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The Supplementary Value of Choline and Methionine in a Corn and Soybean Oil Meal Chick Ration1,2
The Supplementary Value of Choline and Methionine in a Corn and Soybean Oil Meal Chick Ration12 JAMES A. MARVEL, C. W. CARRICK, ROY E. ROBERTS, AND S. M. HAUGE
(Received for Publication December 29, 1943)
source of protein, but no increased growth resulted. An addition of methionine to the diet produced a growth response similar to that of heating the soybeans. The failure of choline to induce a growth increase was attributed to the lack of homocystine for the synthesis of methionine from choline and homocystine. An increase in growth resulting from the addition of choline to chick rations composed largely of corn and soybean oil meal has been reported by Synold (1942) and Betty et al. (1943). This is in contrast to the work of Jukes (1940, 1941), which had indicated that high levels of soybean oil meal in the chick diet completely protected against perosis and promoted marked growth. In investigations at this station, we have consistently obtained poor growth with rations containing approximately 35 percent soybean oil meal unless choline chloride was added. We might suspect from the findings of Almquist et al. (1942) that there may have been insufficient methionine in our diets, even with the high level of soybean oil meal used. We can not explain the growth-promoting action of choline additions, in view of the results obtained by Klose and Almquist (1941), which indi1 Journal Paper No. 144 of the Purdue University cated that chicks could not synthesize Agricultural Experiment Station. 2 This investigation was supported in part by a methionine from the choline of soybean oil meal (or choline additions) because of the grant from Joseph E. Seagram & Sons, Inc.
/^"•HOLINE has been shown by Jukes ^ - ^ (1940, 1941) to prevent perosis and promote growth in turkeys and chicks. Soybean oil meal was observed to have perosis-preventing and growth-promoting properties. These were ascribed at least in part to the choline content of the meal. Confirmatory evidence of the role of choline in preventing perosis and promoting growth in chicks has been reported by Hegsted and others (1941) and Record and Bethke (1942). Methionine was shown by Jukes (1941) to have no growth-promoting or perosis-preventing properties when added to a basal ration containing 30 percent casein. This suggested that methionine does not act as a precursor of choline for the chick. Conversely, it has been demonstrated by Griffith and Wade (1939) that methionine may act as a precursor of choline for the rat. On the other hand, choline has been reported by Klose and Almquist (1941) to act as a precursor of methionine for the chicks if homocystine is added to the diet. The same has been shown to be true for the rat (Du Vigneaud and coworkers, 1939). Almquist et al. (1942) fed choline to chicks receiving raw soybeans as the sole
[294]
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Departments of Poultry Husbandry and Agricultural Chemistry, Purdue University Agricultural Experiment Station, Lafayette, Indiana
VALUE OF CHOLINE AND METHIONINE IN CORN AND SOYBEAN OIL MEAL RATION
295
TABLE 1.—The effect upon growth from additions of choline and methionine to rations high in com and soybean oil meal Lot 1
Lot 2
Lot 3
Lot 4
Lot 5
Lot 6
Ground yellow corn Soybean oil meal Alfalfa leaf meal Distillers' dried solubles Wheat bran Wheat midds Meat and bone scraps Dried skimmilk Steamed bone meal Ground limestone Salt mixture 1 Riboflavin concentrate 2 Calcium pantothenate 3 Pyridoxin hydrochloride 4 Choline chloride d-1 methionine D-activated animal sterol6
48.0 26.0 5.0
52.5 34.5 5.0 5.0
52.35 34.50 5.00 5.00
52.0 34.5 5.0 5.0
52.88 34.00 5.00 5.00
52.73 34.00 5.00 5.00
1.5 0.5 0.5 0.5
1.50 0.50 0.50 0.50
1.5 0.5 0.5 0.5
1.50 0.50 0.50 0.50
1.50 0.50 0.50 0.50
.15
0.5
.12
.15 .12
Total Percentage protein Average four-week weights Average six-week weights Standard deviations! Coefficients of variation! Grams feed per gram gainf Number of chicks at end
5.0 5.0 5.0 5.0 0.5 0.5
+ +
+ +
+ +
+ +
+ +
+
+
+
100.0
100.0
100.00
100.00
100.00
100.00
22.7 223 399 73 18.3 2.23 23
22.8 155 282 73 25.9 3.12 24
22.8 216 367 53 14.4 2.72 '25
22.7 207 400 82 20.5 2.15 25
22.7 178 375 62 16.5 2.70 19
22.7 203 397 71 17.9 2.50 20
+
+'
1
95 percent iodized salt; 5 percent manganese sulfate. 280 micrograms of riboflavin per gram of concentrate. 1.95 milligrams per 100 grams feed. * 400 micrograms per 100 grams feed. 6 Supplies 100 D units per 100 grams feed, t Six-week weights.
