THE OCCURRENCE OP A PELLAGRA-LIKE SYNDROME IN CHICKS*

Transcription

1 THE OCCURRENCE OP A PELLAGRA-LIKE SYNDROME IN CHICKS* A. T. RINGROSE, L. C. NORRIS AND G. P. HEUSER From the Laboratory of Poultry Nutrition, Department of Poultry Husbandry, Cornell University Agricultural Experiment Station, Ithaca, New York (Received for Publication ) This laboratory has been attempting to obtain further information concerning the cause of a peculiar paralysis 1, 2 of nutritional origin involving the legs and feet of chicks. In continuing the study of this paralysis it was decided to purify the casein used not only to obtain a more uniform product in respect to growth-promoting characteristics but also to determine whether this might not be a means of increasing the amount of paralysis. Commercial casein was therefore extracted with 0.2 per cent acetic-acid solution for 6 days. This acetic-acid solution was changed twice every 24 hours. On the seventh day the casein was washed four times with tap water and finally dried over a steam radiator under a stream of air. It was also decided to study egg albumin, Merck's impalpable powder, with the thought that this material might be deficient in the anti-paralytic factor, although reported rich in vitamin GP. White Leghorn chicks of the Cornell strain were used in this experiment. These were selected for size and vitality approximately 24 hours after hatching, divided into 7 lots of 25 chicks each, and individually weighed into the experiment. These chicks were each weighed weekly thereafter at which time observations were made of any signs of nutritional disturbances. A weekly record of feed consumption was also made. The experimental rations fed these lots are presented in Table I. The ingredients used in these rations were analyzed for protein and moisture and combined in such a way that the protein content of each ration was 25 per cent dry basis. This was neces- * Read at the annual meeting of POULTKY SCIENCE ASSOCIATION held at JWacdonald College, St. Anne de Bellevue, Quebec, July 9-11,

2 A PELLAGRA.-LIKE SYNDROME 167 sary because of the high moisture content of the milk vitamin concentrate* used in lot A. TABLE I EXPERIMENTAL RATIONS Ingredients A B C D B F G~~* % % % % % % %~ Yellow corn meal Flour wheat middlings Commercial casein _ Purified casein Egg albumin _ Autoclaved yeast Milk vitamin concentrate Steamed bone meal Cod-liver oil Salt At the conclusion of this study it was found that there was sufficient feed remaining from lots E, F and G to check the results obtained. Therefore, these lots were repeated for a like period of time. The lots in the second experiment were designated E 2, F 2 and G 2. The results of experiment 1 were checked a second time by using a different supply of dried egg albumin from the same source and purified casein prepared similarly to that used in the first two experiments except that the washed casein was dried more rapidly without the aid of a stream of air. The lots in this experiment were designated E 3, F 3 and G 3. The rations fed these lots were the same as those used previously except for minor adjustments to maintain the protein content at 25 per cent dry basis, made necessary by natural variations in the composition of the ingredients used. Lot * XX Liquor, manufactured by the Dry Milk Co., New York City.

3 .168 POULTRY SCIENCE The growth results of these experiments are presented in figure 1. These results are weighed for sex influence. The growth results of lots G and G 2, given commercial casein, as compared with lots E and E 2, given purified casein, show that extraction with 0.2 per cent acetic-acid solution and sub- FlGUBE 1 Growth Results sequent drying in a stream of air materially reduced the growth-promoting properties of the commercial casein used in experiments 1 and 2. Little effect of this nature, however, was obtained in the third experiment. Whether or not this was due to difference in method of drying the extracted casein or to difference in method of manufacture can not be determined. The casein used in the first two experiments was of American manufacture while that used in the third experiment was imported from Argentina. Supplementing the purified casein, experiment 1, with 2.5, 5 and 10 per cent of autoclaved yeast caused remarkable growth responses. At the conclusion of the experiment lot B, which received 10 per cent of autoclaved yeast, possessed an average weight approximately 4 times that of lot E, fed purified casein only, and exceeded the generally accepted normal weight of this species. The use of 10 per cent of milk vitamin concentrate caused lot A to develop at a rate approximately normal and nearly equal

