Avian Nutrition Research. Its Complexities and Interrelationship to Other Disciplines

Avian Nutrition Research. Its Complexities and Interrelationship to Other Disciplines R A L P H B. N E S T L E R State Experiment Stations Division, Agrictdtural Research Service, U. S. Department of Agriculture, Washington 25, D. C. (Received for publication May 20, 1960)

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THE INTERRELATIONSHIPS OF ALL DISCIPLINES

From the infinitesimal neutrons and protons of the atom to the enormous galaxies of the heavens, all nature appears to be interdependent and interrelated. All parts are made to synchronize into a perfectly ordered universe. The balance in nature is extremely delicate, as scientists are recognizing more and more. Disease, abnormalities, waste—such are symptoms of an imbalance. Consider, for example, the interrelationships involved in human disease. Snyder (1959) declares: "The term 'genetic disease' is used in this paper to apply

broadly to any deviation from the usual or normal condition, for which a genetic basis can be established. I shall attempt to show that there are reasons for believing that genetics is involved in one way or another in the development of all disease." (italics mine). He then points out that "there are unnumerable potential interactions between hereditary and environmental influences," as he brings nutrition into the limelight: "Although the protein core is determined by a gene, the coenzyme is often of vitamin origin, and is thus, in man at least, environmentally conditioned. . . . For example, nutritional siderosis appears to be a phenocopy of genetic hemochromatosis. For many years I have predicted to my classes in medical diseases that the vitamin-deficiency diseases resulting from the nutritional lack of coenzymes would some day be found to be paralleled by similar diseases due to genetic deficiencies of the corresponding apoenzymes." The vast field of Agriculture is not only closely interrelated with other fields such as the one just discussed, which includes human biology and pathology, but also manifests an internal mechanism of wellcoordinated parts. Both the external and internal interrelationships must be recognized and taken into consideration if truth is to be uncovered. In a recent statement prepared by the Experiment Station Section of the American Association of Land-Grant Colleges and State Universities (1958) regarding "Recommenda-

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FACT as self-evident as the importance of consideration of intra- and interrelationships in the experimental design of biological research, seemingly should not warrant an article devoted exclusively to it. Unfortunately, however, even the most obvious principles are often obscured by our absorption in the immediate problem before us or in a particularly interesting subject of research. This myopia may become evident, however, to former researchers assigned to administrative duties which force them out of the channels of their specialities to survey the broad expanses of agricultural science. All segments of agriculture are so closely interrelated and interdependent, that a scientist cannot leave out of his reckoning the other disciplines beside his own.

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The interrelationship of genetics, environment and nutrition to livestock diseases, as was found true in human disease, is noted by Hutt (1958) who presents considerable evidence to prove that animals differ genetically in ability to resist invasion by bacteria, viruses, fungi, parasitic protozoa, and worms. Moreover, he gives examples to illustrate how the genotype can interact with the environment to determine the ultimate fate of animals. Then he introduces a chapter on "Aberrant Metabolism" with these words: "Diseases resulting from genetic defects in the normal processes for utilizing nutrients in the diets are sufficiently numerous and important to warrant a special chapter." THE INTERRELATIONSHIPS AND INTRARELATIONSHIP OF POULTRY SCIENCE

Researchers in Poultry Industry must constantly recognize the fact that Poultry Science is very dependent in many ways on the other facets of Agriculture, Agronomy, for an example. Without corn, soybeans, and alfalfa leaf meal, or similar