2
3
lack of homocystine. Therefore, we conducted an experiment with chicks to study the role of choline and methionine in the corn and soybean oil meal ration. EXPERIMENTAL The experiment was started April 24, 1943. Barred Plymouth Rock male chicks (sexed by the method of Quinn and Knox, 1939) hatched from experiment station stock were wing-banded and distributed into six lots of 40 chicks each according to weight. Weights were recorded at two, four, and six weeks. At four weeks, the numbers of chicks were reduced to 25 per lot, except lots 5 and 6, which were reduced to 20 because of the expense of the methionine^ by discarding all chicks except the median chick and the appropriate
number on each side of the median. Standard brooder table equipment (Carrick, 1925) was used. The chicks were fed ad libitum on all-mash rations. The calcium pantothenate, pyridoxin hydrochloride, and choline chloride* used were mixed into the mashes in the form of premixes prepared by dissolving the crystals in 50 percent ethyl alcohol and drying the solution on ground yellow corn. The methionine was premixed dry on small amounts of ground yellow corn. Fresh tap water was supplied daily. RESULTS AND DISCUSSION T h e p e r c e n t a g e c o m p o s i t i o n of t h e r a * We are indebted to Lederle Laboratories, Pearl River, New York, for the choline chloride used in the experiment.
Downloaded from http://ps.oxfordjournals.org/ at UNIVERSITY OF AUCKLAND LIBRARY on May 17, 2015
Ingredients
296
J. A. MARVEL, C. W. CARRICK, R. E. ROBERTS, AND S. M. HATJGE
is the deficiency, then it must be assumed that the chick is synthesizing methionine from the added choline chloride when the 3 4 6 5 Lot No. 2 ration contains a limited amount of me32 1 117* 24 1 2 thionine. However, if choline is the defi85* 118* 93* 115* 2 —. ciency, then methionine may act as a 30 33 8 3 — —. 3 4 —. — —. 25 precursor of choline for the chick. In 22 5 — — either case, the addition of the deficient * Least significant difference at the 1 percent entity does not prove the synthesis of the level is 51 grams. At the 5 percent level the required other. Nevertheless, if the deficiency indifference is only 39 grams. volves both choline and methionine, our tions fed and the resulting data are given results would indicate that these two are in Table 1. A statistical analysis of these data is given in Table 2. Lot 1, which interchangeable. The data suggest that the quantitative served as a practical control, was fed a diet of known growth-promoting value. choline requirement of chicks is dependLot 2 received the corn and soybean oil ent upon the methionine and cystine conmeal diet without choline addition and tent of the diet. Therefore, any specificashowed the characteristic poor growth. In tion for requirements must take into lot 3, the addition of 0.15 percent choline consideration the amounts of these amino chloride gave the usual marked growth acids in the diet. response. In lot 4, a larger addition of Since the available methionine content choline chloride, 0.5 percent, produced of soybean oil meal has been shown by additional, but not significantly, increased Almquist et al. (1942) to be influenced by growth. The addition of methionine, lot 5, heat treatment of the raw soybeans, the likewise increased growth. While the addi- importance of the effect of the processing tion of both choline chloride and methio- of the soybeans upon the methionine connine (lot 6) gave better growth than either tent of the meal must be taken into accholine or methionine alone, with the count. Meals relatively low in available amounts added, the differences were not methionine may require the presence of statistically