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Effect of Choline on the Methionine Requirements of Broiler Chickens1
Effect of Choline on the Methionine Requirements of Broiler Chickens 1 E. C. QUILLIN 2 , G. F. COMBS, R. D. CREEK AND G. L. ROMOSER3
Department of Poultry Husbandry, University of Maryland, College Park, Maryland (Received for publication July 11, 1960)
Scientific Article No. A8S7. Contribution No. 3155 of The Maryland Agricultural Experiment Station (Department of Poultry Husbandry). 2 Present address: Berlin Milling Company, Berlin, Md. 'Present address: Monsanto Chemical Company, St. Louis, Mo.
reduced the total fat in rats maintained on a high fat diet. The present study was conducted to determine the extent to which choline additions might spare the methionine requirement for broilers fed practical rations high in fat. EXPERIMENTAL Three experiments were conducted with day-old male White Rock chicks maintained in floor pens. Each experimental group consisted of 50 chicks. These were provided feed and water ad libitum during an experimental period of 8 weeks in experiment 1 and 7}/2 weeks in experiments 2 and 3. Basal rations used, shown in Table 1, contained 10 or 12% added fat. Ration 1 was used in experiments 1 and 2; ration 2 was used in experiment 3. Both basal rations were formulated to contain 700 mg. of choline per pound and to be suboptimal in methionine level. DL-methionine and choline chloride (100%) were used as supplements. The experimental design in each of the three experiments involved three levels of choline with each of three or four levels of dietary methionine. In experiment 2 various levels of added vitamin B12 also were used at each choline and methionine level. The levels of added vitamin B 12 were 1.5, 7.5, and 13.5 meg. per pound. The weight data from the three levels of choline and the three levels of methionine common to all experiments were combined and statistically treated by analysis of variance. RESULTS AND DISCUSSION
639
The growth and feed efficiency data ob-
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T J OTH methionine and choline are known -*-* to furnish labile methyl groups as well as assume independent essential roles in nutrition. The work of Klose and Almquist (1941), Gillis and Norris (1951), Jukes and Stockstad (1952), Stekol et al. (1953) and others has established the common role of these compounds as methyl donors Young, Norris and Heuser (1954), using vitamin B12 depleted chicks and a basal diet deficient in choline, methionine and vitamin B12, found supplementation with mono-methylaminoethanol, homocystine, and betaine, both in the presence and absence of vitamin B12, to be utilized as effectively as equimolar concentrations of dietary choline and methionine. In the absence of sufficient betaine to methylate adequately the precursors of choline or methionine, vitamin B i 2 improved the growth rate indicating that it is required in the synthesis of methyl groups in chicks, but not for transmethylation reactions. Choline is also known to be associated with fat metabolism. Best (1934), Deuel et al. (1937), Best and Ridout (1938), and Abbott and DeMasters (1940) found that choline prevented fatty infiltration of the liver. More recent work by Snyder et al. (1957) showed that choline and betaine supplementation of the diet substantially
640
E. C. QUILLIN, G. F. COMBS, R. D. CREEK AND G. L. ROMOSER TABLE 1.—Composition of basal rations Ingredients
Ration 1
Ration 2
Percent Ground yellow corn Fat, stab, animal grease Fish solubles (50% solids) Meat and bone scrap (50% protein) Soybean oil meal (50% protein) Dhy. alfalfa meal (20% protein) Dried whey prod. (65% lactose) Distillers dr. solubles (corn) Limestone, ground Di-calcium phosphate Trace mineral mix1 Sodium sulfate Glycine Salt, iodized Special mix
44.85 12.0
2.5
2.5
3.75 35.0
— 36.0 — — —1.4
2.0 1.25 1.25
1.0 1.0 0.1
— —0.3 0.52
,493 25.2 59.4 12.5
.37 .38 700
2.25
0.1 0.1 0.25 0.35 0.23
1,562 23.5 66.6 14.1
.34 .36 700
1 Supplied to final feed: manganese, 60 ppm.; iodine, 1.2 ppm.; iron, 20 ppm.; copper, 2.0 ppm.; zinc, 20 ppm.; cobalt, 0.2 2ppm. Supplied per ton: Nicarbazin, 0.25 lbs.; B.H.T., 0.25 lbs.; arsanilic acid, 0.2 lbs.; procaine penicillin, 4 gms.; riboflavin, 4 gms.; niacin, 20 gms.: pantothenic acid, 8 gms.; choline, 92 gms.; vitamin B12, 3 mg.; menadione sodium bisulfite, 1 gm.; vitamin A, 33 million I.U.; vitamin D3, 0.8 million I.C.U. Supplied per ton: B.H.T., 0.25 lbs.; vitamin A, 7.2 million I.U.; vitamin D3, 1.3 million I.C.U.; vitamin E, 8,000 I.U.; pyridoxine (vitamin B6), 2 gms.; riboflavin, 5 gms.; niacin, 30 gms.; pantothenic acid, 15 gms.; menadione sodium bisulfite, 2 gms.; vitamin B12, 30 mg.; Glycamide, 1 lb.; Oleandomycin, 2 gms.; choline, 143 gms. 1 Calories of metabolizable energy per pound divided by percent5 crude protein. Determined by microbiological assay by Dr. Hans Rosenberg, E. I. duPont de Nemours, Wilmington, Del.
