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 rumen‐protected methionine on milk production in early lactation dairy cattle fed with a diet containing 14.5% crude protein

We evaluated the influence on milk production of feeding early lactation cows a diet that included 14.5% crude protein (CP) and that did not meet methionine (Met) requirements or that met them by supplying rumen‐protected Met (RPMet). Thirty‐nine multiparous Holstein cows were allocated into two groups. For 15 weeks after calving, each group was fed one of the two total mixed rations, Control (n = 20) or Treatment (n = 19). The Treatment group received added RPMet at 0.034% (8 g/day) of the Control diet on dry matter basis. The adequacies of Met for the Control and Treatment groups were 96% and 106%, respectively, and for other amino acids, >110%. The CP level (14.5%) was 1 percentage point lower than that recommended by the Japanese Feeding Standard (2006). No between‐group differences were found in milk yield (40 kg/day), milk composition, plasma profile, rumen fermentation, nitrogen balance, or cow health. Met intake and the amount of rumen‐undegradable feed Met were higher in the Treatment group (p < 0.05). Microbial Met and total metabolizable Met did not differ between groups. Supplying RPMet in a 14.5% CP diet during early lactation did not dramatically affect milk production, because the amount of total metabolizable Met was unchanged.     

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.     

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 dietary methionine supplementation on growth performance,intestinal morphology,antioxidant capacity and immune function in intra‐uterine growth‐retarded suckling piglets

This study investigated the effects of dietary supplementation with _L‐methionine (_L‐Met), _DL‐methionine (_DL‐Met) and calcium salt of the methionine hydroxyl analog (MHA‐Ca) on growth performance, intestinal morphology, antioxidant capacity and immune function in intra‐uterine growth‐retarded (IUGR) suckling piglets. Six normal birthweight (NBW) female piglets and 24 same‐sex IUGR piglets were selected at birth. Piglets were fed nutrient adequate basal diet supplemented with 0.08% _L‐alanine (NBW‐CON), 0.08% _L‐alanine (IUGR‐CON), 0.12% _L‐Met (IUGR‐LM), 0.12% _DL‐Met (IUGR‐DLM) and 0.16% MHA‐Ca (IUGR‐MHA‐Ca) from 7 to 21 days of age respectively (n = 6). The results indicated that IUGR decreased average daily milk (dry matter) intake and average daily gain and increased feed conversion ratio of suckling piglets (p < 0.05). Compared with the NBW‐CON piglets, IUGR also impaired villus morphology and reduced antioxidant capacity and immune homeostasis in the intestine of IUGR‐CON piglets (p < 0.05). Supplementation with _L‐Met enhanced jejunal villus height (VH) and villus area and ileal VH of IUGR piglets compared with IUGR‐CON piglets (p < 0.05). Similarly, _DL‐Met supplementation increased VH and the ratio of VH to crypt depth in the jejunum compared with IUGR‐CON pigs (p < 0.05). Supplementation with _L‐Met and _DL‐Met (0.12%) tended to increase reduced glutathione content and reduced glutathione: oxidized glutathione ratio and decrease protein carbonyl concentration in the jejunum of piglets when compared with the IUGR‐CON group (p < 0.10). However, supplementation with MHA‐Ca had no effect on the intestinal redox status of IUGR piglets (p > 0.10). In conclusion, supplementation with either _L‐Met or _DL‐Met has a beneficial effect on the intestinal morphology and antioxidant capacity of IUGR suckling piglets.     

Supplementation of diets for lactating sows with zinc amino acid complex and gastric nutriment‐intubation of suckling pigs with zinc methionine on mineral status,intestinal morphology and bacterial translocation in lipopolysaccharide‐challenged weaned pigs