plant products to feed the birds, there would be no poultry industry. Moreover, the genetic research on plants can have a significant effect on the nutrition of poultry, as was well demonstrated at the Beltsville Research Center during the thirties when hybrid corn was replacing openpollinated corn. The difference in the composition of the two was sufficient to throw the nutrition research askew until the scientists stopped using the old, longaccepted standard analysis of corn from "Feeds and Feeding" in developing formulas. But more astounding than these relationships is the fact that principles uncovered in one discipline may be keys to research doors in other disciplines, such as the genetic truths brought to light in hybridization of corn, and the breeding of insects (Drosophila and Tribolium)— all of which are helpful to poultry geneticists. Poultry Science is composed of the following parts: genetics, nutrition, physiology, management, housing, pathology, egg and meat technology, economics, and marketing. Less than thirty years ago, most of the scientists concerned with poultry apparently looked upon their respective areas of specialization as all important to the industry. Close cooperation either "vertically" or "horizontally" with those of like specialty was given scant consideration. Although there is still much room for improvement this "prima donna" attitude is changing, per force. Poultry scientists are beginning to see more clearly the interrelationship of the various facets, as have scientists in other fields. According to McBride (1958), "In animal breeding problems, genotype-environment interactions are likely to influence breeding plans." Then he defines four types of interactions with poultry: A. intra-population, micro-environmental (growth stimulation with antibiotics; peck

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tions with Respect to a Policy in Providing Additional Facilities for Soil and Water Conservation Research" is this significant paragraph: "Educational institutions have outstanding scientists trained in many different disciplines. Those working in the area of soil and water conservation would unquestionably benefit a great deal from close association with these scientists and could draw effectively on their special talents and knowledge. The proposed enlarged program of soil and water conservation research will be greatly enhanced by team work among soil scientists, crop scientists, biologists, physicists, engineers, livestock scientists, foresters, conservationists, economists, etc." (italics mine)

NUTRITION RESEARCH

THE CHALLENGE TO AVIAN NUTRITIONISTS

Are the nutritionists giving in turn proper consideration to the other facets of Poultry Husbandry? Here is the striking testimony of three prominent scientists. Under the startling caption of "What Has Happened to Nutrition," Schneider (1959) makes this forceful statement about the interdependence of nutrition and genetics: "In present day terminology the science of nutrition has found itself theoretically bankrupt because in its

theoretical outlook it has failed to come to grips with the problem of genotype. In a very real way, nutrition has run head on into a collision with the science of genetics . . . certain genotypes are suitable for certain kinds of nutritional experimentation. This immediately suggests that we take cognizance of the role of genotype not only between species but also within species: thus within an animal species popularly used for nutritional experimentation—such as rats, chickens, or mice—-we must expect that in our samples of these species we shall encounter suitable genotypes and unsuitable, nutritionally implastic genotypes. It is obvious that we are now entering the bailiwick of the geneticist. Historically the nutritionist has drawn back from doing this because of the tradition that if some biological character is genetically determined, then there is little that nutrition can do about it. Here again, it would seem, the efficacy of nutrition is compressed and restricted by the very real consequences of genetics. I suggest that the time has come to take the pragmatic view that 'if you can't lick 'em, join 'em.' Precisely what I propose is that nutritionists become, first of all, geneticists. If this is done, one will soon find that, viewed from the 'inside,' the world of the geneticist is not so allpowerful or, for that matter, so tidy as it seemed from the 'outside.' To pay proper tribute to it, however, I would say that the science of genetics is at present more sophisticated biologically than is the science of nutrition, if only for the following reason. Geneticists are more selfconsciously aware of the effects of environment (which includes nutrition) in their genetic experiments, and have statistical tools to assess it, than nutritionists are self-consciously aware of the necessity of measuring the role of genotype in their experiments."

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order); B. intra-population, macro-en virronmental (nutrition, batteries vs. floor, date of hatch, brooding temperature, different attendants); C. inter-population, micro-environmental (different strains intermingled or kept in separate pens with other conditions equal); and D. interpopulation, macro-environmental. All four of the current regional poultry breeding projects, W-7, NE-6, NC-47, and S-41, representing coordinated avian genetic research by the State Agricultural Experiment Stations and the U. S. Department of Agriculture, are now studying genotype environmental interactions, and Project NC-43, Physiologic Responses of Laying Fowl to Their Environment, which is conducted by the States of the North Central Region, declares in its outline under Justification: "One station could not provide the variety of expensive facilities and personnel to effectively conduct all the needed research. The many scientific disciplines such as agricultural engineers, physiologists, geneticists, and poultry scientists needed can best be supplied through joint efforts of several states since not all of these talents are available in every state." (italics mine). Although "nutritionist" is not mentioned here, he is definitely in the picture, for nutrition is a part of "environment."