significant. The corn, soybean relatively high levels of choline in order oil meal, alfalfa leaf meal and distillers' to permit the most rapid growth. In a subdried solubles ration supplemented with sequent paper, data will be presented to minerals and vitamins, including choline, show that meals vary in their growth-proproduced growth equal to that obtained moting value when no additional choline from the meat and bone scrap practical is added to the diet. control ration. In all of these experiments perosis failed The results of this experiment show to appear, indicating that the soybean that this particular deficiency in the corn meal at high levels may supply sufficient and soybean oil meal ration can be cor- choline to prevent perosis, yet growth rected by either choline chloride or me- may be increased by additional choline. thionine. It must be recognized that this While an increase in growth may be obration already contains considerable tained by additional methionine as well as amounts of both choline and methionine, by additional choline, the present relaalthough they may not be sufficiently tively low price of choline would suggest available to meet the particular require- that choline would be the more practical ments for one or the other. If methionine of the two supplements.
TABLE 2.—Differences between the average weights of the chickens of lots 1 to 6
—
•
—
•
Downloaded from http://ps.oxfordjournals.org/ at UNIVERSITY OF AUCKLAND LIBRARY on May 17, 2015
VALUE OF CHOLINE AND METHIONINE IN CORN AND SOYBEAN OIL MEAL RATION SUMMARY
REFERENCES
Almquist, H. J., E. Mecchi, F. H. Kratzer, and C. R. Grau, 1942. Soybean protein as a source of amino acids for the chick. J. Nutrition 24:385-391. Berry, E. P., C. W. Carrick, Roy E. Roberts, and S. M. Hauge, 1943. Whey solubles as a source of growth factors in chick rations. Poultry Sci. 22: 252-263. Carrick, C. W., 1925. A brooder for indoor use in nutritional studies. Poultry Sci. 5:98-101. du Vigneaud, V., J. P. Chandler, A. W. Moyer, and D. M. Keppel, 1939. The effect of choline on the
ability of homocystine to replace methionine in the diet. Jour. Biol. Chem. 131:57-76. Griffith, W. L., and N. J. Wade, 1939. Relation of methionine, cystine and choline to renal lesions occurring on low choline diets. Proc. Soc. Exp. Biol, and Med. 41:333-334. Hegsted, M. D., R. C. Mills, C. A. Elvehjem, and E. B. Hart, 1941. Choline in the nutrition of the chick. Jour. Biol. Chem. 138:459-466. Jukes, T. H., 1940. Effect of choline and other supplements on perosis. J. Nutrition 20:445-448. , 1941. Studies of perosis in Turkeys. I. Experiments related to choline. Poultry Sci. 20:251254. Klose, A. A., and H. J. Almquist, 1941. Methionine in the diet of the chick. Jour. Biol. Chem. 138: 467-469. Record, P. R., and R. M. Bethke, 1942. Further observations on choline and yeast in poultry nutrition. Poultry Sci. 21:271-276. Synold, R. E., 1942. Distillers' dried solubles as a vitamin supplement for chick rations. Master's Thesis, Purdue University, West Lafayette, Indiana. Quinn, J. P., and C. W. Knox, 1939. Sex identification of Barred Plymouth Rock baby chicks by down, shank, and beak characteristics. Poultry Sci. 18:259-267.
Downloaded from http://ps.oxfordjournals.org/ at UNIVERSITY OF AUCKLAND LIBRARY on May 17, 2015
1. Choline and methionine exerted an interchangeable supplementary action in a chick ration consisting largely of corn and soybean oil meal. 2. The corn, soybean oil meal, alfalfa leaf meal and distillers' dried solubles ration supplemented with minerals and vitamins, including choline, produced growth equal to that obtained from the meat and bone scrap practical control ration.
