THE National Research Council's

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1 Lysine, Arginine and Methionine Balance of Diets for Turkeys to 4 Weeks of Age* S. L. BALLOUN Department of Poultry Science, Iowa State University, Ames THE National Research Council's Nutrient Requirements of Poultry (960) states that the lysine requirement of the poult in the starting period is.% of the diet; that of arginine.6% and that of methionine %. No recommendations are set for the concentrations of these amino acids in the diet beyond the age of 8 weeks. However, the studies of Almquist (949), Grau and Kamei (90) and, more recently, the work of Nelson et al. (960) all show that the needs of poultry for specific amino acids, when expressed as percentages of the dietary protein, tend to remain constant. Almquist (9), Balloun and Phillips (97) and Waibel (99) have adequately reviewed the work with lysine and methionine supplementation of rations for turkey poults. Waibel's work indicates that increasing dietary protein from 8 to percent was not effective in producing faster growth in either Broad Breasted Bronze or White turkey poults in the first six weeks. Using high-energy diets containing % of fat, methionine was shown to be a limiting amino acid at the highest protein level. Both methionine and lysine were deficient in a 0% protein ration. Lysine and methionine supplementation was continued to only six weeks of age, but growth differences at six weeks were maintained to weeks, even though all birds received a common growing ration in the period from 6 to weeks. * Journal Paper No. J-48 of the Iowa Agricultural and Home Economics Experiment Station, Ames, Iowa. Project No. 7. (Received for publication May, 96) 47 Balloun and Phillips (97) fed practical diets varying from to % protein to BB Bronze poults in the period to six weeks of age, and from to % in the period from 6 to weeks. In the 0 to 6 weeks period, lysine improved growth of females when the diet contained % protein or less, and, in the case of males, when the diet contained 7% protein or less. Females responded to lysine supplementation in the 6 to weeks period when the diet contained 9% or less protein. Males responded in this period whenever the diet contained % or less protein. Sherman et al. (960) added lysine to rations of different protein content and quality in four turkey experiments extending over starting, growing, and finishing periods. Supplements of 0.0% or 0.0% L-lysine improved the growth and feed efficiency of poults receiving a 8% practical protein starter and a 0% protein grower. When 0% of the soybean protein was replaced by protein of either cottonseed or sesame meal, growth and feed efficiency were depressed. In another experiment, with turkeys raised to 4 weeks of age on corn-soybean oil meal rations, starting diets contained, 8, 4 or 0% protein and corresponding growing and finishing diets were proportionately lower in protein. Lysine additions improved performance with all protein levels. A corn-soybean oil meal diet containing 0% of protein fed during the 9 to 6 weeks period was improved by supplementation with lysine, methionine and glycine. Downloaded from at Penn State University (Paterno Lib) on September 8, 06

2 48 S. L. BALLOUN TABLE.Composition of high-protein dietsexperiment Ingredients Gr. yellow corn Soybean oil meal (0% prot.) Wheat middlings Meat scrap (0% prot.) Dried whole cheese whey Dr. fish solubles (4% prot.) Dehyd. alf. meal (0% prot.) Torula yeast Dical. phosphate Gr. oyster shell Salt+tr. min. Animal fat Gr. oat hulls Mix No. 476 Calc. protein (%) Calc. prod, energy (Cal. per lb.) , , ,00 Relative energy concentration of diet. Trace minerals provided (mg. per pound diet) were: Mn 8; Zn 6; Fe ; Cu.6. Adds per pound diet: riboflavin. mg.; calcium pantothenate.0 mg.; niacin 0 mg.; choline 40 mg.; vit. B meg.; folic acid mg.; menadione mg.; vit. E 0 I.U.; vit. A.,000 I.U.; vit. D,000 I.C.U.; and methionine 0.0%. Considerable information appears in the literature in regard to long-term feeding of methionine, particularly the work of Donovan et al. (9) and Carter et al. (96). Donovan et al. observed small, but statistically significant, improvements in weights and feed efficiency at 4 weeks as a result of methionine TABLE.Composition of high-protein dietsexperiment Ingredients Gr. yellow corn Soybean oil meal (0% prot.) Wheat middlings Meat scrap (0% prot.) Dried whole cheese whey Dr. fish solubles (4% prot.) Dehyd. alf. meal (0% prot.) Torula yeast Defl. rock phosphate Gr. oyster shell Salt-j- tr. min. Animal fat Mix No. S0 Calc. protein (%) Calc. prod, energy (Cal. per lb.) Downloaded from at Penn State University (Paterno Lib) on September 8, 06 Trace minerals provided (mg. per pound diet) were: Mn 8; Zn 6; Fe ; Cu.6. Adds per pound diet: riboflavin 4 mg.; calcium pantothenate 4 mg.; niacin 0 mg; choline 00 mg.; vit. B 0 meg.; menadione mg.; vit. E. 0 I.U.; vitamin A, 4,000 I.U.; vitamin D,,000 I.C.U.; and methionine, 0.%.