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Combs (1959) sums up his strong convictions regarding the desirability of greater consideration being given to the interrelationship of nutrition with other disciplines, as follows: "Perhaps more today than ever before, it is alarmingly clear that the science of physiology, nutrition, biochemistry, genetics, pathology and food technology overlap in many places. Any a t t e m p t to administer research on practical problems within only one of these areas seems somewhat questionable. There seem to be more and more unsolved problems which fail to fall clearly into b u t one of these 'defined disciplines,' without requiring the concern of another. Joint effort and more cooperation among scientists engaged in different research areas should greatly speed the solution of many of the complex problems in the future." (italics mine) Moreover the nutritionist needs to p a y more attention to the dietary background of his experimental stock, the effect of his treatments on the progeny, and the cumulative effect over several generations. Declares Kurnick et at. (1958): "Especially, when the nutritional problem is concerned with the investigation of

'micronutrients' the aspect of the maternal diet is of particular importance. I t is not sufficient to conduct the investigation with the aid of purified diets . . . , the source of the chicks with respect to maternal diet must be considered. This requires maintenance of sizeable flocks of mature hens for which the dietary regime is controlled with respect to the micronutrient under study. . . . If the nutrient is required, how then is the requirement related to the dietary regime of the dam, and to the resulting progeny." In Nestler's (1946) research on the vitamin A requirements of bob-white quail, the effect of the parents' diet on the offspring was apparent even during the latter's breeding season a year later, despite the vitamin A content of the offspring's diet. Survival even then was directly influenced by the parents' diet. The vitamin A content of the breeders' diet affected the storage of vitamin A by their offspring; and the vitamin A content of the growth diet affected the storage of vitamin A during winter. Moreover, on all levels of vitamin A, less of the nutrient was supplied to the last hatch of chicks t h a n to the first hatch. I n fact there was less of the vitamin in the livers of the last chicks from parents on 8,000 I.U. of vitamin A than there was in those of the first chicks from parents without vitamin A. This fact m a y account for the reputed poor success of second and third clutches of eggs from quail in the wild. THE INTERDEPENDENCE OF T H E VARIOUS NUTRIENTS

According to Elvehjem and Krehl (1955): "Emphasis in the study of nutritionalbiochemistry has fluctuated among calories, proteins, minerals, vitamins, carbohydrates and amino acids. Each shift of focus brings new knowledge, not only about the subject of investigation,

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Hill (1958), in an article entitled " N e w viewpoints in the nutrition of layers: looking at the current knowledge of layer nutrition in the light of recent developm e n t s , " gives this testimony: " I n recent years, 3 dominant trends have changed our concepts of feeding layers. T h e y are: (1) the use of rations progressively higher in energy content, (2) the development of smaller strains of high production layers, and (3) the intensification of management methods leading to the use of larger flocks, cage equipment, automatic feeders, limited floor space, etc. I t is interesting to look at the current knowledge of layer nutrition in the light of these developments." (italics mine)

NUTRITION RESEARCH

Within the past decade and a half, however, avian nutrition research has been delving deeper into the complexities of this science, and the close interrelationship of its various facets is now being highlighted. A review of nutrition research since 1945 quickly reveals the intricate mechanism entitled Poultry Nutrition. One might logically start with calcium and phosphorus, the minerals t h a t encouraged a penetration of the maze back in the twenties. Phosphorus and Protein: According to Patrick and Bacon (1957) bone ash, as related to the source of phosphorus, is influenced by protein source. Blood fibrin

