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On the Arginine Requirement of the Growing Chick
ARGININE REQUIREMENT OF THE CHICK
eggs and dietary free gossypol supplied by different cottonseed products. Poultry Sci. 34: 81-90. Lillie, R. J., and H. R. Bird, 1950. Effect of oral administration of pure gossypol and of pigment glands on mortality and growth of chicks. Poultry Sci. 29: 390-393. Lorenz, F. W., 1939. Egg deterioration due to ingestion by hens of malvaceous materials. Poultry Sci. 18: 295-300. Milligan, J. L., L. J. Machlin, H. R. Bird and B. W. Heywang, 1951. Lysine and methionine requirement of chicks fed practical diets. Poultry Sci. 30: 578-586. Withers, W. A., and F. E. Carruth, 1915. Gossypol, the toxic substance of cottonseed meal. J. Agr. Res. 5:261.288.
On the Arginine Requirement of the Growing Chick P. GRIMINGER, 1 H. FISHER 2 AND H. M. SCOTT Illinois Agricultural Experiment Station, Urbana, Illinois (Received for publication March 9, 1955)
U
SING a semi-purified diet and 4week-old chicks, Almquist and Merritt (1950) found the arginine requirement of the chick for optimum growth to be 6 percent of the dietary protein, or 1.2 percent of a diet containing 20 percent protein. Using day-old chicks and a purified diet, Snyder el al. (1954) found that arginine supplementation increased chick growth steadily up to a level of 1.68 percent arginine of the total diet. Wietlake et al. (1954) also found the arginine requirement to be higher than the level given by Almquist. Similarly, the work of Young et al. (1953) can be interpreted to mean, that, using day-old chicks, optimum growth with a purified diet containing 25 percent casein is reached only when approximately 1.7 percent arginine is supplied. 1
Present address: Poultry Department, University of Nebraska, Lincoln, Nebraska. 2 Present address: Poultry Department, Rutgers University, New Brunswick, New Jersey.
Using acid-hydrolyzed casein3 as the only source of protein, Fisher (1954) found that a level of 6 percent arginine of the total protein would limit the growth of day-old chicks, and supplementation to a level of nearly 9 percent arginine would, as with non-hydrolyzed casein, improve growth considerably. The hydrolyzed casein, containing 12.3 percent nitrogen, was analyzed microbiologically without further hydrolysis and found to contain 2.95 percent arginine, or 3.84 percent on a 16 percent nitrogen basis. This shows, that nonavailability of arginine in casein could not be a decisive factor in causing the high requirement observed by other investigators. Henry and Kon (1950) found that storage of casein with glucose at 37° and 70 percent relative humidity reduced the nutritive value of casein for young rats. The reaction of casein with glucose 3 HY-CASE, a salt-free product of Sheffield Chemical Company, Inc., Norwich, N. Y.
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on May 24, 2015
1949. The effect of the dietary level of cottonseed meal on hatchability. Poultry Sci. 28: 610-717. Heywang, B. W., H. R. Bird and A. M. Altschul, 1950. The effect of pure gossypol on egg hatchability and weight. Poultry Sci. 29:916-920. Heywang, B. W. and H. R. Bird, 1950. Supplements for cottonseed meal in diets for chickens. Poultry Sci. 29: 486^495. Heywang, B. W., H. R. Bird and R. P. Kupperman, 1952. The loss or inactivation of pure gossypol in a mixed diet. Poultry Sci. 31:35-39. Heywang, B. W., and H. R. Bird, 1952. Failure of several amino compounds and of sardine meal to inactivate gossypol in diets of breeding chickens. Poultry Sci. 31:805-809. Heywang, B. W., H. R. Bird and A. M. Altschul, 1955. Relationship between discolorations in
1247
1248
P. GRIMINGER, H. FISHER AND H. M. SCOTT
TABLE 1.—Composition of diet containing 1.25 percent arginine Ingredient
Percent
Casein, vitamin-free Refined com oil Solka floes Salt mixture 1 Choline-Cl Glycine DL-methionine L-arginine HCl 2 Cerelose or dextrin
22.00 2.00 3.00 5.34 0.20 1.00 0.30 0.50 65.66
Total 3
100.00
TABLE 2.—Average weights of chicks
Lot
Carbohydrate
1 2 3 4 5 6
Cerelose Cerelose Dextrin Dextrin Cerelose2 Cerelose2
Arginine level
Av. weights (gm.) at three w,eeks of age
1.75 1.25 1.75 1.25 1.75 1.25
283 (9)1 213 (10) 267 (10) 169 (9) 265 (10) 221 (10)
1