tained in experiments 1 and 2 are shown in Tables 2 and 3, and those of experiment 3 in Table 4. The data reveal that the minimal levels of vitamin B 12 were adequate, and higher levels did not influence the response to choline or methionine; therefore, vitamin B12 levels were ignored and the data combined on the basis of choline and methionine levels. In experiments 1 and 2 choline additions stimulated growth only on the lower levels of methionine, and, likewise, methionine supplementation was effective only on the lower levels of choline. In experiment 3 choline stimulated growth at all levels of methionine. No consistent differences were noted in feed conversion as a result of methionine or choline additions. Table 5 summarizes the average weights
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Calculated Analysis: Metabolizable energy, Cal./lb. Crude protein, % (NX6.25) Calorie-protein ratio* Fat, crude, % Methionine, % (calc.) 5 Methionine, % (assay) Choline, mg./lb. (calc.)
41.35 10.0
obtained when the data of all three experiments are combined on the basis of similar choline and methionine levels. Since the methionine and energy levels were slightly different for the two basal rations, the range in grams of methionine per therm of metabolizable energy for various groups combined is given in Table 5 and Figure 1. The combined data were subjected to statistical analyses which revealed the choline response to be significant at the 1% level, and that of methionine significant at the 5% level. The data (Table 5) reveal that the growth increase from choline was greatest in the low-methionine rations, and the response to methionine greatest in the lowcholine rations. The single degree of freedom test (Snedecor, 1946) verified these observations in that the methionine growth response was significant only with lowcholine rations, and the improvement in growth which accompanied the addition of choline approached significance only with the low-methionine rations. These results indicate that methionine and choline exerted sparing effects on each other in these highfat rations. Marvel et al. (1944) and McKittrick (1947) have demonstrated that either choline or methionine additions to a corn-soy ration improved growth. Such responses would be expected only in rations which are limiting in total methyl donors but not in essential choline, methionine or methionine and cystine. These rations are considered to be adequate in cystine, inorganic sulfur and vitamin B12 as the results of other unpublished data obtained at the same time as those of experiment 1 revealed no response to the inclusion of 1.5 and 3% hydrolyzed feather meal (high in cystine), 0.2% sodium sulfate or 6 micrograms vitamin B12 per pound in basal ration 1.
641
C H O L I N E AND M E T H I O N I N E REQUIREMENTS
TABLE 2.—Effect
of dietary methionine and choline levels on body weights of male broilers Average body weight, gms.1
Added choline, mg./lb.
Added vit. B i2 mcg./lb.
Methionine level, % 0.38
0.418
0.45
0.494
1,426 1,348 1,385
1,421 1,435 1,453
Experiment 1 (S weeks) 1,344 1,394 1,412
1,303 1,426 1,421
None 100 200
1.5 1.5 1.5
None 100 200
1.5 1.5 1.5
1,332 1,431 1,352
1,447 1,435 1,477
1,384 1,422 1,377
None 100 200
7.5 7.5 7.5
1,388 1,353 1,489
1,308 1,369 1,474
1,404 1,472 1,434
None 100 200
13.5 13.5 13.5
1,361 1,396 1,404
1,424 1,360 1,403
1,484 1,422 1,436
E
None 100 200
All levels
1,356 1,394 1,414
1,425 1,416 1,408
—
Experiment 2 ( 7 | weeks')
Average {4 series)
1
1,396 1,398 1,444
Each value based on approximately 50 White Rock males. TABLE 3.—Effect
of dietary methionine and choline
on feed conversion of male broilers Feed required/unit: weight1
Added choline, mg./lb.
Added vit. B12, mcg./lb.
Methionine level, % 0.38
0.418
0.45
0.494
2.14 2.01 2.08
2.13 2.03 2.18
Experiment 1 (8 weeks) 2.30 2.18 2.11
2.34 2.10 2.08
None 100 200
1.5 1.5 1.5
None 100 200
1.5 1.5 1.5
2.12 2.02 2.04
2.03 2.03 2.06
2.07 2.12 2.01
None 100 200
7.5 7.5 7.5
2.02 2.17 2.08
2.00 2.04 2.05
2.06 2.10 1.99
—
None 100 200
13.5 13.5 13.5
2.07 2.08 2.02
2.00 2.04 2.05
2.03 2.06 2.09
—
None 100 200
All levels
2.07 2.09 2.05
Experiment 2 (7j weeks)
Average (4 series)
1
Each value based on approximately 50 White Rock males.