ration gave the best bone ash and growth response; casein, the poorest. Phosphorus and Estrogen-Tranquilizer Combination: Wolterink et al. (1958) found that phosphorus was affected by an oral estrogen-tranquilizer combination. Phosphorus in the intestine was absorbed 44 percent faster in treated birds t h a n in the controls, and the former were heavier and fatter. Phosphorus and Manganese: Various tests at the Ontario Agricultural College (1950) have revealed the need for additional manganese in diets rich in phosphorus; the sparing effect of manganese on phosphorus requirements for growth and feed efficiency; the role of manganese in bone formation and as a regulator of plasma phosphatase activity in relation to the calcium and phosphorus content of the diet; and the marked sparing effects of niacin on the requirement of manganese for perosis prevention. Calcium and Calorie/Protein Ratio: Regarding calcium of the calcium/phosphorus ratio, when rate of growth was the criterion for determining the requirement Edwards et al. (1958) found t h a t chickens receiving 840 Calories/20 percent protein required not over 0.8 percent calcium in the diet, those receiving 1,050 Calories/25 percent protein required at least 0.8 percent calcium, and those receiving the 1,260 Calories/30 percent protein required at least 1.2 percent calcium. The d a t a indicate t h a t high levels of fat, per se, as exist in the 1,260/30 diet may cause poor utilization of calcium by chickens. Potassium and Calorie/Protein Ratio: At the same time if the diet contains suboptimum amounts of another mineral, potassium, an increase of the protein level, according to Leach and Norris (1958) will depress growth and increase mortality; and an increase of the energy content will also increase the need of potassium.

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but about the interrelationships of this new fact with the others. Our expanding knowledge makes it clear t h a t indeed biology is a continuum and one cannot tamper with one area without producing shifts and changes in another. Such is the effect of imbalance in nutrition." Confirmation of these interrelationships can be quickly made by a review of the literature, starting with the first strong pronouncements on the subject. Calcium, Phosphorus and Vitamin D: McCollum et al. (1921) and Sherman and Pappenheimer (1921) pointed out t h a t three dietary components, calcium, phosphorus, and vitamin D or its equivalent, are especially concerned in normal bone formation. Less than a decade later Bethke et al. (1929) showed t h a t the optimum calcium:phosphorus ratio lies between 3:1 and 4 : 1 for chick growth when low levels of vitamin D are fed. This observation is an important milestone in the recognition of the interrelationship of nutrients in avian nutrition. Such relationships must have been known before this in a general way, for their presence is very evident, b u t their pattern, importance, and significance, seemingly were given scant attention.

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is increased, provided that the energy content of the diet is not limiting the full utilization of the supplemented protein for growth purposes. . . . If some of the protein is used as a source of energy the methionine requirement would not be expected to increase proportionately to an increase in protein level." Experiments confirmed these theories. The results obtained with isocaloric diets at different protein levels indicate that the energy content of the diet governs the methionine requirement. Methionine, Vitamin E, and Selenium: A high incidence of muscular dystrophy was found by Machlin and Shalkop (1956) to occur in chicks receiving a purified diet deficient in both methionine and vitamin E. A problem element, long considered only as a deadly poison, namely selenium, as sodium selenite at the low level of 1-5 mg. per kg. of basal diet, produced a marked decrease in the incidence of muscular dystrophy but did not completely prevent it when the diet was deficient in both methionine and vitamin E. Research by Nesheim and Scott (1958) has revealed that the addition of either methionine or vitamin E prevents dystrophy, a fact which indicates an interrelationship in metabolism between vitamin E, selenium, and methionine. Methionine, Choline, and Vitamin B^: A relationship between the methionine requirement of chicks and the "animal protein factor" was indicated by Patton el al. (1946) who noted that the growth promoting effect of methionine as a supplement to a corn-soybean diet was no longer observed when two percent of sardine meal was added to the basal diet. Three years later Shive (1949) found that methionine and vitamin B12 could function interchangeably in enabling the growth of Escherichia coli to take place on a medium containing sulfanilamide; and Jukes et al.