Fisher et al. (1954). Additional arginine in the 1.75 percent arginine lots added at the expense of cerelose or dextrin respectively. 3 Plus the following vitamins (milligrams per kilogram diet). Thiamine HCl 25.0, riboflavin 16.0, niacin 200.0, calcium pantothenate 20.0, pyridoxine HCl 6.0, folic acid 4.0, biotin 0.6, B, 2 0.02, para-aminobenzoic acid 2.0, inositol 100.0, ascorbic acid 250.0, alpha-tocopherol acetate 20.0, menadione 5.0, 10,000 T.U. vitamin A and 600 I.U. vitamin D3. 2
caused greater losses and with greater rapidity than those experienced with lactose in milkpowder. Considerable work has been done since that time on the "browning reaction" and the concomitant destruction or nonavailability of certain amino acids. It was thus felt worth while to investigate whether amino acid losses of this nature might be a factor in causing an apparent increase of the arginine requirement. Sixty day-old crossbred male chicks (New Hampshire c? X Columbian 9 ) were allocated at random into twelve lots of five chicks each. The basal diet, shown in Table 1, was fed in one case with cerelose added as the carbohydrate, in another with dextrin, and in a third the cerelose was added to the basal diet before each feeding only. These three diets were each fed with a total level of arginine of 1.25 percent and 1.75 percent, respectively, giving six treatments, replicated twice. Feed and water were kept before the chicks at all times. In the
lots where the cerelose was combined with the basal diet immediately before feeding, the feed was renewed every 48 hours and any left-over feed discarded. All diets were stored at 3°C. The chicks were raised in electrically heated batteries with wire floors, situated in an air-conditioned room kept at 25°C. and 45 percent relative humidity. The weight of the chicks was recorded at weekly intervals and the experiment terminated at three weeks. Table 2 shows the average weights of the chicks at three weeks of age. While there was no statistically significant difference between the three high-arginine treatments, the average weight of each high-arginine lot was significantly greater than its low-arginine. counterpart; between lots 1 and 2, and 3 and 4 respectively, this significance goes beyond the 1 percent level, and between 5 and 6 beyond the 5 percent level. Though there is no difference between lots 2 and 6, lot 4 is significantly different from either one of these. If, as a result of the reaction between cerelose and amino acids, the arginine in the completely mixed diet was subject to partial destruction, then the use of dextrin, having a much smaller number of reactive endgroups than an equal amount of cerelose, should have spared arginine. A similar effect could have been
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on May 24, 2015
; Number in parenthesis indicates the number of chicks surviving. 2 Cerelose added to the diet before each feeding.
NEWS AND NOTES
quist and Merritt (1950) and the much higher one proposed by Snyder et al. (1954). We wish to thank Carol Promislow for carrying out the microbiological assay for arginine. REFERENCES Almquist, H. J., and J. B. Merritt, 1950. Protein and arginine levels in chick diets. Proc. Soc. Exp. Biol. Med. 73: 136. Fisher, H., H. M. Scott and R. G. Hansen, 1954. Further studies on the alfalfa factor and its relation to the liver and whey factors. J. Nutrition, 52: 13-24. Fisher, H., 1954. Nicotinic acid and tryptophan metabolism in the chick. Doctoral thesis, University of Illinois, Urbana, Illinois. Fisher, H., H. M. Scott and B. C. Johnson, 1955. The role of glycine in chick nutrition. J. Nutrition, 55: 415-430. Henry, K. M., and S. K. Kon, 1950. Effect of reactions with glucose on the nutritive value of casein. Biochem. Biophys. Acta 5: 455-456. Lea, C. H., and R. S. Hannan, 1950. Studies of the reaction between proteins and reducing sugars in the "dry" state. III. Nature of the protein groups reacting. Biochem. Biophys. Acta 5: 433454. Lea, C. H., 1951. Non-enzymatic browning. Proc. 2nd Intern. Congress on Canned Food. Paris 1951, XXXVII 1-9. Snyder, J. M., W. D. Morrison and H. M. Scott, 1954. A re-evaluation of the arginine requirement of the chick fed a purified diet. Poultry Sci. 33: 1082. Wietlake, A. W., A. G. Hogan, B. L. O'Dell and H. L. Kempster, 1954. Amino acid deficiencies of casein as a source of protein for the chick. J. Nutrition, 52:311-324. Young, R. J., M. B. Gillis and L. C. Norris, 1953. An unidentified factor in peanut meal required by the chick. J. Nutrition, 50: 291-298.