297
(Received for Publication December 29, 1943)
source of protein, but no increased growth resulted. An addition of methionine to the diet produced a growth response similar to that of heating the soybeans. The failure of choline to induce a growth increase was attributed to the lack of homocystine for the synthesis of methionine from choline and homocystine. An increase in growth resulting from the addition of choline to chick rations composed largely of corn and soybean oil meal has been reported by Synold (1942) and Betty et al. (1943). This is in contrast to the work of Jukes (1940, 1941), which had indicated that high levels of soybean oil meal in the chick diet completely protected against perosis and promoted marked growth. In investigations at this station, we have consistently obtained poor growth with rations containing approximately 35 percent soybean oil meal unless choline chloride was added. We might suspect from the findings of Almquist et al. (1942) that there may have been insufficient methionine in our diets, even with the high level of soybean oil meal used. We can not explain the growth-promoting action of choline additions, in view of the results obtained by Klose and Almquist (1941), which indi1 Journal Paper No. 144 of the Purdue University cated that chicks could not synthesize Agricultural Experiment Station. 2 This investigation was supported in part by a methionine from the choline of soybean oil meal (or choline additions) because of the grant from Joseph E. Seagram & Sons, Inc.
/^"•HOLINE has been shown by Jukes ^ - ^ (1940, 1941) to prevent perosis and promote growth in turkeys and chicks. Soybean oil meal was observed to have perosis-preventing and growth-promoting properties. These were ascribed at least in part to the choline content of the meal. Confirmatory evidence of the role of choline in preventing perosis and promoting growth in chicks has been reported by Hegsted and others (1941) and Record and Bethke (1942). Methionine was shown by Jukes (1941) to have no growth-promoting or perosis-preventing properties when added to a basal ration containing 30 percent casein. This suggested that methionine does not act as a precursor of choline for the chick. Conversely, it has been demonstrated by Griffith and Wade (1939) that methionine may act as a precursor of choline for the rat. On the other hand, choline has been reported by Klose and Almquist (1941) to act as a precursor of methionine for the chicks if homocystine is added to the diet. The same has been shown to be true for the rat (Du Vigneaud and coworkers, 1939). Almquist et al. (1942) fed choline to chicks receiving raw soybeans as the sole
[294]
Downloaded from http://ps.oxfordjournals.org/ at UNIVERSITY OF AUCKLAND LIBRARY on May 17, 2015
Departments of Poultry Husbandry and Agricultural Chemistry, Purdue University Agricultural Experiment Station, Lafayette, Indiana
VALUE OF CHOLINE AND METHIONINE IN CORN AND SOYBEAN OIL MEAL RATION
295
TABLE 1.—The effect upon growth from additions of choline and methionine to rations high in com and soybean oil meal Lot 1
Lot 2
Lot 3
Lot 4
Lot 5
Lot 6
Ground yellow corn Soybean oil meal Alfalfa leaf meal Distillers' dried solubles Wheat bran Wheat midds Meat and bone scraps Dried skimmilk Steamed bone meal Ground limestone Salt mixture 1 Riboflavin concentrate 2 Calcium pantothenate 3 Pyridoxin hydrochloride 4 Choline chloride d-1 methionine D-activated animal sterol6
48.0 26.0 5.0
52.5 34.5 5.0 5.0
52.35 34.50 5.00 5.00
52.0 34.5 5.0 5.0
52.88 34.00 5.00 5.00
52.73 34.00 5.00 5.00
1.5 0.5 0.5 0.5
1.50 0.50 0.50 0.50
1.5 0.5 0.5 0.5
1.50 0.50 0.50 0.50
1.50 0.50 0.50 0.50
.15
0.5
.12
.15 .12
Total Percentage protein Average four-week weights Average six-week weights Standard deviations! Coefficients of variation! Grams feed per gram gainf Number of chicks at end
5.0 5.0 5.0 5.0 0.5 0.5
+ +
+ +
+ +
+ +
+ +
+
+
+
100.0
100.0
100.00
100.00
100.00
100.00
22.7 223 399 73 18.3 2.23 23
22.8 155 282 73 25.9 3.12 24
22.8 216 367 53 14.4 2.72 '25
22.7 207 400 82 20.5 2.15 25
22.7 178 375 62 16.5 2.70 19
22.7 203 397 71 17.9 2.50 20
+
+'
1
95 percent iodized salt; 5 percent manganese sulfate. 280 micrograms of riboflavin per gram of concentrate. 1.95 milligrams per 100 grams feed. * 400 micrograms per 100 grams feed. 6 Supplies 100 D units per 100 grams feed, t Six-week weights.