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Calcium and Lysine-Arginine: Wasserman et al. (1957) found in rats that certain amino acids, L-lysine and L-arginine, will increase the calcium absorption in the digestive tract. Such effect has not been demonstrated yet in chickens. The New Calorie/Protein Approach in Feed Formulation: The new facet introduced in the above-given review, namely, Calorie/protein ratio is a relationship that was given meager consideration until about four years ago. At that time the attention of the nutritionist was turned away to a great extent from antibiotics and surfactants and focused on this new interest. Combs (1955) presented a paper on "The new energy-protein approach for poultry feed formulation." States this report : "A ratio of approximately 42 Calories of productive energy per pound of ration for each percent protein is satisfactory for broiler starting rations; 45-50 Calories for broiler finishing rations. It is estimated that hens laying at the rate of 55 percent production (200 eggs per year) should be supplied one percent crude protein each 62 Calories of productive energy per pound of feed." Complexity of Calorie/Protein Ratio: This report is definite and clear cut. But protein is made up of amino acids—at least 22—in various combinations, and the Calories can come from starch, sugar, fat, or even the protein. Furthermore, there are a variety of starches, sugars, and fats as well as of proteins. Then there are the intestinal microflora that can be changed by the type of diet or cleanliness of the environment. Let us follow one of the paths leading from this large central area. Calorie/Protein Ratio and Methionine: Rosenberg and Baldwin (1957) postulated: " . . . that the methionine requirement of the chick, expressed as percent of diet, will increase as dietary protein level

NUTRITION RESEARCH

Choline, Cholesterol, Protein, Vitamins A, B\i, and E: On a low protein diet a marked hyper-cholesteremia was observed by Leveille and Fisher (1958) and Kokatnur et al. (1958), which was not affected by type of fat studied or level of

supplementation. On high protein diet, plasma cholesterol levels were essentially normal regardless of source and level of dietary fat. Mayfield and Roehm (1958) found that the feeding of 3,000 millimicrograms of vitamin B12 to female rats (but not males) receiving 250 mg. of choline and a synthetic-vitamin-mix diet caused a significant lowering of serum cholesterol when compared with that of females receiving the same amount of choline but no vitamin B12. Moreover, there are factors present in yeast which, together with vitamin B12, but not alone, bring about greater utilization of carotene and larger storage of vitamin A in the livers and kidneys than when vitamin B12 is fed with a synthetic-vitamin-mix diet. Vitamin E also, according to Hebert and Morgan (1953), is needed for utilization of carotene and vitamin A. Other Vitamin Bu Linkages: Thyroid glands in embryos from vitamin Bi2-deficient hens exhibit reduced ability to concentrate iodine, according to Ferguson et al. (1957). Vitamin B12 is required in the methylation of homocystine to form methionine. Nevertheless, Young et al. (1954) found that vitamin Bi2-depleted chicks utilized monomethylaminoethanol and homocystine plus betaine sufficiently to methylate these compounds, when they were fed in place of dietary choline and methionine. Furthermore, research by Doctor et al. (1953) indicates that there is a metabolic interrelationship between vitamin B12, folic acid and the citrovorum factor; and studies by Dinning et al. (1955) show a biochemical basis for an interrelationship of methionine with pantothenic acid. Spivey-Fox et al. (1956) found that the incorporation of 20 percent fat into a corn-soybean oil meal diet increased the vitamin B12 requirement of nondepleted chicks ten to twenty-fold for optimal growth at four weeks of age.

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(1950) showed that vitamin B12 may be needed for the transformation of homocystine to methionine in chicks. At the same time, the interplay between choline and vitamin B12 was recorded by Gillis and Norris (1949), and Schaefer et al. (1949). It appeared that dietary choline has a significant sparing action on vitamin B12. More recently Spivey-Fox et al. (1957) published results to show that only methionine, and to a small degree, choline, have significant vitamin Bi2-sparing activity. Actually 0.15 percent methionine completely replaced vitamin B12. Free methionine was more effective than that present in casein. Methionine, Choline, Betaine, and Cholesterol: Methionine and choline now lead us into another area of nutrition of particular importance to human health, an area that has received considerable publicity in recent years because of its association with heart diseases. Severe aortic atherosclerosis, believed to be the cause of about two-thirds of all human deaths, directly or indirectly, can be induced in chickens by the addition of 5-10 percent animal fat to laying rations. Corn oil and particularly the whole corn germ restricts the hypercholesterolemic response to cholesterol feeding. Kurnick et al. (1958) found that either methionine, choline, or betaine added to a diet containing added cholesterol prevented a further increase in the previously elevated serum cholesterol. When either substance was added to a diet not containing added cholesterol, serum cholesterol levels were decreased 15.7, 11.4, and 10.7 percent respectively.

