THE UNIVERSITY OF ILLINOIS LIBRARY 6? SO . *" ( G NON CIRCULAR CHECK FOR UNBOUND CIRCULA Vitamin- B and Vitamin- G Content of Cereals By ROSSLEENE ARNOLD HETLER, CLARA ROCKE MEYER, and DOROTHY HUSSEMANN UNIVERSITY OF ILLINOIS AGRICULTURAL EXPERIMENT STATION BULLETIN 369 CONTENTS PAGE INTRODUCTION 167 REVIEW OF LITERATURE 168 I. VITAMIN-B COMPLEX IN EMBRYO AND ENDOSPERM ENDS OF HAND-DISSECTED OAT KERNELS 170 II. VITAMIN-B VALUE OF WHOLE HULL-LESS OATS, WHOLE HULLED OATS, OAT PRODUCTS, WHOLE YELLOW CORN, AND WHOLE WHEAT 176 III. VITAMIN-G CONTENT OF WHOLE HULL-LESS OATS, WHOLE YELLOW CORN, WHOLE WINTER WHEAT, AND RICE POLISHINGS 188 IV. VITAMIN-B AND VITAMIN-G VALUE OF WHOLE HULL- LESS OATS FOR LACTATION 201 SUMMARY 208 LITERATURE CITED . . . 210 Urbana, Illinois May, 1931 Publications in the Bulletin series report the results of investigations made or sponsored by the Experiment Station Vitamin-B and Vitamin-G Content of Cereals ROSSLEENE ARNOLD HETLER, CLARA ROCKE MEYER, and DOROTHY HUSSEMAXN' /'U^HE FACT that cereals play a large part in the nutrition of both man and animals makes the vitamin content of this class of foods of basic importance. The data recorded in the present investigations have been obtained from observations made in the Home Economics laboratory at the University of Illinois during the years 1927 to 1930. A series of studies have been in progress concerning the nutritive condition of animals when corn, oats, wheat, or rice polish- ings have been included in the diet, either as a sole source of vitamins B and G or as the sole source of one or the other of these two vitamins. When the work was begun in 1927, Sherman's standard method 22 * was used for the determination of vitamin B, and no distinction was made between the two factors of the "vitamin-B complex." During the progress of the investigation, however, definite recognition of vitamins B and G was made, and thus it was found necessary to introduce changes in the methods of procedure. A tentative method, both for the determination of vitamin B and of vitamin G, was developed and applied to the determination of the content of these vitamins in some of the cereals and cereal products. It was found that the inclusion of 25 percent oats, corn, or wheat in the diet probably provides an ade- quate supply of vitamin B for growth if plenty of vitamin G is in- corporated in the basal diet. On the other hand, at least 50 percent of the diet must be made up of any one of these cereals if sufficient vita- min G is to be supplied for approximately normal growth. This ex- plains the fact that in the earlier work it was found necessary to in- clude the cereal to the extent of 50 or 60 percent of the diet if the diet was to supply sufficient "vitamin-B complex" for normal growth. The investigation was extended to include observations on the nutritive requirements of lactation. Whereas a diet made up of 25 percent of any of the cereals supplied adequate vitamin B for growth and maintenance, it was necessary to include in the diet of a lactating rat at least 50 percent cereal in order to supply adequate vitamin B for the growth of the young. Sufficient vitamin G could be furnished for successful lactation if, in addition to the cereal, the diet included 15 percent autoclaved yeast. It seems justifiable to conclude from the ROSSLEE.VE ARNOLD HETLER, formerly Assistant Chief in Nutrition; CLARA ROCKE ME YER, Research Assistant in Nutrition; and DOROTHY HUSSEMAXN. formerly Assistant in Nutrition. 167 168 BULLETIN No. 369 [May, above observations that more of the B and G vitamins are necessary for lactation than for growth and maintenance. It is of particular interest to note the relative amounts of vitamins B and G in the different cereals. Since the diet must include at least 25 percent of either oats, wheat, or corn if it is to provide an adequate supply of vitamin B, it is clear that the vitamin-B content of cereals is rather low. Twice as much cereal, or 50 percent, is necessary, how- ever, to supply sufficient vitamin G for approximately normal growth, and it thus appears that vitamin G is the limiting factor of the "vita- min-B complex" of cereals. Whether there is a real difference in the content of vitamins B and G in the cereals, or whether the relative amounts of vitamins B and G needed by the rat differ, is a subject for further investigation. It is very possible that an interrelationship exists in the needs of the rat for these two vitamins, and thus that we are testing this factor rather than the content of the vitamins in the cereals and cereal products. REVIEW OF LITERATURE Not only the discovery of the antineuritic factor, vitamin B, but also the recognition of the pellagra-preventive factor, vitamin G, has come thru the study of deficiency diseases occurring on a restricted cereal diet. In 1897 Eijkman 7 * first attributed the disease beriberi to the deficiency occurring when the diet was made up largely of polished rice. Goldberger 11 * in 1925, studying the prevention of pellagra, found that the diets of pellagrins are composed largely of wheat flour, rice flour, and corn meal. In 1916 McCollum, Simmonds, and Pitz, 16 * using whole wheat as a source of vitamin B in the diet of growing animals, showed that this vitamin is contained in wheat in a very high concentration. The next year 17 * these investigators discovered that: "The oat kernel, like unpol- ished rice, wheat, wheat germs, maize kernel, alfalfa leaves, cabbage, and clover leaves, contains a liberal supply of water-soluble B, the preparations of which induce relief from polyneuritis." In 1918 Steen- bock, Kent, and Gross 25 * concluded that there is an abundance of water-soluble vitamin B in barley. Bell and Mendel 2 * in 1922 decided that wheat is comparatively low in vitamin B, for about 40 percent of this cereal in the diet was found necessary for normal growth in the rat. Steenbock, Sell, and Xelson 26 * in 1923 confirmed this observation but found that even more than 40 percent of the diet of a rat must consist of cereals if normal growth is to be produced. The rats were housed in raised-bottom cages so that they could not supplement their diets by consumption of excreta. Under such conditions 60 percent of 1931 \ VITAMIN- B AND VITAMIX-G CONTEXT OF CEREALS 169 the diet, it was shown, must be furnished by any of these grains in order to supply enough vitamin B for maximum growth. Croll and Mendel 6 * in 1925 induced normal growth in rats when the only source of vitamin B was supplied by a ration which included 25 to 30 percent white or yellow maize. Recent work by Hunt 13 * shows that growth occurs when rats are fed on a vitamin- B- free