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

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

Supplemental methionine sources have a neutral impact on oxidative status in broiler chickens

To determine the effect of different dietary Met sources on oxidative status, male Cobb 500 broiler chickens were fed from day of hatch to 26 days of age (d26) a diet deficient in sulfur amino acids (control) or a diet containing 0.22% DL‐Met, 0.22% L‐Met or 0.31% Met precursor, DL‐2‐hydroxy‐4‐(methylthio) butanoic acid (DL‐HMTBA) to meet the Met + Cys requirements. Liver, breast muscle, duodenum, jejunum, and ileum were collected at day 10 (d10) and d26 to assay markers of oxidative stress, including total glutathione (TGSH), oxidized glutathione (GSSG), reduced glutathione (rGSH), protein carbonyls, thiobarbituric acid reactive substances (TBARS) and ferric reducing/antioxidant power (FRAP). In breast muscle, TGSH and rGSH were greater in L‐Met and DL‐HMTBA groups than in the control group (p < 0.05). An interaction of treatment and age was observed for TGSH in ileum (p = 0.01) and jejunum (p = 0.01), for GSSG in jejunum (p < 0.001), and for rGSH in ileum (p = 0.02). The ratios of rGSH to GSSG and GSSG to TGSH, which define oxidative status, were not affected by Met source. Protein carbonyls varied among groups in jejunum (p = 0.05) and breast muscle (p < 0.001), but were in the normal physiological range. No difference among treatment groups was observed for TBARS and FRAP in different tissues. Age effects were observed in all tissues for multiple oxidative stress markers. In conclusion, consuming different sources of supplementary Met did not alter the oxidative status in several tissues of broilers. Met + Cys deficiency did not compromise antioxidant capacity of chickens although growth was retarded.     

Digestibility,ruminal fermentation,blood metabolites and antioxidant status in ewes supplemented with DL‐methionine or hydroxy‐4 (methylthio) butanoic acid isopropyl ester

The effects of supplementing ewe diets with either DL‐methionine (DL‐Met) or 2‐hydroxy‐4 (methylthio) butanoic acid isopropyl ester (HMBi) were investigated on ruminal in situ degradability of grain and forage diets, in vivo digestibility, rumen fermentation, blood metabolites and antioxidant status. Six ruminally cannulated ewes were used in a replicated 3 × 3 Latin square design with 28‐day periods. The dietary treatments were as follows: (i) no supplemental Met (control; CON), (ii) DL‐Met at 1.2 g/kg DM intake and (iii) HMBi at 1.8 g/kg dry matter (DM) intake. Corn grain, barley grain and alfalfa hay were evaluated for their ruminal degradability by both in situ incubation and effective degradability measurements of DM, neutral detergent fibre (NDF) and acid detergent fibre (ADF). Compared to other treatments, HMBi supplementation increased (p < 0.05) the digestibility of organic matter, crude protein and NDF and also tended (p = 0.08) to increase the digestibility of DM and ADF. Moreover, HMBi supplementation increased (p < 0.01) total VFA concentrations, the molar proportions of valerate and iso‐butyrate in the rumen. Compared to the CON treatment, DL‐Met and HMBi treatments tended (p = 0.08) to increase the molar proportion of acetate but decreased (p < 0.05) ruminal ammonia‐N concentration. Ewes supplemented with HMBi and DL‐Met recorded greater (p < 0.05) serum concentrations of glutathione peroxidase, total antioxidant capacity and superoxide dismutase than the CON treatment. Serum concentrations of glucose, total protein, albumin, high‐density lipoprotein and very low‐density lipoprotein were greater (p < 0.01) and serum urea nitrogen (p < 0.05), malonyl dialdehyde and triglyceride were lower (p < 0.02) in the HMBi and DL‐Met animals than in the CON ewes. The results concluded that HMBi is a very effective form of dietary Met supplementation for ewes with a positive effect on digestion, rumen fermentation and serum antioxidant function.     