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THE FUTURE OF AVIAN NUTRITION

Combs et al. (1958), using semi-purified rations, raised broilers to three pounds in 51 days with a feed conversion factor of 1.04! The first pound of weight in some cases was achieved in three weeks on only 0.82 pounds of feed. T h e best lifetime laying record as of 1949 was a little more than 1,500 eggs, fewer than half the maximum number of oocytes found in an ovary. In the light of these facts, the potentiality of greater efficiency in growth and reproduction of poultry being achieved still exists. Moreover, the battle against disease and parasites is still indecisive. Team work by the various disciplines, as well as within each one, is essential to the accomplishment of the objective. Bird (1958), after reviewing the progress during the past 50 years, made this statement regarding the future of poultry nutrition: " I t appears possible to go back and do some work on energy metabolism on a much sounder basis than 50 years ago. However, in the meantime nutritionists have also discovered the environment. They have discovered the intestinal microorganisms have nutritional requirements as real and exacting as those of their hosts, t h a t enzymes and hormones m a y be

useful dietary supplements, t h a t balance or unbalance among some nutrients profoundly affects efficiency of feed utilization, and that there are substances on the borderline between vitamins and metaboloids that may be desirable components of the diet, under certain conditions. These discoveries presage some further diversion in the next 50 years which m a y be as interesting as amino acids, minerals and vitamins." Of the 567 current Federal-grant projects in poultry husbandry a t the beginning of 1960, the largest number, 132 or 23 percent, was nutrition, whereas the next was breeding, 93 or 16 percent, and the third was poultry diseases, 88 or 15 percent. Nevertheless, of the 16 regional projects directly concerned with poultry, all areas of poultry science, except nutrition are represented. (In fact, there are only two regional nutrition projects for all animal science.) So there is plenty of room for horizontal integration as well as for vertical integration in poultry science if greater progress is to be made. REFERENCES Bethke, R. M., D. C. Kennard, C. H. Kick and G. Zinzalian, 1929. The calcium-phosphorus relationship in the nutrition of the growing chick. Poultry Sci. 8: 257-265. Bird, H. R., 1958. Amino acids, minerals and vitamins were an interesting diversion. What's next? Poultry Sci. 37: 1185. Combs, G. F., 1955. The new energy-protein approach for poultry feed formulation. Feedstuffs, 27(29): 50-52. Combs, G. F., 1959. Going "like 60" for 50 years in broiler nutrition. Feedstuffs, 31(12): 34-37. Combs, G. F., E. C. Quillin, N. V. Helbacka and C. D. Caskey, 1958. Studies on high efficiency broiler rations. Feedstuffs, 30(28): 18-20, 22. Dinning, J. S., R. Neatrour and P. L. Day, 1955. A biochemical basis for the interrelationship of pantothenic acid and methionine. J. Nutr. 56: 431-435. Doctor, V. M., B. E. Welch, R. W. Perrett, C. L. Brown, S. Gabay and J. R. Couch, 1953. Metabolic interrelationship between folic acid, vitamin

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Vitamins Bn and D: Finally, in an att e m p t to bring this brief review of the nutritional complexities to a satisfactory termination, the author presents the research of Frolich (1954) which reveals a close interrelationship of vitamin B12 to the nutrient t h a t held the limelight in the twenties for its role in the calcium/phosphorus relationship, namely vitamin D. High vitamin D supplements were found to have significant growth-depressing effects on birds receiving low vitamin B12. Supplementing the diet with vitamin B12 eliminated these effects.

