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PEROSIS, an anatomical deformity of the leg bones of young chicks, was first described by Hunter and Funk (1930). The condition is associated with a high intake of calcium and phosphorus, and a low intake of manganese (Hunter et al., 1931; Milby, 1934; Wilgus et al., 1936–1937; Clifcorn et al., 1938). Later, the work of Wiese et al. (1939) showed that the blood and bone phosphatase activity of perotic birds was lower than that of chicks receiving an adequate amount of manganese in their diet. As the leg bones of chicks with perosis are bent and twisted from their normal shape it was thought that there might be an abnormal distribution of the enzyme phosphatase in the bones of these chicks. In this paper we wish to report the results we have observed on the distribution of this enzyme in the leg bones of perotic and normal chicks. EXPERIMENTAL

We were interested in the recent report by Norris, Heuser and Wilgus1 on the occurrence of a peculiar type of paralysis in poultry involving the legs and feet, the cause of which has been attributed by them to the lack of some hitherto unrecognized factor occurring in milk, vitamin in nature. In 1925 we had begun a series of experiments on the effect of varying calcium-phosphorus ratios on the growth and skeleton development of the chick. Three series of experiments were started using Barred Rock chicks, each group receiving the Wisconsin Baby Chick ration as such or somewhat modified in respect to its calcium-phosphorus content, but each series receiving no or different sources of vitamin D. One series was kept out-of-doors in sunlight; another series started at the same time was kept in the attic of our laboratory but given 10 minutes’ daily irradiation with the emanations from a quartz . . .

AS Hou (1928) has pointed out in his review of the literature, the preen gland in birds has long been a subject of discussion; but, as yet, no specific indispensable function has been assigned to it. Hou, on the basis of his own work, concluded that the removal of the preen gland from adult birds caused a disturbance of the feather growth and an impairment of general health, while in young birds its removal caused rickets, in spite of normal feeding and sunshine treatment. One of the functions of this gland, as proposed by Hou, is the production of a secretion which the bird, during the act of preening, rubs over its feathers. Activation of some of the constituents of this material, during its exposure to the sun, leads to the formation of vitamin D. The vitamin is then taken in by the bird, via the mouth, at a subsequent . . .

THE essential nature of vitamin B6 for the chick was first shown by Carter and O’Brien (1939), Hegsted, Oleson, Elvehjem, and Hart (1939), and Jukes (1939). No attempt was made by these workers to establish the actual requirement of this vitamin, probably because their rations were low in other necessary factors as evidenced by the poor growth obtained at three weeks when vitamin B6 was added to their rations. Stokstad, Manning, and Rogers (1940) reported that chicks receiving 250 micrograms of vitamin B6 per 100 grams of ration weighed 99 grams at three weeks. Increasing the level of vitamin B6 produced no increase in growth. These workers state that their basal ration was also low in factor U. The first observations on the actual requirement of the chick for vitamin B6 were made by Hogan, Richardson, Patrick, O’Dell, and Kempster (1941) while studying anemia and perosis in chicks. These workers . . .

IN a previous paper by Deobald, Lease, Hart, and Halpin (1936) it was shown that the blood calcium of the laying pullet does not show any marked variation during an entire laying cycle and that approximately 10 percent of the calcium in the bones of a laying pullet is available for egg shell formation when the calcium is removed from the ration. Feinberg, Hughes, and Scott (1937) and Common (1936) have published data in substantial agreement with these findings. Common (1936) has shown quite wide variations in calcium content of the blood of individual hens over a four-month period when samples for analysis were taken at approximately monthly intervals. Riddle (1926) working with pigeons, and Roepke and Hughes (1935) working with laying pullets, have reported a marked rise in total serum phosphorus when the birds approach sexual maturity. The rise in blood calcium during this period is also well known . . .

