Varying the ratio of Lys:Met while maintaining the ratios of Thr:Phe, Lys:Thr, Lys:His, and Lys:Val alters mammary cellular metabolites, mammalian target of rapamycin signaling, and gene transcription.

Amino acids are not only precursors for but also signaling molecules regulating protein synthesis. Regulation of protein synthesis via AA occurs at least in part by alterations in the phosphorylation status of mammalian target of rapamycin (mTOR) pathway proteins. Although the ideal profile of Lys:Met to promote milk protein synthesis during established lactation in dairy cows has been proposed to be 3:1, aside from being the most-limiting AA for milk protein synthesis, the role of Met in other key biologic pathways such as methylation is not well characterized in the bovine. The objective of this study was to determine the influence of increasing supplemental Met, based on the ideal 3:1 ratio of Lys to Met, on intracellular metabolism related to protein synthesis and mTOR pathway phosphorylation status. MAC-T cells, an immortalized bovine mammary epithelial cell line, were incubated (n = 5 replicates/treatment) for 12 h with 3 incremental doses of Met while holding Lys concentration constant to achieve the following: Lys:Met 2.9:1 (ideal AA ratio; IPAA), Lys:Met 2.5:1 (LM2.5), and Lys:Met 2.0:1 (LM2.0). The ratios of Thr:Phe (1.05:1), Lys:Thr (1.8:1), Lys:His (2.38:1), and Lys:Val (1.23:1) were the same across the 3 treatments. Applying gas chromatography-mass spectrometry metabolomics revealed distinct clusters of differentially concentrated metabolites in response to Lys:Met. Lower Phe, branched-chain AA, and putrescine concentrations were observed with LM2.5 compared with IPAA. Apart from greater intracellular Met concentrations, further elevations in Met level (LM2.0) led to greater intracellular concentrations of nonessential AA (Pro, Glu, Gln, and Gly) compared with IPAA and greater essential AA (EAA; Met, Ile, and Leu) and nonessential AA (Pro, Gly, Ala, Gln, and Glu) compared with LM2.5. However, compared with IPAA, mRNA expression of ß-casein and AA transporters (SLC7A5, SLC36A1, SLC38A2, SLC38A9, and SLC43A1) and mTOR phosphorylation were lower in response to LM2.5 and LM2.0. Overall, the results of this study provide evidence that increasing Met while Lys and the ratios of Phe, Thr, His, and Val relative to Lys were held constant could increase the concentration and utilization of intracellular EAA, in particular branched-chain AA, potentially through improving the activity of AA transporters partly controlled by mTOR signaling. Because EAA likely are metabolized by other tissues upon absorption, a question for future in vivo studies is whether formulating diets for optimal ratios of EAA in the metabolizable protein is sufficient to provide the desired levels of these AA to the mammary cells.

Increasing the availability of threonine, isoleucine, valine, and leucine relative to lysine while maintaining an ideal ratio of lysine:methionine alters mammary cellular metabolites, mammalian target of rapamycin signaling, and gene transcription.

