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Methionine is an essential amino critical to many cell functions including the synthesis of proteins. Supplementation of methionine in vivo is typically through L-methionine, DL-methionine, or a methionine hydroxy analog (MHA). The goal of this study was to compare the function of L-methionine, DL-methionine, and an MHA as a source of methionine to myoblasts in vitro. Avian myoblasts isolated from turkey embryos were plated in media containing varying concentrations of DL-methionine (DLM; 1.125 mg/mL or 0.56 mg/mL) or methionine hydroxy analog (MHA; 1.28 mg/mL or 0.64 mg/mL) as well as a methionine deficient negative control group and an L-methionine supplemented positive control group. The results of the proliferation assay exhibited cell division in the absence of methionine which was not significantly different than the positive control group. Results from the myoblast fusion assay revealed significantly greater myotube diameter between methionine supplemented groups compared to the methionine deficient negative control. The findings of this study show an ability for avian myoblasts to proliferate in the absence of methionine, the significance of which is discussed. Additionally, findings from the fusion assay suggest that DL-methionine and MHA are potential cost-effective substitutes for methionine supplementation during terminal differentiation of avian myoblasts.

Wooden breast (WB) is a degenerative myopathy seen in modern broiler birds resulting in quality downgrade of breast fillets. Affected filets show increased toughness both before as well as after cooking and have decreased water holding capacity and marinade pick up compared to normal fillets. Although the exact etiology is unknown, the circulatory insufficiency and increased oxidative stress in the breast muscles of modern broiler birds could be resulting in damage and degeneration of muscle fibers leading to myopathies. Three independent experiments were conducted to evaluate the effect of various dietary interventions on the incidence of WB when birds are exposed to oxidative stress associated with feeding oxidized fat and mild heat stress. Feed additives such as dietary antioxidant [Ethoxyquin (ETX)], mineral methionine hydroxy analog chelate (MMHAC) of Zn, Cu, and Mn, and organic selenium (Org Se) were tested at recommended levels. In experiment 1, ETX reduced (P < 0.05) the incidence of severe WB induced by oxidized fat diet. The magnitude of improvement in percentage of normal (no WB) filets and reduction in muscle lipid peroxidation was greater (P < 0.05) when ETX and MMHAC were fed together as shown by experiment 2. In birds exposed to mild heat stress (Experiment 3), feeding MMHAC by itself reduced (P < 0.05) tissue damage by reducing incidence of tibial head lesions, skin scratches, breast blisters, in addition to increasing the incidence of normal (no WB) fillets. When MMHAC was combined with ETX and Org Se, further improvement (P < 0.05) in normal (no WB) filets was observed. In summary, under different oxidative stress conditions, dietary intervention programs that contain ETX, MMHA-Zn, -Cu, and -Mn and Org Se can improve performance and increase carcass integrity, reducing problems, such as WB, either independently or with additive effect. This effect is most likely attained by simultaneously improving the exogenous and endogenous antioxidant status, reducing oxidative stress, and improving tissue healing process of the bird.

The present study investigated the interactive effects of copper sources and a high level of phytase on growth performance, nutrient digestibility, tissue mineral concentrations, and plasma parameters in nursery pigs. Weaning piglets (N = 192; 6.06 ± 0.99 kg), blocked by body weight, were randomly allotted to 1 of 4 dietary treatments, with 12 pens per treatment and 4 pigs per pen. A basal diet for each phase was formulated to meet nutrient requirements for nursery pigs with the exception that standardized total tract digestibility (STTD) P was reduced by 0.12% and Ca was adjusted to achieve Ca/STTD P = 2.15. The 4 dietary treatments were arranged in a 2 × 2 factorial design, with 2 Cu sources (125 mg/kg Cu from copper methionine hydroxy analogue chelate (Cu-MHAC) or copper sulfate (CuSO4)) and 2 phytase levels (0 or 1500 phytase units (FTU)/kg). Results showed that there was an interaction (P P 4. Pigs fed Cu-MHAC had greater apparent total tract digestibility (ATTD) of neutral and acid detergent fiber and STTD of P than those fed CuSO4. Phytase increased (P 4 when phytase was supplemented at 1500 FTU/kg. Cu-MHAC enhanced fiber and P digestibility regardless of phytase, compared with CuSO4. Phytase addition in P-deficient diets was effective in improving growth performance, Ca and P digestibility, and plasma inositol and growth hormone concentrations.

This study investigated the interactive effects of copper (Cu) sources and superdose phytase on growth performance, nutrient digestibility and tissue mineral concentrations in nursery pigs. A total of 288 weaning barrows (PIC 337 × 1050, Hendersonville, TN; BW = 6.06 ± 0.99 kg), blocked by BW, were randomly allotted to 1 of 6 dietary treatments, with 12 pens/treatments and 4 pigs/pen. Six dietary treatments were arranged in 3 × 2 factorial design, with 3 Cu sources [125 ppm Cu from copper sulfate (CuSO4), tribasic copper chloride (TBCC) or copper methionine hydroxy analogue chelate (Cu-MHAC, MINTREX® Cu, Novus International, Inc.), respectively] and 2 phytase levels [0 or 1500 FTU/kg from phytase PHYTAVERSE® G10 (Novus International, Inc.)]. There was an interaction (P = 0.05) between Cu sources and superdose phytase on ADG during d 0–41, with the improvement by superdose phytase being 25.84, 33.96 and 40.85% when the Cu sources were CuSO4, TBCC and Cu-MHAC, respectively. Superdose phytase significantly (P < 0.01) reduced F:G during d 0 to 41. Cu-MHAC led to greater ATTD of neutral and acid detergent fiber (NDF, ADF, respectively; P = 0.01) than CuSO4 and TBCC, respectively. Cu-MHAC led to greater (P = 0.03) ATTD of P than CuSO4, with TBCC intermediate. There was no interaction (P > 0.10) between Cu sources and superdose phytase in terms of nutrient digestibility and tissue mineral concentrations. Cu source did not (P > 0.10) affect tissue mineral concentrations; however, superdose phytase increased (P ≤ 0.05) bone Ca and P concentrations. In conclusion, Cu-MHAC may improve the efficacy of superdose phytase in terms of growth performance, most likely by improving phytate P hydrolysis and therefore providing more P for absorption. Superdose phytase is effective in improving growth performance of nursery pigs, mainly driven by increased Ca and P absorption and utilization.

The study was conducted to determine the effects of mineral methionine hydroxy analog chelate (MMHAC) partially replacing inorganic trace minerals in sow diets on epigenetic and transcriptional changes in the muscle and jejunum of progeny. The MMHAC is zinc (Zn), manganese (Mn), and copper (Cu) chelated with methionine hydroxy analog (Zn-, Mn-, and Cu-methionine hydroxy analog chelate [MHAC]). On day 35 of gestation, 60 pregnant sows were allotted to two dietary treatments in a randomized completed block design using parity as a block: 1) ITM: inorganic trace minerals with zinc sulfate (ZnSO4), manganese oxide (MnO), and copper sulfate (CuSO4) and 2) CTM: 50% of ITM was replaced with MMHAC (MINTREX trace minerals, Novus International Inc., St Charles, MO). Gestation and lactation diets were formulated to meet or exceed NRC requirements. On days 1 and 18 of lactation, milk samples from 16 sows per treatment were collected to measure immunoglobulins (immunoglobulin G, immunoglobulin A, and immunoglobulin M) and micromineral concentrations. Two pigs per litter were selected to collect blood to measure the concentration of immunoglobulins in the serum, and then euthanized to collect jejunal mucosa, jejunum tissues, and longissimus muscle to measure global deoxyribonucleic acid methylation, histone acetylation, cytokines, and jejunal histomorphology at birth and day 18 of lactation. Data were analyzed using Proc MIXED of SAS. Supplementation of MMHAC tended to decrease (P = 0.059) body weight (BW) loss of sows during lactation and tended to increase (P = 0.098) piglet BW on day 18 of lactation. Supplementation of MMHAC increased (P < 0.05) global histone acetylation and tended to decrease myogenic regulatory factor 4 messenger ribonucleic acid (mRNA; P = 0.068) and delta 4-desaturase sphingolipid1 (DEGS1) mRNA (P = 0.086) in longissimus muscle of piglets at birth. Supplementation of MMHAC decreased (P < 0.05) nuclear factor kappa B mRNA in the jejunum and DEGS1 mRNA in longissimus muscle and tended to decrease mucin-2 (MUC2) mRNA (P = 0.057) and transforming growth factor-beta 1 (TGF-β1) mRNA (P = 0.057) in the jejunum of piglets on day 18 of lactation. There were, however, no changes in the amounts of tumor necrosis factor-alpha, interleukin-8, TGF-β, MUC2, and myogenic factor 6 in the tissues by MMHAC. In conclusion, maternal supplementation of MMHAC could contribute to histone acetylation and programming in the fetus, which potentially regulates intestinal health and skeletal muscle development of piglets at birth and weaning, possibly leading to enhanced growth of their piglets.