4 A PELLAGRA-LIKE SYNDROME 169 to lot C which received 5 per cent of autoclaved yeast. This indicates that the milk vitamin concentrate was nearly as strongly growth-promoting as 5 per cent of autoclaved yeast. Lots G, Gr 2 and G 3 which received commercial casein developed at a rate intermediate between lots E, E 2, F, F 2 and F 3 and lot D showing that, while commercial casein was more growth-promoting than purified casein and dried egg albumin, the effect was not equal to that obtained by the use of 2.5 per cent of autoclaved yeast. Lots F, F, and F 3, fed dried egg albumin, grew somewhat faster at the start than lots B, E 2 and B 3, given purified casein, but later fell behind these lots owing to the development of external lesions which will be described subsequently. These results show that the chick has an intense requirement for a heat-stable growth-promoting factor present in yeast and other foot materials and corroborate the results of Hauge and Carrick 4, reported in 1926, which indicated that the chick requires a growth-promoting as well as an antineuritic factor for maintenance of life and growth. The grams of feed required to produce a gram of gain are given in table II. Lot B, which received 10 per cent of autoclaved yeast, made the most efficient utilization of food while the poorest food utilization was made by lots F, F 2 and F 3, which were given dried egg albumin. The utilization of food by lots E, E 2 and E 3, which received purified casein, was very poor but much better than the lots given dried egg albumin. More food per gram of gain was required in the last week of the experimental period than in the first week in all lots except those which were given commercial casein. This casein appeared to contain just enough of the growth-promoting factor to maintain the ratio of food required for maintenance and food required for growth relatively constant for the experimental period of 12 weeks. Had the experiments been conducted for a longer period of time, no doubt the efficiency of food utilization would have decreased in accordance with the laws of growth, as in the other lots. The efficiency of food utilization of lots F, F 2 and F 3, given dried egg albumin, was better during the first week of growth than the comparable lots E, E, and E 3 which received purified

5 170 POULTRY SCIENCE casein and in two instances better than the comparable lots which received commercial casein, but decidedly poorer than these lots during the twelfth week of growth. Lot A B C D E, E 2) F, F 2, G.G,, E s F 3 G 3 TABLE II GRAMS FEED REQUIRED PER GRAM OF GAIN First Week Twelfth Week All 12 Weeks Nutritional paralysis 1, 2 appeared in all lots except lot B which was given 10 per cent of autoclaved yeast and lots F, F 2 and F 3 which received dried egg albumin. The paralysis, however, affected only a very few chicks in lot A which received the milk vitamin concentrate and in lot C which was given 5 per cent of autoclaved yeast. In the other lots, although not all the chicks were paralyzed, the number affected was quite large. The data show in general that the amount of paralysis decreased with increase of the autoclaved-yeast supplement. In addition to the results already described external lesions of a distinctly pellagrous character 5, e, 7, 8, 9 appeared at about 3 weeks of age in the chicks of lots F, F 3 and F 3, which received egg albumin, followed shortly by the appearance of similar lesions among the chicks of lots E, E 2 and E 3, given purified casein, G, G 2 and G s, given commercial casein, and D, given purified casein plus 2.5 per cent of autoclaved yeast. These lesions developed at the eyes, at the mouth corners and on the feet. The margins of the eyelids became granular and contracted so that vision was restricted. A viscous exudate was produced which frequently caused the eyelids to stick firmly together. Crusty scabs appeared at the corners of the mouth. These gradually enlarged so as to involve the margins of the skin around the nostrils and underneath the lower mandible. The outer layers of the skin on the bottoms of the feet and between 3.97

6 A PELLAGRA-LIKE SYNDROME m the toes peeled off. Afterwards, small cracks and fissures appeared at these points. These enlarged and deepened so that chicks affected were sensitive to walking. In the meantime, the remaining skin layers thickened and cornified and in some cases wart-like protuberances developed on the balls of the feet. Figures 2, 3, 4 and 5 are good representations of these pellagra-like lesions and of their effect on chicks. None of these lesions appeared in lots C and B which received 5 and 10 per cent of autoclaved yeast, respectively, nor in lot A which received 10 per cent of milk vitamin concentrate. The lesions, therefore, could not be laid to the fineness of subdivision of the rations used or to the floor of hardware cloth upon which the chicks were confined. PlGL'liE 2 Chick 7 weeks old, showing pellagra-like granulation of the eyelids and scabby encrustation of the mouth corners. Feathering was retarded and the feathers produced were rough and staring. At first no loss of down or feathers comparable to the loss of hair found in pellagra in rats was obtained but later, in a group of chicks 18 weeks of age suffering from an insufficiency of the antipellagric factor, complete loss of feathers occurred in limited areas on the head and neck. Post-mortem examination of chicks that died frequently showed the presence of a pus-like substance in the mouth and of an opaque, grayish-white exudate in the proventriculus. Eroded lesions were found in the gizzard and the duodenum usually showed severe enteritis. The entire intestinal tract was almost entirely devoid of food residues. The small intestines lacked tonicity and appeared atrophic. In many cases portions of the small intestines and the caeca were distended with gas. The liver frequently possessed an abnormal color, varying from a faint yellow to a deep dirty yellow. In a few instances