2.01 2.04 2.05
2.05 2.09 2.03
—
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—
642
E. C. QUILLIN, G. F. COMBS, R. D. CREEK AND G. L. ROMOSER TABLE 4. Effect of dietary methionine and choline levels on body weight and feed conversion of male broilers {Experiment 3)1
Added choline mg./lb.
Methionine level, % 0.36
None Average 100 Average 200
0.50
Average 7\ week weight, gms. 1,319 1,339 1,309 1,362
1,378 1,354
1,404 1,416
1,314
1,350
1,366
1,410
1,331 1,481
1,405 1,417
1,414 1,550
1,491 1,508
1,375
1,411
1,482
1,499
1,360 1,362
1,504 1,503
1,470 1,466
1,497 1,474
1,468
1,485
1.97 1.99
2.01 2.03
1,361 1,504 Feed requin'A/unit weight (0 -7\ wk.) 2.06 2.08 2.01 2.06
None Average 100 Average 200 Average 1
0.465
2.03
2.07
1.98
2.02
2.18 2.16
1.99 2.09
1.99 1.99
1.88 1.97
2.17
2.04
1.99
1.92
1.95 2.12
1.99 2.00
1.97 2.00
1.85 2.05
2.03
1.99
1.98
1.95
Each value based on approximately 50 White Rock males.
In order to approximate the relative value of supplemental choline and methionine in correcting deficiencies of labile methyl groups in high-fat diets, two apTABLE 5.—Summary of average weights obtained in experiments 1, 2, and 3 with similar methionine and choline additions Methionine level Percent
gms./Therm M.E.
.36 - . 3 8 .418-.43 .45 -.465
1.04-1.15 1.25-1.27 1.35-1.36
Added choline, mg./lb. 0
100
200
Av . wt., gms} 1,342 1,388 1,396 1,381 1,402 1,464 1,405 1,438 1,428
1 Each value based on 6 groups of approximately 50 White Rock males each. The weights were taken at 8 weeks in Experiment 1 and 7 J weeks in Experiments 2 and 3.
proaches have been used. First by plotting the data involving growth against methionine additions for each level of choline, and a similar plot involving the effect of choline supplementation on growth at each level of methionine, the actual weight increase resulting from each increment of methionine or choline, respectively, were determined. These calculations revealed that comparable body gains were derived from the addition of either 100 mg. of choline per pound or 0.053% DL-methionine, indicating that 1 gram of choline was equivalent to 2.4 grams of DL-methionine. Secondly, the apparent methionine requirement for each level of choline was estimated by extrapolation from the data in
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Average
0.43
C H O L I N E AND M E T H I O N I N E
643
REQUIREMENTS
Av. 7, methionine in ration
3.68
3.84
.401
.417
.434
.451
.468
.434
.501
1460 High c h o l i n e
1450
1440 f
1430
Mediuai c h o l i n e
^ *
^*
+•*' *
S
<*'
1420
1410
X
^+
Lo» choline
Jft
1400
*****
1380
S
1370
1360
)3S0
1340
1330 1.05
1.10
1.15
1.20
1.25
1.30
1.35
1.40
1.45
1.50
1.55
Av. nx'thlonlnt.. level, gms./therm meeeboltzablc energy (x)
FIG. 1. Effect of added choline on methionine requirement of male broiler chickens as measured by body weights. Low, medium and high choline levels refer to feeds containing approx. 700, 800 and 900 mg. of choline per pound, respectively.
Fig. 1. These were found to be 1.23, l.S and 1.S4 grams of methionine per therm of metabolizable energy or 0.411, 0.484 and 0.514% for the rations containing 200, 100 and 0 mg. of added choline per pound, respectively. These then were plotted as shown in Figure 2, and a regression line determined. From this, 1 gram of choline chloride appeared to have an effect equal to that of 2.3 grams of DL-methionine. Both approaches suggest then that 1 gram of choline can replace 2.3-2.4 grams of methionine in practical rations containing high levels of fat where the deficiency is one only of labile methyl groups.
SUMMARY Three experiments involving male White Rock chicks reared on litter have been conducted to determine the effect of dietary choline level on the requirement of methionine. Three levels of choline and three or four levels of DL-methionine were added to practical rations containing 10 or 12% fat in all combinations. The addition of methionine produced a significant growth response only with low-choline rations; similarly, the improvement in growth which accompanied the addition of choline approached statistical significance only with methionine-low rations. The addition of
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1390
644
E. C. QUILLIN, G. F. COMBS, R. D. CREEK AND G. L. ROMOSER 7. m e t h i o n i n e required In r a t i o n (y)
.41
• 45
• 43
• 49
• 47
• 51
-i.
20 40 60
80
•
120 140
160
180 200
•
y
0
41
220
240 260
131
222
313
404
494
Additional methionine, mgs. per pound
FIG. 2. Quantitative sparing effect of choline and methionine on the apparent requirements of each other, based on data obtained with high fat broiler rations which were limiting primarily in sources of labile methyl groups.
both was not significantly better than the addition of either alone at the highest levels used. Based on the results obtained, 1 gram of choline was calculated to be equivalent to approximately 2.3-2.4 grams of DL-methionine as methyl donors in these rations.