ration of which 15 per- cent is wheat or corn if this ration is supplemented with autoclaved yeast. Hunt's work followed observations by Osborne and Mendel, 1919, 20 * Mitchell, 1919, 18 * Emmett and Luros, 1920, 8 * and others, who had expressed the opinion that vitamin B might be made up of two parts. Hunt believed the cereals supplied plenty of the antineuritic vitamin when fed at a 15-percent level but that autoclaved yeast was necessary to supply the limiting factor, vitamin G. In 1926 Smith and Hendrick 24 * attempted to correct the deficiency of a diet composed of oats, casein, vitamin A, and inorganic salts. These investigators found that the above diet was materially remedied by a small addition of dried brewers' yeast. It was not clear whether the improvement was due to yeast protein, water-soluble vitamin, or to some unknown factor. In attempting to rule out vitamin B, the basal ration was supplemented with dried brewers' yeast, autoclaved for six hours at 15 pounds pres- sure, which procedure, as was shown by tests, completely destroyed vitamin B. Rats responded with growth. It was therefore demon- strated that autoclaving brewers' yeast, tho destroying its vitamin-B content, did not impair its efficiency in supplementing the oat deficiency. That the oat kernel was not deficient in vitamin B but in some other factor which was present in brewers' yeast and which withstood pro- longed autoclaving, was the logical conclusion. In 1927 Chick and Roscoe 3 * tested wheat embryo, autoclaved wheat embryo, alcoholic extracts of wheat, and other substances for the two factors called by them B x and B 2 . They concluded that there is an antineuritic vitamin which prevents or cures collapse, including pa- ralysis, and a second factor which is pellagra-preventive. The two taken together are growth-promoting. Salmon, Guerrant, and Hays in 1928, 21 * using the velvet bean, found that by controlling the hydrogen- ion concentration, the B-P factor, vitamin B, could be adsorbed on fuller's earth more completely than the P-P factor, vitamin G. In 1928 Evans and Burr 10 * again differentiated between the two factors by using an alcoholic extract of rice polishings in addition to a purified diet, and thereby produced no growth, yet no neuritis. By adding yeast, growth resulted. Munsell 19 * in 1929 found in testing rice pol- ishings that it takes more of the rice polishings to supply the amount of vitamin G required by the rat for optimum growth than it does to 170 BULLETIN No. 369 [May, supply the needed amount of the antineuritic vitamin. Hauge and Car- rick 12 * showed corn to be rich in the antineuritic substance but poor in the growth-promoting substance. Aykroyd and Roscoe 1 * in 1929 re- ported data concerning the B 2 (G) content of several foods. They found that wheat and maize are poor in vitamin B,, "the maize being on the whole the lower in the samples examined ; in wheat the germ and bran are better sources than the endosperm, and about equal to each other; in maize the germ is not so rich a source, but whole maize is better than maize endosperm." I. VITAMIN-B COMPLEX IN EMBRYO AND ENDOSPERM ENDS OF HAND-DISSECTED OAT KERNELS 3 In studying the vitamin-B complex in embryo and endosperm ends of hand-dissected oat kernels, whole hull-less oats obtained from the Agronomy Department of the College of Agriculture were carefully sorted by hand to eliminate all foreign matter. The kernels were then cut with a scalpel into two sections, there being left at the embryo end about one-third of the whole kernel by weight and at the endosperm Fine dissection cut (VJ - V4 By wt. emtoryo) Endosperm Roufh dissection cut (V2-V3 by wt. embryo) FIG. 1. HAND-DISSECTED EMBRYO AND ENDOSPERM SECTIONS OF OAT KERNEL By using the fine dissection cut, an embryo fraction was obtained which was almost, if not entirely, free from endosperm. end about two-thirds of the whole kernel. This dissection gave an embryo fraction which was far from being free from adhering endo- sperm. Later it was found necessary to make a more careful dissec- tion of the two structures, embryo and endosperm. By a curved cut an embryo fraction was obtained which was almost, if not entirely, free from endosperm. This dissection resulted in an embryo fraction which was approximately one-fifth to one-fourth of the whole kernel by weight. The fractions were finely ground and stored in glass jars. In the vitamin test an attempt was made to follow the technic recommended by Sherman and MacArthur, 1927, 23 * for the quantitative determination of vitamin B. Young rats from the laboratory stock "This work was carried on in 1927-28. VlTAMIN-B AND VlTAMIX-G CONTENT OF CEREALS 171 TABLE 1. EFFECT OF YEAST AS SOURCE OF VITAMIN- B COMPLEX IN DIET Rat No. and sex Average daily basal food intake Average daily total food intake Test period Average daily gain or loss in weight Control animals, .4 gram yeast daily 7799. 787 cf. 754 9 . 723 cf. 626 cf. 625 9 . 616cf . 607 cf. 5999 . gms. 8.2 7.4 6.8 9.0 7.2 5.7 6.8 7.5 7.4 gms. 8.6 7.8 7.2 9.4 7.6 6.1 7.2 7.9 7.8 days 32 32 56 46 43 46 43 50 56 gms. +2.5 +4.1 + 1.8 +3.0 +2.6 +2.1 +2.2 +2.3 +20 Control animals, no yeast 771 cf . 763 9 . 755 cf . 747 9 . 724 cf. 627 cf. 624 cf. 617cf . 609 cf. 601 9 . 3.6 4.4 3.0 3.0 3.5 2.4 2.6 33 31 2.7 No supplement 30 30 45 33 31 35 46 37 37 37 TABLE 2. EFFECT OF WHOLE OATS AS SOURCE OF VlTAMiN-B COMPLEX IN DIET Rat No. and sex Daily amount whole oats Average daily basal food intake Average daily total food intake Percent whole oats was of total food intake Average daily gain or loss in weight gms. 740cf 3 739 cf 3 542 cf 1 541 9 1 619cf. 618 9. 598 cf. 597 9. 596cf ^ 595 9. 594 9. 765 cf H 764 cf. 7599. 758 cf. 781 9. 7809. 773cf. 772 9. 749 cf. 748 cf. 742 cf. 741 9. 7349. 