaccinated gilts became carriers was not demonstrated in a single vaccinated animal. The vaccine apparently exerted no ill effect upon the prolificacy of the gilts, as an average of 8.6 live pigs per litter were farrowed (Tables 9 and 9A). In the unvaccinated control group three of the eleven gilts aborted. A total of six fetal membranes were examined by direct culture and animal inoculations for Brucella Traum, with negative results. One fetal membrane was too badly contaminated to be examined. Ten fetuses were examined, five of which were obtained from one of the three aborting gilts, and from each of these five fetuses Brucella Traum was isolated by direct culture of the internal organs. In the 222 BULLETIN No. 343 [March, AglS I O 2 &|2 qJ: I!- &!- &!- Or- &!- C .0 o^ "S * .E | 1 *! fr| i?I o c o o o c o u P * 55 33 55 oj 55 "S 55 55 55 55 S < 1 1 ffi > 40 kg M> 1 o CC > i o 10 o M> q !* o S V* fc? 55 'i SZ? 55 1 55 "3 '! ? + z'-i e9 O Ml ~ St E o o o o o - 1 o "o C g o IS CO t _ o CO O O CN CN ,H 00 .2 3'S < o- b ^ _>. >, j* X >> ^ ^ ^ sg 3 3 3 3 3 3 3 3 3 > 0-H 1-5 * >^ >-> <-> 1-5 >-s 1-3 CO CD 1^ "o w 2 S 5 1-3 -! P/ Q D"* ^ >-5 _; _; O Q h " O O , * \ Q Q ci 11 I O c . II ll II II II II II II a C li lj o T _o T s r .2 ^ T ^ ^ *c o o o o 1 o o o S 5 o 0^ 02 02 02 m 02 02 02 02 02 o c M 1 II 111 .2 to a ^c- "S .2 cs S S .a _o O _O 1 O 1 e g E * '8 c .2 "S o c a ll "S^ 5| Ii Hides o iminent S 1 1-5 l-a "i s 1 1 _5 .2 3 of cxami 3k| o g ?|g co .5 0, ^2 fe a g^ co^ 4< tc'S *-f '|]| 3 Il'l^l o 00 00 00 00 L L 1 IQ O "3 5 o 00 J= o 00 d 1 * - ' - *3 in 1 ^ ^"*^ J2 z'i : "- 3 S o s c S o!_0 C3 a e ||i 1 1 si = C3 _o o |1 be C 1 c 1 g o 1 o" 1 Eg 1 3*8 B-g O o O e g 6 " * O.2 o a E i "to * 3 j-_^j >>g ^s O^y O**" g o ^ bD o *'n 1-2 3 S ">> "O ~"S o fe o aS o o a JB J3 ^ ^^o 3 02 0^5 1 CQ^ < 1 + oo 02 O^ 02 Jl ll 1 02 g S . 1 0) o 1 S S 1 c c e s C hfi Cfl c c t k g 1 2 1 I I ' s 'i 1 i .1 i t r S "* E is ll ll ll ll 1 =1 ll 11 11 fe E fa E fa E fa E fa E fa E Q S^ fa E fa E fa E "a Q 1 2 P CO 8 CO CN CM 1 CO i *T V T CN 1 7 i < i 1 fe o 5 8 S CN IO &O t 1 * CN CN CN S CN CO i f- CN CN CN CN CN CN CN CN CN CN CN 1990] STUDIES ON POHCINE INFBCTIOUS ABORTION 228 e 41! ss !! !i ! II! lisls Is |o I II 2 2 m II < 1 g | 3 1 ? n '?} ?J g o is - 3 <7 < 1 | IS t- O t 00 o Ji o 2 e O 1 5 OA 2 r^ ** m 5 fti S O b d .J O - a. a. ft. II II 11 fci ! . g-s ^i5 it 01 01 j ^ 3 i _S< U 3 ^ 2 S c c c | "s'S 1 o* **** M cc 1 X -S = * cS c? t *as | 11 x* 1 la || examination 8 O si 3 *^ Spleen slight d. Spleen fo r prominent f w fit 5 c * Z-5 !i Ii =2 3 Jj !jii c 25^ 3 sM " s = M iJ ' 3 ilg a ^,j| 4-11 4 ax's i 1 1 - JS1 ?! ?> ?> 4 C-S3 2? SK! Direct culture Q+rods Streptococci Diploid forms Heart 1 tube, sterile 1 tube, saprophytes and staphylococcus. Stomach Staphylo- coccut albui Heart 1, tube, sterile 1 tube, staphylococci. Stomach Staphylo- eoecut nurrut. G + rod ; acid and gas in dextrose and lactose 3 tubes, spreader. I tube, StapAyfoeoecui 0) > a 2'S. CO >! o o o o o < Zl 'Z Zl 'Z o a ^ d ^ R o Q &H Q A aS B| II II II II II II II 3culations Cultures (spleen) 02 02 S CO ' "g sIJT O U uT3 MC xC o c "3 c o *o ~B Guinea-pig in Autopsy 1922 So o S^ 03 S 'S3 o 'S 'w 2 8 il P e 1& II 1^ "i i ^- S 2 "i g^ gf-l * 1 6 III & U U" 2 -U !? ^1*^5? ^ 8|| M | g 2 lo|I 1 __:_ ^ L^ o "So "EJ2 10 1 ||| M ^ M ^ td > ^ c3 ^ cj "8! Direct culture cfi J .2 2* O^fl * fl |p 1^=1 B . || | !& P 8-0 feO "Sol ^.S ^ ^ V C S_ js Is;- "jj jj *i e 1 | |.|1 1 1^11 oil 11 1 111 1 i III! i i a | i o> o o o o CUM C 3 M 2 a ' 2222 'S 3|"g "S 3|3|3|3s "g "S Q 04 d H i I ^H CN"COC^M CN co cJ, ro co 4- 4. ^ J. S i b* CO 00 I s * ^ 00 ^* *O O O*!? O^" CO kQ ^ CO CO ^ t-s* - ^ M -^ ?O . ... 1 Inoculation of guinea pig* (concluded) Staphylococcut alfnu 1 Gram-negative large rod 1 Injected with fetal blood and stomach con- tents 6 Died following injection 1 Showing no lesions characteristic of abor- tion Spleens sterile to cultural methods Organisms isolated from guinea pigs' spleens by cultural methods: Gram-positive large coccus 1 B. eoli 1 Srroloffic examination Gilts not vaccinated, negative to agglutina- tion test at time of farrowing: Blood negative . Milk negative 7 Gilt* positive to agglutination test nt time of farrowing: Blood positive 4 Milk positive 2 Gilt* for which samples were not obtained: Blood 2 Milk 2 Guinea pigs injected with fetal membrane emulsions, negative to agglutination test Fetuses examined, negative to agglutination test 10 Fetuses examined, positive to agglutination test Guinea pigs injected with: Fetal blood and stomach contents, nega- tive to agglutination test 4 Fetal blood and stomach contents, posi- tive to agglutination teat 1 226 BULLETIN No. 343 [March, i i^l Mg , M^2 M^2 M> M> > M>^ ' e *|| iz'iet %{1 SSH ^84 fclS 55 '"iS 55 ? 