Amino acids not only serve as precursors for protein synthesis but also function as signaling molecules that can regulate the mammalian target of rapamycin (mTOR) pathway. Methionine and Lys are the most-limiting AA for milk production and a ratio of ∼3:1 Lys:Met in the metabolizable protein has been determined to be ideal. Besides Met and Lys, recent studies have evaluated Ile, Leu, Val, and Thr as potentially limiting for milk protein synthesis. The objective of this experiment was to determine if varying the ratio of Lys:Thr, Lys:Ile, Lys:Val, and Lys:Leu while maintaining an ideal ratio of Lys:Met and fixed ratio of other essential AA (IPAA) elicits changes in intracellular metabolites, gene transcription related to protein synthesis, and phosphorylation status of mTOR pathway proteins. Immortalized bovine mammary epithelial cell line (MAC-T) cells were incubated for 12 h (n = 5 replicates/treatment) with IPAA (2.9:1 Lys:Met; 1.8:1 Lys:Thr; 2.38:1 Lys:His; 1.23:1 Lys:Val; 1.45:1 Lys:Ile; 0.85:1 Lys:Leu; 2.08:1 Lys:Arg) or IPAA supplemented with Thr, Ile, Val, and Leu to achieve a Lys:Thr 1.3:1 (LT1.3), Lys:Ile 1.29:1 (LI1.29), Lys:Val 1.12:1 (LV1.12), or Lys:Leu 0.78:1 (LL0.78). Compared with IPAA, metabolomics via gas chromatography-mass spectrometry revealed that increases in availability of Thr, Ile, Val, and Leu led to greater concentrations of essential AA (Leu, Ile, Thr), nonessential AA (Gly, Glu, Gln, Ser, Pro, Asp), and various metabolites including uric acid, phosphoric acid, N-acetylglutamic acid, and intermediates of glycolysis and the tricarboxylic acid cycle. Compared with other treatments, LV1.12 led to greater phosphorylation status of serine/threonine kinase B (Akt), mTORC1, and ribosomal protein S6 and lower phosphorylation of α subunit of eukaryotic translation initiation factor 2. In addition, LV1.12 upregulated abundance of CSN2 and both the abundance and promoter methylation of CSN1S1. Although LI1.29 led to the second highest response in mTORC1 phosphorylation status, it resulted in the lowest phosphorylation of Akt and eEF2 and mRNA abundance of CSN2 and various AA transporters (SLC7A5, SLC36A1, SLC38A2, SLC38A9, SLC43A1). Overall, data indicate that an increase in Val at an ideal ratio of Lys:Met could further enhance milk protein synthesis by altering intracellular concentrations of essential AA and metabolites that could play a regulatory role, increasing phosphorylation status of mTORC1 and key signaling proteins, and upregulation of AA transporters.

Hepatic phosphorylation status of serine/threonine kinase 1, mammalian target of rapamycin signaling proteins, and growth rate in Holstein heifer calves in response to maternal supply of methionine.

The study investigated whether methionine supply during late pregnancy is associated with liver mammalian target of rapamycin (MTOR) pathway phosphorylation, plasma biomarkers, and growth in heifer calves born to cows fed a control diet (CON) or the control diet plus ethylcellulose rumen-protected methionine (MET; 0.09% of dry matter intake) for the last 28 d prepartum. Calves were fed and managed similarly during the first 56 d of age. Plasma was harvested at birth and 2, 7, 21, 42, and 50 d of age and was used for biomarker profiling. Liver biopsies were harvested at 4, 14, 28, and 50 d of age and used for protein expression. Body weight, hip height, hip width, wither height, body length, rectal temperature, fecal score, and respiratory score were measured weekly. Starter intake was measured daily, and average daily gain was calculated during the first 8 wk of age. During the first 7 wk of age, compared with calves in the CON group, calves in the MET group had greater body weight, hip height, wither height, and average daily gain despite similar daily starter intake. Concentration of methionine in plasma was lower at birth but increased markedly at 2 and 7 d of age in MET calves. Plasma insulin, glucose, free fatty acids, and hydroxybutyrate did not differ. A greater ratio of phosphorylated α-serine/threonine kinase (AKT):total AKT protein expression was detected in MET calves, namely due to differences at 4 d of age. The phosphorylated MTOR:total MTOR ratio also was greater in MET calves due to differences at 28 and 50 d (8 d postweaning). The decrease in phosphorylated MTOR:total MTOR between 14 and 28 d in CON calves agreed with the increase in phosphorylated eukaryotic translation initiation factor 4E binding protein 1 (EIF4EBP1):total EIF4EBP1 ratio during the same time frame. The overall expression of phosphorylated ribosomal protein S6 kinase B1 (RPS6KB1):total RPS6KB1 and phosphorylated eukaryotic translation elongation factor 2 (EEF2):total EEF2 was lower in MET calves. Regardless of methionine supply prepartum, there was an 11-fold temporal decrease from 4 to 50 d in phosphorylated AKT:total AKT. Similarly, regardless of methionine supply, there were overall decreases in phosphorylation ratios of AKT, MTOR, RPS6KB1, and eukaryotic translation initiation factor 2A (EIF2A) over time. Data provide evidence of a positive effect of methionine supply during the last month of pregnancy on rates of growth during the first 7 wk of age. Phosphorylation status of some components of the MTOR pathway in neonatal calf liver also was associated with greater maternal supply of methionine. Thus, the data suggest that molecular mechanisms in the liver might be programmed by supply of methionine during late pregnancy. The exact mechanisms coordinating the observed responses remain to be determined.