Our objective was to evaluate the lactational responses of dairy cows to methionine provided from 2 ruminally protected sources of methionine activity. Twenty-one Holstein dairy cows [11 primiparous (634 kg of body weight, 140 d in milk) and 10 second-parity (670 kg of body weight, 142 d in milk)] were assigned to a treatment sequence in 4 replicated 5 × 5 Latin squares plus 1 cow, with 14-d periods. Treatments were as follows: control; 7.5 or 15 g/d of a ruminally protected product of 2-hydoxy-4-methylthio-butyric acid (NTP-1401; Novus International Inc., St. Charles, MO); or 7.5 or 15 g/d of a ruminally protected dl-methionine product (Smartamine M; Adisseo, Alpharetta, GA). The diet was predicted to meet metabolizable protein and energy requirements. Diets contained 16.1% crude protein, and the control diet was predicted to be deficient in metabolizable methionine (1.85% of metabolizable protein) but sufficient in lysine (6.8% of metabolizable protein). Feed intake and milk yield were measured on d 11 to 14. Blood was collected on d 14. Dry matter intake, milk yield, energy-corrected milk, milk fat yield and percentage, and efficiencies of milk and energy-corrected milk yield were not affected by treatment. Milk protein percentage and milk protein yield increased linearly with supplementation, without differences between methionine sources or interactions between source and level. Linear regressions of milk protein percentage and milk protein yield against supplement amount within source led to slope ratios (NTP-1401:Smartamine M) of 95% for protein percentage and 84% for protein yield, with no differences between sources for increasing milk protein. Plasma methionine concentrations were increased linearly by methionine supplementation; the increase was greater for Smartamine M than for NTP-1401. Plasma d-methionine was increased only by Smartamine M. Plasma 2-hydoxy-4-methylthio-butyric acid was increased only by NTP-1401. Our data demonstrated that supplementation with these methionine sources can improve milk protein percentage and yield, and the 2 methionine sources did not differ in their effect on lactation performance or milk composition.

This study investigated the interactive effects of zinc (Zn) and copper (Cu) sources and phytase on growth performance, oxidative status, mineral digestibility, tissue mineral concentrations, and gut morphology in nursery pigs. A total of 288 weaning barrows [body weight (BW) = 5.71 ± 0.81 kg], blocked by initial BW, were randomly allotted to one of eight dietary treatments, with nine pens per treatment and four pigs per pen. The eight dietary treatments were arranged in 2 × 2 × 2 factorial design, with two Zn sources [2,000, 2,000, and 100 mg/kg Zn from zinc oxide (ZnO) during phase 1 (days 1–14) and phase 2 (days 15–28), and phase 3 (days 29–42), respectively; 100 mg/kg Zn from zinc methionine hydroxy analogue chelate (Zn-MHAC) from phases 1 to 3], two Cu sources [150, 80, and 80 mg/kg Cu from copper sulfate (CuSO4) or copper methionine hydroxy analogue chelate (Cu-MHAC) during phases 1–3, respectively], and two phytase inclusion levels (0 or 500 FTU/kg). Results showed that ZnO supplementation at 2,000 mg/kg Zn significantly increased average daily feed intake (ADFI; P = 0.01) and average daily gain (ADG; P = 0.03) during phase 1 compared to Zn-MHAC group; however, Zn-MHAC supplementation tended (P = 0.06) to improve gain to feed ratio (G:F) during phase 2 compared to ZnO group. There were no differences (P > 0.10) between ZnO and Zn-MHAC groups in terms of ADG, ADFI, and G:F during the entire nursery period. Compared with CuSO4, Cu-MHAC tended to increase ADG (P = 0.07) and G:F (P = 0.08) during the entire nursery period. Phytase supplementation significantly increased ADG (P < 0.01), ADFI (P < 0.01), and G:F (P < 0.01) during the entire nursery period compared with no phytase supplementation. There was a significant interaction (P < 0.01) between Zn source and phytase on standardized total tract digestibility (STTD) of phosphorus (P), whereas there was no interaction (P = 0.21) between Cu sources and phytase on STTD of P. However, there was a significant interaction between Cu sources and phytase on calcium (Ca; P = 0.02) and P (P = 0.03) concentrations in metacarpal bones and G:F in phase 2 (P = 0.09). Furthermore, pigs fed diets containing Zn-MHAC tended to have lower ileum villus width (P = 0.07), compared with those fed diets containing ZnO, and pigs fed diets containing Cu-MHAC tended to have lower plasma malondialdehyde concentration (P = 0.10) compared with those fed diets containing CuSO4. In conclusion, under the conditions of the current study, ZnO supplementation at 2,000 mg/kg Zn was only effective in the first 2 wk postweaning, whereas Zn-MHAC supplementation at 100 mg/kg Zn could achieve better feed efficiency during phase 2 compared to pharmacological levels of ZnO, therefore, leading to no difference of growth performance in the entire nursery period. Low levels of Zn-MHAC may improve phytase efficacy on degrading phytate P compared to pharmacological levels of ZnO. Cu-MHAC may be more effective to promote growth compared to CuSO4, which may be partially driven by reduced oxidative stress. Results also indicated that Cu-MHAC might exert a synergistic effect with phytase on improving feed efficiency and bone mineralization.

Trace minerals have important roles in immune function and oxidative metabolism; however, little is known about the relationships between supplementation level and source with outcomes in dairy cattle. Multiparous Holstein cows (n=48) beginning at 60 to 140 days in milk were utilized to determine the effects of trace mineral amount and source on aspects of oxidative metabolism and responses to intramammary lipopolysaccharide (LPS) challenge. Cows were fed a basal diet meeting National Research Council (NRC) requirements except for no added zinc (Zn), copper (Cu) or manganese (Mn). After a 4-week preliminary period, cows were assigned to one of four topdress treatments in a randomized complete block design with a 2×2 factorial arrangement of treatments: (1) NRC inorganic (NRC levels using inorganic (sulfate-based) trace mineral supplements only); (2) NRC organic (NRC levels using organic trace mineral supplements (metals chelated to 2-hydroxy-4-(methythio)-butanoic acid); (3) commercial inorganic (approximately 2×NRC levels using inorganic trace mineral supplements only; and (4) commercial organic (commercial levels using organic trace mineral supplements only). Cows were fed the respective mineral treatments for 6 weeks. Treatment effects were level, source and their interaction. Activities of super oxide dismutase and glutathione peroxidase in erythrocyte lysate and concentrations of thiobarbituric acid reactive substances (TBARS) and total antioxidant capacity (TAC) in plasma were measured as indices of oxidative metabolism. Effects of treatment on those indices were not significant when evaluated across the entire experimental period. Plasma immunoglobulin G level was higher in cows supplemented with organic trace minerals over the entire treatment period; responses assessed as differences of before and after Escherichia coli J5 bacterin vaccination at the end of week 2 of treatment period were not significant. Cows were administered an intramammary LPS challenge during week 5; during week 6 cows fed commercial levels of Zn, Cu and Mn tended to have higher plasma TAC and cows fed organic sources had decreased plasma TBARS. After the LPS challenge, the extent and pattern of response of plasma cortisol concentrations and clinical indices (rectal temperature and heart rate) were not affected by trace mineral level and source. Productive performance including dry matter intake and milk yield and composition were not affected by treatment. Overall, results suggest that the varying level and source of dietary trace minerals do not have significant short-term effects on oxidative metabolism indices and clinical responses to intramammary LPS challenge in midlactation cows.

The present study was conducted to evaluate the efficacy of organic acid blends as dietary antibiotic replacer in marine fish olive flounder, Paralichthys olivaceus. Fish averaging 3.5 ± 0.05 g (mean ± SD) were fed one of the four experimental diets: (1) without antibiotic or organic acid (Control/CON); (2) with antibiotic—50 mg oxytetracycline per kg body weight per day (OTC); (3) with organic acid blend A—4 g/kg diet (OAA); and (4) with organic acid blend B—4 g/kg diet (OAB), for 10 weeks. At the end of the experiment, total intestinal bacterial counts in fish‐fed OAA, OAB and OTC were significantly lower than that of fish‐fed CON diet (p

This study aimed to investigate the effect of increasing the standardized ileal digestible (SID) total sulfur amino acid to lysine (TSAA:Lys) on the growth performance of nursery pigs raised with or without antibiotics (AGP) and to determine the optimal SID TSAA:Lys in nursery pigs raised without AGP. In Exp. 1, 924 nursery pigs (7.9 ± 1.3 kg), blocked by initial BW and sex, were randomly allotted to one of six treatments, with seven pens per treatment and twenty-two pigs per pen. The treatments were arranged in a 2 × 3 factorial design, with two AGP levels (0 or 50 mg/kg Carbodox) and three levels of SID TSAA:Lys (51.0, 58.5 or 66.0%). In Exp. 2, 990 weaned piglets (5.1 ± 0.9 kg), blocked by initial BW and sex, were randomly allotted to one of five dietary treatments (SID TSAA:Lys at 51, 58, 65, 72 or 79%) in the absence of AGP, with nine pens per treatment and twenty-two pigs per pen. Competing heteroskedastic models including broken-line linear (BLL), broken-line quadratic (BLQ), and quadratic polynomial (QP) were fitted for the growth performance data to estimate the optimal TSAA:Lys. In Exp. 1, AGP supplementation increased (p &lt, 0.05) ADG and ADFI during the 21 d period. Increasing SID TSAA:Lys in the diets with AGP did not affect growth performance, however, increasing SID TSAA:Lys in the diets without AGP resulted in a linear increase (p &lt, 0.05) in ADG and G:F. In Exp. 2, the best-fitting models for ADG and G:F from d 0 to 21 post-weaning were BLL, which yielded the optimal SID TSAA:Lys of 62% and 72%, respectively. The best-fitting models for ADG and G:F from d 21 to 42 post-weaning were BLL, which yielded the optimal SID TSAA:Lys of 59% and 58%, respectively. In conclusion, SID TSAA to Lys requirements under an antibiotic-free feeding regime during the first 21 d post-weaning were 62% and 72% in terms of ADG and G:F, respectively, whereas an SID TSAA:Lys of approximately 58% was required to maximize ADG and G:F for the late nursery phase.