ACKNOWLEDGMENTS
The authors are indebted to Dr. Hans Rosenberg, E. I. duPont de Nemours & Co., Wilmington, Del., for conducting the methionine assay. Appreciation is also expressed to E. I. duPont de Nemours & Co., Wilmington,
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y
100
CHOLINE AND METHIONINE REQUIREMENTS
REFERENCES Abbott, O. D., and C. U. DeMasters, 1940. Choline in the diet of chickens. J. Nutrition, 19: 47-55. Best, C. H., 1934. The role of the liver in the metabolism of carbohydrate and fat. Lancet, 1: 1274-1277. Best, C. H., and J. H. Ridout, 1938. Undernutrition and liver fat. J. Physiol. 94: 47-66. Deuel, H. J., Jr., S. Murray, L. F. Hallman and D. B. Tyler, 1937. Ketosis. XII. The effect of choline on the ketonuria of fasting rats following a high-fat diet. J. Biol. Chem. 120: 277-288.
Gillis, M. B., and L. C. Norris, 1951. Methylation of homocystine by chicks deficient in vitamin B K . Proc. Soc. Exp. Biol. Med. 77: 13-15. Jukes, T. H., and E. L. R. Stockstad, 1952. Further observations on the utilization of homocystine, choline and related compounds by chicks. J. Nutrition, 48: 209-229. Klose, A. A., and H. J. Almquist, 1941. Methionine in the diet of the chick. J. Biol. Chem. 138: 467-469. Marvel, J. A., C. W. Carrick, R. E. Roberts and S. M. Hauge, 1944. The supplementary value of choline and methionine in a corn and soybean oil meal chick ration. Poultry Sci. 23: 294-297. McKittrick, D. S., 1947. The interrelations of choline and methionine in growth and the action of betaine in replacing them. Arch. Biochem. 15: 133-155. Snedecor, G. W., Statistical Methods. (1946), The Iowa State College Press, Ames, Iowa. Snyder, F., W. E. Cornatzer and G. E. Simonson, 1957. Comparative lipotropic and lipid phosphorylating effects of choline, betaine and inositol. Proc. Soc. Exp. Biol. Med. 96: 670-672. Stekol, J. A., P. Hsu, S. Weiss and P. Smith, 1953. Labile methyl groups and its synthesis de novo in relation to growth in chicks. J. Biol. Chem. 203: 763-773. Young, R. J., L. C. Norris and G. F. Heuser, 1954. The utilization by vitamin BJ2 deficient chicks of monomethylaminoethanol, homocystine and betaine as precursors of choline and methionine. J. Nutrition, 53: 233-248.
NEWS AND NOTES (Continued from page 638) Chicago, Illinois; D. Olson, North Hollywood, California; E. B. Olson, Wilmar, Minnesota; E. E. Palmer, Minneapolis, Minnesota; H. Perry II, Topeka, Kansas; F. A. Priebe, Chicago, Illinois; V. Pringle, Broadway, Virginia; R. G. Purnele, Tupelo, Mississippi; J. Ray, Danville, Arkansas; M. H. Simmons, Siloam Springs, Arkansas; A. L. Stevens, Waterville, Maine; H. J. Wendt, Omaha, Nebraska; and L. Woodall, Chicago, Illinois. Members of the Executive Committee are: J. R. Hargreaves, M. H. Simmons, V. Pringle, H. Beyers, C. R. Lofgren, H. C. Carbaugh, R. G. Purnell, and O. A. Day.
covering all phases of turkey cookery in the institutional field, has been published by the National Turkey Federation. The 76-page book contains many pages of colorful photography and art work, illustrating the latest techniques in turkey preparation as well as a wide variety of new serving ideas. There are more than 40 cartoons and drawings, more than 150 photographs and a 16-page recipe section containing 122 of the latest turkey recipes in quantity portions. The book tells where, when, and how to buy turkey; explains how to prepare the bird for roasting; the best ways to roast large and small TURKEY HANDBOOK turkeys; how to cut up turkey for roast-poaching The third edition of the Turkey Handbook, and deep frying; how to make boneless turkey (Continued on page 761)
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Del. for supplying the DL-methionine; A. E. Staley and Co., Decatur, 111., for the soybean oil meal; Limecrest Research Laboratory, Newton, N.J., for the trace mineral mix; Monsanto Chemical Co., St. Louis, Mo., for glycine; Merck and Co., Inc., Rahway, N.J., for vitamins, procaine penicillin, Nicarbazin and glycamide; Chas. Pfizer & Sons, Terre Haute, Ind., for oleandomycin; Nopco Chemical Co., Inc., Harrisonburg, N.J., for vit. A and D 3 concentrates; The Fox Co., Newfield, N.J., for condensed fish solubles; Abbot Laboratories, Chicago, 111., for arsanilic acid and menadione sodium bisulfite; and Distillation Products, Inc., Rochester, N.Y., for alpha tocopherol acetate used in this study.