2 ^'^S : "* SS a S? ?> ^f> > : f> l?> *|*> &> I" 3 s 55 -S ^ !5 ^"3 : 55 IS 55* tt ^ a fl 1 fc| i?| s?| i?| fr.J i?| s?| s-| s pq * * Zo! ZsJ Zqj ZicJ ZoJ el ZsJ _ , .? OOOOOos 1 - 1 OO Q ft g o c p >a U5C5OOOO'-'00 OO O5 06 .2 2'S. 1-1 1-H O * i K II $jfj s 3 3 3 3 3 ca - 1! 5 ^^Sd^^^ ^2 2 JS -j-g i)g cC^ M&: i> M^ Mg M^ g it% " "1 Z^ ^^ Z^ 5518 55^ ^ 55 ZU Z^ s .c n 1 00 c o 1 3 o?c)OOO oo c; o o M^MWW M W02 % g o a ^ H 5 M 'S. SEooooCioco 2 c c o c c c S iT> .2 .2 o ' o o .2 Jj JJJJ^JJ j J^l S2 xamin 3 O ! CQ CO (C CO CO - |MM"M& M ^"g H QJ o ^r^CO^OO i^; C-o < -2 S S M W W^ S-'S 2 V .2 eoco-i-i-ioo Js oc*"f O g 3 ^ -o "S s| *| i i i : ^| ; &j if| B < ^5 * * ^oS/5o! d , o .2 X ft ^H C = 3 "S 8 & l 5 3 JS | !!_ Ji B CO _C i i oo oo oo og3 5j3 o o o S o i+ g i+ i+ ++!+ +M++I+I CQ OO 55 OO OO OOO O.2 OOOO OOO O 2 o & $ & c c c c c c c C3 |-g 3| 3| |-i 3| -|-g 3| j| 3| >l fei l >l feg >l feg s tie o oo o o o O t Q-2 T T V A T ? d OOCON^CO*^ S * -n2^;J:2g SS 1930] STUDIES ON PORCINE INFECTIOUS ABORTION 227 three aborting gilts the presence of the abortion organism was sug- gested by positive agglutination tests of either blood or milk at the time of abortion (Tables 10 and 10A). Examination of Fetal Membranes of Vaccinated and Unvaccinated Gilts, 1922-23. During the month of May, 1922, nine female pigs (Duroc- Jersey and Poland China) approximately two months old were injected subcutaneously with porcine abortion vaccine. Eleven unvaccinated pigs of the same age were allowed to associate with the vaccinated pigs until farrowing time. These gilts, both vaccinated and unvaccinated, were bred to boars of their respective breeds during November and December, 1922. None of the nine vaccinated gilts aborted; ten pigs were born dead. TABLE 11 A. EXAMINATION- OF FETAL MEMBRANES AND VAGINAL SWABS FROM GILTS INJECTED WITH LIVING CULTURE BRUCELLA TRAUM (Vaccinated in May, 1922. Bred in November and December, 1922. Materials ob- tained at time of farrowing in March, April, and May, 1923. Part of data summarized from Table 11) GilU vaccinated 9 Inoculation of guinea p*'g Gilts aborted Injected with emulsions of fetal membrane* 12 Live pics farrowed 65 Showing no grow lesions o f abortion. . 12 (Average 7.2 per litter) Spleens sterile to cultural methods. . . . Dead pics farrowed 10 Injected with emulsions of vaginal swabs. Showing no gross lemons of abortion. . Botterioloffie examination Spleens sterile to cultural methods. . . . Fetal membranes examined 6 Organisms isolated: Fetal membranes not cultured 1 Gram-positive coccus Organisms isolated from fetal membranes: Srroloffic examination Gram-positive coccus 4 Vaccinated gilts: Gram-negative coccus 2 Negative to agglutination test at time >f Gram-positive small rod 1 farrowing Gram-negative rod 4 Giving suspicious reaction to agglutina- Gram-positive large rod 1 tion test at time of farrowing 1 Gram-positive staph> lococcus 1 Guinea pigs injected with: Vaginal swabs examined 3 Fetal membrane emulsions, negative to Organisms isolated from vaginal swabs: agglutination teat 12 Gram-positive coccus 3 Vaginal swab emulsions, negative to ag- Gram-negative rod 2 glutination test 6 Fetal membranes and vaginal swabs from the nine vaccinated gilts were examined for Brucella Traum in March, April, and May, 1923. Direct cultures, as well as guinea-pig inoculations of suspen- sions of these materials, proved negative, while the agglutination tests of the blood and milk of these sows, at time of farrowing, with one exception gave no evidence of the presence of Brucella Traum agglu- tinins. The vaccinated group averaged 7.2 live pigs per litter. The blood of the sows was rechecked thirty days later, with negative results (Tables 11 and 11 A). Similar materials from the control or unvaccinated group, includ- ing blood and milk, were examined at the time of farrowing. One of the eleven unvaccinated gilts, Poland China 41, aborted and gave a positive agglutination test. Poland China 32 at the time of farrow- ing also gave a positive agglutination test with blood and milk sera. Eighty-one live pigs and eight pigs born dead, exclusive of the five 228 BULLETIN No. 343 [March, ill is;?s ig M? i>2 ^>2 i>2 >" > ll z"s2 55 '-2 5 Ss'gJ 5?4 55? 