White Striping (WS) and Woody Breast (WB) are 2 conditions that adversely affect consumer acceptance as well as quality of poultry meat and meat products. Both WS and WB are characterized with degenerative myopathic changes. Previous studies showed that WS and WB in broiler fillets could result in higher ultimate pH, increased drip loss, and decreased marinade uptake. The main objective of the present study was to compare the proteomic profiles of muscle tissue (n = 5 per group) with either NORM (no or few minor myopathic lesions) or SEV (with severe myopathic changes). Proteins were extracted from these samples and analyzed using a hybrid LTQ-OrbitrapXL mass spectrometer (LC-MS/MS). Over 800 proteins were identified in the muscle samples, among which 141 demonstrated differential (P < 0.05) expression between NORM and SEV. The set of differentially (P < 0.05) expressed proteins was uploaded to Ingenuity Pathway Analysis® (IPA) software to determine the associated biological networks and pathways. The IPA analysis showed that eukaryotic initiation factor-2 (eIF-2) signaling, mechanistic target of rapamycin (mTOR) signaling, as well as regulation of eIF4 and p70S6K signaling were the major canonical pathways up-regulated (P < 0.05) in SEV muscle compared to NORM. The up-regulation of these pathways indicate an increase in protein synthesis which could be part of the rapid growth as well as cellular stress associated with ongoing muscle degeneration and the attempt to repair tissue damage in SEV birds. Furthermore, IPA analysis revealed that glycolysis and gluconeogenesis were the major down-regulated (P < 0.05) canonical pathways in SEV with respect to NORM muscle. Down-regulation of these pathways could be the reason for higher ultimate pH seen in SEV muscle samples indicating reduced glycolytic potential. In conclusion, comparison of proteomic profiles of NORM and SEV muscle samples showed differences in protein profile which explains some of the observed differences in meat quality parameters. Future studies based on these differences could provide valuable insights into various cellular changes and identification of biomarkers related to WS and WB.

OBJECTIVE The objective of this study was to evaluate increasing doses of a novel microbial phytase (Cibenza Phytaverse, Novus International, St. Charles, MO, USA) on standardized total tract digestibility (STTD) of P in canola meal (CM), corn, corn-derived distiller's dried grains with solubles (DDGS), rice bran (RB), sorghum, soybean meal (SBM), sunflower meal (SFM), and wheat. METHODS Two cohorts of 36 pigs each (initial body weight = 78.5±3.7 kg) were randomly assigned to 2 rooms, each housing 36 pigs, and then allotted to 6 diets with 6 replicates per diet in a randomized complete block design. Test ingredient was the only dietary source of P and diets contained 6 concentrations of phytase (0, 125, 250, 500, 1,000, or 2,000 phytase units [FTU]/kg) with 0.4% of TiO2 as a digestibility marker. Feeding schedule for each ingredient was 5 d acclimation, 5 d fecal collection, and 4 d washout. The STTD of P increased (linear or exponential p≤0.001) with the inclusion of phytase for all ingredients. RESULTS Basal STTD of P was 37.6% for CM, 37.6% for corn, 68.6% for DDGS, 10.3% for RB, 41.2% for sorghum, 36.7% for SBM, 26.2% for SFM, and 55.1% for wheat. The efficiency of this novel phytase to hydrolyze phytate is best described with a broken-line model for corn, an exponential model for CM, RB, SBM, SFM, and wheat, and a linear model for DDGS and sorghum. Based on best-fit model the phytase dose (FTU/kg) needed for highest STTD of P (%), respectively, was 735 for 64.3% in CM, 550 for 69.4% in corn, 160 for 55.5% in SBM, 1,219 for 57.8% in SFM, and 881 for 64.0% in wheat, whereas a maximum response was not obtained for sorghum, DDGS and RB within the evaluated phytase range of 0 to 2,000 FTU/kg. These differences in the phytase concentration needed to maximize the STTD of P clearly indicate that the enzyme does not have the same hydrolysis efficiency among the evaluated ingredients. CONCLUSION Variations in enzyme efficacy to release P from phytate in various feedstuffs need to be taken into consideration when determining the matrix value for phytase in a mixed diet, which likely depends on the type and inclusion concentration of ingredients used in mixed diets for pigs. The use of a fixed P matrix value across different diet types for a given phytase concentration is discouraged as it may result in inaccurate diet formulation.

Lameness is a welfare issue and is the most common involuntary reason for culling pigs. Study objectives were to compare objective measures of lameness to gait score and assess effects of dietary chelated trace minerals in finisher pigs. The study was a 2 x 2 factorial arrangement consisting of naturally- occurring lame vs healthy pigs and 2 dietary treatments: 1) metal methionine hydroxy analogue chelate (MMHAC) supplied as MINTREX® Zn-Cu-Mn (Novus International, Inc.) supplemented at 80-10-20 mg/kg diet; and 2) sulfates Zn-Cu-Mn supplemented at 120-20-40 mg/kg diet. The study consisted of 4 groups of pigs (50-70 kg; 8 lame/8 non-lame per group) staggered one month apart. Dietary treatments were fed 53d. Measurements included gait scoring (0-4) and a panel of serum biomarkers (P2CP, CTX2, C2C, CTX1, osteocalcin) and were measured at baseline, d28 and d53. Serum biomarkers included bone synthesis (osteocalcin), bone degradation (CTXI), cartilage synthesis (P2CP), and cartilage degradation (C2C, CTX2) as well as the ratio of synthesis/ degradation for a total of 8 biomarkers. Proc Mixed GLM procedure of SAS was used and means were determined using a Tukey test. The main effects of healthy vs lame and dietary treatment were significant, but no diet x lameness interactions were observed. Lame pigs had elevated CTX2 (P< .01), C2C (P=.02) and tended to have reduced ratio of P2CP:CTX2 (P=.09) compared to healthy pigs. MMHAC increased P2CP (P=.01) and P2CP:CTX2 ratio (P=.03) in both healthy and lame pigs compared to sulfates. These findings demonstrate that biomarkers can distinguish mildly lame pigs (average score of 1) from healthy pigs and lower concentrations of MMHAC vs commercial level of sulfates increased cartilage synthesis and the ratio of cartilage synthesis/degradation in both lame and healthy pigs.

The present study aimed to determine the optimal ratio of standardized ileal digestible (SID) total sulfur amino acid (TSAA) to Lys for growth performance in 5 to 22 kg pigs under antibiotic-free regime. Experimental diets were formulated to ensure that lysine was the second limiting AA throughout the experiments. A total of 990 weaning piglets (PIC 337 × 1050, Hendersonville, TN; 5.08 ± 0.86 kg), blocked by initial BW and gender, were randomly allotted to 1 of 5 dietary treatments, with 9 pens per treatment and 22 piglets per pen. Dietary treatments consisted of SID TSAA to Lys ratios of 51%, 58%, 65%, 72% and 79%, with 1.31%, 1.28% and 1.20% SID Lys in phases 1, 2 and 3, respectively. The source of TSAA used in this study was the dry calcium salt of D, L-2-hydroxy-4-(methylthio)butanoic acid (84% Met, MHA®, Novus International, Inc., St. Charles, MO). Data was analyzed using general linear mixed models accounting for heterogeneous residual variances. Competing heteroskedastic models included broken-line linear (BLL), broken-line quadratic (BLQ), and quadratic polynomial (QP). For each response, the best-fitting model was selected using Bayesian information criterion. Results showed that increasing ratios of SID TSAA to Lys linearly (P = 0.01) increased ADG during the entire nursery period. Increasing SID TSAA to Lys ratios tended to increase (P = 0.09) overall ADFI in a quadratic manner. The best-fitting model for overall ADG is BLL in which the maximum ADG was estimated at 60% SID TSAA to Lys ratio, with 95% confidence interval (CI) between 55% and 65%. In conclusion, the optimal ratio of SID TSAA to Lys was 60% for maximum ADG. These results indicate that the NRC (2012) recommendation may underestimate the SID TSAA to Lys requirement for nursery pigs raised under antibiotic-free regime.