Sixty‐four pigs from 16 sows were used to evaluate addition of zinc amino acid complex (ZnAA) to lactating sows and gastric nutriment‐intubation of zinc methionine (ZnMet) to suckling pigs on mineral status, intestinal morphology and bacterial translocation after weaning. Sows were fed a barley‐based diet supplying 120 ppm zinc (Zn; control) or the control diet supplemented with 240 ppm Zn from ZnAA. At birth, day‐10 and day‐21 (weaning) of age, pigs from each litter were nutriment‐intubated with 5 ml of an electrolyte solution without or with 40 mg Zn from ZnMet. At weaning, 24 h prior to the collection of small and large intestinal lymph nodes and sections of the duodenum, jejunum and ileum, the pigs received an intramuscular injection of saline without or with 150 μg/kg body weight of Escherichia coli O26:B6 lipopolysaccharide (LPS). With the exception of a tendency (p = 0.09) for lower serum concentration of copper in pigs at weaning from ZnAA‐supplemented sows, there were no differences (p > 0.1) than for pigs from control‐fed sows for mineral status or intestinal morphology. Nutriment‐intubation of ZnMet increased serum (p = 0.001) and liver (p = 0.003) Zn concentrations, number of goblet cells per 250 μm length of jejunal villous epithelium (p = 0.001) and tended (p = 0.06) to enhance jejunum mucosa thickness. Interactive effects (p < 0.05) for higher jejunal villi height and villi:crypt ratio and increased ileal goblet cell counts were apparent for pigs from ZnAA‐supplemented sows that also received nutriment‐intubation of ZnMet. Challenge with LPS increased (p = 0.05) ileal villous width. Nutriment‐intubation of ZnMet decreased (p = 0.05) anaerobic bacteria colony forming unit counts in the large intestinal mesenteric lymph nodes. In conclusion, nutriment‐intubation of ZnMet increased serum and liver tissue concentrations of Zn and resulted in limited improvement to intestinal morphology of weaned pigs.     

Effects of adding liquid dl‐methionine hydroxy analogue‐free acid to drinking water on growth performance and small intestinal morphology of nursery pigs

This study was conducted to evaluate the effect of adding liquid dl ‐methionine hydroxy analogue free acid (LMA) to drinking water on growth performance, small intestinal morphology and volatile fatty acids in the caecum of nursery pigs. Twenty‐four crossbred pigs (Large White × Landrace, BW ～18 kg) were divided into three groups with four replications of two piglets each. The piglets received drinking water without (control), with 0.05 or 0.10% LMA. The results indicated that adding LMA at 0.10% to drinking water significantly increased their weight gain, average daily feed intake (p < 0.05) and tended to improve the feed conversion ratio. Adding LMA to drinking water significantly increased their water intake and significantly reduced the pH of drinking water (p < 0.01), thus total plate count (p < 0.01) and Escherichia coli in drinking water was reduced (p < 0.05), while the total number of bacteria in the caecum was not significantly affected. Liquid dl ‐methionine hydroxy analogue free acid supplementation in drinking water tended to decrease pH in the stomach, duodenum, jejunum, colon and rectum. Furthermore, adding LMA at 0.10% significantly increased villous height in the duodenum, jejunum and ileum (p < 0.05), and the villous height:crypt depth ratio in the jejunum and ileum (p < 0.01) was higher, whereas acetic acid concentration in the caecum was significantly lower than in the control group. It could be concluded that adding LMA to drinking water improved growth performance of the nursery pigs because of high water quality and high nutrient utilization caused by an improvement of small intestinal morphology (not from nutritional effect of methionine source).     

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.     

The effect of dietary supplementation with rumen‐protected methionine alone or in combination with rumen‐protected choline and betaine on sheep milk and antioxidant capacity