3 AMINO ACID BALANCE 49 supplementation. Carter et al. fed rations varying in protein level to small-type White turkeys during the 8 to 6 weeks period. With a 4% protein diet, lysine appeared to be the first limiting amino acid, while supplementing the 4% diet with methionine alone actually decreased weight gains. Lysine supplementation, or a combination of lysine and methionine, improved growth and feed conversion. In some cases, the results obtained when the 4% protein diet was supplemented with amino acids were equal to those obtained with the 7% protein control ration. Kratzer et al. (947) studied the arginine requirement of turkey poults in the starting period, but little information is available on the poult's requirement for this amino acid in later stages of growth. The trials to be reported were designed to determine the effects of lysine, methionine and arginine supplementation of turkey diets over the entire growing period to market age. EXPERIMENTAL PROCEDURE Two trials have been conducted with Broad Beasted Bronze male poults, utilizing rations varying in protein and energy and in amino acid supplementation. The high-protein diets fed in each experiment are shown in Tables and. Adjustments were made for protein and energy levels by adjustment of corn, wheat by-products, soybean oil meal, stabilized tallow and ground oat-hulls. In calculating the productive energy values, the values of Titus (9) were used. Lysine was added as 9% L-lysine-HCl to provide the levels of L-lysine indicated. Methionine was added as DL-methionine and arginine as L-arginine-HCl. Experiments were conducted in typical battery brooders for the first three weeks, in unheated growing batteries for the to 6 week period and in the floor pens thereafter. The poults were randomly allotted to battery pens and experimental diets were then randomly assigned to the pens in a replicated block design. Experiment was of 6 weeks duration and two replicate pens were fed each experimental diet. Poults in experiment were weighed individually at, 6, 0 and 6 weeks and feed consumption and efficiency were calculated at each weigh period-. In experiment three replications were used and poults were weighed individually at, 6,, 6, 0 and 4 weeks of age. Groups were considered as the experimental units and the methods of analyses as described by Balloun et al. (99) were used for testing the significance of treatment effects. RESULTS Table shows the calculated amino acids of the "low" protein diets used in these trials as compared to published re- TABLE.Calculated amino acids in low-protein diets, and estimated requirements Arginine (%).7.6 Lysine.40. Methionine 0.40 Cystine Protein 4. Calculated diet amino acids and requirements Diet Req. Diet Req. Diet Req Diet Req Diet Req Downloaded from at Penn State University (Paterno Lib) on September 8, 06 Based on NRC (960) published requirements for 0-6 weeks, reduced proportionately to reduction in protein in each age period.