OSBORNE and Mendel (1917), Vestal and Shrewsbury (1932) report that cooking the soybean causes a very significant increase in its growth promoting properties for white rats. Robison (1930), and Vestal and Shrewsbury (1932, 1935) report similar results for swine. Hayward, Steenbock, and Bohstedt (1936) found in experiments with rats that heating the soybean to medium and high temperatures in the commercial methods of oil extraction practically doubles the nutritive value of the protein (grams of growth per gram of protein eaten). Raw beany tasting soybean oil meal prepared at a low temperature, gave about the same nutritive value for the protein as raw soybeans. In nitrogen balance studies these workers found that heating the soybean to high temperatures in the expeller method of oil extraction increased the digestibility of the protein about 3 percent and the biological value about 12 percent. Hayward, Bohstedt, and Fargo (1935) found pigs fed medium . . .

The recent work reported by Elvehjem and Hart1 demonstrating that day-old chicks, when placed on a restricted diet, invariably develop anemia, has introduced the question of the possible presence of anemia in chicks raised under practical conditions. A diet consisting of whole cow’s milk and alcohol extracted rice supplemented with lime and salt produced anemia in chicks in a period of 12 to 15 days when the chicks were kept in pens equipped with wire screen bottoms. The anemia produced on such a diet was readily relieved by additions of iron alone because the rice in the ration contained enough copper to supplement the added iron. However, when greater precautions were taken to eliminate the copper from the basal ration, the addition of pure iron was not effective until minute amounts of copper were also fed. This shows that iron needs to be supplemented with copper for hemoglobin synthesis in . . .

HART, Steenbock, Lepkovsky, and Halpin (1925) demonstrated the synthesis of vitamin C in the chick by using the livers from chicks on vitamin C deficient diets to cure the avitaminosis in scorbutic guinea pigs. The chicken livers were found to contain more vitamin C than could be accounted for on the basis of the vitamin C content of the diet. The role of vitamin C in the metabolism of hens was studied by Bell, Satterfield and Cook (1941) who observed a reduced egg production in heavy laying breeds fed diets low in vitamin C. Injection of vitamin C increased the egg production. Briggs, Luckey, Elvehjem, and Hart (1944) observed a growth stimulation in chicks when vitamin C was added to a purified diet containing crude concentrates of folic acid. Since synthetic folic acid has become available to replace the semi-crude liver preparations used in the earlier work, a consistent growth . . .

THE occurrence of crater lesions in the linings of the gizzards of young chicks has been studied by various investigators (Hoist and Halbrook 1933, Dam 1934, Dam and Schonheyder 1934, Almquist and Stokstad 1935) and has been ascribed to a deficiency in the ration. Studies on the nature of the specific factor involved have been reported by us (1936) and by Almquist and Stokstad (1936). In attempting to extend our investigations on this subject, we encountered a serious obstacle in the development of a second type of abnormality characterized by a very considerable thickening or swelling of the gizzard lining, which not only tended to conceal any crater lesions which might be present, but also led in some cases to the appearance of cracks in the lining which simulated true crater lesions. Such gizzard linings were softer and lighter in color than the normal and showed very marked ridges. The . . .

IN A series of papers; Rose and co-workers (1936); Rose (1937); Womack, Kemmerer and Rose (1937); Womack and Rose (1941), presented evidence proving the dispensability of cystine and the indispensability of methionine in the diet of the rat. These workers also showed that cystine is capable of stimulating growth in rats only if the diet is suboptimal in methionine. Their findings indicate that on a cystine deficient diet, body cystine arises from dietary methionine and that on a suboptimal methionine diet, cystine included in the diet has methionine sparing action. That the sulphur of cystine synthesized in the body actually originates from methionine was established by Tarver and Schmidt (1939) and confirmed by Rose and Wood (1941). The relationship between choline, homocystine and methionine was worked out by du Vigneaud et al. (1939) who showed that in the presence of choline, homocystine is capable of replacing methionine in the diet . . .

A NUMBER of products have been found to supplement chick rations in which the protein was furnished by soybean oil meal, cereal grain products, and alfalfa leaf meal (Christiansen, Deobald, Halpin, and Hart [in press], Hayward, Halpin, Holmes, Bohstedt, and Hart, 1937). The value of the materials used as supplements does not always appear to be consistent with the quality of their protein. A number of vitamin deficient basal rations (Norris, Wilgus, Ringrose, Heiman, and Heuser, 1936; and Jukes, 1937) have a composition similar to that of the basal ration used in these studies of soybean oil meal. The growth trials reported here were devised to test the relative value of the vitamin or protein carried in various crude supplements to the “soybean meal—grain ration.” EXPERIMENTAL Day-old Single-Comb White Leghorn chicks were placed in experimental groups and housed in heated brooder houses which opened onto screened-in sun porches. Heat . . .