733 5 GM. DAILY EMBRYO OF OATS ^OM. DAILY WHOLE OATS )^OM. DAILV ENDOSPERM OF OATS 770? y t GM. DAILY Rouen-CVr jon. DAILY- CLOSE-CUT EMBRYO OF OATS EMBRYO OF OATS FIG. 2. WEIGHT CURVES OF RATS RECEIVING WHOLE OATS, OATS ENDOSPERM, AND ROUGH- AND CLOSE-CUT OATS EMBRYO AS SOURCES OF VITAMIN-B COMPLEX Slightly better growth was obtained with 1/3 gram daily of rough-cut em- bryo or 14 gram daily of close-cut embryo as a source of the vitamin-B com- plex in the diet than with the same amounts of whole oats or oats endosperm. Good growth was obtained with 3 grams daily of whole oats as a source of the vitamin-B complex. Animals receiving even the smallest daily allowance of whole oats (1/4 gram daily) fared better than the animals receiving the basal diet alone. initial gain in weight followed either by a sudden decline and death when very small amounts of the vitamin were given, or by a more gradual gain or loss in weight when more of the supplement was given. It is probable that the experiments were sufficient in number to mini- 176 BULLETIN No. 369 [May, mize the effects of individual variations, especially in the cases where 1/3 or 14 gram of the supplements was given. It may therefore be concluded that the vitamin-B complex is present both in the endosperm and in the embryo of hull-less oats, occurring in greater concentration in the embryo section of the kernel. II. VITAMIN-B VALUE OF WHOLE HULL-LESS OATS, WHOLE HULLED OATS, OAT PRODUCTS, WHOLE YELLOW CORN, AND WHOLE WHEAT Young rats from the stock colony weighing 40 to 50 grams were placed in individual, raised-bottom, screen-mesh cages. The diet, as indicated on growth-curve charts, was given ad libitum. All animals were under observation as to outward signs of nutritional condition, and were weighed every four days. The food intake was recorded simultaneously with the weighing of the animal. The cereals or cereal products were incorporated in a diet that was otherwise free from vitamin B. Autoclaved yeast was used as a source of vitamin G. The yeast, 8 placed in uncovered petri dishes to a depth of about 1/2 inch, was autoclaved for five hours at 120 C. and 15 pounds pressure, dried at room temperature, and then ground to a fine powder. The autoclaved yeast was given in amounts approximating .4 to .5 gram daily. The whole hull-less oats used were carefully cleaned by hand as described on page 170. They were obtained, as were the Reid Yellow Dent corn and Turkey Red Winter wheat, from the Agronomy De- partment of the College of Agriculture. The oat products which were tested are produced from a variety of hulled oats. The hulls were re- moved from these oats by first grinding coarsely, then sifting out the hulls in a sieve. The oat products used were oat groats, "Quaker" oats, and "Quick Quaker" oats. The oat groats, dried and hulled in the com- mercial process, are slightly broken, with an accompanying loss of a portion of the germ. The "Quaker" oats and the "Quick Quaker" oats are subjected to prolonged high temperature in order to mini- mize the time of the cooking period when used for human consump- tion. The data presented in Table 6 and in the weight curves (Figs. 3 and 4) demonstrate that the growth of animals receiving whole oats or oat products in a diet otherwise free from vitamin B is always in- creased by the addition of autoclaved yeast to the diet. The acceler- "Dry powdered yeast foam obtained from the Northwestern Yeast Company, Chicago. 1931} VlTAMIN-B AND VlTAMIN-G CONTENT OF CEREALS 177 ated growth, which occurred in all animals when any level of the whole oats was supplemented by autoclaved yeast, was of short duration in the cases where lower levels of cereal were fed. The data have been DlET PERCENT CASEIN (HARRIS VITAMIN-B FREE)- 18 CRISCO 20 OSBORNE-MENOELSALTS 4 OATS 10, 15 OR 25 COHNSTARCH 45.40oo.30 COD-LIVER OIL 3 DlED EXPERIMENT ENDED 1002? 10% OATS .4 OM. AUTOCLAVED YEAST DAILY 46? 25% OATS .4GM. AUTOCLAVED YE:AST DAILY FIG. 3. WEIGHT CURVES OF RATS RECEIVING (1) OATS AS SOLE SOURCE OF VITAMINS B AND G; (2) OATS AS SOLE SOURCE OF VITAMIN B WITH AUTOCLAVED YEAST FURNISHING VITAMIN G Increased growth always resulted when .4 gram of autoclaved yeast sup- plemented the cereal of the basal diet, which included Crisco and cornstarch. tabulated, therefore, to show the rate of growth and food intake early in the experimental period, or over the first twenty days, as well as during the entire period. A minimum of 25 percent whole oats or oat products in the diet (or an amount approximating li/ grams daily) supplemented by auto- claved yeast for vitamin G is shown by these results to be necessary to 178 BULLETIN No. 369 [May, TABLE 6. EFFECTS OF VARYING LEVELS OF OATS AS SOURCE OF VITAMIN B IN DIET WHEN AUTOCLAVED YEAST WAS USED AS SOURCE OF VITAMIN G (Crisco and cornstarch were used in the diet) During first twenty days During entire test period Rat No. and sex Average daily amount cereal Average daily food intake Average daily gain or loss in weight Average daily amount cereal Average daily food intake Average daily gain or loss in weight Test period Condition at end of test 10 percent oats without autoclaved yeast gms. gms. gms. gms. gms. gms. days 940 cP .5 4.7 + 1.2 .4 3.7 - .3 38 Dead 1001 9... .6 5.6 +1.1 .3 3.5 - .1 48 Dead 980 o .4 3.9 + .8 .4 2.7 - .1 47 Paralyzed 10 percent oats plus .4 gram autoclaved yeast daily 941 cP .6 5.9 +2.3 .5 5.0 + .2 49 Dead 942 9 .6 5.9 +2.5 .5 5.3 + .2 48 Humped, wobbly 943 9 .6 5.8 +2.2 .3 3.5 + .2 53 Convulsive 981 9 .5 4.5 + 1.7 .4 3.9 + .3 54 Convulsive 996 9 .5 5.3 +2.3 .4 3.9 + .5 54 Humped, paralyzed 997 9 .5 5.2 +2.0 .3 3.4 + .2 54 Paralyzed 1002 9 ... .7 7.4 +2.4 .5 4.7 + .4 50 Paralyzed 1003 9 ... .6 6.3 + 1.8 .4 4.2 + .2 49 Convulsive; died 989 9 .5 5.1 + 1.8 .4 4.1 + .1 47 Humped, wobbly; died 15 percent oats without autoclaved yeast 925 cP .6 4.2 + 1.0 .5 3.5 + .1 39 Convulsive 990 9 .6 4.0 + 1.1 .6 4.2 + .1 264 Dead 15 percent oats plus .4 gram autoclaved yeast daily 870 cf. . . 1.0 7.0 +3.0 .9 6.2 + .5 85 Ne untie 871 cP 1.3 9.0 +2.9 .8 5.4 + .4 88 Paralyzed 878 9 1.2 8.3 ' +2.8 .9 6.0 + .4 107 Slightly neuritic 879 9 1.1 7.1 +2.1 .9 6.0 + .8 107 Slightly neuritic 944 cP .9 6.3 +2.7 .7 5.0 + .6 58 Convulsive 945 cP 1.0 6.8 +2.7 .7 4.6 + .3 58 Convulsive 926 9 .8 5.3 + 1.8 .8 5.4 + .7 79 Convulsive 927 9 1.0 6.6 +2.1 .9 6.2 + .7 85 Convulsive 1007 9 ... .9 6.0 +2.0 .6 4.3 + .4 60 Paralyzed 1006 9 ... .7 4.9 + 1.6 .5 3.3 68 Paralyzed 991 9 .8 5.4 +2.3 .7 4.6 + .3 156 Dead 983 9 .8 5.1 + 1.7 .5 3.3 + .3 79 Paralyzed 25 percent oats without autoclaved yeast 922 cP 1.3 5.1 +1.4 1.7 6.7 + .5 268 Very pellagrous, slightly neuritic 998 9 1.2 4.8 +1.3 1.6 6.5 + .5 324 Pellagrous 25 percent oats plus .4 gram autoclaved yeast daily 946 9 1.4 5.5 +2.4 2.0 7.8 + .8 274 Neuritic, symptoms improved 947 cf ... 1.3 5.1 +2.6 1.3 5.0 + .5 84 Paralyzed, dying 923 9 1.4 5.7 +2.3 2.1 8.3 +1.4 108 Good 924 cP 1.5 6.0 +2.3 2.0 8.0 + 1.1 205 Good 999 9 1.7 6.7 +2.5 2.2 8.8 +1.2 162 Fur rough 10009... 1.2 4.9 + 1.6 1.5 5.8 + 1.1 110 Good 25 percent sifted hulled oats 1032 cP. . . 1.5 6.1 + 1.1 1.2 4.8 + .5 136 Humped, pellagrous sores on mouth, priapism 1040 9 ... 1.5 6.1 +1.1 1.5 6.2 + .6 95 Humped, brown fur on head 1048 9 ... 1.8 7.3 + .8 1.3 5.1 + .3 95 Humped, flabby muscles 1931] VlTAMIN-B AND VlTAMIN-G CONTENT OF CEREALS 179 TABLE 6. Concluded During first twenty days During entire test period Rat No. and sex Average daily amount cereal Average daily food intake Average daily gain or loss in weight Average daily amount cereal Average daily food intake Average daily gain or loss in weight Test period Condition at end of test 25 percent sifted Quaker oats plus .4 gram autoclaved yeast daily 1031 cT... 1039 9 ... 