Tissue and plasma antioxidant status in response to dietary methionine concentration and source in broilers.

This study hypothesized that plasma and tissue antioxidant status of broilers is positively influenced when dietary Met concentrations exceed, and negatively when they go below NRC recommendations. In addition, different Met sources are hypothesized to affect the antioxidant defence system differently. Day-old male Cobb-500 broilers (n = 336) were allotted to seven groups and phase-fed three wheat-soya bean meal-based basal diets during days 1-10, 11-21 and 22-35. The basal diets (Met- group, Met + Cys concentration 15% below NRC recommendations) were supplemented with 0.10%, 0.25% or 0.40% Met either as DL-Met (DLM) or DL-2-hydroxy-4-(methylthio) butanoic acid (DL-HMTBA) (equimolar comparison). Growth performance and carcass weights were lower in the Met- group compared to the groups whose diets met or exceeded Met requirements. The antioxidant defence system was not influenced by the Met source. However, in the liver, concentrations of glutathione increased with increasing dietary Met concentrations. Tocopherol concentrations in the liver at days 10 and 21 were lower in the Met- group than in the groups supplemented with Met. However, liver concentrations of thiobarbituric acid reactive substances (TBA-RS) and protein carbonyls (PC) were largely not influenced by dietary Met concentration. Plasma tocopherol concentrations at day 35 were lower, and those of TBA-RS and PC at day 35 were higher in Met- group than in the groups fed the Met-supplemented diets. In jejunum, but not in liver, relative mRNA abundances and activities of superoxide dismutase, catalase and glutathione peroxidase were higher in the Met- group than in the groups fed Met-supplemented diets. These data indicate that suboptimum supply of Met results in decreased antioxidant concentrations in plasma and body tissues, and increases oxidative stress in the jejunum mucosa. However, supplementation of Met in excess of the requirements (based on NRC) compared to diets adequate in Met + Cys did not influence the antioxidant defence system.

Effect of glycine supplementation in low protein diets with amino acids from soy protein isolate or free amino acids on broiler growth and nitrogen utilisation.

Here, it was investigated whether substitution of amino acids (AA) from soy protein isolate with free AA in low crude protein diets influences the growth performance and N utilisation in broilers, and whether interactions with dietary glycine equivalent (Glyequi) concentration exist. Birds were distributed in two 2 × 2 factorial arrangements of 48 floor pens containing 10 birds each, plus 48 metabolism cages containing two birds each. Experimental feed was provided for ad libitum consumption from d 7 to 22. Diets contained either a soy protein isolate at 79 g/kg or a mix of free AA, which supplied the same amount of 18 proteinogenic AA. A mix of free glycine and l-serine was used to obtain low and high (12.0 and 20.5 g/kg dry matter) levels of dietary Glyequi. Substitution of soy protein isolate with free AA reduced the average daily gain and feed efficiency, mainly due to reduced feed intake. Efficiency of N accretion was not influenced by the AA source or Glyequi concentration on d 21, possibly due to the lower AA digestibility of soy protein isolate and higher urinary excretion of nitrogenous substances in the treatments with the AA mix. The average daily weight gain of the treatments with high Glyequi concentration was higher for both AA sources. This increase was due to higher average daily feed intake by broilers in the treatments with soy protein isolate and due to the increased feed efficiency in the treatments with the AA mix. Broilers exhibited different growth responses to dietary Glyequi between the AA sources; however, these responses could not be attributed to the different utilisation of Glyequi for uric acid synthesis.

A quantitative study of the interactive effects of glycine and serine with threonine and choline on growth performance in broilers.

