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Dietary valine needs of commercial broilers M. T. Kidd,1 S. Dridi,1 J. Bai,1,3 and E. Diehl2 1 Center of Excellence for Poultry Science, Division of Agriculture, University of Arkansas System, Fayetteville, Arkansas; 2Cobb-Vantress, Inc., Siloam Springs, Arkansas Summary 1) The global acceptance of L-Thr has resulted in several research laboratories focusing on the needs of BCAA for broilers. 2) Val is typically the fourth limiting amino acid in corn-soybean meal based diets, excluding animal by-product meals, after TSAA, Lys, and Thr. 3) The dietary inclusion of animal by-product meals to diets based on corn-soybean meal renders Ile as typically fourth limiting, or co-limiting with Val, but over formulation of Ile or Arg in diets containing a variety of cereals (i.e., corn, sorghum or wheat) can easily render these amino acids as fourth limiting. 4) Feeding Cobb 500 female broilers varying levels of BCAA resulted in digestible Val and Ile to Lys of 77 and 69, respectively, improving 14 to 28 d weight gain, but 28 to 42 d BCAA levels failed to impact live performance. Broilers (42 d) fed the 72 Val/Lys ratio had the highest leg yields. Molecular assessment of genes indicated that low BCAA inhibit hepatic fatty acid synthesis and enhance hepatic fatty acid β-oxidation. 5) Feeding Cobb 500 female broilers a digestible Val/Lys dose response failed to yield quadratic responses from 14-28 d, 28-42 d, and 42-56 d of age. The digestible Val/Lys dose responses used in the work were 64, 67, 70, 73, 76, 79, and 82 from 14-28 d of age and 65, 68, 71, 74, 77, 80, and 83 in both finishing periods. 6) More research should be conducted with the Cobb 500 female broiler regarding BCAA needs as our work points to potential dietary BCAA needs considerably lower than currently fed. 7) Future research with Val should be conducted in stress versus non-stress environments as the amount of body Val converted to mucin Gln could be substantial. Abbreviations: Arginine = Arg; branched-chain amino acids = BCAA; glutamine = Gln; glycine = Gly; Isoleucine = Ile; day = d; leucine = Leu; lysine = Lys; total sulfur amino acids = TSAA; threonine = Thr; tryptophan = Trp; valine = Val. 3 Current address: Jiangsu Tyson Foods Co., Ltd. Jiangsu Province China. Kidd et al. 2015 Arkansas Nutrition Conference Proceedings, Rogers, AR. DIETARY VAL FOR BROILERS Introduction In a previous venue of the Arkansas Nutrition Conference historical acceptance of feed grade amino acids in the United States and their subsequent feed formulation pressure on downstream amino acids was discussed (Kidd, 2013). Clearly, the widespread acceptance of L-Thr has placed pressure on the fourth limiting amino acid nutrient minimum in linear programming. In order to assess L-Thr dietary opportunities, formulation projections with corn-soybean meal based diets, excluding meat meals, revealed that Val is typically fourth limiting in a broiler grower diet containing 1.0 or 1.1 % digestible Lys (Kidd and Hackenhaar, 2005). In the former formulation examples, downstream amino acid ratios to Lys (i.e., Ile, Arg, and Trp) were set at 65, 105, and 17, respectively, and Val was set at 77. Corzo et al. (2007a) began a series of studies to assess Val marginality in 21 to 42 d old Ross 708 male broilers fed on corn and soybean meal diets devoid of animal by-product meals. Val, Ile, Arg, or Gly were added at surfeit levels to a negative control diet and a corn-peanut meal diet was used to create Val marginality whereby L-Val was titrated. Val was confirmed to be fourth limiting and the dose response experiment established a Val/Lys ratio of 78 for average performance and processing measurements (Corzo et al. 2007a). These proceedings provide an overview of dietary Val needs and L-Val inclusion considerations, as well as recent broiler research conducted at the Center of Excellence for Poultry Science with Cobb 500 females. Broiler studies with L-Val: Early literature Early published reports in the United States (Dobson et al., 1964; Dean and Scott, 1965; Thomas et al., 1981) and the United Kingdom (Hewitt and Lewis, 1972 and D’Mello, 1974) established BCAA needs in chicks by adding one or a combination