2‐Hydroxy‐4‐methylthio butanoic acid and DL‐methionine for Japanese quails in production

An experiment was performed using 1,000 laying Japanese quails to assess the availability of two alternative dietary methionine sources. Treatment 01 = Basal Feed that is deficient in digestible methionine + cystine (Met + Cys). The other treatments were constituted by Met + Cys levels of 0.8, 1.60 and 2.40 g/kg, supplemented with DL‐Methionine‐99%, HMTBA‐88% and HMTBA‐84%, being 10 treatments in total. The following characteristics were studied: feed intake (g/bird/day), egg production (egg/day × 100), egg weight (g/egg), egg mass (g/egg), feed conversion per egg dozen (kg feed/dozen eggs), feed conversion per egg mass (kg feed/kg eggs), relative yolk weight (g/100 g of egg), relative albumen weight (g/100 g of egg), relative shell weight (g/100 g of egg), shell thickness (mm) and specific gravity (g/cm³). In general result comment, supplemental methionine sources must be included in the poultry diet. The different methionine sources affect the performance of quails, and the increase in the levels within each source improves the performance variables. Significant effect was observable on performance variables and egg quality variables, being that DLM‐99% is superior to the other sources. The HMTBA‐88% source is superior to the HMTBA‐84% source for the same aforementioned variables. In conclusion, the bioefficacy values of the HMTBA‐88% and HMTBA‐84% sources compared to the DLM‐99% source on an equimolar basis were 81 and 79%, respectively, for the performance variables, and 83 and 74 while the methionine sources were equivalent for the variables related to egg quality.     

Gene expression differences in the methionine remethylation and transsulphuration pathways under methionine restriction and recovery with D,L‐methionine or D,L‐HMTBA in meat‐type chickens

This study examined the molecular mechanisms of methionine pathways in meat‐type chickens where birds were provided with a diet deficient in methionine from 3 to 5 weeks of age. The birds on the deficient diet were then provided with a diet supplemented with either D,L‐methionine or D,L‐HMTBA from 5 to 7 weeks. The diet of the control birds was supplemented with L‐methionine from hatch till 7 weeks of age. We studied the mRNA expression of methionine adenosyltransferase 1, alpha, methionine adenosyltransferase 1, beta, 5‐methyltetrahydrofolate‐homocysteine methyltransferase, 5‐methyltetrahydrofolate‐homocysteine methyltransferase reductase, betaine‐homocysteine S‐methyltransferase, glycine N‐methyltransferase, S‐adenosyl‐L‐homocysteine hydrolase and cystathionine beta synthase genes in the liver, duodenum, Pectoralis (P.) major and the gastrocnemius muscle at 5 and 7 weeks. Feeding a diet deficient in dietary methionine affected body composition. Birds that were fed a methionine‐deficient diet expressed genes that indicated that remethylation occurred via the one‐carbon pathway in the liver and duodenum; however, in the P. major and the gastrocnemius muscles, gene expression levels suggested that homocysteine received methyl from both folate and betaine for remethylation. Birds who were switched from a methionine deficiency diet to one supplemented with either D,L‐methionine or D,L‐HMTBA showed a downregulation of all the genes studied in the liver. However, depending on the tissue or methionine form, either folate or betaine was elicited for remethylation. Thus, mRNA expressions show that genes in the remethylation and transsulphuration pathways were regulated according to tissue need, and there were some differences in the methionine form.     

Effects of L‐methionine on performance,gut morphology and antioxidant status in gut and liver of piglets in relation to DL‐methionine

This study investigated the hypothesis that dietary supplementation of L‐methionine (L‐Met) in weaned piglets in relation to DL‐methionine (DL‐Met) results in a higher antioxidant status and lower need for antioxidant enzyme activation in intestinal epithelium and body tissues, and improves gut morphology and gut barrier function as well as performance. A total of 99 early‐weaned 21‐day old piglets were allotted to six groups and fed a semi‐synthetic wheat–barley‐based basal diet supplemented with 0.067%, 0.107% and 0.147% of either DL‐Met (MetAmino; Evonik, Hanau, Germany) or L‐Met (L‐Met100; CJ Europe, Schwalbach am Taunus, Germany) to reach dietary Met concentrations of 0.16%, 0.20% and 0.24%, of which the latter met the requirements for maintenance and growth based on a pre‐experiment. Feed intake and body weights were recorded weekly, and samples of plasma, liver and duodenum and jejunum mucosa were collected after 3 weeks at slaughter. Plasma concentrations of L‐Met were similar, and those of D‐Met and total Met were higher in piglets fed DL‐Met in relation to those fed L‐Met. Feed intake, daily gains and feed:gain ratio, and the relative bio‐efficacy based on gains and feed:gain ratio were similar for both groups. Likewise, villi length, crypt depth, the villi length:crypt depth ratio in duodenum and jejunum and gene expression of tight junction proteins in the jejunum did not differ. Concentrations of antioxidants like glutathione and tocopherol, the total antioxidant capacity, the mRNA abundance or activity of antioxidant enzymes like superoxide dismutase and glutathione peroxidase, concentrations of thiobarbituric acid reactive substances, markers for oxidative damage of lipids and the expression of inflammatory genes were similar in liver and jejunum mucosa. These data indicate that the effects of L‐Met and DL‐Met supplementation are comparable considering both piglet performance and parameters of gut health and function like gut morphology and the intestinal antioxidant status.     