7 172 POULTRY SCIENCE the liver was hypertrophied. The spleen, on the other hand, seemed small and atrophic. The kidneys revealed a tendency to enlargement and appeared grayish-white or inflamed and hemorrhagic. Boas 10 in 1927 reported the production of symptoms in rats by feeding air-dried egg albumin similar to those just described, but did not obtain these symptoms on egg elbumiu, coagulated before drying. In the experiments reported here the experi- PlGURE 3 Chick 5 weeks old, showing pellagra-like cornification of the skin between the toes and development of Assures. ments reported here the egg albumin used was air-dried. This may explain why results were obtained contrary to those of Aykroyde and Roscoe 3, as they no doubt used coagulated fresh egg albumin. Boas, however, did not believe that the symptoms in her rats were pellagrous but her description of these lesions is similar to pellagra in rats as described by Salmon' 0, who used purified casein in his basal diet. The pellagra-like syndrome in chicks produced at this laboratory also corresponds closely to Salmon's description of pellagra in rats. This pellagra-like syndrome in chicks was produced both by means of dried egg albumin and by means of purified casein. The symptoms in both cases were almost exactly the same except that on dried egg albumin they

8 A PELLAGRA-LIKE SYXDROME I73 were more severe, of longer duration, and involved a larger number of chicks. The parts of the body affected were identical on both protein substances. It is probable, therefore, that the syndrome in rats produced by Boas was true pellagra but of a particularly severe character. Because of the work of Boas and that of this laboratory the possibility suggests itself that pellagra in rats as obtained by the use of puriled casein is in general brought about not by a FlGUBK 4 Chick 5 weeks old, showing pellagra-like cornification of the skin on the bottoms of the feet and development of deep fissures. (Same chick as in figure 3.) complete deficiency of the antipellagric factor but rather by an insufficiency. Moreover, the results obtained here indicate that this factor is so strongly absorbed by casein as to make its complete removal extremely difficult. The mortality record in the different lots of these experiments is presented in table III. The heaviest mortality occurred in lots F, F 2 and F g, which rceived egg albumin, amounting to 90.9 per cent, 84.0 per cent and 78.3 per cent, respectively. However, mortaility was also heavy in lots B, E 2 and E 8, which received purified casein, being 58.3 per cent, 56.0 per cent and 45.8 per cent,

9 174 POULTRY SCIENCE respectively. In lots G, G 2 and G 3, given commercial casein, the mortality was 16.7 per cent, 8.0 per cent and 16.7 per cent, respectively. In the other lots there was either no mortality or it was insignificant. Period (4 wks.) (each) > B C D B TABLE III REOOED OP MORTALITY 1 Lot E = E 3 F F F 3 G G, G , Total Per Cent * Chicks which died the first week of these experiments or from accidental causes or were killed for experimental purposes are not considered as being in the experiments in calculating per cent mortality. From the results of these experiments it is difficult to believe that growth-promoting vitamin G and antipellagric vitamin G are a single entity. The egg-albumin ration was more strongly growth promoting at the start than the purified-casein ration. At 3 weeks of age the average weight of lots F, F 2 and F 3, which received dried egg albumin, exceeded the average weight of the comparable lots E, E 2 and E 3, receiving purified casein, to such an extent that the differences in the mean weights of the lots were respectively 2.84, and 3.99 times greater than the probable error of the differences. The difference in the mean weight for all these experiments combined was times its probable error. The difference in growth between these lots at 3 weeks of age is therefore highly significant. This greater growth was not due to greater food consumption during this period but to- more efficient food utilization, particularly during the first week of growth as already pointed out. Therefore, the greater growth can not be explained on the basis of greater palatability of the egg-albumin ration. Yet when the chicks' body reserve of the antipellogrie factor was reduced by this more rapid growth, the development of the pellagrous symp-