Lameness in gilts and sows has an economic impact on pig production and is a major welfare concern. The objective of the present project was to evaluate and quantify lameness in pigs. The following methods were compared to visual gait scoring: force-plate, hoof hardness and hoof lesions. Eight barrows and seven gilts (114 ± 15 kg) were selected using visual gait scoring (0-4, 4 most severe). From this group there were 4-0’s, 4-1’s, 5-2’s, and 2-3’s. Factorial analyses, (site: front vs rear) and lameness score (0-3) and their interaction was used to evaluate effect of treatment. Consistent with published literature, rear legs were more affected by lameness than front legs (73% rear, 27% front). Only two categories for hoof lesions (white line lesion and heel erosion) were prevalent in this population. White line lesion severity increased with increasing lameness score and differed between site (P < 0.05). In contrast with literature, more white line lesions were prevalent in front vs rear. Hoof hardness was scored at 5 locations on the bottom of the hoof. At the toe tip only, there was a trend (P = 0.052) for hoof hardness to decrease with increasing lameness score, but no effect of site (P = 0.39) or interaction. Among force-plate measurements, there was a significant effect of lameness (P < 0.05) for maximum force, range (95th percentile-5th percentile), standard deviation of force, amplitude of weight-bearing (average of all positive peaks), amplitude of weight-removing (average of all negative peaks), but no significant effect observed for average force, weight shift (# of peaks/min) or contralateral ratio. On average, more weight was applied to front vs rear [30.4 vs 19.6%; (P < 0.05)]. Although further validation is warranted, these results indicate that these objective measures of lameness were sensitive enough to detect differences between lame vs non-lame pigs.

The present experiment was conducted to evaluate the effects of trace mineral (Cu, Zn, Mn and Fe) premixes from inorganic and chelated (chelated to 2-hydroxy-4-methylthiobutanoic acid or hydroxy analog of methionine; Mintrex™, Novus International Inc., St. Charles, MO, USA) sources, in rockfish, (Sebastes schlegeli). fed diets containing mineral inhibitor phytic acid. Seven isonitrogenous (46% crude protein/CP) and isocaloric (16.63 KJ available energy per g diet) semi-purified diets comprising of a Basal control and diets supplemented either with the inorganic (I) or chelated premix (M) at three different levels of 0.75 (I0.75/M0.75), 1.5 (I1.5/M1.5) and 3 (I3/M3) g kg−1 diets were formulated. Twenty-four fish averaging individual wet body weight of 9 ± 0.2 g (Mean ± SD) were fed one of the experimental diets in quadratic group for 16 weeks. Average weight gain (WG) of fish fed M3 was significantly higher than that of fish fed Basal control and I1.5 diets (P

The present experiment was conducted to evaluate the efficacy of trace minerals (Cu, Zn and Mn) premixes from inorganic and chelated (chelated to 2-hydroxy-4-methylthiobutanoic acid or hydroxy analog of methionine; Mintrex™) sources, in juvenile Pacific white shrimp, ( Litopenaeus vannamei ) fed plant protein based practical diets. Eight experimental diets comprising a trace minerals (Cu, Zn and Mn) deficient Basal control, and diets supplemented with the trace mineral premixes at four different levels of 2.5 (M 2.5 ), 5 (M 5 ), 7.5 (M 7.5 ) and 8.5 g/kg (M 8.5 ) from chelated source and at three different levels of 5 (I 5 ), 8.5 (I 8.5 ) and 20 g/kg (I 20 ) from inorganic source were formulated. Eleven numbers of juvenile shrimp averaging 0.6 ± 0.01 g (mean ± SD) were fed one of the eight experimental diets in triplicate groups for 8 weeks. At the end of the feeding trial, shrimp fed M 2.5 and I 8.5 diets exhibited the similar final weight (FW) and weight gain (WG) ( P 5 compared to I 8.5 diet ( P Zn super oxide dismutase (Cu Zn SOD) from the serum and hepatopancreas tissue was recorded to be peaked for the group of shrimp fed M 5 diet. Moreover, plasma protein and glucose levels were recorded to be similar between the groups of shrimp fed M 2.5 and I 20 diets ( P L. vannamei ) fed plant protein based diets. Statement of relevance The significance of trace mineral bioavailability has become more important as the composition of majority of commercial fish feed has been changing and there is an increased use of dietary plant protein. As a result, the bioavailability of trace minerals is being adversely affected by the presence of antagonistic factor such as phytic acid in plant protein. Even though, our knowledge in fish nutrition has advanced significantly, the information on trace minerals requirement is still limited and fragmentary. Inorganic form (sulfate/nitrate) of trace mineral has traditionally been used in aquafeed formulation. However, the limited bioavailability of inorganic source of trace mineral due to its higher affinity to antinutrients has hastened the search for alternative form of inorganic trace minerals. Overall performance observed in the present experiments vouched the potential benefit of using chelated trace mineral, Cu, Zn & Mn premix in marine shrimp, Pacific white shrimp to promote the optimum growth, trace minerals saturation in shrimp body and tissue as well as to ensure the optimum enzyme activity and health of cultured shrimp. The present experiment opens a new avenue to compare the efficacy of inorganic and chelated source of trace minerals in other commercially important marine and fresh water fish species. Whereas, the sustainability issue has put a new dimension in aquafeed formulation with a wide array of new ingredients and additives, on the other hand, the importance of basic nutrient such as trace minerals is still in sideline. Substantial investment and integrated scientific efforts are warranted to bridge the knowledge gap and further improve our understandings on the significance of dietary trace mineral in fish nutrition and health at the least cost to environment.

Research has indicated that trace mineral (TM) supplementation may alter immune function and reduce morbidity associated with bovine respiratory disease. The objective of this experiment was to determine the influence of dietary Cu, Mn, and Zn supplementation on the performance, clinical signs, and TM balance of calves following a bovine viral diarrhea virus (BVDV) and (MH) combination respiratory pathogen challenge. Steers ( = 16; 225 ± 20 kg BW) from a single ranch were processed, weaned, and randomly pairwise assigned to either the TM-supplemented (MIN) or the control (CON) experimental treatments. The MIN calves received an additional 150 mg of Cu, 130 mg of Mn, and 320 mg of Zn daily and the CON calves received the basal diet with no additional Cu, Mn, or Zn supplementation. The basal diet contained sufficient Mn and Zn but inadequate Cu based on published nutrient requirements. After 46 d on the experimental treatments, all calves were naturally exposed to a heifer persistently infected with BVDV type 1b for 4 d and then subsequently intratracheally challenged with MH. Data were analyzed using the GLIMMIX procedure of SAS with sampling time serving as a repeated measure and calf serving as the experimental unit. The respiratory challenge was validated via increased BVDV type 1b antibody concentrations, MH whole cell and leukotoxin antibody concentrations, rectal temperatures (TEMP), and subjective clinical severity scores (CS). Calf performance ( ≥ 0.48) was not affected by TM supplementation. Mineral supplementation also did not impact the CS or TEMP of calves ( ≥ 0.53). There was a treatment × time ( < 0.001) interaction observed for liver Cu concentrations. The concentrations of Cu, Mn, Zn, and Fe within the liver; Cu, Mn, and Zn within the muscle; and Cu, Zn, and Fe within the serum were all impacted by time ( ≤ 0.03). Calves receiving the MIN treatment had greater ( < 0.01) liver Cu and Mn concentrations compared with CON calves. In contrast, serum Cu and Fe concentrations were increased ( ≤ 0.05) in CON calves compared with MIN calves. Mineral supplementation did not impact TM concentrations within the muscle ( ≥ 0.38). The supplementation of Cu, Mn, and Zn can improve the Cu and Mn status within the liver and serum of calves in response to a BVDV and MH challenge. When Cu is supplemented to calves receiving a marginally Cu-deficient diet, Cu status within the body is significantly improved.

Lameness in gilts and sows has an economic impact on pig production and is a major welfare concern. Study objectives were to compare subjective and objective lameness measurements and assess efficacy of chelated trace minerals in finisher pigs. Two dietary treatments: metal methionine hydroxy analogue chelate (MMHAC) supplied as MINTREX® Zn-Cu-Mn (Novus International, Inc.) supplemented at 80-10-20 mg/kg diet; and sulfates Zn-Cu-Mn supplemented at 120-20-40 mg/kg diet were fed. Four groups of pigs (50-70 kg; 8 pigs/grp) were fed dietary treatments for a duration of 8 wk prior to injection of 2 mg sodium urate crystals into the right rear distal interphalangeal joint. Data collection occurred at baseline, 6, 12, 24, 48, 72 and 144 hr post-urate injection. Measurements included gait scoring (0-4), a panel of serum biomarkers for synthesis and degradation of cartilage (P2CP, CTX2, C2C), bone (osteocalcin, CTX1) and static force-plate. A Proc Mixed GLM procedure of SAS was used and means were determined using a Tukey test. Urate injections resulted in elevated gait scores, peaking at 12 hr and similar to baseline at 72 hr; pigs fed MMHAC had lower gait scores (P< 0.01) compared to sulfate. C2C (P=0.09), CTX2 (P=0.10) and osteocalcin (P< 0.01) paralleled closely to changes in gait score, peaking at 12 h and returning to baseline at 24 to 72 hr. Osteocalcin (P=0.07) and osteocalcin: CTX1 ratio (P=0.05) were increased with MMHAC. These findings demonstrate that biomarkers can distinguish between healthy vs lame pigs and MMHAC, at lower concentrations vs sulfates, reduced gait score and increased osteocalcin.