2
3
lack of homocystine. Therefore, we conducted an experiment with chicks to study the role of choline and methionine in the corn and soybean oil meal ration. EXPERIMENTAL The experiment was started April 24, 1943. Barred Plymouth Rock male chicks (sexed by the method of Quinn and Knox, 1939) hatched from experiment station stock were wing-banded and distributed into six lots of 40 chicks each according to weight. Weights were recorded at two, four, and six weeks. At four weeks, the numbers of chicks were reduced to 25 per lot, except lots 5 and 6, which were reduced to 20 because of the expense of the methionine^ by discarding all chicks except the median chick and the appropriate
number on each side of the median. Standard brooder table equipment (Carrick, 1925) was used. The chicks were fed ad libitum on all-mash rations. The calcium pantothenate, pyridoxin hydrochloride, and choline chloride* used were mixed into the mashes in the form of premixes prepared by dissolving the crystals in 50 percent ethyl alcohol and drying the solution on ground yellow corn. The methionine was premixed dry on small amounts of ground yellow corn. Fresh tap water was supplied daily. RESULTS AND DISCUSSION T h e p e r c e n t a g e c o m p o s i t i o n of t h e r a * We are indebted to Lederle Laboratories, Pearl River, New York, for the choline chloride used in the experiment.
Downloaded from http://ps.oxfordjournals.org/ at UNIVERSITY OF AUCKLAND LIBRARY on May 17, 2015
Ingredients
296
J. A. MARVEL, C. W. CARRICK, R. E. ROBERTS, AND S. M. HATJGE
is the deficiency, then it must be assumed that the chick is synthesizing methionine from the added choline chloride when the 3 4 6 5 Lot No. 2 ration contains a limited amount of me32 1 117* 24 1 2 thionine. However, if choline is the defi85* 118* 93* 115* 2 —. ciency, then methionine may act as a 30 33 8 3 — —. 3 4 —. — —. 25 precursor of choline for the chick. In 22 5 — — either case, the addition of the deficient * Least significant difference at the 1 percent entity does not prove the synthesis of the level is 51 grams. At the 5 percent level the required other. Nevertheless, if the deficiency indifference is only 39 grams. volves both choline and methionine, our tions fed and the resulting data are given results would indicate that these two are in Table 1. A statistical analysis of these data is given in Table 2. Lot 1, which interchangeable. The data suggest that the quantitative served as a practical control, was fed a diet of known growth-promoting value. choline requirement of chicks is dependLot 2 received the corn and soybean oil ent upon the methionine and cystine conmeal diet without choline addition and tent of the diet. Therefore, any specificashowed the characteristic poor growth. In tion for requirements must take into lot 3, the addition of 0.15 percent choline consideration the amounts of these amino chloride gave the usual marked growth acids in the diet. response. In lot 4, a larger addition of Since the available methionine content choline chloride, 0.5 percent, produced of soybean oil meal has been shown by additional, but not significantly, increased Almquist et al. (1942) to be influenced by growth. The addition of methionine, lot 5, heat treatment of the raw soybeans, the likewise increased growth. While the addi- importance of the effect of the processing tion of both choline chloride and methio- of the soybeans upon the methionine connine (lot 6) gave better growth than either tent of the meal must be taken into accholine or methionine alone, with the count. Meals relatively low in available amounts added, the differences were not methionine may require the presence of statistically significant. The corn, soybean relatively high levels of choline in order oil meal, alfalfa leaf meal and distillers' to permit the most rapid growth. In a subdried solubles ration supplemented with sequent paper, data will be presented to minerals and vitamins, including choline, show that meals vary in their growth-proproduced growth equal to that obtained moting value when no additional choline from the meat and bone scrap practical is added to the diet. control ration. In all of these experiments perosis failed The results of this experiment show to appear, indicating that the soybean that this particular deficiency in the corn meal at high levels may supply sufficient and soybean oil meal ration can be cor- choline to prevent perosis, yet growth rected by either choline chloride or me- may be increased by additional choline. thionine. It must be recognized that this While an increase in growth may be obration already contains considerable tained by additional methionine as well as amounts of both choline and methionine, by additional choline, the present relaalthough they may not be sufficiently tively low price of choline would suggest available to meet the particular require- that choline would be the more practical ments for one or the other. If methionine of the two supplements.
