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The goal of this review article, based on a systematic literature search, is to critically assess the state of knowledge and experimental methodologies used to delineate the conversion and metabolism of the 2 methionine (Met) sources DL-methionine (DL-Met) and DL-2-hydroxy-4-(methylthio) butanoic acid (HMTBa). The difference in the chemical structures of HMTBa and DL-Met indicates that these molecules are absorbed and metabolized differently in animals. This review explores the methodologies used to describe the 2-step enzymatic conversion of the 3 enantiomers (D-HMTBa, L-HMTBa and D-Met) to L-Met, as well as the site of conversion at the organ and tissue levels. Extensive work was published documenting the conversion of HMTBa and D-Met into L-Met and, consequently, the incorporation into protein using a variety of in vitro techniques, such as tissue homogenates, cell lines, primary cell lines, and everted gut sacs of individual tissues. These studies illustrated the role of the liver, kidney, and intestine in the conversion of Met precursors into L-Met. A combination of in vivo studies using stable isotopes and infusions provided evidence of the wide conversion of HMTBa to L-Met by all tissues and how some tissues are net users of HMTBa, whereas others are net secreters of L-Met derived from HMTBa. Conversion of D-Met to L-Met in organs other than the liver and kidney is poorly documented. The methodology cited in the literature to determine conversion efficiency ranged from measurements of urinary, fecal, and respiratory excretion to plasma concentration and tissue incorporation of isotopes after intraperitoneal and oral infusions. Differences observed between these methodologies reflect differences in the metabolism of Met sources rather than differences in conversion efficiency. The factors affecting conversion efficiency are explored in this paper and are mostly associated with extreme dietary conditions, such as noncommercial crystalline diets that are very deficient in total sulfur amino acids with respect to requirements. Implications in the diversion of the 2 Met sources toward transsulfuration over transmethylation pathways are discussed. The strengths and weaknesses of some methodologies used are discussed in this review. From this review, it can be concluded that due to the inherent differences in conversion and metabolism of the 2 Met sources, the experimental methodologies (e.g., selecting different organs at different time points or using diets severely deficient in Met and cysteine) can impact the conclusions of the study and may explain the apparent divergences of conclusion found in the literature. It is recommended when conducting studies or reviewing the literature to properly select the experimental models that allow for differences in how the 2 Met precursors are converted to L-Met and metabolized by the animal to enable a proper comparison of their bioefficacy.

This literature review evaluates the absorption of methionine (Met) sources such as 2-hydroxy-4-methylthiobutyric acid (HMTBa), its calcium salts (HMTBa-Ca), and DL-methionine (DL-Met) by focusing on the state of knowledge regarding the absorption mechanism, experimental methodology, and factors affecting their absorption. The 2 Met sources differ in mechanism and site of absorption due to differences in their chemical characteristics and enzymatic conversion. This review addresses diffusion- and transport-mediated absorption systems for amino acids and carboxylic compounds, best elucidated by in vitro, ex vivo, and in vivo experimental models. Opportunities and limitations in the use of radioisotopes to depict absorption sites as well as host and microbial metabolism are described. Physiological and environmental conditions that lead to changes in gut absorptive capacity and the impact of Met source absorption are also evaluated. This review concludes that any comparison between HMTBa and DL-Met should consider their different behaviors during the absorption phase. Hence, the chemical characteristics of these 2 molecules entail different absorption sites and mechanisms, from passive absorption in the case of HMTBa and HMTBa-Ca to active transporters for DL-Met, HMTBa, and HMTBa-Ca. In addition, the different conversion modes of these 2 molecules further differentiate their absorption modes. Considering these important differences, it is easier to understand the apparent divergence between the conclusions of existing publications. When comparing these 2 molecules, it is recommended to properly adapt to the conditions under which the absorption of Met sources is evaluated.

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.

Abstract 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.