Three experiments were conducted to quantify the efficacy of exogenous protease in poultry diets containing soybeans with varying trypsin inhibitor (TI) concentrations. Test diets contained 4 g/kg of TiO2 as a digestibility marker and protease (CIBENZA® DP100 at 300 U/g diet, Novus International). A randomized complete block design was analyzed using ANOVA with LSD multiple pair-wise comparison used to compare treatment means. For Exp 2 and 3, a common starter diet was fed to broiler chickens until d 16 of life and then switched to test diets. Exp 1 was a 2×6 factorial arrangement of 2 protease levels (0 and 300 U/g) and 6 dietary TI levels (0.72, 1.16, 1.61, 2.06, 2.50, and 2.95 mg/g) with 6 replicates/treatment; 8 chicks/pen. In Exp 2 (12 replicates/treatment; 8 chicks/pen), apparent ileal AA digestibility (AIAAD) was evaluated in diets containing 250 g/kg full fat extruded soybeans (FFES)+SBM (negative control; 5.8 mg/g TI) with or without protease or SBM (positive control; 3.1 mg/g TI). Test diets were fed from d 16–21 of age, AIAAD and relative pancreas weight was measured on d 21. In Exp. 3, broilers were allocated to a 2×2 factorial arrangement of treatments with 9 replicates/treatment and 8 chicks/pen. The dietary factors were SBM inclusion (420 or 960 g/kg; SBM TI = 3.85 mg/g), or protease inclusion (0 or 300 U/g). Test diets were fed from d 22–30, ileal digesta was collected on d 30. With increasing TI, pancreas weight increased (P

This study investigated the interactive effects of Zn sources, Cu sources and phytase on growth performance in nursery pigs. A total of 288 weaning barrows (BW = 5.71 ± 0.81 kg), blocked by initial body weight, were randomly allotted to 1 of 8 dietary treatments, with 9 pens per treatment and 4 pigs per pen. The 8 dietary treatments were arranged in 2 × 2 × 2 factorial design, with 2 Zn sources [2000 ppm Zn from ZnO during phase 1 (d 1 to 14) and 2 (d 15 to 28), and 100 ppm Zn from ZnO during phase 3 (d 29 to 42); 100 ppm Zn from Zn methionine hydroxy analogue chelate (Zn-MHAC, MINTREX® Zn, Novus International Inc., St. Charles, MO) during phase 1 to 3], 2 Cu sources [150 ppm Cu from CuSO4 or Cu-MHAC (MINTREX® Cu) during phase 1, and 80 ppm Cu from CuSO4 or Cu-MHAC during phase 2 and 3], and 2 phytase inclusion levels (0, 500 FTU/kg). Results showed that there were no differences (P > 0.10) between ZnO and Zn-MHAC groups in terms of growth performance during the entire nursery period. Compared with CuSO4, Cu-MHAC tended to increase ADG (P = 0.07) and G:F (P = 0.08) during the entire nursery period. There tended to be an interaction (P = 0.09) between Cu sources and phytase on G:F during phase 2. Specifically, without phytase supplementation, there was no difference (P = 0.63) on G:F during phase 2 between Cu-MHAC and CuSO4, whereas with phytase supplementation, Cu-MHAC increased G:F during phase 2 (P = 0.05) compared with CuSO4. In conclusion, Zn-MHAC supplementation at 100 ppm could replace pharmacological level of ZnO without affecting growth performance. Cu-MHAC could improve growth performance compared with CuSO4. Cu-MHAC exerted synergistic effect with phytase on improving feed efficiency.

A multitrial analysis was conducted to evaluate the effect of different levels of Cu from either Cu(2-hydroxy-4-(methylthio)butanoic acid [HMTBa])2 or CuSO4 on growth performance in nursery pigs. Six nursery trials were conducted from 2007 to 2012 under the same commercial conditions with initial BW of 5.75 ± 0.41 kg at 21 ± 3 d of age; the trials lasted for 42 d with a 3-phase feeding program (7 d in Phase I, 14 d in Phase II, and 21 d in Phase III). Diets were medicated with antibiotics and supplemented with 3,000 mg/kg Zn as ZnO during phases I and/or II. Treatments included a basal diet without added Cu or according to the NRC (1998) and supplemental levels of Cu (50 to 250 mg Cu/kg diet) from either Cu(HMTBa)2 or CuSO4; HMTBa was supplemented to make diets isomethionine. Treatments from each trial included 6 or 9 replicate pens/treatment with 22 to 25 piglets/pen. Mixed model analysis was conducted in which trial was considered a random effect, Cu level was considered a continuous fixed effect, and Cu source was a fixed effect. The basal diet within trial and statistical tests of the intercept between sources were not different, resulting in fitting a common intercept mixed model to the overall responses across phases. Cumulative ADG and ADFI quadratically responded (P < 0.05) with increasing Cu supplementation; predicted optimal ADG and ADFI occurred at 174 and 119 mg/kg, respectively. Increasing Cu supplementation linearly improved G:F (P = 0.054). No differences between sources were observed in ADG or ADFI. Numerically, pigs fed Cu(HMTBa)2 had higher ADG and lower ADFI compared to pigs fed CuSO4, resulting in better G:F for pigs supplemented with Cu(HMTBa)2 compared to pigs supplemented with CuSO4 (P < 0.01). The linear slope for increasing Cu supplementation on G:F was 2.1-fold higher for Cu(HMTBa)2 than that of CuSO4, with larger differences occurring in Phase II. In conclusion, Cu supplementation in nursery diets resulted in improved performance and Cu(HMTBa)2 is more effective than CuSO4 in improving feed efficiency.

The objective of this experiment was to test the effect of a Met analog, 2-hydroxy-4-methylthio-butanoic acid (HMTBa), on ruminal fermentation and microbial protein synthesis, nutrient digestibility, urinary N losses, and performance of dairy cows. Eight multiparous lactating Holstein dairy cows were assigned to 4 levels of HMTBa [0 (control), 0.05, 0.10, and 0.15% (dry matter basis)] in a replicated 4×4 Latin square trial. Experimental periods were 28 d, including 21 d for adaptation. Ruminal ammonia and microbial N were labeled through a 6-d intraruminal infusion of (15)NH4Cl, and microbial protein synthesis in the rumen was estimated using the reticular sampling technique. Treatment had no effect on dry matter intake (28.4 to 29.8kg/d), milk yield (44.1 to 45.3kg/d), feed efficiency, and milk composition. Total-tract apparent digestibility of nutrients was generally not affected by treatment, except digestibility of crude protein and starch decreased quadratically with HMTBa supplementation. Fecal, but not urinary, and total excreta N losses were increased quadratically by HMTBa. Ruminal pH, ammonia concentration, protozoal counts, and the major volatile fatty acids were not affected by treatment. Microbial N outflow from the rumen was linearly increased by HMTBa. 2-Hydroxy-4-methylthio-butanoic acid linearly increased the proportion of Fecalibacterium and quadratically decreased the proportion of Eubacterium in ruminal contents. Of the individual bacterial species, HMTBa increased or tended to increase Prevotella loescheii and Prevotella oralis. 2-Hydroxy-4-methylthio-butanoic acid linearly increased the concentration (and yield) of 15:0 in milk fat. In the conditions of this crossover experiment, HMTBa had no effect on feed intake and performance of dairy cows, decreased dietary crude protein digestibility, and increased microbial N outflow from the rumen.

Footpad dermatitis (FPD) is used in the poultry industry as an animal welfare criterion to determine stocking density. Trace minerals (TM) play a role in skin integrity and wound healing. This study evaluated the impact of TM on FPD and consisted of 3 treatments supplemented with 0 (NTM), low (LTM) and high (HTM) TM levels in the same basal diet. On d21, 71% birds in all treatments developed mild FPD and pens were top-dressed with dry litter to promote FPD healing. Compared to NTM, LTM reduced area under the curve (AUC) of FPD lesion scores during d21–42, HTM reduced the AUC of FPD lesion scores during d7–21 and d21–42. LTM improved growth performance on d14, HTM improved growth performance on d14 and d28. LTM and/or HTM increased gene expression of VEGF, TIMP3, TIMP4, MMP13, ITGA2, ITGA3 and CD40, which promoted collagen synthesis, deposition and organization; cell migration, matrix remodeling, and angiogenesis. LTM and/or HTM increased inflammation by upregulating TNFα and IL-1β during the early wound healing phase and reduced inflammation by downregulating IL-1β during the late wound healing phase. Our findings showed that TM not only improved growth performance but also reduced FPD development by promoting FPD wound healing.

The objectives of our study were to evaluate the productive response to methionine supplementation in lactating dairy cows and to define a relationship between metabolizable Met (MP Met) intake and production. A database of 64 papers meeting the selection criteria was developed evaluating postruminally infused dl-methionine (9 papers with 18 control diets and 35 treatment comparisons), 2-hydroxy-4-methylthio butanoic acid (HMTBa) provided as either a liquid or Ca salt form (17 papers with 34 control diets and 46 treatment comparisons), Mepron (Evonik Industries, Essen, Germany; 18 papers with 35 control diets and 42 treatment comparisons), and Smartamine (Adisseo Inc., Antony, France; 20 papers with 30 control diets and 39 treatment comparisons). Dietary ingredients and their accompanying nutritional compositions as described in the reports were entered into the Cornell-Penn-Miner software to model the diets and to predict nutrients that were not reported in the original publication. Data were analyzed using a weighted analysis of response to supplementation compared with the intraexperiment control, as well as through a regression analysis to changing dietary MP Met. Data included in the analysis were from experiments published between 1970 and 2011 with cows supplemented with between 3.5 and 67.9 g of Met or its equivalent from HMTBa. Cows supplemented with Smartamine consumed more, whereas cows supplemented with Mepron consumed less DM compared with controls. Milk yield did not significantly respond to Met supplementation, although it tended to increase for cows supplemented with HMTBa and Mepron. Milk protein yield was increased due to supplementation from all sources or from infusion, and protein concentration was greater for all supplements or infusion of dl-Met, except for cows supplemented with HMTBa. Irrespective of Met source, milk protein yield increased 2.23 g of protein/g of MP Met until reaching the breakpoint. Milk fat yield was increased for Mepron and HMTBa, whereas milk fat concentration was increased for infused dl-Met and for cows supplemented with HMTBa. Based on regression analysis, response of milk fat yield to Met supplementation was not different for infused dl-Met, Mepron, and Smartamine (1.87 g of fat/g of MP Met), whereas the response to HMTBa was significantly greater at 5.38 g of fat/g of MP Met.