1047 d"... 1.9 2.0 2.8 7.8 8.0 11.0 +2.5 +2.4 +2.6 2.5 2.6 2.4 10.1 10.4 9.7 + 1.7 +1.2 + 1.6 162 154 146 Good Good Good 25 percent Quaker oat groats 1034 9 . . 1042 d 1 . .. 1050 d 1 ... .9 1.0 1.1 3.6 3.9 4.4 +1.0 +1.1 +1.3 1.0 1.1 1.1 4.1 4.3 4.5 + .6 + .3 + .6 100 166 95 Pellagrous Pellagrous, neuritic Slightly pellagrous and slightly neuritic 25 percent Quaker oat groats plus .4 gram autoclaved yeast daily 1033d 1 ... 1041 9... 1049 d 1 ... 1.7 1.5 1.6 6.7 6.1 6.3 +3.4 +2.1 +2.5 2.1 1.8 1.9 8.6 7.2 7.7 +1.8 + 1.2 + 1.2 133 127 161 Flabby and neuritic Good, except lids red Good 25 percent Quaker oats without autoclaved yeast 1036 OR QUICK QUAKER OATS J ^EXPERIMENT ENDED 25K.QUAKER 25% QUAKEP OATS 5%Quicn ZS% QUICK QUAKER OATS OATS .4oH.AuToci.AVED QUAKERUATS .4oM. AUTOCLAVED YEAST DAILY - /x* t s\ /' /^ i*' ' 180 s IS* / r**/'*" / ^ 160 HO ISO jfru / / *tf />' y / i * /f / ^ ICO /.a / / /i f / i 10 // !/ i, ' / 1 S 1 1 f>0 ' - *j ! / t i o '/^'"Jr l * 7 ^& / isy j \>' KzTl' AV"^ 1 ' 6DROP5TIKITIKI iDtOPi TIKITIKI DAILY IZ D ^Jfi'5^"" T " DAILY Ao IOAAUTOCLAVCD "~"- T , IE DPOP5 TIKITIKI DAILY AND .4on AUTOCLAVEO YEAJT DAILY YIAST DAILY to FIG. 8. WEIGHT CURVES OF RATS RECEIVING (1) TIKITIKI AS SOLE SOURCE OF VITAMINS B AND G; (2) TIKITIKI AS SOLE SOURCE OF VITAMIN B WITH AUTOCLAVED YEAST FURNISHING VITAMIN G Good growth was obtained when 6 or 12 drops daily of tikitiki were used as the sole source of vitamins B and G in the diet. Improved growth always resulted when autoclaved yeast was given along with the tikitiki. sole source of vitamins B and G, developed neuritic symptoms which were not alleviated even when autoclaved cereal to the extent of 50 percent of the diet was fed. Six or 12 drops of tikitiki daily, in addi- tion to a diet otherwise free from vitamins B and G, produced growth somewhat below normal. The growth thus produced was stimulated either by the inclusion in the diet of 25 or 50 percent autoclaved oats, wheat, or corn, or by the addition of autoclaved yeast. The stimula- 1931] VlTAMIN-B AND VlTAMIN-G CONTENT OF CEREALS 191 SED A that a W bl pa AVED YEA tn Q>H "g w g EJ a J Q8 t th go u I B> H b B 3 "g H 8 u S 1 Condition at end of test o 1 T3 - ^1 3__"g_C js "3 g, 1 " j= MX : " M J= M 3 M >. BC 3 g 3 .ti 3 O " O g 3 O >- ^j ^ M cr * _>. _ ej_X - -j* 9T3jZ~Se-O C^'S'Si O O "3> 3 O g'S D 3^i O O - O O 3 .S o O -S ooeKOKaKKn Tail scaly, yellow, oily, neuritic; killed Rough, small, woolly fur; killed Pellagrous, priapism; killed Fur woolly Fur woolly, nervous, scaly tail and feet Woolly, humped, swollen jaws Rough, scaly tail Woolly, small, humped Woolly Woolly and humped, swollen jaws Flabby, tail rough 138 days respectively with little |1 O ?s Ov ** ^f t^ O IC>O f) *O oo m TJ-VOO oo*'t~'Ot^ ~- o c a S 1* o T) O a 8 d- 3 o to -(StNWJ^t-OOTj.^ | CMTfCN OOOOt~t>O> 00 o 3 JH i s i.s il B g 1 ++++++++++ 3n of tik +++ +++++ + D. u s 01 * S >>-oJ< autoc -5 ct 00 CN vOOi/5 >00N *l* 3 P Wj 01 J3 3 S ooxt r~Ot>O-H tNO he later s^z;2 b ili-S* a A r-" 00 Hrt 3 o >, = 3 'l a _0 V QE^ * a - o. o U O s ftllf a c 2 -OOMOMO "3 e rt T3 S"3 o -2 >T3-i. C Tj. V) *f VO V5 I 3 . O i| 5 || ______ co O (8 3 1 fiBS^R ee -- |8 3 u 3 Q i-l-i | 2a - _ o^ S2 s tiki and autoc ige in growth n c - O^O^O^O^ l b l bO^'b'bO^ o>- o 'b'b'b 'boo'b'b *b -^ ^ c n is 192 BULLETIN No. 369 ) J3 3 M 1 3 O M 3 j a fe 3 } C V o * J J "S >> 5 " _>, j CJ "^ A fin j S 4 5 o 4 O ) ^ 1 if d &!> , vO C Ov jH ^ a 13 o * 'c 3 Q SO 4 S j K 'C bfl ^? j- *> CS - t 3 rt ^* "^ ^ ^ P <' l ^ CO 1 + jUn : C gj : g V u OJ <^ * 5 S JH *- ^ O c3 clli a tr> 5 ^ tw > (-1 *^ a P c 4 w 5 5 w ^"c-- 15 rt ,j? ^ ^ 15 to ^* - HH S"3 8 S '^3 S H IE < C J3 =w '! C O JQ 5 ' >T)'3 > <; M IS Si "E u P 00 1 + 3 < 1U ^ ^ rt ^"^Jfi 4 "^ j S^ 8 -2 $ 6 CO > ^ I4_l g 00 *H j rt < 3 Z T3 ?H U HI j i M >> C "c3 ^o < >-a|S 1-1 00 C "C 3 H J P '3 o^ l l| Q S!PM a 4 rt w J H X d l-y 4; f-< m S rt C b in < o H CN O O OO V o 11 >, o .S 3 "0 J35- J3 ^-1 ff Slf|2| . e >. o i."*" U *i^ l^.l^ c^- 3 ^'a-G 111|III1 Oca^w^K^OO Slightly rough, dark tail Slightly rough Thin, nervous Rough, fur stiff and coarse Humped, rough, yellow o -Hr^Ooi'*O'* < 'tO O-* -*00\O OiO S eO(SOO>O'JOOC^ percent -HO IN IT) 00 cs^ HI 2 S vO^'OOO-H'^OC-* o T3 S -HCSOVO-H -na then 2 XOiO^t^OOOCNOO 15 13 \oo>c f>a tf, N 'S. !5 --l-H^(SCSfO<5CS(S V a 10 \O rh i/} "0 Tjlt^ X! "S lOOOOO^<5 ON <*)0) ++ epletion HCS -H TH (S f> < T- a P ** W WJ o o)O\ o oo vO^OvO^O-H-^OvH o\ o* -H *o t-~ *rsi t^t^.NOf^'-^OtOf^OO t^. so 00 -H O* (N 04 -H-n-S-#f>f>CNCN TfTj r*5 f^ f^(^)rO^OvO\OfNfS?N " O"b VOl^ o'b'bo'b'b'boo ON'JiTjiOvrtOOOv't 'b'b'b'bo 00 \C O^ O 1/5 O *O ^O r* t^ 1931} VlTAMIN-B AND VlTAMIN-G CONTENT OF CEREALS 193 8 Condition at end of test 1) -C s I ||?1 | a. ^ 01 2 o> 2-C.C : - : g3^Q> M 13j-C2 * tfrfj llllll I ^ cu O K PH ^ O O c^ O cfi F T I r T t O c^ r^j iX iX ;ntire test peri ll O O*t^-OlOlO' 'O* 1 ^^ 1 ^ OfNCOOOOOCS^^O ^ t ^2 r ^ ON 2^ZI ovc> ^ * "* ^^ ^ N * ^ be _c "C ^e twenty a^- 5 a Q Q Z b] < K Si K Oi a D a ., H 2 - a w z > o 2< >0 b Q O Z en < W 1 o o Condition at end of test Yellow, rough Good, dark tail Thin, humped, nervous 1 |l -jli O O'S 3 g O O"S o a i -= > 3 Q. ^ ^J3 5 0.13 bc"M o %"bc o. ~r, 331.53 *|. R 2 2-| 2 3 5 Very pellagrous Pellagrous, scaly tail o o c- V .^ "5 a r 5 O S A a z Si , t^NOO (MtsoOOOO-O - - be - .t E "I a B u be B o S? B - tt 3 Q fl.si-3 ^s^^ )le wheat 1 + ole wheat ?:il T3 C3 W+WPW .aved wh < I CS (N fS 0 _ oom O s *J^ a S oo 60 K a HOJS Q Q 00 V "3, 01 Q 1 X o 1^1 o. 10 ^ ^ V e o E e; a^^- _ 0000000* XX B C 3 -c be c l^p E re main 3 Q III r ,_,o^. , w ,. aag Id" and 1213 c? w< 11 o oo o-oo c*co*cx l b l bo'b fJOOUTOrMI'Tl'l/) ssssssssss: b-b 1931] VlTAMIN-B AND VlTAMIN-G CONTENT OF CEREALS 195 8 Condition at end of test Slightly rough Yellow but good Slightly rough Good i 'a .2 0. 1 n = o ^ 2 5 so Jr U S 9 3 ^ S.O J3 g 111 tt^ if b|||l||||| I 8 1 x -2 c t ** C .Sf j*> -0-0 - S.i J=J3 J-J3 ^ Bt-t !! l! f ompletely ceased after the 90th day. 1 ll . *3 H ^ 1 I C u oo -< e i Q |lli! e 8 !++ c o ++ clavcd co +++ !ji+!il+++ claved co ++++T+??W? i ?*** 8 iom percen 00 6 rt 3 cd *fOO O 000 & >-o v 2 c \r> a N \n u j: -Hrt 1 2 1 Mi 3 15 ^ - "! 