Effects of dietary amino acid supplementation on measures of whole‐body and muscle protein metabolism in aged horses

The objective of this study was to examine markers of whole‐body and muscle protein metabolism in aged horses fed a diet typical for North American aged horses, supplemented with amino acids. In a replicated Latin square design, six aged horses (20 ± 1.1 years) were studied while receiving each of three isocaloric, isonitrogenous diets, a control treatment concentrate (CON; 100 mg/kg^?1 BW day^?1 lysine, 84 mg kg^?1 day^?1 threonine, 51 mg kg^?1 day^?1 methionine), LYS/THR (134 mg kg^?1 BW day^?1 lysine, 110 mg kg^?1 BW day^?1 threonine, 52 mg kg^?1 BW day^?1 methionine) and LYS/THR/MET (132 mg kg^?1 BW day^?1 lysine, 112 mg kg^?1 BW day^?1 threonine, 62 mg kg^?1 BW day^?1 methionine). In each 15‐days period, urine and faeces were collected for assessment of nitrogen balance. Blood samples were collected before and after feeding for analysis of plasma urea nitrogen (PUN), glucose, insulin and plasma amino acid concentrations. Skeletal muscle samples were collected for measurement of proteins associated with muscle protein synthesis and degradation, and horses underwent stable isotope infusion procedures for comparison of differences in whole‐body rates of protein synthesis and degradation. There was no effect of treatment on relative abundance of proteins involved in protein synthesis, nitrogen retention or phenylalanine kinetics. PUN concentrations tended to be higher for LYS/THR (p = 0.054) and were higher for LYS/THR/MET (p = 0.0056) than for CON. Atrogin‐1 abundance tended to be higher in the post‐absorptive state for the CON treatment (p = 0.07), indicating that amino acid supplementation resulted in less muscle protein degradation when horses were in the post‐absorptive state. However, lack of differences in nitrogen retention and phenylalanine kinetics indicated that whole‐body protein metabolism was not improved, and higher PUN concentrations in the supplemented diets suggest that the supplemented amino acids may have been catabolized. Amino acid availability was not limiting protein synthesis in the sedentary aged horses in this study when fed the CON diet.     

Dietary methionine source and level affect hepatic sulfur amino acid metabolism of broiler breeder hens

A study was carried out to investigate the effects of dietary methionine source and level on plasma free amino acids patterns and the expression of genes involved in hepatic methionine metabolism in broiler breeders. A total of 2184 broiler breeders were assigned to 13 dietary treatments, with eight replicates per treatment. The 13 treatments included one control group and 12 additional treatments employing two sources and six levels (0.05, 0.10, 0.15, 0.20, 0.25 and 1.00%). Higher plasma methionine concentration was measured for DL‐methionine (DLM) treated hens. Plasma alanine concentration was linearly increased as DLM or 2‐hydroxy‐4‐(methylthio) butanoic acid (HMTBA) supplementation level increased. There was a linear increase in concentrations of tyrosine, valine, glycine and serine as dietary DLM supplementation level increased. Hens treated with DLM had higher relative expression of ADA than those fed HMTBA. The expression of MS, ADA, SAHH and MAT2A changed quadratically as HMTBA supplementation level increased, while the expression of GNMT and SAHH changed quadratically as DLM supplementation level increased. In conclusion, the effects of HMTBA on plasma free amino acid patterns and the expression of hepatic genes involved with methionine are different from DLM.     