Differences in the optimal dietary concentrations of Gly (glycine) and Ser (serine) in broiler diets may be due to levels of endogenous Gly precursors that differ in literature. Therefore, we measured the extent of the interactive effects between equivalents of Gly and Ser (Glyequi) and the endogenous Gly precursors choline and Thr (threonine) on growth performance. A fractional central composite design included concentrations of 15-25 g/kg DM, 0.6-2.0 g/kg DM, and 6.4-10.4 g/kg DM for Glyequi, choline, and Thr, respectively, in 5 levels each. The various concentrations were achieved by adding Gly, choline chloride, and l-Thr to a basal mix. Except for 20 replicates of the central diet, all treatments were tested with 5 replicates, each with 10 birds. Food was provided for ad libitum consumption throughout the experiment. The data were evaluated using artificial neural networks. Digestibility was studied for selected diets using separate birds. Since average daily feed intake (ADFI) varied between replicates, the intake of prececal digestible Glyequi, choline, and prececal digestible Thr were more adequate independent variables than the dietary concentration of each amino acid. From d 1 to d 7, no treatment effects on G:F and average daily gain (ADG) were detected; subsequent results refer to the period from d 7 to d 21. Increasing prececal digestible Thr intake considerably decreased the need for prececal digestible Glyequi to achieve certain levels of G:F and ADG. The extent of this effect cannot be explained only by the endogenous metabolism of Thr to Gly. Since essential amino acids were present above the recommended levels, Thr probably limited performance, and excessive intake of other essential amino acids prompted a Gly-dissipating process. Choline exerted a considerable effect on the required intake of prececal digestible Glyequi and prececal digestible Thr to achieve certain levels of G:F and ADG. The results of this study partly explain the previously reported variations in response to dietary Thr, Gly, and Ser.

Evaluation of the Valine requirement of small-framed first cycle laying hens.

An experiment was conducted to evaluate the total Valine (Val) requirement of first cycle laying hens from 41 to 60 wk of age. A total of 270 Hy-line W-36 laying hens were randomly assigned to 6 treatments with 15 replicate groups of 3 birds for each experimental unit. A Val deficient basal diet was formulated with corn and peanut meal with analyzed Val, Lys and crude protein concentrations of 0.515, 0.875, and 13.38%, respectively. Synthetic L-Val was supplemented to the basal diet in 0.070% increments to generate experimental diets containing 0.515, 0.585, 0.655, 0.725, 0.795, and 0.865% Val respectively. A controlled feeding program was applied during the experiment resulting in approximately 95 g feed intake per hen per day. Linear broken line, quadratic broken line, quadratic polynomial and exponential models were used to estimate the Val requirement of the hens based on hen-housed egg production (HHEP), egg mass (EM), and feed conversion ratio (FCR). Hen-housed egg production ranged from 48.3 to 81.4%, dependent upon dietary concentration of Val. Val requirements estimated by linear broken line, quadratic broken line, quadratic polynomial and exponential models were reported. Using the linear broken line model, the Val requirement was highest for egg mass, 597.3 mg/d, followed by egg production, 591.9 mg/d and lowest for FCR, 500.5 mg/d.

Evaluation of the tryptophan requirement of small-framed first cycle laying hens.

An experiment was conducted to evaluate the total Trp requirement of first cycle laying hens from 41 to 60 wk of age. In total, 270 Hy-Line W36 laying hens were randomly allocated to six treatments with 15 replicate groups of three birds for each experimental unit. A Trp-deficient basal diet was formulated with corn, corn gluten meal, and soybean meal with determined Trp, Lys, and crude protein concentrations of 0.096%, 0.873%, and 15.0%, respectively. Synthetic L-Trp was supplemented to the basal diet in 0.020% increments to generate experimental diets containing 0.096%, 0.116%, 0.136%, 0.156%, 0.176%, and 0.196% Trp, respectively. Hens were provided a controlled amount of feed of approximately 95 g/d. The diet containing the lowest concentration of Trp resulted in reduced egg production and was halted at 45 wk of age due to low performance with all other dietary treatments reaching the conclusion of the experiment at 60 wk of age. Plasma serotonin responded to dietary Trp concentration, but was not a good candidate for Trp requirement estimation. Linear broken line, quadratic broken line, quadratic polynomial, and exponential models were used to estimate Trp requirement based on hen-housed egg production (HHEP), egg mass (EM), and feed conversion ratio (FCR). HHEP ranged from 50.7% to 81.0%, dependent upon dietary concentration of Trp. Tryptophan requirements estimated by linear broken line, quadratic broken line, quadratic polynomial, and exponential models were reported. Using the linear broken line model, Trp requirement was highest for EM, 155.8 mg/d, followed by egg production, 153.2 mg/d and lowest for FCR, 140.4 mg/d.

Influence of a specific amino acid pattern in the diet on the course of an experimental Campylobacter jejuni infection in broilers.