4 40 S. L. BALLOUN TABLE 4.-Eject of added dietary amino acids on poult weight gains and feed efficiency. Experiment l Diet Weight gains by periods (lbs.) 6 weeks Prot. P.E. A.A weight F/G None None L L+M L+A L+A+M None None L L+M L+A L+A+M , Each figure in the table is the rounded average of groups of Bronze males started per group. protein8, 4, 0% in the age periods respectively. protein4, 0, 6% in the age periods. energy,000,,000,,00 prod. Cal./lb. in the age periods. energy800, 800, 900 prod. Cal./lb. in the age periods. AA=added amino acids; L = 0.% L-lysine; M=DL-methionine 0.0%; A = L-arginine 0.%. quirements. In cases where the National Research Council does not list the quantitative requirement for an amino acid, a calculated requirement has been determined which reflects a constant ratio of each amino acid to the protein level of the diet. The high-protein diets essentially met the NRC requirements for methionine, lysine and arginine in all periods, although there was a tendency toward more adequate methionine and less adequate lysine in the later periods. The low-protein diets were deficient in these amino acids in all periods, but not equally so for each of the three amino acids studied. This shift of amino acid balance, as a greater proportion of the total protein was obtained from the cereal portion of the diet in the later periods, is shown in Table. It can be seen that the low-protein diets became markedly more deficient in lysine, while sulfur amino acid balance was actually improved in the later periods. Experiment. Table 4 summarizes the results obtained in experiment. In this experiment, arginine, methionine and lysine were added to the low-protein diets in various combinations, but only methionine (0.0%) was added to the high-protein diets. All diets were supplemented with 0.0% DL-methionine, so when "methionine supplementation" is referred to, this means an additional 0.0% DLmethionine. As was to be expected, the high-protein diets produced greater weight gains in all three periods than did the lowprotein diets. Increased energy did not always increase weight gains. Poults fed the high-protein, low-energy diet were as heavy at 6 weeks as those fed the highprotein, high-energy diet. However, it should be noted that, in the starting period, the 8% protein high-energy diet produced poults considerably heavier than those fed the 8% protein, lowenergy diet. In the starting period (to 6 weeks of age), lysine alone did not improve weight gains of poults fed either the high- or lowenergy diets. Lysine plus arginine resulted in small but non-significant improvements in gains with both high- and low-energy diets and supplementing the low-protein diets with all three amino acids significantly improved gains with both high- and Downloaded from at Penn State University (Paterno Lib) on September 8, 06

5 AMINO ACID BALANCE 4 low-energy diets. When the low-protein diets were supplemented with all three amino acids, weight gains, with either high- or low-energy diets, were essentially the same as those achieved on the 8% protein control diets. In the 6 to 0 week period, weight gain improvements resulting from amino acid additions were significant with both lowand high-energy diets. Adding lysine alone resulted in an 8.8% improvement in weight gains in this period when poults were fed the high-energy diets. Adding additional methionine to the lysine supplementation resulted in no further improvement, but adding all three amino acids produced a 6% improvement in weight gains over the unsupplemented low-protein diet. At 0 weeks of age, poults fed the low-protein, high-energy diets, supplemented with three amino acids, were fully as heavy as those which had been fed the high-protein, highenergy diets supplemented with methionine alone. With low-energy diets, lysine supplementation improved gains by nearly 7%, but adding methionine or arginine, or both, to the lysine-supplemented diet did not further improve weight gains. In the third period (0 to 6 weeks), supplementing the low-protein, highenergy diets with lysine improved weight gains by %, added methionine did not further improve weight gains, but arginine added to the lysine-supplemented diets resulted in a total improvement of gains of.% over the low-protein, highenergy basal diet. As a total for the three periods, to 6 weeks of age, weight gains of poults fed low-protein, high-energy diets were improved 7.8% by lysine, adding additional methionine did not further improve weights, and supplementing this diet with all three amino acids resulted in weights % greater than those of poults fed the low-protein, high-energy basal diet. With low-energy diets, results in the third period were very similar to those obtained with the high-energy diets. Lysine improved weight gains % in this period, but supplementation with additional methionine and (or) arginine did not further improve weight gains. Lysine added to a low-protein, low-energy diet resulted in poults 8.7% heavier at 6 weeks as compared to those fed this diet without added lysine, but still % less than those poults fed the high-protein, low-energy diet. Over the 6-week experimental period, feed efficiency generally was influenced by amino acid additions as were weight gains. Lysine reduced the feed required per pound of gain 8.% on high-energy diets and.