of BCAA. Val needs varied among published reports, and much of the variation was attributed to diet and broiler chick strain. However, confusion existed with regard to BCAA dependence, antagonism, and co-limitation. Farran and Thomas (1990) used a central composite design to assess simultaneous BCAA addition in broiler chicks from 7 to 21 d. Responses to Leu were minimal suggesting the requirement was no higher than 1.16% of diet, whereas Val and Ile were optimized (99% of asymptote) at 0.90 and 0.80%, respectively (Farran and Thomas, 1990). Additional work at the University of Maryland was conducted to investigate the effect of Val deficiency on broiler chick performance, bone quality, and feather structure and it was determined that dietary Val reductions alone were more detrimental than reducing the three BCAA simultaneously (Farran and Thomas, 1992). Burnham et al. (1992) conducted three experiments from 8 to 21 d of age to assess Ile needs and potential BCAA antagonisms via diet dilution methodology. Ile needs were Kidd et al. 2015 Arkansas Nutrition Conference Proceedings, Rogers, AR. DIETARY VAL FOR BROILERS generated, but minimal impact of Leu and no impact from Val were noted on Ile needs (Burnham et al. 1992). Hence, this work diminished the practicality of Val and Ile antagonism. In addition, subsequent work by Farran et al. (2003) showed that excessive levels of Leu can decrease broiler performance, in low, but not adequate, CP diets. Leclercq (1998) of the INRA Poultry Research Station in France, in collaboration with Drs. Schutte (The Netherlands) and de Groote (Belgium), conducted a series of experiments with the most limiting amino acids (TSAA, Lys, Thr, Val, Ile, Arg, and Trp) in 20 to 40 d old broilers. All three laboratories used the same level of Val (0.61, 0.67, 0.72, 0.77, 0.83, and 0.88% of diet) and the results were summarized at the 1997 Annual Poultry Science Association meeting at the University of Georgia (Table 1). Male broilers from 20 to 40 d of age responded to Val/Lys needs from 61 to 78 (broken line model) and 75 to 85 (monomolecular model). Table 1. Digestible Val/Lys needs averaged across experiments with two statistical approaches1 Parameter Broken line Val/Lys Monomolecular Val/Lys Weight gain 78 85 Feed conversion 61 75 Breast meat yield 73 81 1 Data from Leclercq (1998). BioKyowa research was conducted in low protein diets in 22 to 42 d old broilers to assess BCAA needs (Kidd et al. 2000). Diets were formulated to be marginally deficient in BCAA (90% NRC, 1994, Val and Ile) and provide surfeit BCAA with crystalline L-Val and L-Ile. A positive breast meat yield response was noted in birds fed supplemental Ile. Interestingly, this work was conducted with corn-soybean meal all vegetable diets, suggesting Ile was more limiting than Val for breast meat yield, but other parameters were not affected by BCAA (Kidd et al. 2000). Broiler studies with L-Val: Recent literature The dietary acceptance of L-Thr, especially the exponential dietary increase in the late 2000’s, has increased pressure on the fourth limiting amino acid, typically Val, in broiler diets. Dr. Tillman of Poultry Technical Nutrition Services in Georgia summarized recent literature on Val needs for broilers in his proceedings for the Brazilian Tables (Tillman, 2011). These references on Val needs of broilers, in addition to more recent publications, are listed in Table 2. With respect to age and strain, digestible Val estimates can serve as a basis of comparison to feed formulation Val “nutrient” minimums. Expression of Val needs are typically expressed as an ideal protein ratio, but calculation in some references must be taken with caution because ratio expression in experiments without marginal Lys will underestimate the Val ratio (Table 2). One of the most relevant studies to assess Val needs and commercial dietary acceptance of L-Val is the work of Corzo et al. (2011) where L-Val was titrated in diets that only included Kidd et al. 2015 Arkansas Nutrition Conference Proceedings, Rogers, AR. DIETARY VAL FOR BROILERS feed-grade amino acids currently being used by poultry nutritionists (i.e., synthetic Met and crystalline Lys and Thr). Hence, the design allowed for blending of