4. zl4- 55?2 J ^5 10) V Id 1 o 1 c 1 V B S" a" s 55? 1 55? 55? 55? 55? 1 1 1 iS N " ecS iS tS et" u ' 1 5 S Zos Ze8 ZflS ZoS^aeZeS o _ F i| O U5O O -H CM eo Qft S 4. C O -H 2 . _p 1 3 a SOCDCOCO 00000 OD a 5 B. ! c O OO O O OOO ZZZZZ ZZZ " 03 3 S 5co - 9 S 1 O OO C C COO M S.2 .2 .2 ;2 c 'S. - ^ J fi o i CO S 88""oo "IJoiS P c^ a a 1 C 3 O "S 1 O OO * M OO *7>77- o O O"7^ 5 z z z 2 tT O M 88 '-H t.o o < w H.S fii i i 111 > i 1 1 ^00 CCCO 2?CO OiC CCO L'iO o m i? 1 11 11 I ! 11 i 11 71 a 1-1 < '"" rect cultu ill. fill Jll H h iii Q 1 +&U " l-fOcs" u ?o^ +1 +1 +1 + 2 OOtoo ftj OOww 05 J fro OO OO OOO PQ a 2 S 8"2 3 C = c Materi w S _^ _^ -O*! -2 '& M *l *! -2 l-o a '"S 31 -g-S - ", ^ ^ o fj o B 9 X 9 ~ ^ c; * aS ^ >l <^ fcS 9 fai& > fee >I i! I 1 00 O "^ C3 * ' d POCO CO CO COCO'i' 6 O ^H *H CO CQW z S l^S S 2 S I"'' \ STUDIES ON PORCINE INFECTIOUS ABORTION g 4! xi^ xil 2'3J jj i 8 4 i *{ *l *i 25 2? 2? | 1 s II II II 5 H e o ei 1 I it 00 10 4 w> *C o o o 2 22 3 5 S* "' O> ft o> a do 31 II 1 I ^1- 0- = c TABLE 12. Con 8 of examination Guinea-pig inocu * a A Js I ^ i " 4- 22 _ / ._ M ^ f. o -; 3 if II ! II 1 5 1 1+4- +1 +1| ooo oo 082 11 I SI 1' 1 1' llii If llii 11 i 1 i j ! i i i i 230 BULLETIN No. 343 [March, TABLE 12A. EXAMINATION OF FETAL MEMBRANES, VAGINAL SWABS, AND DEAD PIGS FROM UNVACCINATED GILTS ON INFECTED PREMISES IN ASSOCI- ATION WITH VACCINATED GILTS OF TABLE 4 (Materials obtained in 1923. Part of data summarized from Table 12) Gilts not vaccinated 11 Inoculation of guinea pigs (concluded) Gilts aborted I Spleens sterile on culturing 7 Live pigs farrowed 81 Organisms isolated: (Average 8.1 per litter) Gram-positive coccus 2 Dead pigs farrowed 8 Gram-negative small rod 2 Pigs aborted 5 Rods in long chains 1 Bactrriologic examination Injected with emulsions of vaginal swabs. . 5 Fetal membranes examined 7 Showing no gross lesions of abortion. ... 5 Organisms isolated from fetal membranes: Spleens sterile on culturing 5 Gram-negative rod I Injected with composite organs of fetuses. . 2 Gram-positive rod 2 Showing no gross lesions of abortion. ... 2 B. coli 1 Spleens sterile on culturing 2 Gram-positive coccus 4 _ . . . . Gram-negative coccus 1 berologic examination Gram-positive staphylococcus 1 L "vaccinated gilts: Gram-negative small bacillus 1 Blood negative to agglutination test at Aerobic 1 time * 'arrowing 9 Spore-fonmng'clostridiumY. '.'.'.'.'.'.'.'.'.'. 1 Milk negative to agglutination test at Spore-bearing rod 1 ., time of farrowing . 8 Staphylococcus 1 From which milk samples were not ob- Other spore forms. . . 1 _. tained. ...... 2 Vaginal swabs examined 3 Blood positive to agglutination test at Organisms isolated from 2 vaginal swabs: . * lme of farrowing . . . . . . 2 Gram-negative rod 1 Blood positive to agglutination test at Gram-positive coccus'.'.'. '. 2 time of farrowing 2 Gram-positive rod 1 Milk positive to agglutination test at Staphylococcus aur'eus. '.'.'.'.'.'.'.'.'.'.'.'. '.'.'. 1 time of farrowing 1 Fetuses examined 1 Guinea pigs injected with: Organisms isolated: Fetal membrane emulsions, negative to B. coli 1 agglutination test 10 Inoculation of guinea pigs Vaginal swab emulsions, negative to ag- Injected with emulsions of fetal membranes 12 glutination test 5 Showing no gross lesions of abortion. ... 