This study investigated the effects of dietary inclusion of rumen‐protected methionine alone or in combination with rumen‐protected choline and betaine on: (i) milk yield, chemical composition and fatty acids (FA) profile and (ii) blood plasma glutathione transferase (GST) activity of periparturient ewes. Furthermore, the oxidative stress indicators for measuring total antioxidant and free radical scavenging activity [ferric reducing ability of plasma (FRAP) and 2,2′‐azino‐bis (3‐ethylbenzothiazoline‐6‐sulphonic acid) (ABTS) assays] were also determined in plasma and milk of ewes. Thus, 45 ewes were divided into three equal groups. Each animal of the control group fed daily with a basal diet. The same diet was offered also in each animal of the other two groups. However, the concentrate fed to M group was supplemented with 2.5 g/kg rumen‐protected methionine, while the concentrate fed to MCB group with 5 g/kg of a commercial product which contained a combination of methionine, choline and betaine, all three in rumen‐protected form. The results showed that the M diet, compared with the control, increased significantly the ewe's milk fat and the total solids content. Likewise, a tendency for higher milk fat and total solids content in ewes fed the MCB diet was also observed. Both M and MCB diets had not noticeable impact on ewes milk FA profile. Significantly higher FRAP values in the blood plasma of ewes fed the MCB and in the milk of ewes fed with the M diet compared with the control were found. Additionally, significantly higher GST activity in the blood plasma of ewes fed the M diet, compared with the control, was observed. Moreover, a significant increase (by 20%) and a tendency for increase (by 16.72%) in the growth rate of lambs nursing ewes fed with M and MCB diets, respectively, compared to controls, were found.     

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.     

Effect of methionine hydroxy analog-free acid on growth performance and chemical composition of liver of broiler chicks fed a corn–soybean based diet from 0 to 6 weeks of age

The experiment was carried out to determine the effects of liquid DL‐methionine hydroxy analog‐free acid (LMA) and dry DL‐methionine (DLM) on growth performance, carcass quality and chemical composition of the livers of broiler chicks during 0–6 weeks of age. Four hundred and fifty male commercial broiler chicks (Ross strain) were used. The chicks were divided into three groups, and each group consisted of six replicates of 25 chicks each. The chicks were kept in floor pens, and water and feed were supplied ad libitum throughout the experiment. Three experimental diets were provided as follows: (i) corn–soybean‐based diet deficient in methionine; (ii) methionine‐deficient corn–soybean‐based diet supplemented with DLM to meet the methionine requirements of broiler chicks; and (iii) methionine‐deficient corn–soybean‐based diet supplemented with LMA (1.25‐fold (w/w) the amount of DLM supplied to the second group, given an assumption that 100 units of liquid LMA can be replaced by 80 units DLM to give similar performance results). During the starter period, the weight gains of chicks fed LMA or DLM were significantly greater than those in chicks receiving the methionine‐deficient diet (P < 0.05), and the addition of LMA significantly improved weight gain compared with the addition of DLM. Adding DLM or LMA significantly improved the feed conversion ratio (FCR) and percentage of uniformity (P < 0.05). No significant differences between the effects of DLM and LMA on these parameters were found. During the grower period (3–6 weeks of age), weight gain, FCR, uniformity and feed intake of chicks that received diet supplemented with DLM or LMA were superior to those of the methionine‐deficient group (P < 0.05). Chicks fed LMA had the same bodyweight gain and uniformity as those fed DLM. However, adding LMA resulted in a significant increase of FCR resulting from excess feed consumption. Outer breast meat yields were significantly improved and abdominal fat was significantly decreased when methionine sources were added (P < 0.05), and adding LMA tended to promote edible meat growth better than did adding DLM. Although no significant effects of methionine sources on the chemical composition of the liver were seen, adding methionine sources tended to increase liver fat content. In conclusion, it seems that the bioefficacy of LMA relative to DLM is not less than 80%. Therefore, chicks fed with diet supplemented with 1.25‐fold (w/w) as much LMA as DLM might exceed requirements for growth performance, while meeting requirements for meat production. Moreover, the relative bioefficacies of LMA and DLM between the starter and grower periods may perhaps be different.     

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.     

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.     

Effect of dietary methionine on growth performance and insulin‐like growth factor‐I mRNA expression of growing meat rabbits