13 13 tN 1 *OOOOOOv^t^-H>OTf e JS ooHtse>iiOTfO^ l ooo__ Ovf t^oo t~o>r-f- -Oio Oooi~ aoov 1 a * I-P i 3 Q ~ o *' ra w I" n , n , ssaa CT n ,, sss >o t^ c*c o XX s-ssisss^ SsiS:SS =r 196 BULLETIN No. 369 [May, tion of growth was much more marked when these autoclaved foods were given along with the 3 drops of tikitiki daily than when the larger amounts of tikitiki were used. The food consumption of animals that received cereal along with tikitiki in the diet was usually greater than that of animals that re- ceived only tikitiki for vitamins B and G (Tables 13 to 16). In the cases of control animals that received the tikitiki alone, the food intake increased when more tikitiki was given, or after the first twenty days of the experiment, when the animals were older and somewhat larger. The average daily food consumption of the control animals receiving autoclaved yeast and tikitiki and of the comparable experimental ani- mals receiving autoclaved cereals and tikitiki was about the same, both during the first twenty days of the experiment and over the entire ex- perimental period. Where the larger amounts of tikitiki were given daily along with the cereal, the food intake of animals receiving 12 drops of tikitiki was only slightly more than that of animals receiving 6 drops, and the average daily gain in weight was very similar. The animals receiving 3 drops of tikitiki daily and 25 or 50 percent auto- claved cereal, however, ingested considerably less food and grew at a slower rate than the animals with diets that differed only by the in- clusion of larger amounts of tikitiki. Considerably better growth was obtained when the diet included 25 percent untreated cereal with 3 drops of tikitiki daily than when the same amount of autoclaved cereal was given with 3 drops of tikitiki. Marked increase in food intake accompanied the use of untreated cereal, and for this reason it was difficult to determine whether in- creased growth was due to the larger intake of food, to the fact that more vitamin B was included in the diet, or to the fact that more vitamin G may be present in untreated than in autoclaved cereal. The use of autoclaved cereal eliminated the factors of increased ingestion of food and the inclusion of increased vitamin B in the diet since the appetites did not increase appreciably when autoclaved cereal free from vitamin B was given along with tikitiki. Animals whose diets consisted of 50 percent untreated oats and 3 drops of tikitiki daily grew at about the same rate as animals receiving 50 percent of oats without tikitiki, and somewhat better than animals receiving the same amount of tikitiki with 50 percent autoclaved oats. The animals receiving the autoclaved cereal, however, ingested a some- what lower amount of food over the entire experimental period and exhibited outwardly a somewhat better nutritive condition than did animals whose diets differed only in that untreated cereal was used. It 1931] VlTAMIN-B AND VlTAMIN-G CONTENT OF CEREALS 197 GMS. 100 100 80 60 40 100 eo 160 HO ieo 100 80 Z4 zzo 200 180 160 MO ISO 100 80 60 n,,-, GLUCOSE BASAL EXPERIMENTAL PERCENT PERCENT CASEIN (HABisViTAHiN-8FBtt) 18 18 OSBORN- MENDEL SALTS 4 4 GLUCOSE 78 63,-JJoREB OATS OR AUTOCLAVED OATS \S, 2^op50 Ice. COD-LIVER OIL GLUCOSE BASAU Di I TIKITIKI GIVEN DAILY OATS IN THE DIET + EXPERIMENT ENDED o DIED 3 DROPS TIKITIKI TIKITIKI TIKITIKI DAILY DAILY AND Z5% , UDm> . ........... AND 25V. AUTOCLAVED DAILY AND 50% AND 15% WHOLEOATS OATS OATS I v*|^-^ UAT5 yk^ /,,/ ,^i.^\^ M > 1162?- 50% AUTOCL ^X FIG. 9. WEIGHT CURVES OF RATS RECEIVING UNTREATED OR AUTOCLAVED OATS AS SOURCE OF VITAMIN G WITH TIKITIKI AS SOURCE OF VITAMIN B Good growth was obtained when 6 or 12 drops of tikitiki were used daily as the sole source of vitamins B and G in the diet. Improved growth always occurred when untreated or autoclaved oats were added to the diet. 198 BULLETIN No. 369 [May, n, FT GLUCOSE BASAL EXPERIMENTAL _ ,"' ' PERCENT PERCENT CASEIN (HARRIS VITAMIN -B FREE) 18 |fi OSBORNE-MCNOEL SALT 3 4 4 GLUCOSE -78 63,5Joe38 WHEAT OR AUTOCLAVED WHEAT I5'25ou50 ICC.COD-UIVEP OIL ..... OLUCOSC BASAL DIET ^ TIKITIKI GIVEN DAILY WHEAT IN THE DIET + EXPERIMENT ENDED DIED t "v^N/IDnop TIKITIKI DAILY *l DROPS TIKITIKI DAILY 2DROPST.KIT1 AND JOSAu-roc WHEAT 25%AuTocLAvrt> WHEAT FIG. 10. WEIGHT CURVES OF RATS RECEIVING UNTREATED OR AUTOCLAVED WHEAT AS SOURCE OF VITAMIN G WITH TIKITIKI SUPPLYING VITAMIN B Improved growth always occurred when untreated or autoclaved wheat was fed. 1931] VlTAMIN-B AND VlTAMIN-G CONTENT OF CEREALS 199 GLUCOSE BASAL EXPERIMENTAL DIET CASEIN (HARP u VITAMIN -B FREE) 18 18 OSBOHNE-MENDELSALT 4 4 GLUCOSE 78 63.530R38 CORN OR AUTOCLAVED CORN IS.iSoaSO Ice. COD-LIVER OIL .....GLUCOSE BASAL DIET * TIKITIKI GIVEN DAILY * AUTOCLAVEO YEAST GIVEN DAILY COON IN THE DIET + EXPERIMENT ENDED e Oico FIG. 11. WEIGHT CURVES OF RATS RECEIVING UNTREATED OR AUTOCLAVED CORN AS SOURCE OF VITAMIN G WITH TIKITIKI AS SOURCE OF VITAMIN B Improved growth occurred whenever the untreated or autoclaved cereal was added to the diet. 200 BULLETIN No. 369 [May, is thus indicated that a diet consisting of 50 percent autoclaved oats furnishes sufficient vitamin G for growth when sufficient vitamin B is included. The same was found to be true for wheat and for corn. Oats, wheat, and corn, judging from these results, are comparable in vitamin G content; rice polishings contain a considerable amount 6ns. 200 180 IfaO 140 UO 100 80 &0 GLUCOSE BASAL PCQCENT CASEIN (MAORIS VITAMIN -B FREE) 18 OSBOBNE- MENDEL SALTS 4 GLUCOSE 78 AUTOCLAVED RICE. POLISHINGS Ice. COD-LIVER OIL GIVEN EVEPY FOUR DAYS GLUCOSE BASALDIET AUTOCLAVED RICE POLISHINGS IN THE DIET ^TIKITIKI GIVEN DAILV EXPERIMENTAL PERCENT IB -28 -50 "\/* + 50%AL)TOCLAVEO RICE POLISHINGS 3 DROPS TIKITIKI DAILY AND 50% AUTOCLAVED RICEPOLISHINOS FIG. 12. WEIGHT CURVES OF RATS RECEIVING TIKITIKI, AUTOCLAVED RICE POLISHINGS, AND TIKITIKI AND AUTOCLAVED RICE POLISHINGS AS SOURCES OF VITAMINS B AND G The good growth obtained with tikitiki and autoclaved rice polishings in- dicates the presence of vitamin G in the rice polishings. of vitamin G. This last finding may be of considerable significance since tikitiki was used thruout the present investigation as a source of vitamin B. Evans and Burr 10 * considered tikitiki a potent source of vitamin B but almost totally lacking in vitamin G. According to Chick and Roscoe 5 * vitamin B! (B), the antineuritic vitamin, is soluble in all concentrations of alcohol while vitamin B 2 (G) is soluble only in alcohol concentrations of less than 83 to 93 percent by weight. In view of the fact that tikitiki is a dilute alcoholic extract of rice polishings, it seems possible that it may contain a considerable amount of vitamin G, particularly since it has been shown here that rice polishings contain this vitamin. The fairly good growth obtained when large amounts of tikitiki were fed as the sole source of vitamins B and G support the observation that tikitiki contains vitamin G. Further work is in prog- ress in an attempt to determine the vitamin-G content of tikitiki. 