10 Composite fetal organs emulsion, negative Died following injection 2 to agglutination test 2 pigs aborted by one sow, were farrowed by the sows in this group, an average of 8.1 live pigs per litter (Tables 12 and 12A) . Examination of Fetal Membranes of Vaccinated and Unvaccinated Gilts, 1923-24. On June 2, 1923, sixteen gilts (Poland China, Duroc- Jersey, Hampshire, Chester White, and Berkshire breeds) two to four months old were vaccinated. The live porcine abortion vaccine was injected subcutaneously. Fifteen unvaccinated gilts of the same age and breeds were allowed to associate with the vaccinated pigs. The gilts were bred in November and December, 1923. Because of an outbreak of bronchitis in the herd during the winter, eight of the vac- cinated gilts and three of the unvaccinated gilts died. In March and April, 1924, three of the vaccinated gilts farrowed a total of 24 pigs. The fetal membranes of these gilts, the four fetuses and ovaries of Chester White 3, which died, and the uterus and ovaries of Duroc- Jersey 46 were examined for Brucella Traum by inoculation of guinea pigs. The results were negative (Tables 13 and 13A). Similar materials from the unvaccinated group were injected into guinea pigs, w r hile the blood and milk of the sows were subjected to the agglutination test, with negative results. One gilt in this group aborted, but no evidence of the presence of Brucella Traum could be found. Seventy-one live pigs and three dead pigs were farrowed by 1990] STUDIES ON PORCINE INFECTIOUS ABORTION I o H x sf s~ So < - a -9 221 3 S I 1 1 I I ! I H i 2J g g S 's -a -a 0000 s| o o 'is 22 22 2 33 11 4. 4* S3 88 2 5J> 2 Sg S3S? e B 11 i! i! il !i C; | 1 1 2 oc - 4. J 4- I 232 BULLETIN No. 343 [March, O H^ 8 *lf ^ a-s 2 5-g X M OS 5! K "* ^ O S 2 J= S r <^ S5 ^ i H i fi H4 1 > * 5M 5Z- 2 K S 2 S r a3 <* T3 - as < s _ *_ o-rt a> o 5j ^3 c I i 1 OS I o 1 I 3 _ e e p > E jH '5 w ts e !S B 'S s a _E 1 1 s? "Z 1 Z 3 1 1 .1 10 1 c 1 s c B p a a at a i S ? Z. ? S? rt J5 2; K | ll o o o - - i 00 15 o 00 CO j a 6 o o o o o .S z * 'Z Z a s CO a * J co S 5 Q U U P-' Q 55 O ' ^ > > > il a ''i "S "ol <3 s? Sf S | 8 ? | o w * rv S 1 J* OQ C 00 *** C C C B B B a .3 o o 3 S C o B ' * ji ft) O 00 O o o o o o o z z z z z z o o o C, b C^ fe"" S M 8 ! "S 7 T8 24- $ <"O It 8*8 ,45 ~ 339 II If |l ll il Pe = U.B = U.E !! J i O tj * 234 BULLETIN No. 343 [March, TABLE 13A. EXAMINATION OF FETAL MEMBRANES, FETUSES, UTERI, AND OVARIES FROM GILTS INJECTED WITH LIVING CULTURE BRUCELLA TRAUM (Vaccinated in September, 1923. Bred in December, 1923, and farrowed in April, 1924. Part of data summarized from Table 13) Gilts vaccinated 5 Gilts aborted Live pigs farrowed 24 (Average 4.8 per litter) Dead pigs farrowed Fetuses in uterus at death of sow 4 Bacteriologic examination Feta} membranes examined 3 Uteri examined 1 Ovaries examjned 2 Fetuses examined 4 Inoculation of guinea pigs Injected with fetal membrane emulsion 6 Died following injection 4 Showing no gross lesions of abortion. ... 6 Injected with fetal liver emulsion 2 Showing no gross lesions of abortion. ... 2 Injected with uterine and ovarian emulsion 1 Showing no gross lesions of abortion. ... 1 Inoculation of guinea pigs (concluded) Injected with ovarian emulsion 2 Died following injection 1 Showing no gross lesions of abortion 2 Spleen cultures showing no growth 2 Serologic examination Guinea pigs injected with: Fetal membrane emulsions, negative to agglutination test None made Uterine and ovarian emulsions, negative to agglutination test 1 Ovarian emulsion, negative to agglutina- tion test 1 Fetal liver emulsions, negative to aggluti- nation test 2 Vaccinated gilts: Negative to agglutination test at time of farrowing 4 Not tested 1 TABLE 14A. EXAMINATION OF FETAL MEMBRANES, VAGINAL SWABS, UTERI, OVARIES, AND FETUSES OF UNVACCINATED GILTS ON INFECTED PREMISES IN ASSOCIATION WITH VACCINATED GILTS OF TABLE 6 (Farrowed in 1924. Part of data summarized from Table 14) Gilts not vaccinated ...................... 13 Gilts aborted ............................. 1 Live pigs farrowed ........................ 71 (Average 5.9 per litter) Dead pigs farrowed ....................... 3 Fetuses in uterus at death of sow ........... 10 Bacteriologic examination Fetal membranes examined .............. 9 Vaginal swabs examined ................. 3 Uteri examined ......................... 1 Organisms isolated from above sources: B. coli ............................... 1 Pasteurella ........................... 1 Ovaries examined ....................... 2 Fetuses examined ....................... 10 Inoculation of guinea pigs Injected with fetal membrane emulsions. . . 