10 A PELLAGRA-LIKE SYNDROME 175 loins caused tremendous mortality and eventually brought about a rate of growth either no better or less than that of the chicks which received the purified casein ration and a utilization of food much less efficient. On the other hand, the intensity of the pellagra-like condition was much more severe in the egg-albumin lots than in the purified casein lots. After weighting the results of the individual weekly observations of the the chicks in these lots, if the degree of pellagra, in lots F, F 2 and F 8, given dried egg elbumin, is assumed as 100, the intensity of the pellagrous condition in lots E, E.. and E,, given purified casein, was 25.9, 18.2 and 20.1, respectively, and in lots G, 6 2 and G. given commercial casein, 13.5, 14.9 and 9.9, respectively. FIGUHE 5 Chick 18 weeks old, showing loss of feathers on face and neck due to a marked insufficiency of the antipellagric factor in its ration. If the egg-albumin ration had more of the growth-promoting factor and less of the antipellagric factor than the purified casein ration, then the faster early growth and the severer pellagrous symptoms of the egg-albumin lot are explainable. This reasoning points to the conclusion already suggested by Salmon 8, that growth-promoting and antipellagric vitamin G are two different factors. If further experimental work substantiates this conclusion, then it is probable that this growth-promoting factor will be found to be the same as the Reader 1 ' vitamin B 3. Objection may be raised to this conclusion on the basis of a possible superiority in the quality of proteins in the egg-albumin as compared with the purified-casein ration. A study of the amino-acid composition of the proteins in the ingredients used in these two rations does not indicate any such superiority. Moreover, it appears that 25 per cent protein dry basis, approximately 22 per cent air-dry basis, is somewhat in excess of the re-

11 176 POULTRY SCIENCE quirement of the chick for maximum growth during early life, as this laboratory has not been able to demonstrate a requirement as great as this nor have reports appeared in the literature of such a requirement. Whether or not the factor which prevents nutritional paralysis in chicks and so-called growth-promoting vitamin G are identical can not be determined from the evidence. However, no paralysis was obtained on the egg-albumin ration while considerable was obtained on the purified casein ration. If they are not identical, then these factors appear to accompany each other. SUMMARY 1. Chicks are subject to a pellagra-like syndrome strikingly similar to pellagra in human beings and in rats which develops at an early age and causes heavy mortality. 2. The requirement of the chick for a relatively heat-stable growth-promoting factor present in yeast and in other food materials is extremely large. moting factor present in yeast and in other food materials is extremely large. 3. Preliminary results indicate the possibility that vitamin G is not a single entity but consists of two factors. BIBLIOGRAPHY i NORRIS, L. C, HEUSER, G. F., AND WILGUS, H. S. JR. ; Is the chief value of milk for feeding poultry due to the presence of a new vitamin; Poultry Science, 9, 1930, NORRIS, L. C, HEUSER, G. F., WILGTJS, H. S. JR., AND RINGROSE, A. T.»- The occurrence of a paralysis in chicks of nutritive origin; Poultry Science, 10, 1930, 93. s AYKROYD, W. R., AND ROSCOE, MARGARET H.; The distribution of vitamin B 5 in certain foods; Biochem. Jour., 23, 1929, 483. *HAUGEI S. M., AND CARRICK, C. W.; A differentiation between the water-soluble growth-promoting and anti-neuritic substances; Jour. Biol. Chem., 69, 1926, GOLDBEKGER, JOSEPH, AND TANNER, W. F.; A study of the treatment and prevention of pellagra; U. S. Pub. Health Rep., 39, 1924, 87. s GOLDBERGER, JOSEPH, AND LILLIE, R. D.; A note on an experimental pellagra-like condition in the albino rat; V. S. Pub. Health Rep., 41, 1926, i CHICK, HARRIETTE, AND ROSCOE, MAGRABET H.; The dual nature of water-soluble vitamin B. II. The effect upon young rats of vitamin-b^

12 A PELLAGRA-LIKE SYNDROME 177 deficiency and a method for the biological assay of vitamin B 2 ; Biochem. Jour., 22, 1928, 790. s SALMON, W. D., HAYES, I. M., AND GTIERBANT, N. B.; Etiology of dermatitis of experimental pellagra in rats; Jour. Inf. Dis., 43, 1928, NOBRIS, L. C, AND RINGBOSE, A. T.; The occurrence of a pellagralike syndrome in chicks; Science, New Series, 71, 1930, BOAS, MARGARET A.; The effect of desication upon the nutritive properties of egg-white; Biochem. Jour., 21, 1927, READER, VERA; A second thermolabile water-soluble accessory factory necessary for the nutrition of the rat; Biochem. Jour., 23, 1929, 689; Further evidence for a third accessory "B" factor; Biochem. Jour., 24, 1930, 77.