Campylobacter jejuni (C. jejuni) is one of the most important zoonotic pathogens worldwide. In Europe, the majority of the cases are caused by consuming contaminated poultry meat. The objective of the present study was to investigate potential effects of different crude protein levels in complete diets for broilers on infection dynamics of C. jejuni after experimental infection. In total, 300 commercial broilers line Ross 308 were divided into 4 different groups, including 5 replications of 15 chickens each. The chickens were fed a conventional diet (212 g CP/kg DM) and a protein-reduced test diet (190 g CP/kg DM) supplemented with essential amino acids. This resulted simultaneously in lower amino-acid concentrations preferentially utilized by C. jejuni, such as aspartate, glutamate, proline, and serine. One group of each feeding concept was infected artificially with C. jejuni at day 21 by applying an oral C. jejuni inoculum containing 4.17 ± 0.09 log10 cfu of C. jejuni to 3 of 15 chickens, called "seeders." Feeding the test diet resulted in a significant reduction (P < 0.001) in CP intake (31.5 ± 1.20 g CP/broiler/day and 27.7 ± 0.71 g CP/broiler/day, respectively), a significant decrease (P < 0.05) in crude mucin in excreta (55.7 ± 8.23 g/kg DM and 51.9 ± 7.62 g/kg DM, respectively), and in goblet cell number in cecal crypts (P < 0.05; 15.1 ± 5.71 vs. 13.6 ± 5.91 goblet cells/crypt). In groups receiving the test diet, the excretion of C. jejuni was significantly reduced in seeders by 1.9 log10 cfu/g excreta at day 23 (3.38a ± 2.55 vs. 1.47b ± 2.20; P = 0.033). At day 25, prevalence of C. jejuni in cloacal swabs amounted to 53.3% in the group fed the test diet and 75.7% in the control group, respectively (P < 0.05). In summary, a definite amino acid pattern in the broiler diets could contribute to a development of an effective feeding strategy to reduce the prevalence of C. jejuni infection in chickens (Patent No 17187659.2-1106).

Comparison of amino acid digestibility in full-fat soybean, two soybean meals, and peanut flour between broiler chickens and growing pigs.

The aim of this study was to determine the apparent ileal digestibility (AID) and the standardized ileal digestibility (SID) of CP and AA in full-fat soybean (FFSB), solvent-extracted soybean meal containing 430 g/kg CP (SBM-43), solvent-extracted soybean meal containing 470 g/kg CP (SBM-47), and peanut flour (PNF) fed to broiler chickens and pigs and to compare the digestibility of CP and AA between broiler chickens and pigs. Four diets were prepared to contain FFSB, SBM-43, SBM-47, and PNF as the sole source of N. A N-free diet was formulated to estimate the basal ileal endogenous losses of CP and AA for broiler chickens and pigs. In Exp. 1, a total of four hundred sixteen 21-d-old male broiler chickens (922.1 ± 79.9 g initial BW) were assigned to 5 experimental diets in a randomized complete block design with BW as a blocking factor. After 5 d of adaptation, ileal digesta samples were collected after birds were euthanized by CO asphyxiation. In Exp. 2, twenty barrows (62.0 ± 6.9 kg initial BW) surgically fitted with T-cannulas at the distal ileum were allotted to 5 experimental diets with 2 consecutive 7-d experimental periods. After 5 d of adaptation, ileal digesta samples were collected at d 6 and 7. For statistical analysis, treatments were considered a 2 × 4 factorial arrangement with effects of species and experimental diets (Exp. 1 vs. Exp. 2). There were no interactions between species and diets for the digestibility of CP and AA except for Cys ( < 0.01). The AID of CP and indispensable AA in pigs were greater ( < 0.01) than in broiler chickens. In both broiler chickens and pigs, the AID of CP and indispensable AA in SBM-47 were greater ( < 0.05) than in FFSB, and the AID of Lys in PNF was the least ( < 0.05) among ingredients. The SID of CP and indispensable AA in pigs were greater ( < 0.05) than in broiler chickens except for Trp. In both broiler chickens and pigs, the SID of Lys in PNF was the least ( < 0.05) among ingredients. In broiler chickens, the SID of CP, Ile, Leu, Thr, Trp, and Val in FFSB were less ( < 0.05) than that in SBM-43, SBM-47, and PNF. In pigs, the SID of Arg, Ile, Leu, Met, Phe, and Val in FFSB were less ( < 0.05) than in SBM-43, SBM-47, and PNF. In conclusion, the digestibility of CP and most AA were less in broiler chickens than in pigs, but the pattern of differences in the AA digestibility among ingredients was similar between broiler chickens and pigs.