18 Died following injection ............... 10 Showing no gross lesions of abortion. ... 18 Injected with vaginal swab emulsion ...... 6 Died following injection ............... 3 Showing no gross lesions of abortion. 6 Injected with uterine and ovarian emulsion 2 Showing no gross lesions of abortion. ... 2 Injected with uterine mucosa emulsion. ... 2 Showing no gross lesions of abortion. ... 2 Injected with ovarian emulsion ........... 2 Inoculation of guinea pigs (concluded) Died following injection 1 Showing no gross lesions of abortion. ... 2 Injected with fetal liver and stomach-con- tent emulsion 2 Showing no gross lesions of abortion .... 2 Spleen cultures sterile 3 Serologic examination Guinea pigs injected with: Fetal membrane emulsions, negative to agglutination test 5 Vaginal swab emulsions, negative to ag- glutination test 2 Uterine and ovarian emulsions, negative to agglutination test 2 Uterine mucosa emulsions, negative to agglutination test 2 Ovarian emulsions, negative to agglutina- tion test 2 Fetal liver and stomach-content emul- sions, negative to agglutination test. 2 Unvaccinated gilts: Negative to agglutination test at time of farrowing 11 Giving suspicious reaction 1 Not tosted. . . 1 these sows, while ten healthy fetuses were found in the uterus of one sow at the time of slaughter (Tables 14 and 14A). Summary of Results in Vaccinated and Unvaccinated Gilts, 1921- 24. The carrier feature of Brucella Traum in 24 gilts given porcine abortion vaccine was not detected by direct culture of the fetal mem- branes and fetuses, or by guinea-pig inoculation at time of farrowing. In cases where fetal membranes were not available vaginal swabs, uteri, and ovaries were examined. The negative findings suggest that 1930] STUDIES ON PORCINE INFECTIOUS ABORTION 235 young pigs possess considerable resistance to the Brucella Traum vaccine. None of the vaccinated gilts aborted. The unvaccinated or control animals included a total of 35 gilts that were kept with the vaccinated gilts. Five of the unvaccinated control gilts exposed in this manner aborted. Therefore, it appears in the Illinois experiments that the vaccine administered to pigs two to four months of age, two or more months before breeding, had a tendency to reduce the number of abortions during the first preg- nancy, tho continuous association of unvaccinated gilts with vac- cinated gilts tended to perpetuate abortion in the unvaccinated group. The animals in these experiments were not available for observation during the second or third pregnancies, and the abortion rate over a period of years in the vaccinated and unvaccinated animals was not determined. Vaccinated and unvaccinated gilts were not examined at the time of slaughter for Brucella Traum. The permanent carrier feature in these animals, therefore, could not be determined. Al- tho the value of vaccination as a means of controlling infectious porcine abortion is suggested, the results herein reported do not justify the use of living culture vaccine in the control of this disease except for experimental purposes. In fact it seems undesirable to em- ploy living vaccine in a herd of aborting swine unless a large portion of the herd has previously suffered from the infectious type of the dis- ease as established by accurate laboratory methods. SUMMARY AND CONCLUSIONS 1. Abortion in swine on Illinois farms has occurred sporadically over a period of many years. The infectious type of the disease was recognized in Illinois in 1920. Following initial outbreaks in differ- ent herds in which sows and gilts abort, the incidence of abortion generally subsides. In other words, the disease loses rather than gains momentum after the initial storm, and infected sows generally far- row normally in subsequent pregnancies. The benign clinical mani- festation of the disease does not necessarily imply recovery, for in- fected animals may harbor the infection indefinitely and serve in the capacity of spreaders at time of farrowing. Infected boars are also a potential source of danger at time of breeding. 2. Field evidence of a convincing character has failed to show that abortion disease in cattle spreads to swine. By experimentally ex- posing swine, this supposition was not materially altered. On the other hand, the susceptibility of a pregnant heifer, as judged by abor- tion and positive agglutination test, suggests the danger of Brucella Traum invading cattle. 