1931] VlTAMIN-B AND VlTAMIN-G CONTENT OF CEREALS 201 IV. VITAMIN-B AND VITAMIN-G VALUE OF WHOLE HULL-LESS OATS FOR LACTATION In testing the vitamin-B and vitamin-G value of whole hull-less oats for lactation, adult albino rats were maintained on the diets described in Table 17, in most cases during and for a month preceding preg- nancy, and in all cases during the lactation period. Before remating, the rats were kept on the test diet for one month following lactation ; as a rule each animal was used for not more than three lactation tests. The animals were mated for four-day periods; the lactation period TABLE 17. PERCENTAGE COMPOSITION OF DIETS USED IN RAT LACTATION TEST (All diets included cod-liver oil given separately in 1-cc. amounts semi-weekly) Diet Whole hull-less oats Corn- starch Osborne- Mendel salt mixture Casein (Harris vitamin-B free) Auto- claved yeast Tikitiki daily 1 perct. 50 percl. 28 percl. 4 perct. 18 percl. drops 2 78 4 18 3 50 13 4 18 15 4 50 28 4 18 6 5 . 25 38 4 18 15 6 25 23 4 18 30 7 25 38 4 18 15 6 terminated when the young were 21 days old. On the third day after the birth of the young, the litters were reduced to six. Raised-bottom cages were used thruout the entire experiment. Spe- cially designed cages, as shown in Fig. 13, were used during the lacta- tion periods. The weights and food intake of the mothers were recorded at four-day intervals during the entire experimental period. During lactation the young were weighed daily; they were considered normal if they weighed 40 grams when they were 21 days old. The diets that contained whole hull-less oats as a sole source of vitamins B and G proved inadequate for successful lactation (Table 18, Fig. 14). The young of mothers on diets that included 50 or 78 percent oats (Diets 1 and 2) grew and developed at a rate slightly below normal for a period of 10 to 14 days, at which time there was a cessation of growth followed by decreased activity and loss of weight. The young animals sometimes cried out and ran blindly about the cage. Sure 27 * describes this condition as "screaming, running fits." The muscles twitched or became flabby. A typical animal is pictured in Fig. 15. Death usually resulted before the rats were three weeks old. Success in lactation was obtained with Diet 3, which differed from Diet 1 in that autoclaved yeast, a potent source of vitamin G and 202 BULLETIN No. 369 [May, totally lacking in vitamin B, was included with the oats (Table 19, Fig. 16). Litters of rats whose mothers had this diet for the first time were not always normal in weight, possibly because the effect of preceding diets had not been fully compensated by Diet 3 during the time of its ingestion. In the cases of Rats 971 and 1223 the second litters on Diet 3 were heavier at three weeks of age than were the young of the first litter raised on this diet. Young of mothers receiv- ing Diet 3 grew rapidly after weaning, so that even in the litters where the young were below normal weight at three weeks, they soon made up for the weight deficiency. In all cases the young of mothers on this diet 50 percent oats and 15 percent autoclaved yeast for vitamins B and G appeared to be in better nutritive condition than the young of rats on any of the other diets tested. When Diet 4 (6 drops daily of tikitiki and 50 percent oats for vita- mins B and G) was used, there was improvement in lactation over the re- sults when Diets 1 and 2 were used (Table 20). The young raised by mothers on Diet 4 did not appear to be as healthy nor did they gain as rapidly after weaning as the young of mothers on Diet 3. Results reported in Part III of this bulletin showing good growth with 6 drops of tikitiki daily as the sole source of vita- mins B and G, indicate that vitamin G is present in tikitiki. In Diet 4 it seems likely that it was the vitamin G of tikitiki which supplemented the oats sufficiently to provide for successful lactation. This conclusion is strengthened by the fact that successful lactation occurred with Diet 3 when autoclaved yeast, a source of vitamin G free from vitamin B, was added to the oats diet. Diets including 25 percent oats, even when supplemented by 15 or 30 percent autoclaved yeast (Diets 5 and 6), were inadequate for suc- cessful lactation (Table 21). When 6 drops of tikitiki were given daily with 25 percent oats and autoclaved yeast (Diet 7), two litters grew almost normally, but again they were not so well developed as the young of mothers on Diet 3. No tests were made using diets con- FIG. 13. TYPE OF CAGE USED DURING LACTATION PERIOD The animals were left in small cages until just a few days before the young were born. At this time they were transferred to the large cage and allowed to remain thruout the period of lactation. 1931] VlTAMIN-B AND VlTAMIN-G CONTENT OF CEREALS 203 TABLE 18. EFFECTS OF DIETS 1 AND 2 ON LACTATION IN THE RAT, WHOLE OATS ALONE SUPPLYING VITAMINS B AND G Average weight Condition of Rat No. Litter of young young at at 21st 21st day day Diet 1; 50 percent oats 973 1st gms. Dead All dead by 1 7th day after ap- 891 1st Dead parently normal initial growth 1227.. . 1st Dead sive muscles and collapse preceded death All dead by 3d day 913 1st Dead All but 2 dead by 17th day 2 1039 1st Dead lived 3 weeks, then died All dead by 16th day 1035 1st Dead All dead by 16th day 1059 1st Dead All dead or dying by 17th day 971 1st Dead 1223.. . 1st Dead in week following All dead by 17th day 895 2d 26 5 l 895 3d Dead All dead or dying by 17th day Diet 2; 78 percent oats 1035 1 2d 30 Small but good 1035 3d Dead 1227 2d Dead All dead by 16th day 1227 3d Dead All dead by 18th day __ 'First litter with Diet 1 and egg yolk. 'First litter with Diet 1. taining oats at levels between 25 and 50 percent for the sole source of vitamin B, but it is probable that 50 percent oats is the least that will provide for optimum lactation since even 6 drops of tikitiki daily, in addition to Diets 5 and 6, did not furnish enough vitamin B to make these diets entirely comparable with the 50 percent oats and 15 percent autoclaved yeast diet (Diet 3). TABLE 19. EFFECT OF DIET 3 ON LACTATION IN THE RAT, WHOLE OATS (50 PERCENT) AND AUTOCLAVED YEAST (15 PERCENT) SUPPLYING VITAMINS B AND G Average weight Condition of Rat No. Litter of young young at at 2 1st 21st day day gms. 1000 1 . . 2d 32 971* 2d 37 971 3d 40 1223' 2d 24 1223 3d 37 1225' 3d 34 1036* 3d 41 1222 5 3d 40 1463 1st 32 Small but very good 'After 1 unsuccessful litter with Diet 5. 'After 1 unsuccessful litter with Diet 1. 'After 2 unsuc- cessful litters with Diet 1 and lettuce. 