Effects of different dietary protein levels and DL‐methionine supplementation on hair growth and pelt quality in mink (Neovision vision)

The effect of different dietary protein levels and DL‐methionine (Met) supplementation on hair growth and the resulting pelt quality in mink was studied. Four groups of male mink were fed with four isocaloric diets containing 32% (P32), 24% (P24), 16% (P16) or P24+Met (0.8%) crude protein of dry matter (DM) from September to December. Skin biopsies were taken at the pelting. Histological techniques and computer‐assisted light microscopy were used to determine the ratio of activity (ROA) of under hairs and guard hairs respectively. The results showed that when the dietary protein level reduced from 32% to 16%, body length, number and diameter of under hairs and guard hairs of minks declined, and pelt length and pelt weight of minks decreased significantly (p < 0.05). These parameters were similar between P32 and P24 with Met supplementation (p > 0.05). The hair follicle density of the winter coat was not influenced by the dietary protein levels and Met supplementation (p > 0.05). Low‐protein diets content led to a reduction of hair follicle developing to next phase. It was documented that 24% crude protein of DM with Met supplementation during growing‐furring period was sufficient for minks to express their genetic capacity to develop hair follicles and achieve the prime fur characteristics. Overall this study demonstrated that hair growth and hair properties in pelts are very dependent on the dietary protein and Met supply in the growing‐furring period of minks.     

Marginal deficiencies of dietary arginine and methionine could suppress growth performance and immunological responses in broiler chickens

The present study was conducted to investigate the effects of different levels of dietary arginine (Arg) and methionine (Met) on performance, immune responses, and carcass characteristics of broiler chickens. A total of 540 day‐old Ross 308 broiler chicks were randomly assigned into the nine experimental diets, consisting five replicates of 12 birds each. Dietary treatments included three different levels (90%, 100%, and 110% of National Research Council [NRC] specifications) of either dietary Arg or Met, which were fed to the birds according to a 3 × 3 factorial arrangement of treatments during a 42 days feeding trial. Results showed that supplementation of Arg and Met into the deficient‐diets increased (p < .01) weight gains during all trial periods. Although average daily feed intake (ADFI) was not influenced by dietary treatments, increasing Arg up to 100% of NRC recommendations improved (p < .05) feed conversion ratio (FCR) throughout the trial period. Similarly, supplementation of deficient‐diets with Met improved FCR values. There was a significant (p < .01) Arg × Met interaction for ADFI during the starter period; increasing the dietary Arg level increased ADFI when the diets were deficient in Met, while had an opposite effect in diets containing higher dietary Met levels. On the other hand, dietary Met fortification improved (p = .067) FCR values to a greater extent in 110% Arg‐diets during the entire trial period. Although different levels of Arg and Met had no marked effects on carcass yield and abdominal fat percentage, supplemental Arg up to 100% of NRC values increased (p < .01) the relative weights of spleen and bursa of Fabricius. Furthermore, bursa weight was affected by Arg × Met interaction (p < .01), so that supplemental Arg level of 100% of NRC increased the relative bursa weight in birds that were fed diets containing 90% and 110% of Met. Serum uric acid level was decreased (p < .05) as a result of dietary Arg fortification up to 110% of NRC recommended values. Supplementation of deficient‐diets with Met decreased (p < .05) serum cholesterol level. Although Newcastle antibody titer was not affected by dietary Arg or Met levels, Arg fortification of deficient‐diets increased (p < .001) antibody responses against infectious bronchitis (IBV) and bursal (IBD) disease viruses. Similarly, Met supplementation of deficient‐diets increased IBD antibody titer. There were significant (p < .05) Arg × Met interactions for IBV and IBD titers; Met fortification of 110% Arg‐diets was more effective in increasing antibody titers. An increase in dietary Met level up to 100% of NRC values increased (p < .001) serum concentration of γ‐globulins. The present findings imply that supplemental Arg could affect feed efficiency and antibody responses when the diets were already fortified with a sufficient Met level.     