645
Department of Poultry Husbandry, University of Maryland, College Park, Maryland (Received for publication July 11, 1960)
Scientific Article No. A8S7. Contribution No. 3155 of The Maryland Agricultural Experiment Station (Department of Poultry Husbandry). 2 Present address: Berlin Milling Company, Berlin, Md. 'Present address: Monsanto Chemical Company, St. Louis, Mo.
reduced the total fat in rats maintained on a high fat diet. The present study was conducted to determine the extent to which choline additions might spare the methionine requirement for broilers fed practical rations high in fat. EXPERIMENTAL Three experiments were conducted with day-old male White Rock chicks maintained in floor pens. Each experimental group consisted of 50 chicks. These were provided feed and water ad libitum during an experimental period of 8 weeks in experiment 1 and 7}/2 weeks in experiments 2 and 3. Basal rations used, shown in Table 1, contained 10 or 12% added fat. Ration 1 was used in experiments 1 and 2; ration 2 was used in experiment 3. Both basal rations were formulated to contain 700 mg. of choline per pound and to be suboptimal in methionine level. DL-methionine and choline chloride (100%) were used as supplements. The experimental design in each of the three experiments involved three levels of choline with each of three or four levels of dietary methionine. In experiment 2 various levels of added vitamin B12 also were used at each choline and methionine level. The levels of added vitamin B 12 were 1.5, 7.5, and 13.5 meg. per pound. The weight data from the three levels of choline and the three levels of methionine common to all experiments were combined and statistically treated by analysis of variance. RESULTS AND DISCUSSION
639
The growth and feed efficiency data ob-
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T J OTH methionine and choline are known -*-* to furnish labile methyl groups as well as assume independent essential roles in nutrition. The work of Klose and Almquist (1941), Gillis and Norris (1951), Jukes and Stockstad (1952), Stekol et al. (1953) and others has established the common role of these compounds as methyl donors Young, Norris and Heuser (1954), using vitamin B12 depleted chicks and a basal diet deficient in choline, methionine and vitamin B12, found supplementation with mono-methylaminoethanol, homocystine, and betaine, both in the presence and absence of vitamin B12, to be utilized as effectively as equimolar concentrations of dietary choline and methionine. In the absence of sufficient betaine to methylate adequately the precursors of choline or methionine, vitamin B i 2 improved the growth rate indicating that it is required in the synthesis of methyl groups in chicks, but not for transmethylation reactions. Choline is also known to be associated with fat metabolism. Best (1934), Deuel et al. (1937), Best and Ridout (1938), and Abbott and DeMasters (1940) found that choline prevented fatty infiltration of the liver. More recent work by Snyder et al. (1957) showed that choline and betaine supplementation of the diet substantially
640
E. C. QUILLIN, G. F. COMBS, R. D. CREEK AND G. L. ROMOSER TABLE 1.—Composition of basal rations Ingredients
Ration 1
Ration 2
Percent Ground yellow corn Fat, stab, animal grease Fish solubles (50% solids) Meat and bone scrap (50% protein) Soybean oil meal (50% protein) Dhy. alfalfa meal (20% protein) Dried whey prod. (65% lactose) Distillers dr. solubles (corn) Limestone, ground Di-calcium phosphate Trace mineral mix1 Sodium sulfate Glycine Salt, iodized Special mix
44.85 12.0
2.5
2.5
3.75 35.0
— 36.0 — — —1.4
2.0 1.25 1.25
1.0 1.0 0.1
— —0.3 0.52
,493 25.2 59.4 12.5
.37 .38 700
2.25
0.1 0.1 0.25 0.35 0.23
1,562 23.5 66.6 14.1
.34 .36 700
1 Supplied to final feed: manganese, 60 ppm.; iodine, 1.2 ppm.; iron, 20 ppm.; copper, 2.0 ppm.; zinc, 20 ppm.; cobalt, 0.2 2ppm. Supplied per ton: Nicarbazin, 0.25 lbs.; B.H.T., 0.25 lbs.; arsanilic acid, 0.2 lbs.; procaine penicillin, 4 gms.; riboflavin, 4 gms.; niacin, 20 gms.: pantothenic acid, 8 gms.; choline, 92 gms.; vitamin B12, 3 mg.; menadione sodium bisulfite, 1 gm.; vitamin A, 33 million I.U.; vitamin D3, 0.8 million I.C.U. Supplied per ton: B.H.T., 0.25 lbs.; vitamin A, 7.2 million I.U.; vitamin D3, 1.3 million I.C.U.; vitamin E, 8,000 I.U.; pyridoxine (vitamin B6), 2 gms.; riboflavin, 5 gms.; niacin, 30 gms.; pantothenic acid, 15 gms.; menadione sodium bisulfite, 2 gms.; vitamin B12, 30 mg.; Glycamide, 1 lb.; Oleandomycin, 2 gms.; choline, 143 gms. 1 Calories of metabolizable energy per pound divided by percent5 crude protein. Determined by microbiological assay by Dr. Hans Rosenberg, E. I. duPont de Nemours, Wilmington, Del.