TABLE 2.—Differences between the average weights of the chickens of lots 1 to 6
—
•
—
•
Downloaded from http://ps.oxfordjournals.org/ at UNIVERSITY OF AUCKLAND LIBRARY on May 17, 2015
VALUE OF CHOLINE AND METHIONINE IN CORN AND SOYBEAN OIL MEAL RATION SUMMARY
REFERENCES
Almquist, H. J., E. Mecchi, F. H. Kratzer, and C. R. Grau, 1942. Soybean protein as a source of amino acids for the chick. J. Nutrition 24:385-391. Berry, E. P., C. W. Carrick, Roy E. Roberts, and S. M. Hauge, 1943. Whey solubles as a source of growth factors in chick rations. Poultry Sci. 22: 252-263. Carrick, C. W., 1925. A brooder for indoor use in nutritional studies. Poultry Sci. 5:98-101. du Vigneaud, V., J. P. Chandler, A. W. Moyer, and D. M. Keppel, 1939. The effect of choline on the
ability of homocystine to replace methionine in the diet. Jour. Biol. Chem. 131:57-76. Griffith, W. L., and N. J. Wade, 1939. Relation of methionine, cystine and choline to renal lesions occurring on low choline diets. Proc. Soc. Exp. Biol, and Med. 41:333-334. Hegsted, M. D., R. C. Mills, C. A. Elvehjem, and E. B. Hart, 1941. Choline in the nutrition of the chick. Jour. Biol. Chem. 138:459-466. Jukes, T. H., 1940. Effect of choline and other supplements on perosis. J. Nutrition 20:445-448. , 1941. Studies of perosis in Turkeys. I. Experiments related to choline. Poultry Sci. 20:251254. Klose, A. A., and H. J. Almquist, 1941. Methionine in the diet of the chick. Jour. Biol. Chem. 138: 467-469. Record, P. R., and R. M. Bethke, 1942. Further observations on choline and yeast in poultry nutrition. Poultry Sci. 21:271-276. Synold, R. E., 1942. Distillers' dried solubles as a vitamin supplement for chick rations. Master's Thesis, Purdue University, West Lafayette, Indiana. Quinn, J. P., and C. W. Knox, 1939. Sex identification of Barred Plymouth Rock baby chicks by down, shank, and beak characteristics. Poultry Sci. 18:259-267.
Downloaded from http://ps.oxfordjournals.org/ at UNIVERSITY OF AUCKLAND LIBRARY on May 17, 2015
1. Choline and methionine exerted an interchangeable supplementary action in a chick ration consisting largely of corn and soybean oil meal. 2. The corn, soybean oil meal, alfalfa leaf meal and distillers' dried solubles ration supplemented with minerals and vitamins, including choline, produced growth equal to that obtained from the meat and bone scrap practical control ration.
297