Effects of dietary supplemental methionine source and betaine replacement on the growth performance and activity of mitochondrial respiratory chain enzymes in normal and heat‐stressed broiler chickens

This study aimed to evaluate the effects of dietary supplemental methionine (Met) source and betaine (Bet) replacement for Met on performance and activity of mitochondrial respiratory chain enzymes (MRCEs) in normal and heat‐stressed broiler chickens. Total of 1,200‐day‐old Ross 308 chicks were allocated to two houses, each consisted of 12 treatments, five replicates of 10 birds each with 2 × 2×3 × 2 (temperature × Met source × Met level × Bet, respectively) split‐plot factorial arrangement. Met level in the basal diets was 70% requirements (Req) that was increased to the requirement or 130% by supplemental dl ‐ or l ‐Met. Bet was or was not substituted at the rate of 30% supplemental dl ‐ or l ‐Met. Feed conversion ratio (FCR) in chicks fed 70% l ‐Met was lower than those fed 70% dl ‐Met diet during 1–10 days (p = 0.04). Broilers fed diets containing requirement or 130% Met, regardless of its source, showed higher weight gain (WG) than those received 70% Met diet during 11–42 days (p < 0.001). Feed intake (FI) of broilers fed 130% Met diet was decreased compared to other two groups during 11–42 days (p < 0.05). One hundred thirty percent Met requirement diet resulted in lower FCR comparing to other two groups during 11–42 days (p < 0.001). Heat‐stressed birds grew less than those under normal condition (p < 0.05). Broilers fed Req Met diet under normal temperature exhibited higher activities of complexes (Cox) I and III (p < 0.05). Cox I activity in heat‐stressed birds fed Bet + diet was similar to those fed Bet‐diet under normal temperature (p = 0.046). It is concluded that performance and the activities of Cox I and III were increased as the level of Met increased. Bet replacement for 30% supplemental Met resulted in similar consequences comparing to non‐Bet replacement diets on performance, but increased the activity of Cox III. l ‐Met was effective than dl ‐Met at the cellular level. High ambient temperature depressed performance and MRCE activity.     

Bioavailability of methionine hydroxy analog-free acid relative to DL-methionine in broilers

Three experiments were conducted to determine and validate the relative bioefficacy of the liquid form of hydroxy analog (liquid MHA‐FA) to that of DL‐methionine (Met) in male broilers. In experiments 1 and 2, 945 and 550 male broilers were fed either a Met‐deficient basal diet, or the basal diet supplemented with four and five equimolar levels of each Met source, respectively. In experiment 3, 1232 male broilers were fed either a Met‐deficient basal diet, or the basal diet with three levels (low, moderate, or high) of liquid MHA‐FA or DL‐Met at 65% of the liquid MHA‐FA level. Growth performance improved in all trials, regardless of the Met source, relative to those broilers fed the basal diet. In experiments 1 and 2, the bioefficacy estimates for liquid MHA‐FA relative to DL‐Met on a product basis were: 50% and 64% for weight gain, 51% and 59% for the feed conversion ratio, and 54% and 48% for breast‐meat yield, respectively. In experiment 3, there were no differences between the DL‐Met or liquid MHA‐FA treatments, and the broilers fed liquid MHA‐FA were 68% as efficient on a product basis as those fed DL‐methionine for weight gain. The results from these three trials indicate that the bioefficacy of liquid MHA‐FA relative to DL‐methionine is 57% on a product basis on average across all criteria tested.     

Dose–responses of zinc–methionine supplements on growth,blood metabolites and gastrointestinal development in sheep

The effects of zinc–methionine (Zn‐Met) supplementation on growth, blood metabolites and gastrointestinal development were investigated in two experiments with sheep. The objective of Experiment 1 was to determine the effects of Zn‐Met supplementation on hormones and metabolites involved in growth and energy balance regulation, while Experiment 2 aimed to determine the effects of Zn‐Met on feed intake, body weight, gastrointestinal development and liver glycogen concentration in lamb. The animals were assigned to groups with different concentrations of dietary Zn‐Met (0, 0.4, 0.8 and 1.2 g/day) in both experiments. In Experiment 1, feeding different doses of Zn‐Met increased plasma insulin‐like growth factor 1 (IGF‐1) concentration, but it linearly decreased plasma growth hormone (GH). No differences were observed in blood cortisol, insulin and glucose concentrations among the treatments. In Experiment 2, addition of Zn‐Met to the diets did not lead to changes in the body weights of the lambs. Both average daily gain and dry matter intake (DMI) increased linearly with increasing concentrations of dietary Zn‐Met. Lambs receiving Zn‐Met showed higher liver glycogen concentrations than the control. While significant increases were observed in the villus height and crypt depth in the duodenum and jejunum as a result of Zn‐Met supplementation, no change was detected in blood glucose concentrations (p > 0.05). Our findings suggest that dietary Zn‐Met may improve growth, energy balance and gastrointestinal development in sheep.     