% on low-energy diets. At both energy levels, feed utilization was as efficient with low-protein, lysine-supplemented diets as with high-protein diets not supplemented with lysine. As was to be expected, the feed required per pound of gain was significantly less (about 4%) when poults were fed high-energy diets. Experiment. Table summarizes the results obtained in experiment. Triplicate groups of BB Bronze male poults were fed the experimental diets to 4 weeks of age. All diets were supplemented with 0.% DL-methionine to insure that methionine would not be a limiting factor. Lysine supplementation of both high- and low-protein diets significantly improved weight gains. When poults were fed the high-protein dietsstarting at 8% protein and gradually reducing to 4% at 0 weeksno appreciable improvement in weight gains as a result of added lysine was observed until after 6 weeks. In fact, at 0 weeks of age, poults fed the basal high-protein diet weighed as much as those fed the basal diet supplemented with % L-lysine and within 0.4 Downloaded from at Penn State University (Paterno Lib) on September 8, 06

6 4 S. L. BALLOUN TABLE.Effect of added dietary lysine on gains and feed conversion of poults. Experiment l Prot. Diet Weight gains by periods (pounds) 4 weeks Lysine Weight F/G (%) Each figure in the table is the rounded average of groups of Bronze males started per group. protein8, 4, 0, 6 and 4% protein in the age periods respectively. protein4, 0 6, 4 and % protein in the age periods respectively. pound as much as those poults fed 0% L-lysine. However, there was a strong trend toward more advantage from the feeding of lysine in the finishing periods. Even with the high-protein diets, poults receiving % L-lysine were 0.8 pound heavier at 4 weeks, and those receiving the higher level of lysine were. pounds heavier than poults fed the basal highprotein diets. In the 4-week finishing period (0-4 weeks), adding L-lysine increased gains by 0.8 pound, or %, over gains made by poults fed the high-protein diet without supplemental lysine. Lysine additions resulted in greater weight gains in poults fed the low-protein diets in all periods, but, as with the highprotein diets, the greatest effect was obtained in the finishing periods. Adding lysine at % increased 4-week weights 0.8 pound and adding lysine at % increased weights.7 pounds. In the 8-week period after 6 weeks, poults fed % and % added lysine gained 0 and.4% more, respectively, than did poults fed the low-protein diets without added lysine. Until the 6th week, improvements of gains as a result of adding lysine were small and non-significant. Supplementing low-protein diets with lysine improved amino acid balance sufficiently so that greater weights were achieved than with the basal high-protein diet. In fact, the low-protein diets supplemented with % L-lysine resulted in poults as heavy as those which had been fed the high-protein diets supplemented with % L-lysine, and 0.7 pound heavier than those fed the basal high-protein diet. No consistent pattern was evident in the feed efficiency data, except that, in general, high-protein diets were more efficiently converted than were low-protein diets. Since diets were very nearly iso-caloric, the difference in feed efficiency could not be ascribed to energy differences. DISCUSSION In the two experiments reported, the beneficial effects of lysine appeared to be considerably greater in the later periods of the experiments, whereas, when methionine was a variable, the effect of this amino acid was evident only in the beginning periods and not in the finishing periods. An examination of the amino acid balance of the diets fed indicates that the diets were somewhat deficient in methionine in the starting period, but were only slightly deficient, if at all, in lysine in the beginning periods. As the experiments progressed, however, and Downloaded from at Penn State University (Paterno Lib) on September 8, 06

7 AMINO ACID BALANCE 4 protein was lowered by the addition of corn and reduction of soybean oil meal to accommodate the lowered protein needs of the poult, the amino acid balance was changed considerably so that in the finishing periods lysine became quite deficient and methionine balance actually improved. It is thought that this is the reason for the greater benefit from lysine feeding in the later periods. It should not be overlooked, however, that methionine may be more important to the poult in the early periods for the production of large amounts of feathers in proportion to general growth. The greater improvement in weight gains obtained in experiment as a result of amino acid supplementation when the diet contained a high concentration of energy, supports the theory that protein requirements are higher in high-energy diets. This was not so evident with lysine supplementation alone, but was evident when lysine plus methionine, or lysine, methionine and arginine, were added to high- and low-energy diets. With the high-energy diet, supplementation with all three amino acids resulted in improved weight gains at 6 weeks of age of approximately. lbs., or nearly %. With lowenergy diets, supplementation with all three amino acids resulted in weight gain improvements of just over a pound, or 8.