'After 2 unsuccessful litters with Diet 1 and lean beef. 'After 2 unsuccessful litters with Diet 1 and egg white. "This animal was raised on a diet in which 50 percent oats and .4 gram autoclaved yeast daily supplied vitamins B and G. Diet 3 was not given before the animal was pregnant. An abnormal condition of the tail made it necessary to kill the animal before a second litter was produced. 204 BULLETIN No. 369 [May, \ < B I w '-5 S a Q o j2 o Q '* < 2 M P T3 U g < rt H w < E 1 P u . O T3 . C 1931] YlTAMIX-B AXD VlTAMIX-G CONTEXT OF CEREALS 205 It may be concluded from the above observations that it is possible to include enough oats (50 percent) in the diet of a rat to furnish sufficient vitamin B for successful lactation. On the other hand, it FIG. 15. TYPICAL YOUXG OF RAT 913 ox DIET 1 WHICH IXCLUDED 50 PERCEXT OATS FOR VITAMINS B AXD G When the picture was taken, this animal was 17 days old and weighed 16 grams. It died the same day. was found impossible to furnish sufficient vitamin G for successful lactation when oats were the sole source of this vitamin in the diet, as demonstrated by the lack of success with both Diets 1 and 2. In the latter case 78 percent oats were used, or the maximum amount pos- sible without altering the composition of the diet with respect to salt TABLE 20. EFFECT OF DIET 4 ON LACTATION IN THE RAT, WHOLE OATS (50 PERCENT) AXD TIKITIKI (6 DROPS) SUPPLYING VITAMIXS B AND G Average weight Condition of Rat No. Litter of young young at at 21st 21st day day gms. 1039 1 . 2d 29 Small but good 10591 2d 31.5 Small but good 1473 1st 40 Good (5 young) 1643 1st Dead All dead by 7th day >First litter with Diet 1 was unsuccessful. mixture and casein content. The above findings are particularly sig- nificant since the results reported in Part II of this bulletin demon- strate that a diet including 25 percent oats when supplemented with autoclaved yeast is adequate in vitamin B for growth approaching normal, while the results discussed in Part III indicate that a diet in- cluding 50 percent oats is adequate in vitamin G for good growth. Macy, Outhouse, and Long 15 * in 1927 stated that three to five 236 BULLETIN No. 369 [-Way, times as much vitamin B is necessary for lactation as for growth. These investigators used yeast as the source of vitamin B but the work was carried on before the final recognition of vitamin G. Apparently it was the vitamin G of yeast which was an important factor in promot- ing successful lactation. Evans and Burr 9 * in 1928 made the follow- FIG. 16. TYPICAL YOUNG OF RAT 971 ON DIET 3 WHICH INCLUDED 50 PERCENT OATS AND AUTOCLAVED YEAST FOR VITAMINS B AND G When the picture was taken this animal was 21 days old and weighed 40 grams. ing statement: "The indications are that when yeast is the accessory source of vitamin B about five times the usual intake is required dur- ing the latter part of the lactation period." They state further that "the additional yeast needed for lactation is solely due to its addition TABLE 21. EFFECTS OF DIETS 5, 6, AND 7 ON LACTATION IN THE RAT, WHOLE OATS AND AUTOCLAVED YEAST OR WHOLE OATS, TIKITIKI, AND AUTOCLAVED YEAST SUPPLYING VITAMINS B AND G Rat No. Liter Average weight of young at 21st day Condition of young at 2 1 s-t day Diet 5; 25 percent oats, 15 percent autoclaved yeast 1000.. . 1st 1st 1st 2d gms. Dead Dead 20.5 Dead All dead by 16th day All dead by 17th day 4 young, small All dead by 12th day 1041.. 1034 1034 Diet 6; 25 percent oats, 30 percent autoclaved yeast 1041. . 2d 3d Dead Dead Lived 4 days Lived 2 days 1041 Diet 7; 25 percent oats, 15 percent autoclaved yeast, 6 drops tikitiki daily 1034. . . 3d 4th 32.7 25.5 Good Grew slowly 1034 1931] \~ITAM1N-B AND VlTAMIX-G CONTEXT OF CEREALS 207 to the antineuritic vitamin B and not to the growth-promoting vitamin B of the diet, for when tikitiki is given to lactating mothers without increased yeast dosage we can also produce normal lactation." These investigators used tikitiki and yeast as sources of vitamins B and G, but at that time it was thought that tikitiki contained no vitamin G. Sure 27 * in 1928, using rice polishings and autoclaved yeast as sources of vitamins B and G, concluded that both these vitamins are necessary for successful lactation. Hussemann and Hetler 14 * showed that equally successful lactation is possible when 5 percent yeast and 15 percent TABLE 22.- AVERAGE DAILY FOOD INTAKE OF RATS AS AFFECTED BY VARIATIONS IN SOURCE OF VITAMINS B AND G IN DIET DURING RESTING, PREGNANCY, AND LACTATION Rat No. Litter Average daily food intake during resting Average daily food intake during pregnancy Average daily food intake during lactation Diet 1; 50 percent oats gms. gms. gms. 1059 l?t 12.3 15.2 19.3 973 1st 13.8 15.0 14.8 1039 1st 13.5 10.6 13.5 1227 1st 12.8 7.0 971 1st 15.2 895 3d 11.4 10.4 12.6 1223 1st 12.5 15.0' Diet 2; 78 percent oats 1035 2d 15.4 12.5 24.4 1035 3d 13.4 13.5 15.6 1227 2d 11.1 14.3 17.4 1227 3d 13.8 14.4 12.1 Diet 3; 50 percent oats, 15 percent autoclaved yast 971 2d 17.02 132 24.0 971 3d 11.0 12.2 31.0 1223 2d 9.6 13.5 21.0 1223 3d 13.5 13.8 26.2 1086 3d 17.5 17.5 27.6 1000 2d 12.3' 15.3 19.0 Diet 4; 50 percent oats, 6 drops tikitiki daily 1059 2d 17. 3 J 15.2 21.8 1039 2d 14.6' 14.6 16.3 1473 1st 9.7 24 9 Diet 5; 25 percent oats. 15 percent autoclaved yeast 1000 1st 9.8 16.5 22.2 1034 1st 12.8 17.0 20.9 1034 2d 15.0 15.0 12.7 1041 1st 13.5 18.8 Diet 6; 25 percent oats, 30 percent autoclaved yeast 1041 2d 15. 2" 11.4 8.0 1041 3d ! 14.6 15.0 13.3 Diet 7; 25 percent oats, 15 percent autoclaved yeast, 6 drops tikitiki daily 1034 3d 12.1 16.0 25.7 1034 4th 11.2 16.0 25.1 'Diet 3 given at end of first week of lactation. 'Diet 1 during resting period. 'Diet 5 during resting period. 