a diet with no L-Val and a diet with 2.6 pounds of L-Val per short-ton or 1.3 kg of L-Val/metric ton, but both diets contained a digestible Val/Lys of 78. Because practical diets were used, as L-Val increased in the titration, the diets because progressively reduced in the downstream amino acid nutrients (e.g., Ile, Arg, and Trp). Good broiler performance was attained in diets with 1.04 pounds of LVal per short-ton or 0.52 kg of L-Val/metric ton, whereas higher L-Val levels increased feed conversion and reduced meat yields presumably due to Ile and/or Arg limitations (Corzo et al., 2011). Table 2 Recent research on digestible valine requirements in male broilers1 Day of age 1-14 1-21 7-21 8-21 14-28 21-35 21-42 21-42 28-40 28-42 28-42 30-43 42-56 % valine2 0.91 0.90 0.84 774 0.86 0.84 0.65 0.72 0.77 0.7-0.783 0.78 764 0.67 Strain Ross x 308 Cobb x 500 Cobb x 500 Cobb x 500 Ross x 308 Cobb x 500 Ross x 508 Ross x 708 Cobb x 500 Ross x 708 Ross x 308 Cobb x 500 Ross x 308 Reference Corzo et al. (2008) Goodgame et al. (2011) Campos et al. (2009) Tavernari et al. (2013) Corzo et al. (2008) Goodgame et al. (2011) Thornton et al. (2006) Corzo et al. (2007a) Campos et al. (2009) Dozier et al. (2012) Corzo et al. (2008) Tavernari et al. (2013) Corzo et al. (2004) 1 Adapted and modified from Tillman (2011). Average of body weight gain and feed conversion estimates. 3 Dozier et al. (2012) designed this work to assess digestible BCAA ratio interactions and in one experiment Val/Lys improved live performance from increases of 74 to 82. 4 Ratios were established from total Val and Lys contents in test diets after NIR and verified by HPLC. 2 Recent Val research in the Center of Excellence for Poultry Science Two experiments (14 to 28 d and 28 to 42 d) were conducted to evaluate Val and Ile in Cobb 500 female chicks in a factorial array. In both experiments birds were fed on common diets prior to experiment initiation. The levels of Val and Ile as a digestible ratio to Lys were 72 and 77 and 63, 66, and 69 in both experiments, respectively (unpublished). Molecular mechanisms from tissue (liver and thigh) in the 28 to 42 d experiment were collected to assess the role of BCAA in protein and fat metabolism (Bai et al., 2015). In the 14 to 28 d experiment interactions were limited and in the 28 to 42 d experiment interactions did not occur. BW gain interaction response (P = 0.046) resulted in optimal gain in Kidd et al. 2015 Arkansas Nutrition Conference Proceedings, Rogers, AR. DIETARY VAL FOR BROILERS birds fed Val and Ile to Lys of 77 and 69, respectively, as compared to 77 and 63, 72 and 69, and 72 and 66 Val and Ile to Lys, respectively, from 14 to 28 d. Interactions did not occur for feed conversion (P = 0.094) or feed intake (P = 0.619) in either experiment. Thigh meat yield (boneless and skinless) approached significance, similar to wing yield (P = 0.052), but an interaction occurred for drumstick yield (P = 0.029) from 14 to 28 d indicating that optimal yields could be attained with the lowest or highest BCAA levels. Dridi et al. (unpublished) has completed three Val dose response trials from 14-28, 28-42, and 42-56 d of age. Female Cobb 500 broilers were used and the digestible Val/Lys dose responses in the work were 64, 67, 70, 73, 76, 79, and 82 from 14-28 d of age and 65, 68, 71, 74, 77, 80, and 83 in both finishing periods. No linear or quadratic responses were observed pointing to the importance of further work to assess the digestible Val/Lys in Cobb 500 female birds. Bai et al. (2015) removed livers from 1 bird per pen in the 28 to 42 d BCAA factorial study noted above at d 42. Reverse transcription and real-time quantitative PCR were performed. A schematic representation (Figure 1) is presented below which indicates that that low BCAA levels inhibited fatty acid synthesis and enhanced β-oxidation as mediated through the AMPK-mTOR-FoxO1 pathway. Figure 1. Schematic representation of proposed dietary BCAA effect on hepatic fatty acid metabolism (as adapted from Bai et al., 2015). Kidd et al. 2015 Arkansas Nutrition Conference Proceedings, Rogers, AR. DIETARY VAL FOR BROILERS Lessons learned from L-Thr research/implementation and L-Val considerations In the context of amino acid