NUTRITION RESEARCH

degeneration in chickens fed diets low in vitamin E and sulphur. J. Nutr. 60: 87-96. Mayfield, H. L., and R. R. Roehm, 1958. Carotene utilization and cholesterol metabolism as influenced by added choline and vitamin B12 to diets containing yeast or a synthetic vitamin mixture. J. Nutr. 64: 571-586. McBride, G., 1958. The environment and animal breeding problems. Animal Breeding Abstracts, 26: 349-358. McCollum, E. V., N. Simmonds, H. T. Parsons, P. G. Shipley and E. A. Park, 1921. Studies on experimental rickets: I. The production of rachites and similar diseases in the rat by deficient diets. J. Biol. Chem. 45: 333-342. Nesheim, M. C , and M. D. Scott, 1958. Studies on selenium, vitamin E and methionine in the prevention of muscular dystrophy in chicks. Poultry Sci. 37: 1230. Nestler, R. B., 1946. Vitamin A., vital factor in the survival of bobwhites. Trans. 11th N. Am. Wildl. Conf., New York City: 176-195. Patrick, H., and J. A. Bacon, 1957. Relation of protein source to biological value of phosphates. Poultry Sci. 36: 1147. Patton, A. R., J. P. Marvel, H. G. Petering and J. Waddell, 1946. The nutritional significance of animal protein supplements in the diet of the chick. J. Nutr. 31:485. Rosenberg, H. R., and J. T. Baldini, 1957. Effect of dietary protein level on the methionine-energy relationship in broiler diets. Poultry Sci. 36: 247252. Schaefer, A. E., W. L. Salmon and D. R. Strength, 1949. Interrelationship of vitamin B12 and choline. II. Effect on growth of the chick. Proc. Soc. Exp. Biol. Med. 71:202-204. Schneider, H. A., 1958. What has happened to nutrition. Perspectives in Biology and Medicine, 1(3): 278. Schneider, H. A., 1959. Studies from the Rockefeller Institute, 157:363-374. Sherman, H. C , and A. M. Pappenheimer, 1921. A dietetic production of rickets in rats and its prevention by an inorganic salt. Proc. Soc. Exp. Biol. Med. 18: 193-197. Shive, W., 1949. Paper presented at the AntiMetabolite Symposium, New York. Acad. Sci. Snyder, L., 1959. Fifty years of medical genetics. Science, 129(3340): 7-13. Spivey-Fox, M. R., G. M. Briggs and L. O. Ortiz, 1957. Nutrients affecting the vitamin B12 requirements of chicks. J. Nutr. 62: 539-549. Spivey-Fox, M. R., L. O. Ortiz and G. M. Briggs, 1956. Effect of dietary fat on requirement of

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B12 and the citrovorum factor. Proc. Soc. Exp. Biol. Med. 84: 29-32. Edwards, H. M., W. S. Dunahoo and H. L. Fuller, 1958. Effect of protein and calorie content of the diet on the calcium requirement of chickens. Poultry Sci. 37: 1201. Elvehjem, C. A., and W. A. Krehl, 1955. Dietary interrelationships and imbalance in nutrition. Borden's Review of Nutrition Research, 16(5): 69-84. Ferguson, T. M., J. B. Trunnell, B. Dennis, P. Wade and J. R. Couch, 1957. The influence of vitamin B12 deficiency on the uptake of I 131 by the thyroid gland in adult and embryonic chickens. Endocrin. 60: 28-32. Frolich, A., 1954. Relation between vitamin D and vitamin Bi2. Nature, 174(4427): 462-463. Gillis, M. B., and L. C. Norris, 1949. Vitamin B12 and the requirement of the chick for methylating compounds. Poultry Sci. 28: 749-750. Hebert, J. W., and A. F. Morgan, 1953. The influence of alpha-tocopherol upon the utilization of carotene and vitamin A. J. Nutr. 50: 175-190. Hill, F. W., 1958. New viewpoints in the nutrition of layers: Looking at current knowledge of layer nutrition in the light of recent developments. Feedstuffs 30(45): 40-41, 44. Hutt, F. B., 1958. Genetic Resistance to Disease in Domestic Animals. Comstock Pub. Ass'n., Ithaca, N. Y. Jukes, T. H., E. L. R. Stokstad and H. P. Broquist, 1950. Effect of vitamin B12 on the response to homocystine in chicks. Arch. Biochem. 25: 453455. Kokatnur, M., N. J. Rand, F. A. Kummerow and H. M. Scott, 1958. Effect of dietary protein and fat on changes of serum cholesterol in mature birds. J. Nutr. 64: 177-184. Kurnick, A. A., B. L. Reid and J. R. Couch, 1958. Trace elements in poultry nutrition—A review. Soil Sci. 85: 106. Kurnick, A. A., J. B. Sutton, M. W. Pasvogel and A. R. Kemmerer,1958. Effect of betaine, choline, and methionine on the concentration of serum, tissue, and egg yolk cholesterol. Poultry Sci. 37: 1218. Leach, R. M., Jr., and L. C. Norris, 1958. The effect of protein and energy on the potassium requirement of the chick. Poultry Sci. 37: 1220. Leveille, G. A., and H. Fisher, 1958. Plasma cholesterol in growing chicken as influenced by dietary protein and fat. Proc. Soc. Exp. Biol. Med. 98: 630-632. Machlin, L. J., and W. T. Shalkop, 1956. Muscular