NEWS AND NOTES TOM NEWMAN AWARD The Tom Newman International Award for 1955 has been made to Dr. H. Williams Smith of the Animal Health Trust, Houghton Grange, Hunts, England. The Award, consisting of a medal and £50 sterling, is given for the most important contribution
to poultry husbandry research work published in the previous year. The Award was made to Dr. Smith primarily for his work on the control of Salmonella infections, with particular reference to fowl typhoid. Specific papers arising from his research work carried out
(Continued on page 1291)
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on May 24, 2015
expected when cerelose was added only before feeding. Thus, any destruction associated with lengthy storage would have been minimized. It seems, however, that these treatments did not obviate the high arginine requirement. It has also been proposed that lactose, which is present in small amounts in commercial casein, would initiate a browning reaction, as it has been shown to do in casein-glucose mixtures (Lea and Hannan, 1950) and in dried powdered milk (Lea, 1951). This reaction affects mainly lysine, but also arginine and other amino acids to a certain degree. However, this being a stoichiometric reaction, the relatively small amount of lactose in casein, and the high-arginine requirement with hydrolyzed casein, where the arginine was shown to be available by microbiological analysis, makes this appear unlikely. We are of the opinion that the very young chick has a particularly high arginine requirement which is accentuated by its requirement of this amino acid for muscle creatine synthesis. Muscle creatine increases rapidly during the early life of the chick and reaches a level near optimum (3.6 mg./gm.) at 26 days resulting in a decreased need of dietary arginine for creatine synthesis (Fisher et al., 1955). This, and the presence of preformed creatine or suitable precursors in non-purified diets might help to explain the difference between the low arginine requirement established by Alm-
1249
eggs and dietary free gossypol supplied by different cottonseed products. Poultry Sci. 34: 81-90. Lillie, R. J., and H. R. Bird, 1950. Effect of oral administration of pure gossypol and of pigment glands on mortality and growth of chicks. Poultry Sci. 29: 390-393. Lorenz, F. W., 1939. Egg deterioration due to ingestion by hens of malvaceous materials. Poultry Sci. 18: 295-300. Milligan, J. L., L. J. Machlin, H. R. Bird and B. W. Heywang, 1951. Lysine and methionine requirement of chicks fed practical diets. Poultry Sci. 30: 578-586. Withers, W. A., and F. E. Carruth, 1915. Gossypol, the toxic substance of cottonseed meal. J. Agr. Res. 5:261.288.
On the Arginine Requirement of the Growing Chick P. GRIMINGER, 1 H. FISHER 2 AND H. M. SCOTT Illinois Agricultural Experiment Station, Urbana, Illinois (Received for publication March 9, 1955)
U
SING a semi-purified diet and 4week-old chicks, Almquist and Merritt (1950) found the arginine requirement of the chick for optimum growth to be 6 percent of the dietary protein, or 1.2 percent of a diet containing 20 percent protein. Using day-old chicks and a purified diet, Snyder el al. (1954) found that arginine supplementation increased chick growth steadily up to a level of 1.68 percent arginine of the total diet. Wietlake et al. (1954) also found the arginine requirement to be higher than the level given by Almquist. Similarly, the work of Young et al. (1953) can be interpreted to mean, that, using day-old chicks, optimum growth with a purified diet containing 25 percent casein is reached only when approximately 1.7 percent arginine is supplied. 1
Present address: Poultry Department, University of Nebraska, Lincoln, Nebraska. 2 Present address: Poultry Department, Rutgers University, New Brunswick, New Jersey.