limitation and thus dietary balance, upstream adequacy of amino acid limitation is vital for downstream responses. This is especially true with Thr as limitations in Lys inhibit the breast meat yield response (Kidd et al. 1997). This was first noted by Hickling et al. (1990) who showed that Met limited the ability of Lys to optimize breast meat yield. Although expressing the Thr need as a ratio to Lys was accepted, amino acid inclusion in practical formulas in the 1990’s had two main challenges, the residual L-Lys ingredient forced maximum “the Lys cap” and the dearth and variation of Thr estimates. Research in Dr. Ron Ball’s laboratory has established the need of Thr for intestinal mucin (Law et al. 2007). This work clearly pointed to the importance of Thr for intestinal viability at a time when nutritionist were testing ABF programs, coccidiosis vaccinations, and raising nutrient density. Two Thr dose response trials in varying (dirty litter vs new litter) environmental conditions were conducted and demonstrated that Thr needs in dirty environments were increased from 5% (Corzo et al., 2007b) to 12% (Kidd et al., 2003). Star et al. (2012) conducted Thr experiments in birds with necrotic enteritis and found Thr needs relative to Lys increasing from 63 to 67 in non-stressed vs stressed birds, respectively. Could the variation in Val needs observed in broilers be attributable to intestinal needs? Hence, Thr breakdown via Thr dehydrogenase or aldolase yields Gly, an important component of mucin. The BCAA conversion to alpha-ketoglutarate uses the N in BCAA to be converted to Glu and then Gln. Of the mucin needs, both Gly and Gln are high which make upstream precursors of Thr and BCAA, respectively, as potentially limiting during enteric stress. Although Leu would indeed contribute to alpha-ketoglutarate, the need for Val derived alphaketoglutarate during periods of high enteric stress may be attributing to research variations in predicting the Val need. Conclusions 1) Valine estimates in Table 2 can be used as a basis of comparison to Val nutrient minimums in linear programming. 2) Val/Lys of 78 appears to optimize performance of most broiler strains, but may be an overestimation in the Cobb 500 female broiler. 3) An understanding of Ile and Arg minimums in least cost formulation, pending commodity price spreads, should allow for the inclusion of one pound of L-Val per ton of broiler feed. 4) Amino acid work going forward, especially that of L-Val, should focus on stress vs nonstress environments. Kidd et al. 2015 Arkansas Nutrition Conference Proceedings, Rogers, AR. DIETARY VAL FOR BROILERS References Bai, J., E. Greene, W. Li., M. T. Kidd, and S. Dridi, 2015. Branched-chain amino acids modulate the expression of hepatic fatty acid metabolism-related genes in female broiler chickens. Mol. Nutr. Food Res. 00:1-11. Burnham, D., G. C. Emmans, and R. M. Gous, 1992. Isoleucine requirements of the chicken: The effect of excess leucine and valine on the response to isoleucine. Br. Poultry Sci. 33:71-87. Campos, A., E.T. Nogueira, L.F. Albino, and H. S. Rostagno. 2009. Digestible valine:lysine ratios for broilers during the starter and finisher periods. Poult. Sci. 88(E-Suppl. 1): 109. Corzo, A., W.A. Dozier III, and M.T. Kidd. 2008. Valine nutrient recommendations for Ross × Ross 308 broilers. Poult. Sci. 87:335-338. Corzo, A. W. A. Dozier, III, L. Mejia, C. D. Zumwalt, M. T. Kidd, and P. B. Tillman, 2011. Nutritional feasibility of L-valine inclusion in commercial broiler diets. J. Appl. Poult. Res. 20:284-290. Corzo, A., M. T. Kidd, W. A. 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Acknowledgements Feed grade and food grade amino acids for diet formulation in the L-Val research conducted at the Center of Excellence for Poultry Science were provided by Zhirong Jiang of Ajinomoto, Chicago, Illinois. Much of the work cited herein was accomplished through collaborative efforts of Alex Corzo’s and Bill Dozier’s laboratory, which involved vital input from a number of graduate students, and technical and financial support from Ajinomoto: Paul Tillman, James Usry, and Zhirong Jiang. Kidd et al. 2015 Arkansas Nutrition Conference Proceedings, Rogers, AR. DIETARY VAL FOR BROILERS