Effect of 2‐hydroxy‐4‐(methylthio) butanoic acid and acidifier on the performance,chyme pH,and microbiota of broilers

This study was aimed to explore the comparative acidifying properties of 2‐hydroxy‐4‐(methylthio) butanoic acid (HMTBA) and a combination of DL‐methionine (DLM) and acidifier in male broiler production. A total of 480 1‐day‐old broiler chicks were randomly divided into four treatments: A (low HMTBA, 0.057% HMTBA); B (low acidifier, 0.05% DLM + 0.057% acidifier); C (high HMTBA, 0.284% HMTBA); and D (high acidifier, 0.25% DLM + 0.284% acidifier). At 21 d, growth performance, chyme pH, digestive enzyme activities, and intestinal microflora were measured. The pH of crop, gizzard, and ileum contents was higher in the HMTBA treatment group than in DLM + acidifier treatment group. Furthermore, acidifier supplementation promoted growth of butyrate‐producing bacteria such as Faecalibacterium, whereas high HMTBA (0.284%) inhibited the proliferation of acid‐producing bacteria including Roseburia and Collinsella. The chymotrypsin activity was lower in the HMTBA group than in the DLM + acidifier group. In contrast, high‐level HMTBA group showed higher average daily gain and average daily feed intake than the DLM + acidifier group. These results suggested that HMTBA work through different pathways with DLM plus acidifier.     

The site of intestinal disappearance of DL-methionine and methionine hydroxy analog differs in pigs

A study was conducted with pigs to determine the sites of and the influence of dietary wheat middlings (WM) as a fiber source on the absorption of dl-Met (DLM) and dl-2-hydroxy-4-methylthio butanoic acid (HMTBA). Twelve 25-kg barrows were used in a replicated 6 × 4 incomplete Latin-square design with 6 diets and 4 periods per pig. Each pig was surgically fitted with simple T-cannulas in the terminal duodenum and ileum 2 wk before the initiation of the experiment and housed in a stainless-steel metabolism crate. The 6 diets were arranged in a 2 × 3 factorial of WM at 0 (CON) or 8% and Met source including none, DLM, or HMTBA. Supplemental DLM doubled the duodenal free Met concentration from 9.6 to 19.6 g/100 g of DMI. Supplemental DLM also increased the apparent ileal digestibility (AID) of free Met from 52 to 92% (P < 0.05). Adding 8% WM increased (P < 0.01) duodenal NDF concentrations by 2 g/100 g of DMI and decreased (P < 0.05) AID of free Met. No HMTBA was detected in digesta collected from the duodenum or ileum of pigs fed the CON + HMTBA or the WM + HMTBA diets, indicating complete absorption before the terminal duodenum regardless of WM inclusion. Interactions between Met supplementation and WM were observed for the digestibility of all other nutrients (P < 0.05). Fiber and AA digestibility in the ileum were the greatest for the CON + HMTBA diet, and least for the WM + HMTBA diet. The CON + HMTBA diet had greater AID of ADF, NDF, CP, and Thr, and greater AID of ADF, NDF, ash, Arg, Ile, Gly, and Pro when compared with the CON and CON + DLM diets, respectively (P < 0.05). The WM + HMTBA had less AID of ADF, CP, and all AA with the exception of Lys, Trp, and Val when compared with the CON + HMTBA (P < 0.05). In summary, absorption of HMTBA in the pig is complete by the end of the duodenum, but negative interaction between HMTBA and WM can decrease the AID of most AA.     