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vitamin B12 by the chick. Proc. Soc. Exp. Biol. Med. 93: 501-504. Wasserman, R. H., C. L. Comar, J. C. Schooley and F. W. Leugemann, 1957. Interrelated effects of I-lysine and other dietary factors on the gastrointestinal absorption of calcium46 in the rat and chick. J. Nutr. 62: 367-376. Wolterink, L. F., E. Speckman, K. G. Rood and R. K. Ringer, 1958. The effects of an oral estrogen-tranquilizer combination on transit through the digestive tract and on the intestinal absorption of radioactive phosphorus in broilers fed a low protein ration. Poultry Sci. 37: 1254.

Young, R. J., L. C. Norris and G. F. Heuser, 1954. The utilization by vitamin Bi2-deficient chicks of monomethylaminoethanal, homocystine, and betaine as precursors of choline and methionine. J. Nutr. 53: 233-248. Interrelationship of Manganese and Other Nutrients in Poultry, 1950. 75th Ann. Rpt. of Ontario Agr. Col., Guelph: 35. Recommendations with Respect to a Policy in Providing Additional Facilities for Soil and Water Conservation Research, 1958. Proc. Am. Assoc. Land-Grant Colleges and State Universities. 72nd Ann. Convention, Wash., D. C : 164.

T. S. NELSON 2 AND L. C. NORRIS Department of Poultry Husbandry and Graduate School of Nutrition, Cornell University, Ithaca, New York (Received for publication May 20, 1960)

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ITAMIN K synthesis in the intestine of the rat has been reported by Dam, Sch0nheyder and Lewis (1937) and Greaves (1939). Day et al. (1943), comparing intact and cecectomized rats, concluded that the cecum is an important site of vitamin K synthesis but that it could be formed in other parts of the intestine. No deficiency occurred until sulfasuxidine was included in the diet. Barnes and Fiala (1958) were able to produce an uncomplicated vitamin K deficiency in rats when coprophagy was completely prevented. Gustafsson (1959) observed severe hypoprothrombinemia in rats reared in germ-free conditions. 1

The experimental work reported in this paper was supported in part by grants to Cornell University by the Cooperative G. L. F. Exchange, Ithaca, New York and Hoffman-LaRoche, Inc., Nutley, New Jersey. 2 Present Address: International Minerals and Chemical Corp., Skokie, 111.

In contrast, intestinal synthesis of vitamin K by the chick either does not occur or is of little importance. Almquist and Stokstad (1936) reported that vitamin K was present in fresh feces of chicks receiving a vitamin K-free diet. According to these workers the synthesis was in a region where absorption does not ordinarily take place, but under certain conditions, some absorption may occur. Scott (1955) found no significant amount of vitamin K in freshly voided droppings, when care was taken to arrest fermentation immediately. Griminger (1957) observed no vitamin K activity in the feces of turkey poults when precautions were taken to prevent bacterial action after the feces were voided. However, if the feces were allowed to stand for 48 hours without a bacteriostat, vitamin K activity was present. Luckey, Pleasants and Reyniers (1955) reported a spontaneous recovery by chicks to vitamin K deficiency when

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Studies on the Vitamin K Requirement of the Chick. The Effect of Age and Cecectomy on the Vitamin K Requirement of the Chick 1