%. While this difference is not great, it does lend support to the theory that protein, and therefore amino acid, requirements are dependent on the energy level of the diet. It should be noted, however, that the lysine response was not dependent on the energy level. The results of experiment, in which both high- and low-protein diets were supplemented with lysine, gives considerable support to the work of Nelson et al. (960). All of the diets in this experiment were relatively high in energy. Since supplementing the high-protein diets with lysine improved weight gains at 4 weeks of age by. lbs., or about 6%, and adding lysine in comparable amounts to the lowprotein diets improved weight gains at 4 weeks of age by almost exactly.7 lbs., or 7.8%, it would seem that the lysine requirement of poults over the entire experimental period proved to be a constant percentage of the protein fed rather than an absolute percent of the total diet. If lysine were expressed as a percent of the protein it is interesting to note that a remarkable consistency exists in the point at which lysine supplementation improved diets of high- and of low-protein concentration. In the case of the high-protein diets, little improvement in weight gains were obtained until about the 0th week. In the last four weeks of the experimental period, between 0-4 weeks of age, birds gained 0.8 pound more as a result of lysine supplementation. However, with the lowprotein diets, lysine began to manifest its influence at an earlier age, with some improvement, in weight gains being observed in the 6-0 week period, and this increased gain from lysine supplementation continued through the 0-4 week period. Calculations of the diets show that not until the 0th week did the lysine in unsupplemented high-protein diets drop below % of the protein. In the low-protein diets, this point was reached at 6 weeks of age. This could explain the great improvement in weight gains in later periods obtained with either high- or lowprotein diets when supplemented with lysine, and also the fact that, in the case of the high-protein diets, the improvement came later than with low-protein diets. It appears from these data, especially from experiment, that the lysine requirement of the rapidly growing poult, after the starting period, is just over % of the protein content of the diet with a Downloaded from at Penn State University (Paterno Lib) on September 8, 06

8 44 S. L. BALLOUN high-energy diet. Whenever lysine, as a percent of the protein, drops to % substantial benefits may be expected from lysine supplementation. These results confirm those reported in an earlier paper (Balloun and Phillips, 97). SUMMARY Broad Breasted Bronze poults have been fed to 6 weeks in one experiment and to 4 weeks in another on high- and low-protein diets, variously supplemented with lysine, methionine and arginine. Lysine supplementation generally improved weight gains on either high- or low-protein diets when lysine was % or less of the protein in the diet. The effects of methionine, but not lysine, supplementation were slightly greater with highenergy diets. On both high- and lowenergy diets, methionine tended to improve weight gains earlier in the growing period and lysine improved weight gains in later periods. Arginine supplementation improved weights only a small amount at any period. For Broad Breasted Bronze poults, the lysine requirement in growing and finishing diets appears to be at least % and the methionine requirement appears to be about % of the protein of the diet. ACKNOWLEDGMENTS This investigation was supported in part by a grant-in-aid from Merck, Sharp & Dohme, Rahway, New Jersey. Vitamins were supplied through the courtesy of Merck, Sharp & Dohme, Charles Pfizer & Co., Inc., Terre Haute, Indiana, Distillation Products, Rochester, New York and Dawe's Laboratories, Inc., Chicago, Illinois. Animal fat was supplied by Armour and Company, Chicago, Illinois. REFERENCES Almquist, H. J., 949. Amino acid balance at supernormal dietary levels. Proc. Soc. Exp. Biol. Med. 7: Almquist, H. J., 9. Amino acid requirements of chickens and turkeysa review. Poultry Sci. : Balloun, S. L., and R. E. Phillips, 97. Lysine and protein requirements of Bronze turkeys. Poultry Sci. 6: Balloun, S. L., W. J. Owings, J. L. Sell and R. E. Phillips, 99. Energy and protein requirements for turkey starting diets. Poultry Sci. 8: Carter, R. D., E. C. Naber, S. P. Touchbum, J. W. Wyne, V. D. Chamberlin and M. G. McCartney, 96. Amino acid supplementation of low protein turkey growing rations. Poultry Sci.in press. Donovan, G. A., E. L. Johnson, S. L. Balloun and R. E. Phillips, 9. The long range effect of low level methionine supplementation in growing turkey rations. Poultry Sci. 4: -6. Grau, C. R., and M. Kamei, 90. Amino acid imbalance and the growth requirements for lysine and methionine. J. Nutrition, 4: Kratzer, F. H., F. H. Bird, V. S. Asmundson and S. Lepkovsky, 947. The arginine requirement of young turkey poults. J. Nutrition, 4: National Research Council, 960. Nutrient requirements of domestic animals. No.. Nutrient requirements of poultry. Nelson, T. S., R. J. Young, R. B. Bradfield, J. B. Anderson, L. C. Norris, F. W. Hill and M. L. Scott, 960. Studies on the sulfur amino acid requirements of the chick. Poultry Sci. 9: Sherman, W. C, G. A. Donovan and W. M. Reynolds, 960. Evaluation of lysine in turkey rations. Poultry Sci. 9: 9. Titus, H. W., 9. The Scientific Feeding of Chickens. The Interstate Printers, Danville, Illinois. Waibel, P. E., 99. Methionine and lysine in rations for turkey poults under various dietary conditions. Poultry Sci: 8: 7-7. Downloaded from at Penn State University (Paterno Lib) on September 8, 06 SEPTEMBER 8-. FIRST INTERNATIONAL CONGRESS OF FOOD SCIENCE AND TECHNOLOGY, IMPERIAL COLLEGE OF SCIENCE AND TECHNOLOGY, LONDON, ENGLAND