HAYWARD, Steenbock, and Bohstedt (1936) reported that rats required additional cystine in their ration when its protein was supplied by unheated soybean oil meal. Heat treatment of the meal under specified conditions eliminated this deficiency. Hayward, Halpin, Holmes, Bohstedt, and Hart (1937) reported that heated or unheated soybean oil meal was unsatisfactory as the sole protein supplement in poultry rations. The substitution of 2 percent each of dried skimmilk and meat scraps for their protein equivalents of soybean oil meal markedly improved the value of the ration. These two papers included reviews of the previous literature. Very little other work has been published concerning materials which improve the value of soybean oil meal in chick rations. EXPERIMENTAL The following experiments were devised in an attempt to determine the types of commercial products which would be useful as supplements to a soybean oil meal-grain ration. In the experiments to be described, . . .

Since cod liver meal is now coming on the market in considerable quantities as an ingredient for poultry feeds it was believed important to ascertain the vitamin A and vitamin D potency of this material by testing it with chickens. The particular meal used was obtained in 1926 from St. John’s Newfoundland and had a high oil content, viz., 45 per cent; while it also contained 43 per cent of protein. VITAMIN D In testing the vitamin D content, nine groups of chicks (one day old Leghorns) were used; seven birds in six of the groups and six in the remaining three. These birds were housed in pens in an attic where direct light had no access. They were kept on screens in place of shavings. The basal ration used was one that will invariably lead to a rachitic condition in chicks in 5 to 6 weeks . . .

THE availability of several of the vitamins in crystalline form has allowed the use of rather highly purified diets for rats (Oleson et al., 1939; Edgar et al., 1938; Lepkovsky et al., 1936). When chicks are placed on similar purified rations complete failure results. Variation in the nutritional requirements of chicks and rats has long been recognized. Thus in work with chicks it has been necessary to continue to use crude rations fortuitously low in certain factors, or rendered so by proper treatment. However, a few simplified diets containing liver, fishmeal, bran, and other supplements, as the crude materials have been used by some workers with varying degrees of success. Graham et al. (1934) and Van der Hoorn et al. (1935) obtained good growth on a simplified diet containing 10 percent of wheat germ middlings and crude casein, although an “arthritic” condition developed when the casein was extracted (Van der . . .

SEVERAL reports have been made on the value of soybean oil meal as a protein supplement for the production of hatching eggs. These have shown that when the meal alone is used poor hatchability is experienced. Hayward, Halpin, Holmes, Bohstedt, and Hart (1937) reported that soybean oil meal as the sole protein supplement to grain rations fed to hens was unsatisfactory for the production of hatching eggs. They also found that the hatchability of eggs was improved when small amounts of dried milk and meat scraps were included in the rations. Byerly, Titus, Ellis, and Nestler (1937) likewise found the meal to be a poor supplement, and found that the addition of a relatively large amount of meat scraps improved hatchability. They also reported a seasonal fall in hatchability which was more serious during the months of January and February. These reports, then, indicated that soybean oil meal as a . . .

AFTER it had been established (Elvehjem and Hart, 1929) that chickens, like mammals, require both iron and copper for hemoglobin formation, Hart and associates (1930) showed that a practical baby chick ration can supply enough iron and copper to prevent anemia in chicks during the growing period. On the basis of this fact one might expect a practical laying ration to meet the iron and copper demands of the mature non-laying hen, since the mineral requirements of the mature animal are usually smaller than those of the growing organism. However, during the laying period the demands for iron and copper would be increased. The hen is subject to such increased demands for the greater part of the year. During periods of heavy laying the loss of minerals from the body assumes considerable proportions. Although the iron and copper content of eggs is comparatively small, it is conceivable that enough of . . .