Using acid-hydrolyzed casein3 as the only source of protein, Fisher (1954) found that a level of 6 percent arginine of the total protein would limit the growth of day-old chicks, and supplementation to a level of nearly 9 percent arginine would, as with non-hydrolyzed casein, improve growth considerably. The hydrolyzed casein, containing 12.3 percent nitrogen, was analyzed microbiologically without further hydrolysis and found to contain 2.95 percent arginine, or 3.84 percent on a 16 percent nitrogen basis. This shows, that nonavailability of arginine in casein could not be a decisive factor in causing the high requirement observed by other investigators. Henry and Kon (1950) found that storage of casein with glucose at 37° and 70 percent relative humidity reduced the nutritive value of casein for young rats. The reaction of casein with glucose 3 HY-CASE, a salt-free product of Sheffield Chemical Company, Inc., Norwich, N. Y.
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on May 24, 2015
1949. The effect of the dietary level of cottonseed meal on hatchability. Poultry Sci. 28: 610-717. Heywang, B. W., H. R. Bird and A. M. Altschul, 1950. The effect of pure gossypol on egg hatchability and weight. Poultry Sci. 29:916-920. Heywang, B. W. and H. R. Bird, 1950. Supplements for cottonseed meal in diets for chickens. Poultry Sci. 29: 486^495. Heywang, B. W., H. R. Bird and R. P. Kupperman, 1952. The loss or inactivation of pure gossypol in a mixed diet. Poultry Sci. 31:35-39. Heywang, B. W., and H. R. Bird, 1952. Failure of several amino compounds and of sardine meal to inactivate gossypol in diets of breeding chickens. Poultry Sci. 31:805-809. Heywang, B. W., H. R. Bird and A. M. Altschul, 1955. Relationship between discolorations in
1247
1248
P. GRIMINGER, H. FISHER AND H. M. SCOTT
TABLE 1.—Composition of diet containing 1.25 percent arginine Ingredient
Percent
Casein, vitamin-free Refined com oil Solka floes Salt mixture 1 Choline-Cl Glycine DL-methionine L-arginine HCl 2 Cerelose or dextrin
22.00 2.00 3.00 5.34 0.20 1.00 0.30 0.50 65.66
Total 3
100.00
TABLE 2.—Average weights of chicks
Lot
Carbohydrate
1 2 3 4 5 6
Cerelose Cerelose Dextrin Dextrin Cerelose2 Cerelose2
Arginine level
Av. weights (gm.) at three w,eeks of age
1.75 1.25 1.75 1.25 1.75 1.25
283 (9)1 213 (10) 267 (10) 169 (9) 265 (10) 221 (10)
1
Fisher et al. (1954). Additional arginine in the 1.75 percent arginine lots added at the expense of cerelose or dextrin respectively. 3 Plus the following vitamins (milligrams per kilogram diet). Thiamine HCl 25.0, riboflavin 16.0, niacin 200.0, calcium pantothenate 20.0, pyridoxine HCl 6.0, folic acid 4.0, biotin 0.6, B, 2 0.02, para-aminobenzoic acid 2.0, inositol 100.0, ascorbic acid 250.0, alpha-tocopherol acetate 20.0, menadione 5.0, 10,000 T.U. vitamin A and 600 I.U. vitamin D3. 2
caused greater losses and with greater rapidity than those experienced with lactose in milkpowder. Considerable work has been done since that time on the "browning reaction" and the concomitant destruction or nonavailability of certain amino acids. It was thus felt worth while to investigate whether amino acid losses of this nature might be a factor in causing an apparent increase of the arginine requirement. Sixty day-old crossbred male chicks (New Hampshire c? X Columbian 9 ) were allocated at random into twelve lots of five chicks each. The basal diet, shown in Table 1, was fed in one case with cerelose added as the carbohydrate, in another with dextrin, and in a third the cerelose was added to the basal diet before each feeding only. These three diets were each fed with a total level of arginine of 1.25 percent and 1.75 percent, respectively, giving six treatments, replicated twice. Feed and water were kept before the chicks at all times. In the
lots where the cerelose was combined with the basal diet immediately before feeding, the feed was renewed every 48 hours and any left-over feed discarded. All diets were stored at 3°C. The chicks were raised in electrically heated batteries with wire floors, situated in an air-conditioned room kept at 25°C. and 45 percent relative humidity. The weight of the chicks was recorded at weekly intervals and the experiment terminated at three weeks. Table 2 shows the average weights of the chicks at three weeks of age. While there was no statistically significant difference between the three high-arginine treatments, the average weight of each high-arginine lot was significantly greater than its low-arginine. counterpart; between lots 1 and 2, and 3 and 4 respectively, this significance goes beyond the 1 percent level, and between 5 and 6 beyond the 5 percent level. Though there is no difference between lots 2 and 6, lot 4 is significantly different from either one of these. If, as a result of the reaction between cerelose and amino acids, the arginine in the completely mixed diet was subject to partial destruction, then the use of dextrin, having a much smaller number of reactive endgroups than an equal amount of cerelose, should have spared arginine. A similar effect could have been
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on May 24, 2015
; Number in parenthesis indicates the number of chicks surviving. 2 Cerelose added to the diet before each feeding.