Copper (Cu) is widely used at high levels as growth promoter in poultry, the alternative source of Cu to replace the high level of inorganic Cu at poultry farm remains to be determined. Three floor pen experiments were conducted to evaluate the effects of Cu methionine hydroxy-analogue chelate (Cu-MHAC, MINTREX®Cu, Novus International, Inc.) on growth performance and gut health in broilers in comparison to CuSO4 and/or tribasic copper chloride (TBCC). There were 3 treatments in experiment#1 (0, 30 and 75 ppm Cu-MHAC) and experiment#2 (15 and 30 ppm Cu-MHAC, and 125 ppm CuSO4), and 4 treatments in experiment #3 (15 and 30 ppm Cu-MHAC, 125 ppm CuSO4 and 125 ppm TBCC) with nine replicates pens of 10–13 birds in each treatment. The levels of other minerals were equal among all treatments within each experiment. All birds were orally gavaged with a coccidiosis vaccine at 1x recommended dose on d0 in experiment#1 and #2 and 10x recommended dose on d15 in experiment #3. Data were analyzed by one-way ANOVA, means were separated by Fisher’s protected LSD test. A p ≤ 0.05 was considered statistically different. In experiment #1, 30 and 75 ppm Cu-MHAC improved FCR during grower phase, increased jejunal villus height and reduced jejunal crypt depth, 30 ppm Cu-MHAC increased cecal Lactobacillus spp. abundance in 41 days broilers. In experiment #2, compared to CuSO4, 15ppm Cu-MHAC increased cumulative performance index in 28 days broilers, 15 and/or 30 ppm Cu-MHAC improved gut morphometry, and 30 ppm Cu-MHAC reduced the abundance of E. coli and Enterobacteriaceae in cecum in 43 days broilers. In experiment #3, 15 ppm and 30 ppm Cu-MHAC improved FCR vs. CuSO4 during starter phase, reduced the percentage of E. coli of total bacteria vs. TBCC, 30 ppm Cu-MHAC increased the percentages of Lactobacillus acidophilus, Lactobacillus spp. and Clostridium cluster XIVa of total bacteria vs. both CuSO4 and TBCC in the cecum of 27 days broilers. In summary, low doses of Cu-MHAC had comparable growth performance to high dose of TBCC and CuSO4 while improving gut microflora and gut morphometry in broilers subject to coccidiosis vaccination or coccidia challenge, indicating that low doses of bis-chelated Cu could be used as a complimentary strategy to improve animal gut health.

Coccidiosis is a major intestinal challenge that causes economic loss to the broiler industry. Two battery cage studies were conducted to evaluate the effect of trace minerals, source and dose of methionine on growth performance and gut health of broilers subjected to Eimeria challenge. Experiment #1 consisted of 9 treatments of 2 × 2 × 2 factorial design + 1 arrangement with main factors of methionine (Met) sources (DL-Met vs. 2-hydroxy-4-(methylthio)-butanoic acid (HMTBa)), total sulfur amino acid (TSAA) levels (high vs. low; ±5% of recommended level), and sources of trace minerals (TM) Zn:Cu:Mn in the form Inorganic trace minerals (ITM) in sulfates (80:20:100ppm) vs. mineral methionine hydroxy-analogue bis-chelate (MMHAC, 40:10:50ppm), each with 8 pens of 10 birds. Experiment #2 consisted of 2 treatments--ITM [ZnSO4:tribasic copper chloride (TBCC):MnSO4 110:125:120ppm] and MMHAC (Zn:Cu:Mn, 40:30:40ppm), each with 36 pens of 10 birds. All birds except for treatment 9 in experiment #1 were orally gavaged with 1x, 4x and 16x recommended dose of coccidiosis vaccine on d0, d7 and d14, respectively. Data were subjected to one-way and/or three-way ANOVA, and means were separated by Fisher’s protected LSD test with significance at p ≤ 0.05. In experiment #1, factorial analysis revealed the main effects of TSAA level and TM, but not Met source. High TSAA level improved body weight and cumulative feed intake at 14, 20, and 27d. MMHAC improved body weight at 14, and 27d; feed intake at 14, 20, and 27d; and cumulative FCR at 27d vs. sulfates. One-way ANOVA analysis showed that birds fed MMHAC and high levels of TSAA regardless of Met source had similar body weight as unchallenged birds on d27. In experiment #2, MMHAC improved body weight and cumulative FCR, and reduced jejunal IL-17A gene expression on d28. In summary, in broilers subjected to Eimeria challenge, supplementation of the reduced levels of bis-chelated trace minerals MMHAC improved growth performance compared to high levels of ITM (sulfates or TBCC), which might partially result from better immune response, high levels of TSAA improved growth performance, Met source had no effect. Supplementation of both bis-chelated trace minerals MMHAC and high levels of TSAA could overcome the growth performance challenge issue due to coccidiosis.