3. Outbreaks of abortion in swine in Illinois coming to the atten- tion of the authors are also traceable to non-infectious causes. Bru- 236 BULLETIN No. 343 [March, cella Traum is responsible for one type of the disease. Other types of abortion encountered, aside from those due to injury or violence, ap- pear related to febrile diseases; while in other cases the cause or causes of abortion could not be established. 4. Gilts suffering from infectious abortion may, with few excep- tions, continue to react to the agglutination test but farrow normal- ly in subsequent pregnancies. 5. Abortion in healthy gilts or sows may occur following the feed- ing of the porcine abortion organism, but some pregnant sows and gilts following exposure may farrow normally. A positive agglutina- tion reaction was consistently observed in healthy gilts and sows fol- lowing the injection or feeding of Brucella Traum. Males, including barrows, react the same as gilts. 6. Cows, horses, guinea pigs, and rabbits may be artificially in- fected, as judged by the agglutination test. Young pigs are highly resistant, as judged by the mild response and rapid decline of the agglutination titre following exposure. The infection may persist, however, in young pigs for several weeks. 7. Abortion in swine traceable to Brucella Traum can be accu- rately diagnosed by isolation of the infecting agent. Serologic tests are also helpful in diagnosis, provided animals to be tested are not being fed infected cow's milk. 8. The porcine type of Brucella organism resembles the bovine strain morphologically and serologically, but can generally be dis- tinguished by its luxuriant growth and yellow pigment in old agar cultures. Experimental infection of a heifer by intravenous injection with porcine strains was followed by abortion. Field observations and exposure of pigs by feeding naturally and artificially infected cow's milk fail to provide definite proof that Brucella Bang of cattle is a common etiologic factor in infectious abortion in swine. Two different pathogenic types of the abortion or Brucella organism in cattle and swine are thus suggested. 9. In normally farrowing sows in one spontaneously infected herd Brucella Traum could not be demonstrated in the afterbirth or dead fetuses, yet emulsions of these tissues following subcutaneous injec- tion into guinea pigs occasionally produced in these animals specific agglutinins for Brucella Traum. Since the blood sera or colostra in nine of the sows at the time of farrowing completely agglutinated Brucella Traum in a dilution of .01 to .02, the passive transmission of agglutinins by the material injected into guinea pigs seems probable. 10. Direct cultures of the nonlactating mammary glands of sows reacting positively to the agglutination test failed to yield Brucella Traum. Negative results were also obtained by inoculating guinea pigs with emulsions of the mammary tissue. Occasionally the guinea pigs inoculated with the mammary gland emulsion developed specific agglutinins for Brucella Traum. 1930] STUDIES ox PORCINE INFECTIOUS ABOBTION 237 11. Brucella Traum was present in the testicular tissue of young pigs 14 to 45 days after artificial exposure by feeding or intravenous injection. The pigs yielding positive cultures showed slight or no ag- glutinins to Brucella Traum. No gross pathologic lesions in testes harboring Brucella Traum were observed in young pigs experimentally infected. 12. Brucella Traum was demonstrated in the epididymi of pigs 14 to 16 days after feeding the organism. The agglutinins in the blood serum of pigs were slight or imperceptible. Gross lesions were not ob- served in the epididymi yielding positive cultures. 13. Brucella Traum was encountered in the bulbo-urethral glands and seminal vesicles of an actively breeding boar seventeen months after the feeding of the organism. Such findings suggest that males harboring the infection in the reproductive organs might play an active part in the spread of the disease at the time of breeding. 