NEWS AND NOTES
quist and Merritt (1950) and the much higher one proposed by Snyder et al. (1954). We wish to thank Carol Promislow for carrying out the microbiological assay for arginine. REFERENCES Almquist, H. J., and J. B. Merritt, 1950. Protein and arginine levels in chick diets. Proc. Soc. Exp. Biol. Med. 73: 136. Fisher, H., H. M. Scott and R. G. Hansen, 1954. Further studies on the alfalfa factor and its relation to the liver and whey factors. J. Nutrition, 52: 13-24. Fisher, H., 1954. Nicotinic acid and tryptophan metabolism in the chick. Doctoral thesis, University of Illinois, Urbana, Illinois. Fisher, H., H. M. Scott and B. C. Johnson, 1955. The role of glycine in chick nutrition. J. Nutrition, 55: 415-430. Henry, K. M., and S. K. Kon, 1950. Effect of reactions with glucose on the nutritive value of casein. Biochem. Biophys. Acta 5: 455-456. Lea, C. H., and R. S. Hannan, 1950. Studies of the reaction between proteins and reducing sugars in the "dry" state. III. Nature of the protein groups reacting. Biochem. Biophys. Acta 5: 433454. Lea, C. H., 1951. Non-enzymatic browning. Proc. 2nd Intern. Congress on Canned Food. Paris 1951, XXXVII 1-9. Snyder, J. M., W. D. Morrison and H. M. Scott, 1954. A re-evaluation of the arginine requirement of the chick fed a purified diet. Poultry Sci. 33: 1082. Wietlake, A. W., A. G. Hogan, B. L. O'Dell and H. L. Kempster, 1954. Amino acid deficiencies of casein as a source of protein for the chick. J. Nutrition, 52:311-324. Young, R. J., M. B. Gillis and L. C. Norris, 1953. An unidentified factor in peanut meal required by the chick. J. Nutrition, 50: 291-298.
NEWS AND NOTES TOM NEWMAN AWARD The Tom Newman International Award for 1955 has been made to Dr. H. Williams Smith of the Animal Health Trust, Houghton Grange, Hunts, England. The Award, consisting of a medal and £50 sterling, is given for the most important contribution
to poultry husbandry research work published in the previous year. The Award was made to Dr. Smith primarily for his work on the control of Salmonella infections, with particular reference to fowl typhoid. Specific papers arising from his research work carried out
(Continued on page 1291)
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on May 24, 2015
expected when cerelose was added only before feeding. Thus, any destruction associated with lengthy storage would have been minimized. It seems, however, that these treatments did not obviate the high arginine requirement. It has also been proposed that lactose, which is present in small amounts in commercial casein, would initiate a browning reaction, as it has been shown to do in casein-glucose mixtures (Lea and Hannan, 1950) and in dried powdered milk (Lea, 1951). This reaction affects mainly lysine, but also arginine and other amino acids to a certain degree. However, this being a stoichiometric reaction, the relatively small amount of lactose in casein, and the high-arginine requirement with hydrolyzed casein, where the arginine was shown to be available by microbiological analysis, makes this appear unlikely. We are of the opinion that the very young chick has a particularly high arginine requirement which is accentuated by its requirement of this amino acid for muscle creatine synthesis. Muscle creatine increases rapidly during the early life of the chick and reaches a level near optimum (3.6 mg./gm.) at 26 days resulting in a decreased need of dietary arginine for creatine synthesis (Fisher et al., 1955). This, and the presence of preformed creatine or suitable precursors in non-purified diets might help to explain the difference between the low arginine requirement established by Alm-
1249