Trace mineral minerals Zn, Cu, and Mn play important roles in breeder production and progeny performance. The objective of this study was to determine maternal supplementation of trace mineral minerals on breeder production and progeny growth and development. A total of 540 broiler breeders, Cobb 500 (Slow feathering; 0–66 weeks old) were assigned to one of three treatment groups with the same basal diet and three different supplemental trace minerals: ITM–inorganic trace minerals in sulfates: 100, 16, and 100 ppm of Zn, Cu, and Mn respectively; MMHAC -mineral methionine hydroxy analog chelate: 50, 8, and 50 ppm of bis-chelated MINTREX®Zn, Cu and Mn (Novus International, Inc.), and TMAAC - trace minerals amino acid complex: 50, 8, and 50 ppm of Zn, Cu, and Mn. At 28 weeks of age, eggs from breeder treatments were hatched for progeny trial, 10 pens with 6 males and 6 female birds per pen were fed a common diet with ITM for 45 days. Breeder production, egg quality, progeny growth performance, mRNA expression of gut health associated genes in breeder and progeny chicks were measured. Data were analyzed by one-way ANOVA; means were separated by Fisher’s protected LSD test. A p-Value ≤ 0.05 was considered statistically different and 0.1 was considered numerical trend. Breeders on ITM treatment had higher (p < 0.05) body weight (BW), weight gain and lower (p < 0.05) feed conversion ratio (FCR) from 0 to 10 weeks, when compared to birds fed MMHAC. MMHAC significantly improved egg mass by 3 g (p < 0.05) and FCR by 34 points (0.05 < p < 0.1) throughout the reproductive period (26–66 weeks) in comparison to ITM. MMHAC improved (p < 0.01) egg yolk color versus (vs.) ITM and TMAAC in all periods, except 28 weeks, increased (p < 0.01) eggshell thickness and resistance vs. TMAAC at 58 weeks, and reduced (p < 0.05) jejunal NF-κB gene expression vs. TMAAC at 24 weeks. There was a significant reduction in tibial dry matter weight, Seedor index and resistance for the breeders that received MMHAC and/or TMAAC when compared to ITM at 18 weeks. Lower seedor index but numerically wider tibial circumference was seen in hens fed MMHAC at 24 weeks, and wider tibial circumference but lower tibial resistance in hens fed TMAAC at 66 weeks. Maternal supplementation of MMHAC in breeder hens increased (p < 0.0001) BW vs. ITM and TMAAC at hatching, reduced (p < 0.05) feed intake vs. ITM at d14 and d28, and improved (p < 0.01) FCR and performance index vs. TMAAC at d28, reduced (p < 0.01) NF-κB gene expression and increased (p < 0.05) A20 gene expression vs. TMAAC on d0 and vs. ITM on d14, reduced (p < 0.05) TLR2 gene expression vs. ITM on d0 and vs. TMAAC on d14, increased (p < 0.05) MUC2 gene expression vs. both ITM and TMAAC on d45 in progeny jejunum. Overall, these results suggest that supplementation with lower levels of MHA-chelated trace minerals improved breeder production and egg quality and reduced breeder jejunal inflammation while maintaining tibial development in comparison to those receiving higher inorganic mineral supplementation, and it also carried over the benefits to progeny with better growth performance, less jejunal inflammation and better innate immune response and gut barrier function in comparison to ITM and/or TMAAC.

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 < 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 < 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.

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.

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.