tained in experiments 1 and 2 are shown in Tables 2 and 3, and those of experiment 3 in Table 4. The data reveal that the minimal levels of vitamin B 12 were adequate, and higher levels did not influence the response to choline or methionine; therefore, vitamin B12 levels were ignored and the data combined on the basis of choline and methionine levels. In experiments 1 and 2 choline additions stimulated growth only on the lower levels of methionine, and, likewise, methionine supplementation was effective only on the lower levels of choline. In experiment 3 choline stimulated growth at all levels of methionine. No consistent differences were noted in feed conversion as a result of methionine or choline additions. Table 5 summarizes the average weights
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Calculated Analysis: Metabolizable energy, Cal./lb. Crude protein, % (NX6.25) Calorie-protein ratio* Fat, crude, % Methionine, % (calc.) 5 Methionine, % (assay) Choline, mg./lb. (calc.)
41.35 10.0
obtained when the data of all three experiments are combined on the basis of similar choline and methionine levels. Since the methionine and energy levels were slightly different for the two basal rations, the range in grams of methionine per therm of metabolizable energy for various groups combined is given in Table 5 and Figure 1. The combined data were subjected to statistical analyses which revealed the choline response to be significant at the 1% level, and that of methionine significant at the 5% level. The data (Table 5) reveal that the growth increase from choline was greatest in the low-methionine rations, and the response to methionine greatest in the lowcholine rations. The single degree of freedom test (Snedecor, 1946) verified these observations in that the methionine growth response was significant only with lowcholine rations, and the improvement in growth which accompanied the addition of choline approached significance only with the low-methionine rations. These results indicate that methionine and choline exerted sparing effects on each other in these highfat rations. Marvel et al. (1944) and McKittrick (1947) have demonstrated that either choline or methionine additions to a corn-soy ration improved growth. Such responses would be expected only in rations which are limiting in total methyl donors but not in essential choline, methionine or methionine and cystine. These rations are considered to be adequate in cystine, inorganic sulfur and vitamin B12 as the results of other unpublished data obtained at the same time as those of experiment 1 revealed no response to the inclusion of 1.5 and 3% hydrolyzed feather meal (high in cystine), 0.2% sodium sulfate or 6 micrograms vitamin B12 per pound in basal ration 1.
641
C H O L I N E AND M E T H I O N I N E REQUIREMENTS
TABLE 2.—Effect
of dietary methionine and choline levels on body weights of male broilers Average body weight, gms.1
Added choline, mg./lb.
Added vit. B i2 mcg./lb.
Methionine level, % 0.38
0.418
0.45
0.494
1,426 1,348 1,385
1,421 1,435 1,453
Experiment 1 (S weeks) 1,344 1,394 1,412
1,303 1,426 1,421
None 100 200
1.5 1.5 1.5
None 100 200
1.5 1.5 1.5
1,332 1,431 1,352
1,447 1,435 1,477
1,384 1,422 1,377
None 100 200
7.5 7.5 7.5
1,388 1,353 1,489
1,308 1,369 1,474
1,404 1,472 1,434
None 100 200
13.5 13.5 13.5
1,361 1,396 1,404
1,424 1,360 1,403
1,484 1,422 1,436
E
None 100 200
All levels
1,356 1,394 1,414
1,425 1,416 1,408
—
Experiment 2 ( 7 | weeks')
Average {4 series)
1
1,396 1,398 1,444
Each value based on approximately 50 White Rock males. TABLE 3.—Effect
of dietary methionine and choline
on feed conversion of male broilers Feed required/unit: weight1
Added choline, mg./lb.
Added vit. B12, mcg./lb.
Methionine level, % 0.38
0.418
0.45
0.494
2.14 2.01 2.08
2.13 2.03 2.18
Experiment 1 (8 weeks) 2.30 2.18 2.11
2.34 2.10 2.08
None 100 200
1.5 1.5 1.5
None 100 200
1.5 1.5 1.5
2.12 2.02 2.04
2.03 2.03 2.06
2.07 2.12 2.01
None 100 200
7.5 7.5 7.5
2.02 2.17 2.08
2.00 2.04 2.05
2.06 2.10 1.99
—
None 100 200
13.5 13.5 13.5
2.07 2.08 2.02
2.00 2.04 2.05
2.03 2.06 2.09
—
None 100 200
All levels
2.07 2.09 2.05
Experiment 2 (7j weeks)
Average (4 series)
1
Each value based on approximately 50 White Rock males.
2.01 2.04 2.05
2.05 2.09 2.03
—
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—
642
E. C. QUILLIN, G. F. COMBS, R. D. CREEK AND G. L. ROMOSER TABLE 4. Effect of dietary methionine and choline levels on body weight and feed conversion of male broilers {Experiment 3)1
Added choline mg./lb.