208 BULLETIN No. 369 [May, autoclaved yeast furnish vitamins B and G, as when 15 percent yeast is used for these vitamins. They have also demonstrated better success in lactation with 5 percent yeast and tikitiki than with 5 percent yeast alone. But before the results with tikitiki as a supplement can be evaluated in terms of vitamins B and G, it will be necessary to have more exact data concerning the vitamin-G content of this product. The results obtained by Hussemann and Hetler and the results of the present investigation indicate, however, that more vitamin G is neces- sary for successful lactation than for normal growth. Roughly one might estimate that about twice as much vitamin B and three or four times as much vitamin G are required for lactation as for growth. A study of the results here presented of the use of oats as a source of vitamins B and G for lactation is not complete without a considera- tion of the food intake of the animals (Table 22). The average daily food consumption for any one animal was almost constant during the resting and pregnancy periods. A slight increase during pregnancy occurred in some cases, but with other animals less food was eaten during pregnancy than during the resting period. A marked rise in food intake was noted, however, during the lactation period in the cases where success in the development of the young resulted. When Diets 1 or 2, 50 or 78 percent oats, were fed, or when Diets 5 and 6, 25 percent oats with autoclaved yeast, were given, there was little if any increase in food consumption during the lactation period. Lacta- tion was unsuccessful with these diets, in which vitamin B or vitamin G was the limiting factor. Whether the low food intake was entirely responsible for the failure in the nursing of the young, or whether the lack of stimulation to appetite was a direct contributing factor owing to the deficiency of vitamins B and G, is a subject for further investi- gation. Whenever lactation was successful, however, a marked stimu- lation of appetite and rise in food intake of the mother always oc- curred during the nursing period. SUMMARY The vitamin-B and vitamin-G value of oats and oat products, corn, wheat, rice polishings, and tikitiki, was studied by determining the growth rate of albino rats when the cereal or cereal product furnished the sole source of these vitamins in the diet. Vitamins B and G, origi- nally known as vitamin B and later as the vitamin-B complex, were both found to be present in the cereals and cereal products studied. The work of Smith and Hendrick (1926), 24 * of Sherman and Axtmayer, 22: and of Hunt (1928), 13 * showing that vitamin G is the limiting factor 1931} VlTAMIN-B AND VlTAMIN-G CONTENT OF CEREALS 209 of the vitamin-B complex of cereals, was confirmed. It was found, however, that a diet including SO percent of any of the cereals would supply enough of the heat-stabile vitamin G for approximately normal growth, while 25 percent of any of the cereals provided enough vita- min B. It was indicated that vitamin G is present in tikitiki. In earlier work in the Home Economics laboratory at the Univer- sity of Illinois, before the identification of vitamin G, a study was made of the location of the vitamin-B complex in oats. It was found that vitamin B, old nomenclature, is distributed thruout the oat kernel but occurs in greater concentration in the embryo than in the endo- sperm section. Study was made of the vitamin-B and vitamin-G value of oats for lactation in the rat. If a diet containing 50 percent oats was used for vitamin B, successful lactation occurred. However, when oats were the sole source of vitamin G in the diet, it was impossible to furnish suffi- cient vitamin G for successful lactation. Methods are described for testing for vitamin B and for vitamin G. Autoclaved yeast was used to furnish vitamin G in the vitamin-B test, and tikitiki was used to furnish vitamin B in the vitamin-G test. A basal diet which included vitamin- free casein, Osborne-Mendel salt mixture, and glucose was found satisfactory both for vitamin-B and for vitamin-G determinations. Cod-liver oil was fed separately. When a diet containing Crisco and cornstarch instead of glucose was used for the vitamin-B tests, a somewhat higher level of any of the cereals was necessary to furnish sufficient vitamin B for good growth. 210 BULLETIN No. 369 [May, LITERATURE CITED 1. AYKROYD, W. R., and ROSCOE, M. H. LV. The distribution of vitamin B 2 in certain foods. Biochem. 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Are the antineuritic and the growth-promoting water-soluble B vitamines the same? Jour. Biol. Chem. 43, 265. 1920. 9. EVANS, H. M., and BURR, G. O. On the amount of vitamin B required dur- ing lactation. Jour. Biol. Chem. 76, 263. 1928. 10. A new differentiation between the antineuritic vitamin B and the purely growth-promoting vitamin B. Jour. Biol. Chem. 77, 231. 1928. 11. GOLDBERGER, J., and TANNER, W. F. A study of the pellagra-preventive action of dried beans, casein, dried milk, and brewers' yeast, with a considera- tion of the essential preventive factors involved. U.S. Pub. Health Rpts. 40, 54. 1925. 12. HAUGE, S. M., and CARRICK, C. W. A differentiation between the water- soluble growth-promoting and antineuritic substances. Jour. Biol. Chem. 69, 403. 1926. 13. HUNT, C. H. The complex nature of vitamin B as found in wheat and corn. Jour. Biol. Chem. 78, 83. 1928. 14. HUSSEMANN, D. L., and HETLER, R. A. The vitamin B and G requirements of lactation. Jour. Nutrition 4, 127. 1931. 15. MACY, I. G., OUTHOUSE, J., LONG, M. L., and GRAHAM, A. Human milk studies. I. Technique employed in vitamin studies. Jour. Biol. Chem. 73, 153. 1927. 16. McCoLLUM, E. V., SIMMONDS, N., and PITZ, W. The nature of the dietary deficiencies of the wheat embryo. Jour. Biol. Chem. 25, 105. 1916. 17. The nature of the dietary defici- encies of the oat kernel. Jour. Biol. Chem. 29, 341. 1917. 18. MITCHELL, H. H. On the identity of the water-soluble growth-promoting vitamine and the antineuritic vitamine. Jour. Biol. Chem. 40, 399. 1919. 19. MUNSELL, H. E. Rice polishings as a source of vitamin B. Jour. Home Econ. 21, 124. 1929. 20. OSBORNE, T. B., and MENDEL, L. B. The nutritive value of the wheat kernel and its milling products. Jour. Biol. Chem. 37, 557. 1919. 21. SALMON, W. D., GUERRANT, N. B., and HAYS, I. M. On the existence of two active factors in the vitamin B complex. II. Jour. Biol. Chem. 76, 487. 1928. 1931] VlTAMIN-B AND VlTAMIX-G COXTENT OF CEREALS 211 22. SHERMAN, H. C, and AXTMAYER, J. H. A quantitative study of the problem of the multiple nature of vitamin B. Jour. Biol. Chem. 75, 207. 1927. 23. - and MACARTHUR, E. H. A quantitative stud}- of the deter- mination of vitamin B. Jour. Biol. Chem. 74, 107. 1927. 24. SMITH, M. I., and HENDRICK, E. G. Some nutrition experiments with brew- ers' yeast. U.S. Pub. Health Rpts. 41, 201. 1926. 25. STEENBOCK, H., KENT, H. E., and GROSS, E. G. The dietary qualities of barley. Jour. Biol. Chem. 35, 61. 1918. 26. - - SELL, M. T., and NELSON, E. M. Vitamine B. I. A modified technique in the use of the rat for determinations of vitamine B. Jour. Biol. Chem. 55, 399. 1923. 27. SURE, B. Dietary requirements for fertility and lactation. XIV. A quanti- tative biological method for the study of vitamin B requirements for lac- tation. Jour. Biol. Chem. 76, 673. 1928. 28. Dietary requirements for fertility and lactation. XX. A differ- entation of the vitamin B complex in rice polishings as evidenced in studies of lactation. Jour. Biol. Chem. 80, 297. 1928. UNIVERSITY OF ILLINOIS-URBANA Q.630.7IL6B C002 BULLETIN. URBANA 364-3761931-32 2019529202