The Ca-P ratio in cows milk as well as the milk of other mammals is approximately 1:.65. It has been found by Simmonds1 that the optimum amounts of calcium and phosphorus for the rat are approximately .65 gram of calcium and .40 gram of phosphorus per 100 grams of food mixture. These proportions give a Ca-P ratio of 1:.62, a ratio quite similar to that found in milk. Because of the ideal character of milk as a food for the young, and because of the results secured by Simmonds, this ratio has been emphasized as the most suitable one in ration construction for the rat and possibly other mammals. In the hen’s egg—exclusive of shell—the Ca-P ratio is very different from that of milk and shows a preponderance of phosphorus over calcium. The ratio is 1:2.77. These figures are based on analyses of the combined white and yolk . . .

THERE is a well established belief among many practical poultry keepers that salt (sodium chloride) is poisonous to chickens. Many instances have been reported in which chickens were killed by eating salty meat, salty kitchen wastes, and salty bakery wastes. Collier (1892) reported the results of feeding varying amounts of salt to two year old hens. No harmful results were noticed in these experiments involving six hens until the intake of salt reached 0.063 ounces per hen per day. This amount, however, was sufficient to cause diarrhea in two of the hens. Upon reducing the amount of salt to 0.042 ounces per hen per day the trouble disappeared without other treatment. An intake of 0.021 or 0.042 per hen per day was not accompanied by any noticeable symptoms. Wheeler (1908) stated that a safe proportion is 5 ounces of salt in every 100 pounds of feed and that the Station . . .

THE importance of studying the efficiency of different calcium salts in the animal as well as the human diet has long been recognized. Many studies have appeared in the literature concerning the availability of various calcium salts. Much of these data were accumulated, however, before the action of vitamin D on the assimilation of calcium and phosphorus and their relation to bone formation were clearly demonstrated. The results obtained were therefore influenced, if not entirely determined, by the vitamin D content of the diet. Steenbock, Hart, Sell, and Jones (1923) reviewed these data in 1920 and, recognizing the necessity for fat soluble vitamins in the ration, fed rats calcium in the form of the following salts: lactate, carbonate, phosphate, sulfate, and silicate, at a level equivalent to that of .3 percent of calcium. They found no difference in the availability of these salts, basing their conclusions on the rate of . . .

In 1923, Hart and associates1 published data showing that light can play a very important part in the rearing of baby chicks, acting as a supplement or the equivalent to the antirachitic factor of food stuffs. Data in agreement with the above conclusions were also presented by Hughes2. The latter investigator found a condition to develop in chicks identical with mammalian rickets when birds were given adequate food, but received sunlight filtered through glass. Chicks receiving the same food and treatment, but exposed to direct sunlight for a few hours, developed normally. Since common window glass is known to shut off the beneficial ultra violet rays, much attention has been directed toward the manufacture of glass filters which will overcome this deficiency. During the past two years several of these substitutes have been tested in this laboratory for their transmissibility. However, Tisdall and Brown3 at Toronto, Canada, and Cochran and . . .

PEROSIS is a type of leg weakness and deformity which occurs among rapidly growing chicks, particularly when they are confined in battery brooders and on rations high in minerals. The other common name for perosis is “slipped tendon.” High inorganic phosphate additions to the ration are admitted to be a contributing cause of the trouble, although it is also stated in the literature that a high level of calcium carbonate may contribute to the development of perosis. Descriptions of the character of the deformity and attempts to correct it have been reviewed by Milby (1934) and Wilgus, Norris, and Heuser (1935). A variety of rations has been used by different investigators for the production of this deficiency, but in general some form of a practical ration has been used. Since we frequently observed slipped tendon in chicks grown on some of our synthetic diets, we were interested to learn if . . .

WORK leading to the development of a ration for the uniform production of perosis (slipped tendon) in chicks was presented in the previous paper. In this paper we wish to give the results obtained from studies on factors which prevent the development of this syndrome. Jukes (1937) has recently reviewed the meager work available on the perosis-preventing properties of certain feeds. EXPERIMENTAL Day-old White Leghorn chicks were divided into groups of five chicks each and placed in suitable cages. In the majority of the work ration 604 was used as the basal ration, but results obtained with other closely related rations are also included. The supplements tested were added at the expense of the whole ration. The results are tabulated in Table 1. It is readily evident that the addition of 15 percent of rice bran gave complete protection in all trials. The result checks quite well with the conclusion . . .