Interactive effects of dietary arginine and Eimeria acervulina infection on broiler growth performance and metabolism.

The influence of dietary Arg concentration and Eimeria acervulina infection on broiler growth performance and plasma carotenoid, nitric oxide (NO), amino acid, and urea concentrations was evaluated. Male Ross × Ross 308 broilers (384 total) were fed a common diet for 10 d post-hatch and provided experimental diets formulated to contain 1.23 (HA) or 0.74% (LA) standardized ileal digestible Arg from 10 to 28 d. At 21 d, one-half of the broilers were switched to the opposite diet to create 4 dietary regimens where birds were fed the LA diet throughout, the LA diet replaced by the HA diet at 21 d, the HA diet throughout, or the HA diet replaced by the LA diet at 21 d. Broilers were orally inoculated 0 (uninfected) or 3.5 × 105 sporulated E. acervulina oocysts at 15 d, resulting in a factorial arrangement of 4 dietary regimens × 2 infection states (8 replicates/treatment). Overall (10 to 28 d) BW gain and G:F were greatest (P < 0.01) for birds fed the HA diet throughout, regardless of infection status. Eimeria acervulina infection decreased (P < 0.01) BW gain of birds from 15 to 21 d, and G:F was lowest (P < 0.05) for infected birds fed the LA diet during this period. There was no influence (P > 0.05) of E. acervulina on BW gain or G:F of broilers from 21 to 28 d. Plasma Arg, Lys, and Orn levels at 21 d indicated that the LA diet caused an imbalance in the Lys and Arg status of broilers, and E. acervulina infection increased (P < 0.01) the plasma concentration of these 3 amino acids. Diet × infection interactions (P < 0.05) were observed on 21 d for plasma carotenoids and NO, whereby infection decreased plasma carotenoids and increased plasma NO, but dietary Arg concentration only influenced these measures for uninfected birds. Thus, production of NO during E. acervulina infection was not impaired by dietary Arg limitation.

Examining the effect of dietary electrolyte balance, energy source, and length of feeding of nitrogen-free diets on ileal endogenous amino acid losses in broilers.

The effect of dietary electrolyte balance (DEB), energy source (ES), and length of feeding of nitrogen-free diet (NFD) on ileal endogenous amino acid (EAA) loss in mg/kg dry matter intake (DMI) was evaluated in broiler chickens. In Experiment 1, 720 chickens consisting of 15 replicate cages with 6 chickens/replicate were used. Treatments were arranged in a 2 × 2 × 2 factorial and consisted of 4 NFD with 2 levels (low or high) of DEB and 2 ES [corn starch (CS) or dextrose (DX)], and 2 sampling time-points (diets were fed for either 72 h (d 16 to 19) or 120 h (d 16 to 21). Experiment 2 used 360 chickens in a 2 × 2 factorial arrangement of treatments with 2 levels (low or high) of DEB and 2 ES (CS or DX). Diets were fed for 72 h (d 18 to 21). All birds had access to feed and water on an ad libitum basis. Data were analyzed using the GLM procedure of SAS appropriate for a completely randomized design for a factorial arrangement of treatments. For Experiment 1, there were interactions (P < 0.05) between the 3 main factors for nitrogen and all the AA except Trp. Broilers that were fed DX-based NFD with high DEB for 72 h had the highest (P < 0.05) EAA losses. In Experiment 2, there was no interaction between DEB and ES except for His and Lys. When ileal EAA losses from birds fed the low DEB, CS-based NFD were used to standardize apparent ileal digestibility values from a previous study, there was no effect of length of feeding on standardized ileal AA digestibility values. In conclusion, DX-based NFD with high DEB increased endogenous AA loses. Despite differences in ileal EAA losses from CS-based NFD, standardized ileal AA digestibility values were not influenced by the length of feeding of NFD. Based on the results from these studies, NFD could be fed for 72 h without influencing SIAAD values.

Investigation of the effect of coccidial vaccine challenge on apparent and standardized ileal amino acid digestibility in grower and finisher broilers and its evaluation in 21-day-old broilers.