The objective of this experiment was to measure the apparent total tract digestibility (ATTD) and the retention rate of Zn, Cu, Mn, and Fe in pigs fed either inorganic or organic sources of Zn, Cu, Mn, and Fe. The experimental design was a randomized complete block design with a 2 × 3 factorial arrangement of treatments. There were 2 types of diets (corn grits-based or corn-soybean meal [SBM]-based diets) and 3 micromineral treatments (basal micromineral premix [BMM], inorganic micromineral premix [IMM], and organic micromineral premix [OMM]). The BMM contained no added Zn, Cu, Mn, or Fe; the IMM microminerals were provided as sulfates of Zn, Cu, Mn, and Fe at 40, 50, 20, and 100 mg/kg, respectively. The OMM contained the same levels of the 4 microminerals as IMM, but Zn, Cu, Mn, and Fe in this premix were provided by Zn(2-hydroxy-4-methylthio butanoic acid [HMTBa])2, Cu(HMTBa)2, Mn(HMTBa)2, and FeGly, respectively. Forty-eight barrows (initial BW: 31.1 ± 4.2 kg) were housed individually and allowed ad libitum access to the corn grits diet with BMM for 2 wk. All pigs were then moved to metabolism cages and randomly assigned to 1 of the 6 treatment diets with 8 replicates per diet. Fecal and urine samples were collected for 5 d following a 5-d adaptation period. Compared with corn grits diets, pigs fed corn-SBM diets had greater (P < 0.05) absorption and retention of Zn, Cu, and Mn but less (P < 0.05) ATTD of Zn and Cu. Compared with BMM, supplementation of IMM or OMM increased (P < 0.05) absorption, retention, ATTD, and retention rate of Zn, Cu, Mn, and Fe. Compared with IMM, adding OMM to the corn-SBM diet improved (P < 0.05) the absorption and retention of Cu and Mn and the ATTD of Cu, but these differences were not observed in the corn grits diets (interaction, P < 0.05). In addition, adding OMM to the corn-SBM diet increased (P < 0.05) absorption and retention of Zn and Fe and ATTD of Zn, Mn, and Fe compared with adding IMM to the corn-SBM diet. Supplementation of OMM also increased (P < 0.05) the ATTD and retention rate of P in corn-SBM diets. Results indicate that Zn(HMTBa)2 has greater digestibility and Cu(HMTBa)2 and Mn(HMTBa)2 have greater digestibility and retention rates compared with their inorganic sulfates, if included in a corn-SBM diet. Supplementation of organic microminerals also improves the digestibility of P in a corn-SBM diet.

In 2011, aquaculture accounted for almost 64 million MT of fish, crustaceans and mollusks (FAO, 2012). Fish meal has historically been the protein source of choice in aquatic feeds, but global supplies have reached a plateau making it less available and more expensive. As a consequence, the use of cheaper proteins made from processed plant proteins, byproducts from agriculture, fisheries or the slaughtering of terrestrial production animals was popularized within the aquaculture feed industry. Although these alternative ingredients may contain a crude protein (CP) content comparable to fish meal, they may be less digestible and deficient in one or more of the ten essential amino acids (EAAs). This has required the adoption of more modern formulation approaches which take into account nutrient availability, especially in regard to EAAs. EAA requirements have been established for a number of species of fish and shrimp. Methionine (Met) followed by lysine (Lys) are the first limiting amino acids in plant and rendered animal byproducts. Formulating for EAAs by simply increasing the dietary inclusion levels of the feedstuffs that contain intact sources of the targeted EAAs can lead to overformulated feeds with excessive levels of CP and other nutrients. A more rational approach is to supplement the diet with crystalline amino acids (CAAs). A first step to the application of a nutrient-based formulation approach is knowledge of the digestible EAA content of the ingredients available for use in the formula. It is highly desirable to formulate on a CP, EAA and energy digestibility basis in line with current formulation practices of most modern feed companies. Further expansion of current EAA digestibility values across ingredients and farmed aquatic species will further enable nutrient based formulation. By formulating on an EAA basis across farmed aquatic species, nutritionists have realized least cost formulation opportunities with the use of CAAs which are now available throughout the world at accessible prices. The CAAs mostly used by the animal feed industry, in order of usage, are the following: dl -methionine or Met analogs, l -lysine, l -threonine, l -tryptophan, l -isoleucine and l -valine. Met and Lys are the most currently used due to their wide availability and importance as most limiting EAAs in plant protein ingredients, such as soy and corn based meals. There is a relatively wide variation in the dietary Met and Lys requirement values for farmed fish and shrimp. This is due to differences in species requirements, culture systems, developmental stage and composition of experimental diets. The dietary Met requirements for fish and shrimp range from 0.5 to 1.5% and from 0.7 to 0.9% of the diet, respectively (NRC, 2011). The dietary Lys requirements range from 1.2% to 3.3% of the diet for cultured fish and from 1.6 to 2.1% of the diet for cultured shrimp. Appropriate dietary Met and Lys levels improve the use of other EAAs because they have the ability to reduce the oxidation rate of other amino acids. The present review discusses bioavailability, requirements and sources of EAAs and practical considerations for their application in aquaculture feeds. The appropriate supplementation of crystalline amino acids in feeds for fish and shrimp represents an opportunity to reduce formulation costs in the face of the volatile commodity market of protein ingredients and the short supply of fish meal. Competitiveness in the aquatic animal feed industry today depends upon application of modern and environmentally-sound formulation techniques based on nutrient value and supplementation with crystalline EAA to meet animal nutrient requirements.

White striping (WS) and woody breast (WB) are 2 poultry meat quality defects that affect the acceptance of raw breast fillets as well as properties of cooked and further processed products. The present study was intended to evaluate the incidence of these conditions in broilers at different ages and to compare the properties of fillets with different degrees of WS and WB. For this study, 1,920 birds were processed, at 6 and 9 wk of age, in a standard commercial inline processing system. After chilling, carcasses were deboned and butterfly fillets were collected and weighed. Individual fillets were scored for normal, moderate, severe, and very severe degrees of WS and WB, and for petechial hemorrhagic lesions (PHEM, 0 - no lesion to 2 - severe lesion). Representative fillets with NORM-WS/WB, SEV-WS, SEV-WB, and SEV-WS/WB were selected and stored at 4°C. After 24 h, fillet length, width, cranial height, and caudal height, as well as pH, color, and drip loss were recorded. There was an increase in incidence of severe and very severe WS and WB conditions at 9 wk compared to 6 wk of age. The relationship between fillet weight and the myopathies plateaued at 9 wk of age with more fillets showing a higher score. Mean PHEM scores were higher (P < 0.05) in SEV-WS, SEV-WB, and SEV-WS/WB compared to NORM-WS/WB birds, both at 6 and 9 weeks. NORM-WS/WB birds had lower (P < 0.05) live and breast weight, breast yield, and cranial and caudal heights, as well as b* value (yellowness) compared to SEV-WS/WB. NORM-WS/WB had lower (P < 0.05) pH while NORM-WS/WB and SEV-WS samples showed lower (P < 0.05) drip loss when compared to SEV-WB and SEV-WS/WB. The results from this study showed that the severe degrees of WS and WB are associated with heavier and older birds, and thicker breast fillets. Occurrence of severe degrees of WS and/or WB can affect various raw meat quality factors, mainly color and water holding capacity.