14. The body and visceral lymphatic glands and spleen of young pigs artificially infected by feeding yielded positive evidence of Bru- cella Traum 30 to 80 days later. The agglutinin titre for Brucella Traum in the blood sera of these pigs was not characteristic of the infection. 15. The nongravid uteri and ovaries of sows harbored Brucella Traum for a period of six to twenty months following subcutaneous or intravaginal injection. Other animals that yielded positive uterine or ovarian cultures were exposed by feeding or by association with infected animals. The frequency of a uterine or ovarian infection was not determined, but in the animals at the authors' disposal, the nongravid uteri frequently harbored the infection. The colostrum in aborting and normally farrowing infected sows also yielded Brucella Traum. 16. Pigs ten to twelve weeks old injected subcutaneously with Brucella Traum vaccine showed a low average agglutination reac- tion extending over a period of approximately six months, followed by a secondary curve of shorter duration lasting approximately two months. Uninoculated control pigs in the same pen showed a com- parable average primary and secondary agglutination curve with comparable maximum titre to Brucella Traum. The average maxi- mum agglutination titre of the vaccinated pigs preceded the maximum agglutination reaction of the contact control or unvaccinated pigs ap- proximately 90 days. 17. Unvaccinated pigs which were allowed continuous association with pigs of the same age that received live vaccine probably con- tracted abortion infection thru association. This suggests the im- portance of segregation of vaccinated pigs. 18. None of the 24 pigs vaccinated at weaning aborted, nor was evidence regarding the danger of abortion carriers in the pigs injected 238 BULLETIN No. 343 [March, with living culture vaccine found in fetal membranes and dead fetuses of pigs vaccinated at the time the first litters were born. Direct cul- tures and guinea-pig inoculations were negative. The vaccinated gilts also gave a negative agglutination test at the time of farrowing, nine months following vaccination. Materials from the vaccinated and un- vaccinated gilts were examined from the first litters and it is possible that the results of subsequent pregnancies and examinations of the vaccinated animal might not coincide with the findings in the first far- rowing period. 19. Of 35 control, or unvaccinated, pigs of the same age that were kept with the vaccinated pigs, 5 aborted during the first pregnancy, while none of the 24 pigs receiving the vaccine aborted. Further breeding records for the vaccinated and unvaccinated gilts were not available. Since the agglutinin titre of the blood of pigs in each group shows that both vaccinated and unvaccinated pigs become in- fected, it is apparent that the delivery of healthy litters by the vac- cinated group may be related to time of exposure to the virus -of the disease. In the unvaccinated group initial positive agglutinins ap- peared ninety days later than in the vaccinated group, and obviously during pregnancy. The same danger of infection either from natural exposure or by vaccination has been observed in pregnant cattle. 20. The possible danger of living culture vaccine was suggested by the occurrence of abortions among the control, or unvaccinated, gilts that were associated with the vaccinated pigs. Approximately 14 percent of the unvaccinated gilts aborted. The results of the ag- glutination test in the case of the unvaccinated gilts also suggest the possibility of infection being spread by vaccinated animals. It there- fore appears that the use of living vaccine, while of apparent immun- izing value when administered to young pigs, may spread infection to unvaccinated animals. 21. Investigations conducted by various laboratories during the past three years give results which support the possible relation of Brucella Traum to undulant fever in man. Such results prompt the cautious handling of aborted materials by attendants and the seg- regation of infected animals preparatory to fattening for market in order to reduce the danger of direct infection to man, as well as in- directly thru gaining entrance to cattle, where it might become lodged in the udder and thus be transmitted to man. 1930] STUDIES ON PORCINE INFECTIOUS ABORTION 239 LITERATURE CITED 1. BLAKE, F. G. and OARD, H. C. Undulant fever: a report of three cases oc- curring in Connecticut. Yale Jour. Biol. and Med. 1, 128. 