The effect of coccidial vaccine challenge (CVC, Coccivac(®)B; challenged, CHA; or unchallenged, NCH) on ileal endogenous amino acid (IEAA) losses and standardized ileal AA digestibility (SIAAD) in 21- and 42-day-old broilers fed a corn-soybean meal-dried distillers' grains with solubles-poultry by-product meal-based diet (Expt. 1) and the effect of supplemental AA in ameliorating the effect of CVC (Expt. 2) were evaluated. Expt. 1 was designed as a 2 (d 21 or 42) x 2 (NCH or CHA) factorial arrangement of treatments with 8 replicates in a complete randomized design. The CVC birds were gavaged with 12 x coccidial vaccine on d 15 and 36 and were sampled on 6 d post challenge. SIAAD was determined by correcting apparent ileal AA digestibility for IEAA losses. Feed intake (FI) and BW gain were higher (P ≤ 0.05) in 21-day-old NCH birds compared to the 21-day-old CHA birds while 42-day-old birds had higher FI and BW gain than 21-day-old NCH and CHA birds. Ileal endogenous nitrogen loss was higher (P ≤ 0.05) in 42-day-old CHA birds compared to 42-day-old NCH birds. Apparent ileal AA digestibility in 21-day-old CHA birds was lower (P ≤ 0.05) than for 21-day-old NCH and 42-day-old NCH and CHA birds. SIAAD in 21-day-old CHA birds was lower (indispensable AA = 15.2 and dispensable AA = 17.8%-unit; P ≤ 0.05) than for 21-day-old NCH and 42-day-old NCH and CHA birds. Apparent ileal digestibility and total tract utilization of DM, N, and energy were not different between d 21 NCH and d 42 NCH and CHA birds but higher (P ≤ 0.05) than for d 21 CHA birds. Supplemental AA restored feed efficiency, but not BW gain, to that of the positive control (0.715 vs. 0.737). Results from Expt.1 showed that CVC significantly influenced ileal AA digestibility in 21- but not in 42-day-old birds. Expt. 2 showed that consideration of SIAAD in feed formulation may benefit feed efficiency.

Influence of dietary amino acid reductions and Eimeria acervulina infection on growth performance and intestinal cytokine responses of broilers fed low crude protein diets.

Two experiments were conducted to evaluate the influence of Eimeria acervulina infection on growth performance, plasma carotenoids, and intestinal cytokine responses of broilers fed low crude protein (LCP) diets with reduced concentrations of selected amino acids (AA). Experiment 1 was conducted to validate a dietary formulation approach in which broilers were fed 1 of 5 diets including a 19.0% CP corn-soybean meal based (CSBM) diet, a LCP control diet (16.7% CP) that matched the AA profile of the CSBM diet, and 3 LCP diets with 30% reductions in standardized ileal digestible concentrations of TSAA, Lys, or Thr from 14 to 23 d post-hatch. Body weight gain and G:F were greatest (P < 0.05) and similar for broilers fed the CSBM and LCP control diets, whereas reductions in Thr, TSAA, and Lys each decreased (P < 0.05) G:F of broilers. In Experiment 2, birds were allotted to 18 treatment groups in a factorial arrangement of 9 dietary treatments × 2 infection states. Dietary treatments included a LCP control diet similar to that fed in Experiment 1 and 8 LCP diets with 40% individual reductions in TSAA, Lys, Thr, Val, Ile, Arg, Phe + Tyr, or Gly + Ser. Broilers received experimental diets from 10 d to 28 d post-hatch and were inoculated with 0 or 4.0 × 105 sporulated E. acervulina oocysts at 15 d. Decreased (P < 0.05) growth performance (10 to 28 d) of broilers was observed with each AA reduction, except Phe + Tyr, compared with birds fed the LCP control diet. Body weight gain and G:F were lowest (P < 0.05) for birds fed diets reduced in Lys or Val. Eimeria acervulina decreased growth performance and plasma carotenoids of broilers, but effects varied among dietary treatment groups as indicated by diet × infection interactions (P < 0.05). Dietary AA reductions did not alter (P > 0.05) the increase in intestinal gene expression of interferon-γ, interleukin-1ß, or interleukin-10 observed in E. acervulina-infected birds.