An experiment was conducted to characterize a gut health challenge model consisting of a diet containing rye, wheat, and feather meal and a mild mixed-species Eimeria challenge, and to evaluate the effect of carbohydrase and protease on growth performance and gut health of young broilers. The study included 4 treatments: negative control, carbohydrase alone, protease alone, and combination of carbohydrase and protease. Each test diet was fed to 18 battery pens of broilers with 8 male birds per pen from 0 to 22 d of age. Carbohydrase improved body weight, feed intake, and feed conversion ratio (FCR) on d 7, 14, and 21(P < 0.01). Protease increased body weight on d 7 and 21 and improved 0 to 7 d FCR (P < 0.05). More lymphocyte infiltration was observed in small intestine mucosa of negative control birds on d 8, carbohydrase supplementation lessened this. Both carbohydrase and protease reduced digesta viscosity on d 22 with the carbohydrase effect being the greater of the two, and the combination effect was not different from the carbohydrase effect alone (P < 0.01). Ileal Clostridium perfringens of 15-day-old broilers was decreased by carbohydrase, a further reduction was achieved by combining carbohydrase with protease (P = 0.01). Liver vitamin E concentration on d 15 (P < 0.01) and 22 (P = 0.02) was increased by carbohydrase, and the carbohydrase effect was greater in the presence of protease on d 22 (P = 0.04). Plasma α-1-acid glycoprotein level and liver Zn and Cu concentrations of broilers were reduced by carbohydrase on d 15 (P < 0.01). Broilers fed carbohydrase had higher levels of plasma zeaxanthin on d 22 and higher levels of plasma lutein on d 15 and 22 (P < 0.01). In summary, a rye wheat based diet containing feather meal when fed to broilers in addition to a mild Eimeria challenge induced subclinical enteritis characterized by digestion inefficiency, dysbacteriosis, inflammation, and gut barrier failure; carbohydrase and protease could be effective tools to improve growth performance and gut health of broilers suffering from this type of subclinical enteritis.

Background The goal of this study was to compare the antagonism of elevated dietary Cu (250 mg/kg) from CuSO4 on three different Zn sources (ZnSO4 · H2O; [Zn bis(−2-hydroxy-4-(methylthio)butanoic acid)], Zn(HMTBa)2, a chelated Zn methionine hydroxy analogue; and Zn-Methionine), as measured using multiple indices of animal performance in ROSS 308 broilers. Methods Three experiments were conducted in broiler chicks fed a semi-purified diet. All birds were fed a Zn-deficient diet (8.5 mg/kg diet) for 1 wk, and then provided with the experimental diets for 2 wks. Results Experiment 1 was a 2 × 2 factorial design with two levels of Cu (8 vs. 250 mg/kg diet from CuSO4) and two Zn sources at 30 mg/kg [ZnSO4 · H2O vs. Zn(HMTBa)2]. Elevated Cu impaired growth performance only in birds fed ZnSO4. Compared to ZnSO4 · H2O, Zn(HMTBa)2 improved feed intake (12 %; P

SUMMARY Two 54-d grow-out trials were conducted in a commercial broiler facility to determine the effect on performance, carcass yield, footpad lesions, and litter trace mineral concentrations when feeding broiler chicks diets containing reduced levels of chelated Zn-(HMTBA)2 [where HMTBA is 2-hydroxy-4-(methylthio)butanoic acid; trials 1 and 2], Cu-(HMTBA)2 (trial 2), and Mn-(HMTBA)2 (trial 2) in comparison with industry levels of the respective inorganic trace minerals (ITM). A total of 122,400 Ross 708 straight-run chicks were distributed into 8 houses with approximately 15,300 chicks per house for trials 1 and 2, with 4 replicate houses per treatment. Treatments in trial 1 consisted of 30 ppm of Zn as chelated Zn-(HMTBA)2 and 100 ppm of Zn as ZnSO4 (ITM). In trial 2, treatments consisted of 32, 8, and 32 ppm from Zn-, Cu-, and Mn-(HMTBA)2, respectively, and 100, 125, and 90 ppm of Zn, Cu, and Mn, respectively, as sulfates (ITM). Trial 1 was carried out on built-up litter, and trial 2 was on fresh litter. In trials 1 and 2, no differences (P > 0.05) were observed in performance, carcass yield, or tibia Zn over the 54-d period between treatments. On d 54 (trial 1), chicks fed diets supplemented with Zn-(HMTBA)2 had 27.3% more grade 1 and 2 paws than those fed ITM (P < 0.01). On d 54 (trial 2), the chicks fed Zn-, Cu-, and Mn-(HMTBA)2 had 23.3% more grade 1 and 2 paws than those fed ITM (P = 0.07). For trial 2, litter Zn, Cu, and Mn from birds fed Zn-, Cu-, and Mn-(HMTBA)2 were reduced by 40, 74, and 35%, respectively, compared with birds fed the respective ITM. In summary, based on the present data, commercial levels of ITM can effectively be replaced with reduced levels of Zn-, Cu-, and Mn-(HMTBA)2 without compromising bird performance while achieving a significant improvement in footpad health and a reduction in trace mineral concentrations in the litter.

The objectives of the current study were to determine the fate and contribution to Met kinetics of 2-hydroxy-4-(methylthio)butanoate (HMTBA) at the whole-body, splanchnic, and mammary levels. Four multicatheterized cows (31.3 kg of milk/d; 17.7 kg of DMI/d) were used in a crossover design, with two 1-wk periods, to determine the metabolic fate of HMTBA and its effect on Met metabolism. Over the last 2 d of each period, cows were infused, via a jugular vein, with saline or HMTBA (Alimet, Novus International Inc., St. Louis, Mo) at the rate of 36 g/d. During the last 8h, the HMTBA infusion was substituted by equimolar [1-(13)C]HMTBA (8.79 mmol/h) and l[methyl-(2)H(3)]Met (1.31 mmol/h) was infused in all cows. During the last 5h, hourly samples (n=6) were collected to determine plasma flows plus the isotopic enrichments (IE) and concentrations of HMTBA ((13)C) and Met (both (13)C and (2)H(3)) in plasma from an artery plus portal, hepatic, and mammary veins. The IE of [(13)C] and [(2)H(3)]Met were also determined in milk protein taken over the last 1h of infusion in HMTBA-infused cows. The infused HMTBA increased whole-body plasma flux of Met by 6.5 mmol/h (from 17.9 to 24.4 mmol/h). Based on enrichments of (13)C-labeled Met, 3.8 mmol/h of Met flow through plasma was derived directly from HMTBA. These 2 estimates accounted for between 43 to 74% of the HMTBA dose infused, contributing to increased whole-body Met availability. Although the portal-drained viscera, liver, and mammary gland (MG) extracted 11, 37, and 3.4%, respectively, of the infused HMTBA, tissue net Met fluxes were either unchanged (portal-drained viscera, MG) or even reduced (liver: -7.9 vs. -2.4±0.6 mmol/h). Therefore, net postsplanchnic supply of Met decreased from 7.0 to 2.9 mmol/h between control and HMTBA-infused cows, compared with needs for milk protein secretion of 7.6 and 8.1 mmol/h, respectively. The HMTBA provided directly 15% of the Met required for milk protein secretion, with 0.2 mmol/h synthesized within the MG, whereas 1.1 mmol/h originated from Met produced in other tissues and transported to the MG through blood circulation. Most of the remainder needed by the MG arose from unlabeled Met released from protein breakdown in extra-splanchnic tissues and that was not reused to support intracellular protein synthesis, as this function was performed by Met synthesized from HMTBA in situ. Absorbed HMTBA, therefore, both produces and spares Met for use by the MG.

SUMMARY Dietary supplementation of trace minerals with a large safety margin is common practice in the poultry industry to prevent trace mineral deficiency. However, oversupplementation of inorganic trace minerals often results in poor bioavailability and low body retention, and can result in more than 90% of the dietary trace minerals being excreted into the environment. In this trial, litter mineral concentrations at the conclusion of the study had very high concentrations of Ca (2.3%), P (1.7%), Cu (150 ppm), Fe (350 ppm), Mn (420 ppm), and Zn (350 ppm). It has been demonstrated that chelated trace minerals are more bioavailable, perhaps because of reduced loss to antagonisms in the intestinal lumen compared with inorganic sources. In 2 commercial broiler trials, performance, meat yield, and footpad health associated with reducing Zn and Mn supplementation while feeding a 50:50 blend of inorganic (sulfates) and chelated forms of Zn, Cu, and Mn were tested. Birds fed the blended treatment exhibited similar or improved growth performance and similar tissue mineral storage compared with those fed the inorganic trace mineral control. In addition, chelated trace minerals improved breast meat yield and footpad health in one trial. In summary, chelated trace minerals are effective alternative trace mineral sources for satisfying the trace mineral needs of rapidly growing broilers and allowing for a reduction in dietary mineral supplementation, thereby potentially reducing the excretion of minerals into the environment.

The objective of the study was to evaluate the effect of diets supplemented with fatty acids of different degrees of saturation, in the absence or presence of an antioxidant (AOX; Agrado Plus, Novus International Inc., St. Charles, MO), on dairy cow lactation performance. Calcium salts of long-chain fatty acids were supplemented as a source of lower saturation fatty acid, and a palm acid product was supplemented as the higher saturation fatty acid source. Sixty early-lactation Chinese Holstein cows (100+/-23 d in milk) were randomly allocated to 4 dietary treatments in a 2 x 2 factorial design: (1) lower saturation fatty acid (LS), (2) LS and AOX, (3) higher saturation fatty acid (HS), and (4) HS and AOX. The Ca salts of long-chain fatty acids and palm acid product were supplied at 1.8 and 1.5% on a dry matter basis, respectively, to form isoenergetic diets. The AOX was added at 0.025% in the ration. The experiment lasted 9 wk, including 1 wk for adaptation. Lactation performance was recorded and milk was sampled and analyzed weekly. Blood samples were taken from the coccygeal vein to determine metabolism parameters on d 16, 36, and 56 during the experiment. Neither fatty acid type nor AOX supplementation showed a significant effect on dry matter intake during the study. Milk yield was lower in the LS-fed cows compared with the cows fed HS. Milk fat and milk protein concentrations were not affected by fatty acid type or AOX supplementation. Adding AOX increased the yield of milk in the LS-fed cows, but did not affect those fed HS. Activity of plasma superoxide dismutase was significantly lower, plasma glucose tended to be lower, and plasma malondialdehyde was higher in the LS-fed animals compared with those fed HS. Addition of AOX decreased both plasma nonesterified fatty acids and hydrogen peroxide contents and increased total antioxidant capacity across the fatty acid types. Plasma beta-hydroxybutyrate was not affected by fatty acid type or AOX treatment. Cows fed LS had higher cis-9C(18:1) and trans-10, cis-12C(18:2) in milk at the expense of C(18:0), whereas AOX addition increased milk cis-9C(18:1) at the expense of milk C(12:0), C(16:0), and trans-10, cis-12C(18:2). It is inferred that feeding LS resulted in inferior lactation performance, whereas addition of antioxidant partially alleviated these negative effects.