1099. 2. CARPENTER, C. M. Results of injecting pregnant heifers with Brucella abortus isolated from man. Jour. Amer. Vet. Med. Assoc. 70, n.s. 23, 459-468. 1927. 3. and KING, M. J. Brucclla abortus in milk and its relation to undulant fever. Undulant fever symposium, Amer. Pub. Health Assoc. 1929. 4. - and MERRIAM, H. E. I'ndulant fever from Brucella abortus. Jour. Amer. Med. Assoc. 87. 1269-1271. 1996. 5. CONNAWAY, J. W.. DCRANT, A. J., and NEWMAN, H. G. Infectious abortion in swine. Mo. Agr. Exp. Sta. Bui. 187. 1921. 6. COTTON, W. E. The character and possible significance of the Bang abortion bacillus that attacks swine. Jour. Amer. Vet. Med. Assoc. 62, n. s. 15, 179-192. 1922-23. 7. DOYLE, L. P., and SPRAY, ROBB S. Infectious abortion of swine. Jour. Infect. Diseases 27. 165-168. 1920. 8. EVANS, ALICE C. Human infections with organism of contagious abortion of cattle and hogs. Jour. Amer. Med. Assoc. 88, 630-632. 1927. 9. GOOD, E. S., and SMITH, W. V. Bacillus abortus (Bang) as an etiological fac- tor in infectious abortion in swine. Jour. Bact. 1, 415-422. 1916. 10. GRAHAM. ROBERT, BOUGHTON, I. B., and TUNNICLIFF, E. A. The presence of Bact. abortus (Bang) in the bulbo-urethral glands and seminal vesicles of an actively breeding boar. Abs. Bact. 8, 22. 1924. 11. Results of examination of colostra and fetal membranes of gilts previously injected with Bact. abortus vaccine. Abs. Bact. 8, 23. 1924. 12. HADLEY, F. B., and BEACH. B. A. An experimental study of infectious abor- tion in swine. Wis. Agr. Exp. Sta. Res. Bui. 55. 1922. 13. HAGAN, W. A. Cited by Williams, W. L., m The avenue of invasion and the behavior of the infection of contagious abortion in the uterus. Jour. Amer. Vet. Med. Assoc. 52, n. s. 5, 20. 1917. 14. HARDY, A. V. Personal communication. 15. HAYES, FRED. Some studies in swine abortion. Jour. Amer. Vet. Med. Assoc. 60, n. s. 13, 435-452. 1922. 16. HAYES. F. M., and TRAUM, J. Preliminary report on abortion in swine caused by B. abortus (Bang). North Amer. Vet. 1. 1920. 17. HCDDLESON, FOREST. Ann. Rpt. Amer. Med. Milk Comn. 1928. 18. The susceptibility of swine to contagious abortion Bact. abortus (Bang). Mich. Agr. Exp/Sta. Quart. Bui. 2, 4, 43-45. 1921. 19. Is Bacterium abortus pathogenic for human beings? Jour. Amer. Med. Assoc. 86, 943-944. 1926. 20. The differentiation of the species of the genus Brucella. Un- dulant fever symposium, Amer. Pub. Health Assoc. 1929. 21. The isolation of Bacterium abortus from milk. Mich. Agr. Exp. Sta. Tech. Bui. 49. 1920. 22. HULL, THOMAS G. Unpublished paper read at the Ann. Health Officers' Con- ference. 1929. 23. Illinois Undulant Fever Committee. Undulant fever transmission control methods. III. Dept. Pub. Health, Springfield. 1929. 24. McAiriNK. JAMES G., and SLANETZ, CHARLES A. Bact. abortus infection in man and its relation to milk consumption. Interaatl. Assoc. Dairy and Milk Ins,,. 16th Ann. Rpt. 178-183. 1927. 25. SCHLEGEL, M. Infectious abortion in swine. Ztschr. Infektionskrank. d. Haust. 19, 332. 1918; abs. Jour. Amer. Vet. Med. Assoc. 58, 206. 1920. 240 BULLETIN No. 343 26. SCHROEDER, E. C., and COTTON, W. E. Infection of guinea pigs via the con- junctival mucosa with the bacillus of bovine infectious abortion. Jour. Amer. Vet. Med. Assoc. 65, n. s. 18, 211-214. 1924. 27. Recent Bureau of Animal Industry Experiment Station bovine infectious abortion studies. Jour. Amer. Vet. Med. Assoc. 64, n. s. 19, 550. 1925. 28. SMITH, THEOBALD. The relation of Bacillus abortus from bovine sources to Malta fever. Jour Expt. Med. 43, 207. 1926. 29. The truth about undulant fever. Health News, N. Y. State Dept. Health. August 5, 1929. 30. TRAUM, J. Ann. Rpt. Chief of Bureau of Animal Industry, U. S. D. A., for year ending June 30, 1914. 31. VIVIANI, R. Pathogenicity of Bang's Bacillus abortus in the human subject. Policlin., Sez. Prat. Rome, 32, 203. 1925. 32. WEETER, H. M. Infectious abortion in domestic animals. Jour. Infect. Dis- eases 32, 401-416. 1923. UNIVERSITY OF ILLINOIS-URBANA