AS Bell (1908) knew of no method for determining the level of calcium in the blood, he developed a method which he called the calcium index. He precipitated the calcium from the blood as the oxalate and counted the crystals of calcium oxalate on a hemocytometer plate. In the case of a non-laying hen he found the calcium index to vary between 0.9 to 1.3 on successive days, whereas in the laying hen the variation was from 0.5 to 3.1. Pearl and Surface (1909) suggested that the causal factor for the deposition of calcium on the egg was a mechanical one. They sutured the large intestine to the oviduct anterior to the shell gland and found that the faeces were then covered with a calcareous deposit on their passage through the gland. Buckner, Martin, and Peter (1925) showed that there was not enough calcium in the shell gland to account . . .

IN A previous paper, Cravens, Sebesta. Halpin and Hart (1943) demonstrated the essential nature for pyridoxin for egg production, hatchability and the maintenance of body weight in mature fowl. Although 1.0 mg. of added pyridoxin per kilogram of ration appeared to give results comparable to the higher levels fed, no attempt was made from this experiment to define the requirements of laying hens for this vitamin. We wish to report here the results of a further experiment designed to determine the pyridoxin requirements of laying and breeding hens. EXPERIMENTAL Single Comb White Leghorn pullets from the Experiment Station flock were used in this experiment. These birds were reared on range and placed in an individual cage laying battery September 1, 1942. From this time until December 14, 1942, when the experimental feeding was started, the birds were fed a practical breeding mash with grain. Just prior to the experimental period, . . .

THE question has often arisen as to whether the oviduct has any direct influence on the calcium content of the blood of the fowl. Specifically, does the activity of the shell gland or the oviduct have any influence on the observed rise in blood calcium at the onset of maturity in the pullet? The observation has been made in practice that foreign materials in the oviduct are usually calcified before they are expelled. Also in a previous paper by the above authors (in press) evidence has been presented to the effect that the ovaries are the principal mechanism for the control of blood calcium. In order to show whether the oviduct was connected in any way with the blood calcium the oviducts were entirely or partially removed from 20, 6–8 week old White Leghorn pullets 15 of which survived the operation. Five normal pullets were used for controls. The . . .

Carter and O’Brien (1939) and Hegsted, Oleson, Elvehjem, and Hart (1940) demonstrated the essential nature of vitamin B6 for growing chicks. In a more recent paper Briggs, Mills, Hegsted, Elvehjem, and Hart (1942) showed that the young chick requires from 275 to 300 micrograms of vitamin B6 for optimum growth. We wish to report here studies on the essential nature of vitamin B6 for egg production, hatchability, and maintenance of body weight in mature fowl. In preliminary trials the following synthetic basal ration was found to be satisfactory: dextrin, 58; purified casein, 18; gelatin, 5; soybean oil, 5; salts IV, 5; fish oil, 2 (400 A.O.A.C. units vitamin D, 3,000 I.U. vitamin A); kidney residue, 3; solubilized liver extract, 4. Each kilogram of ration also contained thiamin, 2 3 mg.; riboflavin, 5 mg.; calcium pantothenate, 15 mg.; nicotinic acid, 100 mg.; and choline, 2 gm. Oyster shell was supplied ad libitum.

WHILE studying nutritional anemia in 1929, workers in this laboratory found that chicks did not survive when fed milk alone. When whole milk powder, fortified with iron and copper, was fed with the liquid whole milk, the chicks grew poorly (Elvehjem and Hart, 1929). Intermittently, since then, experiments have been set up to try to obtain normal growth of chicks on a milk diet. Two recent experiments are reported here. EXPERIMENT 1 Day-old White Leghorn chicks were put in electrically heated cages with raised screen bottoms. The chicks had a constant supply of whole milk powder and fresh homogenized milk, but no water. The by supplementing each 100 grams with 5 grams of grit, 40 mg. of Fe4(P2 O7)3 · 9H2 O, 2 mg. of CuSO4 · 5H2 O, 0.2 mg. of KI and 30 mg. of MnSO4 · H2 O. In the second experiment vitamins A, D, and E were fed by dropper . . .