Three experiments were conducted to evaluate the effects of feeding 2-hydroxy-4- (methylthio)-butanoic acid (HMTBA) on performance and carcass characteristics of feedlot cattle and on microbial fermentation in a continuous-culture system. In Exp. 1, 160 crossbred steers (initial BW = 385 +/- 10.3 kg) were assigned to 4 treatments consisting of control (0% HMTBA) or 3 diets containing HMTBA (0.069, 0.137, and 0.204%; DM basis) in a randomized complete block design. As the percent of HMTBA increased in the diet, final BW (P = 0.069), final BW adjusted to a constant dressing percent (P = 0.063), and overall ADG (P = 0.099) tended to decrease linearly. Overall DMI decreased linearly (P or = 0.10) were noted for carcass characteristics, except for a tendency (P = 0.078) for a linear increase in the percentage of cattle grading USDA Choice with increasing HMTBA dose. In Exp. 2, 80 crossbred steers (initial BW = 450 +/- 17 kg) in a randomized complete block design were assigned to a control (0% HMTBA) diet or to a diet in which the concentrations of HMTBA were gradually increased from 0.036 to 0.212% of DM over a 50-d period. The HMTBA-containing diet tended to decrease DMI (P = 0.132), but G:F (P = 0.319) for the overall feeding period, carcass measurements, and USDA quality grade (P > or = 0.149) did not differ between treatments. In Exp. 3, continuous culture fermenters (n = 5/treatment) were used to determine the effects of HMTBA (control vs. 0.24% HMTBA) on microbial fermentation. No differences (P > or = 0.31) were detected between treatments in ruminal OM digestibility, microbial N synthesis, pH, ammonia, molar proportions of VFA, or effluent concentration of selected long-chain fatty acids. These results suggest that HMTBA decreased DMI by feedlot steers fed a steam-flaked corn-based diet in a dose-dependent manner; however, gradually increasing the dose over time seemed to moderate effects on DMI. No major changes in microbial fermentation in continuous culture were observed with HMTBA at 0.24% of dietary DM, suggesting effects of HMTBA on DMI were not likely associated with changes in ruminal digestion or fermentation.

The objective of the study was to evaluate the effect of feeding the dietary antioxidant Agrado Plus (AOX; Novus International, St. Louis, MO) in diets that contained 2% fresh fat (FF) or oxidized fat (OF) on milk production and composition and antioxidant status of cows during mid to late lactation. Forty-four mid to late lactating primiparous cows housed in a tie-stall barn were fed a diet that contained 2% FF for 15 d as adaptation period and then randomly allocated to 1 of the 4 dietary treatments (FF, FF+AOX, OF, OF +AOX) for 6 wk. Feeding AOX increased dry matter intake, 3.5% fat-corrected milk, and milk fat yield, and decreased milk protein content but not yield. Feeding OF increased milk fat yield, but decreased dry matter intake and body weight gain. Milk fat composition changed with treatments: AOX increased cis 18:1 and decreased trans-11 18:1, whereas OF decreased trans-9 and trans-11 18:1 and cis-9, trans-11 18:2 in milk. Plasma antioxidant enzymes and status were affected by treatments. Feeding OF increased superoxidase dismutase activity but decreased plasma antioxidant status, whereas AOX supplementation increased glutathione peroxidase activity across fat types and increased the antioxidant status and superoxidase dismutase activity when feeding OF diets. It can be concluded that feeding AOX improved lactation performance and the antioxidant status of the cow across fat types, and feeding OF increased milk fat yield but decreased dry matter intake, body weight gain, and antioxidant status. The negative effects of feeding OF were partially alleviated by AOX.

The objective of this study was to compare the gain-response curve to dietary levels of 2-hydroxy-4(methylthio) butanoic acid (HMTBA) and DL-Met (DLM) across 4 floor pen trials in which different diets were used. Six replicates of 38 or 41 birds per pen (trials 1 to 2 and 3 to 4, respectively) were used in a 2 x 3 factorial arrangement. A control with 12 replicates was also included. The 2 Met sources were fed at 3 equimolar levels equally spaced, with the highest level added at requirements from 1 to 48, 49, 43, or 49 d for trials 1, 2, 3, and 4, respectively. Commercial-type TSAA-deficient control diets contained sorghum, wheat, corn, or corn plus meat and bone meal for trials 1, 2, 3, and 4, respectively. Performance improved at all times for most parameters after supplementing with HMTBA or DLM (P0.05). No differences were found in the birds fed HMTBA or DLM at any age and trial (P0.05), except for trial 1, in which 17-d-old birds performed better when fed HMTBA than DLM (P0.05). In each trial, linear, quadratic, and exponential regressions were conducted upon the gain response of birds fed HMTBA and DLM separately. Equations with better goodness of fit were used to compare the estimated gain responses to feeding HMTBA vs. DLM. In 3 trials, the shape of the gain-response curve differed when feeding HMTBA vs. DLM. In trials 3 and 4, feeding HMTBA at commercial levels resulted in greater gain responses than DLM (P0.05), whereas, in trials 2 and 4, at very deficient levels, DLM-fed birds outperformed those fed HMTBA (P0.05). When the 4 trials were combined, the dose-response curve with the best goodness of fit was linear for HMTBA and quadratic for DLM. It can be concluded that the 2 Met sources have a different dose-response form, HMTBA could outperform DLM at commercial levels, and DLM could outperform HMTBA at deficient levels.

Poultry diets are typically supplemented with DL-2-hydroxy-4(methylthio) butanoic acid (HMTBA, or the hydroxy analog of methionine) or DL-methionine (DLM). Although HMTBA and DLM provide methionine activity, they are structurally distinct molecules with different physiological characteristics until they are converted to L-methionine. The relative rates of intestinal HMTBA vs. DLM absorption have been controversial, and it has been claimed that HMTBA is not fully absorbed. We measured the uptake of HMTBA and DLM in an in vitro everted intestinal slice model. Sections of intestinal slices (jejunum and ileum) were incubated with 0.1 to 50 mM HMTBA that was radiolabeled or DLM that was radiolabeled, and absorption was measured by scintillation counting. The HMTBA uptake was equal to or greater than DLM absorption in each tissue and at every time point with one exception. Furthermore, the rates of HMTBA absorption were always equal to or significantly greater than DLM uptake. In a separate in vivo experiment, absorption of HMTBA and L-methionine was monitored along the entire gastrointestinal (GI) tract. Broilers were fed commercial-type corn-soy diets supplemented with 0.21% HMTBA. Digesta was collected from crop, proventriculus, gizzard, duodenum, jejunum, ileum, large intestine, and cloaca and analyzed for the concentration of free HMTBA and free methionine in each compartment. These studies demonstrated that HMTBA is absorbed completely and along the entire GI tract, especially the upper GI tract. Furthermore, there was a higher concentration of free L-methionine than HMTBA in the digesta from every segment distal to the gizzard.

Controversy about the efficacy of Alimet as a source of methionine arises from the fact that when it is ingested, it is fundamentally different from methionine. Thus, broiler performance studies comparing efficacy of Alimet and DL-methionine (DL-met) continue to be of interest. This report will cover two studies using Alimet Feed Supplement (an 88% aqueous solution of 2-hydroxy-4-(methylthio)butanoic acid, HMTBA). The objective of the first was to compare Alimet with DL-met as sources of supplemental L-methionine (L-met) for broiler production. The sources were fed on an equal dry-matter basis at a level approximating 85% of that used in commercial practice in the UK, where the study was run. A basal diet was used to confirm sensitivity to methionine source addition. No antibiotics were fed. Results confirmed a significant response to supplemental methionine over the 0-42 day study for gain and feed conversion efficiency with no significant difference between Alimet and DL-met. The objective of the second study was to compare performance of Alimet with DL-met when birds were fed high inclusion levels. In this experiment, conducted at Novus International, performance in a basal diet was compared to five supplemental levels of Alimet or DL-met (0.2, 0.5, 1.0, 1.5 and 2.0%). The first level of supplementation was designed to represent an adequate diet with subsequent levels representing excessive addition. Results confirmed that the performance and feed intake of birds fed DL-met was more negatively affected than those fed Alimet, with differences becoming significant above the 0.5% level. Data presented here confirm full molar equivalency of HMTBA to DL-met in broilers fed commercial diets and illustrate the relatively greater safety of Alimet when fed at higher inclusion levels.