Dietary methionine requirement of Jing Brown layer hens from 9 to 17 weeks of age

This study was conducted to evaluate the effect of dietary methionine (Met) supplementation in growth performance and reproductive performance of Jing Brown layer hens. A total of 375 9‐week‐old Jing Brown layer hens were allocated equally to five treatments consisting of 5 replicates with 15 hens. Hens were fed with a diet of corn and soya bean meal supplemented with 0.23%, 0.27%, 0.31%, 0.35% and 0.39% Met respectively. Different Met levels did not significantly affect average daily feed intake (ADFI), average daily gain (ADG) and feed/gain ratio (F/G) (p > 0.05), whereas flock uniformity (FU) and jejunum index were significantly different (p < 0.05), and the largest FU was observed in 0.31% Met. Dietary supplementation of Met significantly affected reproductive system development (p < 0.05), and 0.27–0.31% Met obtained optimal reproductive system development. Different Met levels significantly affected serum uric acid and alkaline phosphatase. Moreover, the relatively higher reproductive hormones in serum were observed in 0.27% Met. Analysis of quadratic curve estimation of flock uniformity, the total number of follicles, the primary follicles and the secondary follicles showed that the optimal Met levels were 0.293%, 0.286%, 0.286% and 0.288%, which could be averaged to 0.288%. These results suggested that the optimal Met requirement for Jing Brown layer hens from 9 to 17 weeks old is 0.29%.     

Effects of guanidinoacetic acid supplementation on nitrogen retention and methionine flux in cattle

Creatine stores high-energy phosphate bonds in muscle and is synthesized in the liver through methylation of guanidinoacetic acid (GAA). Supplementation of GAA may therefore increase methyl group requirements, and this may affect methyl group utilization. Our experiment evaluated the metabolic responses of growing cattle to postruminal supplementation of GAA, in a model where methionine (Met) was deficient, with and without Met supplementation. Seven ruminally cannulated Holstein steers (161 kg initial body weight [BW]) were limit-fed a soybean hull-based diet (2.7 kg/d dry matter) and received continuous abomasal infusions of an essential amino acid (AA) mixture devoid of Met to ensure that no AA besides Met limited animal performance. To provide energy without increasing the microbial protein supply, all steers received ruminal infusions of 200 g/d acetic acid, 200 g/d propionic acid, and 50 g/d butyric acid, as well as abomasal infusions of 300 g/d glucose. Treatments, provided abomasally, were arranged as a 2 × 3 factorial in a split-plot design, and included 0 or 6 g/d of l-Met and 0, 7.5, and 15 g/d of GAA. The experiment included six 10-d periods. Whole body Met flux was measured using continuous jugular infusion of 1-¹³C-l-Met and methyl-²H₃-l-Met. Nitrogen retention was elevated by Met supplementation (P < 0.01). Supplementation with GAA tended to increase N retention when it was supplemented along with Met, but not when it was supplemented without Met. Supplementing GAA linearly increased plasma concentrations of GAA and creatine (P < 0.001), but treatments did not affect urinary excretion of GAA, creatine, or creatinine. Supplementation with Met decreased plasma homocysteine (P < 0.01). Supplementation of GAA tended (P = 0.10) to increase plasma homocysteine when no Met was supplemented, but not when 6 g/d Met was provided. Protein synthesis and protein degradation were both increased by GAA supplementation when no Met was supplemented, but decreased by GAA supplementation when 6 g/d Met were provided. Loss of Met through transsulfuration was increased by Met supplementation, whereas synthesis of Met from remethylation of homocysteine was decreased by Met supplementation. No differences in transmethylation, transsulfuration, or remethylation reactions were observed in response to GAA supplementation. The administration of GAA, when methyl groups are not limiting, has the potential to improve lean tissue deposition and cattle growth.     

Antioxidant status and expression of inflammatory genes in gut and liver of piglets fed different dietary methionine concentrations