Methionine level, % 0.36
None Average 100 Average 200
0.50
Average 7\ week weight, gms. 1,319 1,339 1,309 1,362
1,378 1,354
1,404 1,416
1,314
1,350
1,366
1,410
1,331 1,481
1,405 1,417
1,414 1,550
1,491 1,508
1,375
1,411
1,482
1,499
1,360 1,362
1,504 1,503
1,470 1,466
1,497 1,474
1,468
1,485
1.97 1.99
2.01 2.03
1,361 1,504 Feed requin'A/unit weight (0 -7\ wk.) 2.06 2.08 2.01 2.06
None Average 100 Average 200 Average 1
0.465
2.03
2.07
1.98
2.02
2.18 2.16
1.99 2.09
1.99 1.99
1.88 1.97
2.17
2.04
1.99
1.92
1.95 2.12
1.99 2.00
1.97 2.00
1.85 2.05
2.03
1.99
1.98
1.95
Each value based on approximately 50 White Rock males.
In order to approximate the relative value of supplemental choline and methionine in correcting deficiencies of labile methyl groups in high-fat diets, two apTABLE 5.—Summary of average weights obtained in experiments 1, 2, and 3 with similar methionine and choline additions Methionine level Percent
gms./Therm M.E.
.36 - . 3 8 .418-.43 .45 -.465
1.04-1.15 1.25-1.27 1.35-1.36
Added choline, mg./lb. 0
100
200
Av . wt., gms} 1,342 1,388 1,396 1,381 1,402 1,464 1,405 1,438 1,428
1 Each value based on 6 groups of approximately 50 White Rock males each. The weights were taken at 8 weeks in Experiment 1 and 7 J weeks in Experiments 2 and 3.
proaches have been used. First by plotting the data involving growth against methionine additions for each level of choline, and a similar plot involving the effect of choline supplementation on growth at each level of methionine, the actual weight increase resulting from each increment of methionine or choline, respectively, were determined. These calculations revealed that comparable body gains were derived from the addition of either 100 mg. of choline per pound or 0.053% DL-methionine, indicating that 1 gram of choline was equivalent to 2.4 grams of DL-methionine. Secondly, the apparent methionine requirement for each level of choline was estimated by extrapolation from the data in
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Average
0.43
C H O L I N E AND M E T H I O N I N E
643
REQUIREMENTS
Av. 7, methionine in ration
3.68
3.84
.401
.417
.434
.451
.468
.434
.501
1460 High c h o l i n e
1450
1440 f
1430
Mediuai c h o l i n e
^ *
^*
+•*' *
S
<*'
1420
1410
X
^+
Lo» choline
Jft
1400
*****
1380
S
1370
1360
)3S0
1340
1330 1.05
1.10
1.15
1.20
1.25
1.30
1.35
1.40
1.45
1.50
1.55
Av. nx'thlonlnt.. level, gms./therm meeeboltzablc energy (x)
FIG. 1. Effect of added choline on methionine requirement of male broiler chickens as measured by body weights. Low, medium and high choline levels refer to feeds containing approx. 700, 800 and 900 mg. of choline per pound, respectively.
Fig. 1. These were found to be 1.23, l.S and 1.S4 grams of methionine per therm of metabolizable energy or 0.411, 0.484 and 0.514% for the rations containing 200, 100 and 0 mg. of added choline per pound, respectively. These then were plotted as shown in Figure 2, and a regression line determined. From this, 1 gram of choline chloride appeared to have an effect equal to that of 2.3 grams of DL-methionine. Both approaches suggest then that 1 gram of choline can replace 2.3-2.4 grams of methionine in practical rations containing high levels of fat where the deficiency is one only of labile methyl groups.
SUMMARY Three experiments involving male White Rock chicks reared on litter have been conducted to determine the effect of dietary choline level on the requirement of methionine. Three levels of choline and three or four levels of DL-methionine were added to practical rations containing 10 or 12% fat in all combinations. The addition of methionine produced a significant growth response only with low-choline rations; similarly, the improvement in growth which accompanied the addition of choline approached statistical significance only with methionine-low rations. The addition of
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1390
644
E. C. QUILLIN, G. F. COMBS, R. D. CREEK AND G. L. ROMOSER 7. m e t h i o n i n e required In r a t i o n (y)
.41
• 45
• 43
• 49
• 47
• 51
-i.
20 40 60
80
•
120 140
160
180 200
•
y
0
41
220
240 260
131
222
313
404
494
Additional methionine, mgs. per pound
FIG. 2. Quantitative sparing effect of choline and methionine on the apparent requirements of each other, based on data obtained with high fat broiler rations which were limiting primarily in sources of labile methyl groups.
both was not significantly better than the addition of either alone at the highest levels used. Based on the results obtained, 1 gram of choline was calculated to be equivalent to approximately 2.3-2.4 grams of DL-methionine as methyl donors in these rations.