This study investigated the hypothesis that dietary concentrations of methionine (Met), as a precursor of cysteine which is a constituent of glutathione (GSH), affect tissue antioxidant concentrations and the antioxidant defence system in pigs. Forty‐five piglets (DanZucht × Pietrain) were allotted to three groups of similar mean body weight (11.0 ± 0.9 kg). The basal diet was composed of barley, wheat, corn starch, soybean oil, sucrose, cellulose and a mineral supplement with suboptimal concentrations of Met and was supplemented with dl ‐Met to reach 0.16%, 0.20% and 0.24% of dietary Met and 0.40%, 0.44% and 0.48% of dietary Met and cysteine in groups 0.16, 0.20 and 0.24 respectively. After 3 weeks, at slaughter, samples of liver, jejunum mucosa and plasma were collected. Feed intake and weight gains increased and feed:gain ratio decreased when dietary Met concentrations increased. The Trolox equivalent antioxidant capacity (TEAC), concentrations of GSH and thiobarbituric acid reactive substances (TBA‐RS) and the activity of the glutathione peroxidase (GPx) in liver and jejunum mucosa were similar in all groups (p > 0.05). Relative mRNA concentrations of selected target genes of the nuclear factor (erythroid‐derived 2)‐like 2 (Nrf2), the master regulator of the antioxidant response, and of the nuclear factor ‘kappa‐light‐chain‐enhancer’ of activated B‐cells (NF‐κB), the master regulator of inflammation, were largely unaffected both in jejunum and liver. In conclusion, inflammation‐ and oxidative stress‐related pathways on the molecular level, and concentrations of lipid peroxidation products, of antioxidants and of enzymes involved in the antioxidant defence system were mostly unaffected by dietary Met concentration in gut and liver. These findings suggest that suboptimal dietary Met concentrations did not influence the antioxidant defence system of gut and liver in healthy piglets.     

Methionine promotes the development of hair follicles via the Wnt/β-catenin signalling pathway in Rex rabbits

To investigate the effect and molecular mechanism of methionine (Met) on the growth of hair follicles (HFs) in Rex rabbits. A total of 200 weaning Rex rabbits were divided into four groups and fed varying levels of Met-supplemented diets. We measured the HF density on dorsal skin and the Wnt/β-catenin pathway protein expression level. Meanwhile, whole HFs were isolated from Rex rabbit skins and cultured with Met in vitro to measure hair shaft growth. The relationship between Met and the Wnt/β-catenin signalling pathway was also characterized by using the Wnt/β-catenin signalling inhibitor, XAV-939. The results showed that the addition of dietary Met could significantly increase the HF density on dorsal skin (p < .05) and enhance the protein expression level of Wnt10b (p < .05), β-catenin (p < .05) and DSH (p < .05). Methionine stimulation could also prolong the hair shafts growth in vitro (p < .05). And inhibition of Wnt/β-catenin signalling using XAV-939 could eliminate this phenomenon. In summary, Met can increase the density of HFs on dorsal skin in vitro and prolong the hair shaft growth of HFs in vivo via the Wnt/β-catenin signalling pathway.     

Effects of thermo‐resistant non‐starch polysaccharide degrading multi‐enzyme on growth performance,meat quality,relative weights of body organs and blood profile in broiler chickens

This research was conducted to study the performance and carcass parameters of broiler chickens fed diets supplemented with heat‐treated non‐starch polysaccharide degrading enzyme. A total of 432 one‐day old Ross 308 broiler chickens were allocated to five treatments: (i) CON (basal diet), (ii) E1: CON + 0.05% multi‐enzyme, (iii) E2: CON + 0.1% multi‐enzyme, (iv) E3: CON + 0.05% thermo‐resistant multi‐enzyme and (v) E4: CON + 0.1% thermo‐resistant multi‐enzyme, each treatment consisted of six replications and 12 chickens in each replication. The chickens were housed in three floor battery cages during 28‐day experimental period. On days 1–7, gain in body weight (BWG) improved by feeding the diets supplemented with thermo‐resistant multi‐enzyme. On days 7–21 and 1–28, chickens fed the diets containing thermo‐resistant multi‐enzyme showed improved (p < 0.05) BWG and feed conversion ratio (FCR) compared to CON group. Supplementing the diets with multi‐enzyme or thermo‐resistant multi‐enzyme affected the percentage of drip loss on d 1 (p < 0.05). Drip loss percentage on days 3 and 5 and also meat colour were not affected significantly. Supplementing the diets with multi‐enzyme or thermo‐resistant multi‐enzyme did not affect the relative weights of organs but compared to CON group, relative weight of breast muscle increased and abdominal fat decreased (p < 0.05). Among measured blood constituents, chickens fed supplemented diets with thermo‐resistant multi‐enzyme showed higher (p < 0.05) IgG. Counts of red and white blood cells and lymphocyte percentage were not affected. In conclusion, the results demonstrated that supplementing pelleted diets with thermo‐resistant multi‐enzyme improved performance of broiler chickens.