ACKNOWLEDGMENTS
The authors are indebted to Dr. Hans Rosenberg, E. I. duPont de Nemours & Co., Wilmington, Del., for conducting the methionine assay. Appreciation is also expressed to E. I. duPont de Nemours & Co., Wilmington,
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y
100
CHOLINE AND METHIONINE REQUIREMENTS
REFERENCES Abbott, O. D., and C. U. DeMasters, 1940. Choline in the diet of chickens. J. Nutrition, 19: 47-55. Best, C. H., 1934. The role of the liver in the metabolism of carbohydrate and fat. Lancet, 1: 1274-1277. Best, C. H., and J. H. Ridout, 1938. Undernutrition and liver fat. J. Physiol. 94: 47-66. Deuel, H. J., Jr., S. Murray, L. F. Hallman and D. B. Tyler, 1937. Ketosis. XII. The effect of choline on the ketonuria of fasting rats following a high-fat diet. J. Biol. Chem. 120: 277-288.
Gillis, M. B., and L. C. Norris, 1951. Methylation of homocystine by chicks deficient in vitamin B K . Proc. Soc. Exp. Biol. Med. 77: 13-15. Jukes, T. H., and E. L. R. Stockstad, 1952. Further observations on the utilization of homocystine, choline and related compounds by chicks. J. Nutrition, 48: 209-229. Klose, A. A., and H. J. Almquist, 1941. Methionine in the diet of the chick. J. Biol. Chem. 138: 467-469. Marvel, J. A., C. W. Carrick, R. E. Roberts and S. M. Hauge, 1944. The supplementary value of choline and methionine in a corn and soybean oil meal chick ration. Poultry Sci. 23: 294-297. McKittrick, D. S., 1947. The interrelations of choline and methionine in growth and the action of betaine in replacing them. Arch. Biochem. 15: 133-155. Snedecor, G. W., Statistical Methods. (1946), The Iowa State College Press, Ames, Iowa. Snyder, F., W. E. Cornatzer and G. E. Simonson, 1957. Comparative lipotropic and lipid phosphorylating effects of choline, betaine and inositol. Proc. Soc. Exp. Biol. Med. 96: 670-672. Stekol, J. A., P. Hsu, S. Weiss and P. Smith, 1953. Labile methyl groups and its synthesis de novo in relation to growth in chicks. J. Biol. Chem. 203: 763-773. Young, R. J., L. C. Norris and G. F. Heuser, 1954. The utilization by vitamin BJ2 deficient chicks of monomethylaminoethanol, homocystine and betaine as precursors of choline and methionine. J. Nutrition, 53: 233-248.
NEWS AND NOTES (Continued from page 638) Chicago, Illinois; D. Olson, North Hollywood, California; E. B. Olson, Wilmar, Minnesota; E. E. Palmer, Minneapolis, Minnesota; H. Perry II, Topeka, Kansas; F. A. Priebe, Chicago, Illinois; V. Pringle, Broadway, Virginia; R. G. Purnele, Tupelo, Mississippi; J. Ray, Danville, Arkansas; M. H. Simmons, Siloam Springs, Arkansas; A. L. Stevens, Waterville, Maine; H. J. Wendt, Omaha, Nebraska; and L. Woodall, Chicago, Illinois. Members of the Executive Committee are: J. R. Hargreaves, M. H. Simmons, V. Pringle, H. Beyers, C. R. Lofgren, H. C. Carbaugh, R. G. Purnell, and O. A. Day.
covering all phases of turkey cookery in the institutional field, has been published by the National Turkey Federation. The 76-page book contains many pages of colorful photography and art work, illustrating the latest techniques in turkey preparation as well as a wide variety of new serving ideas. There are more than 40 cartoons and drawings, more than 150 photographs and a 16-page recipe section containing 122 of the latest turkey recipes in quantity portions. The book tells where, when, and how to buy turkey; explains how to prepare the bird for roasting; the best ways to roast large and small TURKEY HANDBOOK turkeys; how to cut up turkey for roast-poaching The third edition of the Turkey Handbook, and deep frying; how to make boneless turkey (Continued on page 761)
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Del. for supplying the DL-methionine; A. E. Staley and Co., Decatur, 111., for the soybean oil meal; Limecrest Research Laboratory, Newton, N.J., for the trace mineral mix; Monsanto Chemical Co., St. Louis, Mo., for glycine; Merck and Co., Inc., Rahway, N.J., for vitamins, procaine penicillin, Nicarbazin and glycamide; Chas. Pfizer & Sons, Terre Haute, Ind., for oleandomycin; Nopco Chemical Co., Inc., Harrisonburg, N.J., for vit. A and D 3 concentrates; The Fox Co., Newfield, N.J., for condensed fish solubles; Abbot Laboratories, Chicago, 111., for arsanilic acid and menadione sodium bisulfite; and Distillation Products, Inc., Rochester, N.Y., for alpha tocopherol acetate used in this study.
645