An experiment was conducted to determine the effects of different amounts of dietary methionine on growth performance, serum protein, growth hormone (GH), insulin‐like growth factor‐I (IGF‐I) concentrations and IGF‐I mRNA expression of growing meat rabbits. One hundred weaned growing meat rabbits were allocated to individual cages and randomly divided into five groups. The methionine addition concentrations of the five groups were 0, 2, 4, 6 and 8 g/kg diet (as‐fed basis) and sulphur amino acids (SAA) concentrations ranging from 3.8 to 11.6 g/kg diet, respectively. The results obtained were as follows: the average daily gain of 2, 4 and 6 g/kg diet groups was higher than that of 0 g/kg diet group (p < 0.01). The feed gain ratio of the 4 g/kg diet group was lower than those of 0 and 8 g/kg diet group (p < 0.01). Methionine concentrations did not affect serum urea nitrogen, total protein, insulin and IGF‐I concentration (p > 0.05). The quadratic effects of methionine on the serum concentration of albumin (Alb) and GH were obtained (p = 0.013, p = 0.018). The quadratic effect of methionine amount on IGF‐I mRNA expression was obtained (p = 0.045). The serum concentration of Alb of the 4 g/kg diet group was higher than those of 0 and 8 g/kg diet group (p < 0.01). The serum concentration of GH of 8 g/kg diet group was higher than that of the 0 g/kg diet group (p < 0.05). The liver IGF‐I mRNA expression of 4 g/kg diet group was higher than those of the 0 and 8 g/kg diet group (p < 0.05). Providing a diet mainly consisted of corn, wheat bran and peanut vine, the optimum dietary methionine addition concentration and SAA concentration for a weaner to 2‐month‐old growing meat rabbits were shown to be 2 and 5.7 g/kg diet respectively.     

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.     

Determination of the methionine requirement of growing double-muscled Belgian blue bulls with a three-step method

The three-step technique was used to determine the requirements of total amino acids (TAA) and the first-limiting amino acid (AA) in growing double-muscled Belgian Blue bulls (BBb). In Exp. 1, three double-muscled BBb weighing initially 306 +/- 28 kg received a basal diet consisting of 30% meadow hay and 70% concentrate that was poor in digestible protein but had adequate NE because of continuous infusion of dextrose into the duodenum. The intestinal apparent digestibility of essential AA (EAA) was defined according to their duodenal and ileal flows. It averaged 72% but varied between 60% for Met and 79% for Arg. In Exp. 2, five double-muscled BBb (334 +/- 22 kg) received the same diet supplemented with duodenal infusions of dextrose and four doses of Na-caseinate (28, 56, 84, and 112% of intestinal digestible dietary AA) in a 4 x 4 Latin square design with one additional animal. Nitrogen retention for the basal diet alone and the four increasing supplements of Na-caseinate reached 49, 61, 70, 80, and 86 g/d, respectively. Nitrogen utilization improved from 34.3% without Na-caseinate supplementation to a maximum of 40.6%, with the third dose supplying 788 g/d of apparently digestible AA. Based on patterns of plasma concentrations, Met, Phe, and Arg were probably the limiting AA when animals optimized N utilization. In Exp. 3, six double-muscled BBb (315 +/- 25 kg) fed the basal diet received duodenal infusions of dextrose and AA, equivalent to the third dose in Exp. 2, except for digestible Met (9.3, 14.4, 18.4, 22.4, 26.4, and 30.4 g/d) in a 6 x 6 Latin square design. The Met requirement was close to 26.4 g/d on the basis of N retention.     

Effects of methionine on muscle protein synthesis and degradation pathways in broilers

This study investigated the hypothesis that supplementation of methionine (Met) to broiler diets increases muscle growth due to regulation of molecular pathways related to protein synthesis and degradation depending on the Met source. Day‐old male Cobb‐500 broilers (n = 240) were phase‐fed three different wheat–soya bean meal‐based basal diets during days 1–10, 11–21 and 22–35. Basal diets (Met‐ group, Met + Cys concentration 15% below NRC recommendations) were supplemented with 0.10% or 0.40% Met either as DL‐Met (DLM) or DL‐2‐hydroxy‐4‐(methylthio) butanoic acid (DL‐HMTBA) (equimolar comparison). Breast muscle weights were lower in the Met‐ group compared to all Met‐supplemented groups and were lower in broilers supplemented with 0.10% of DL‐HMTBA compared to the other groups fed Met‐supplemented diets. However, the expression of genes or relative phosphorylation and thus activation state of proteins involved in the somatotropic axis, the mammalian target of rapamycin (mTOR) pathway of protein synthesis, the ubiquitin–proteasome pathway (UPP) and autophagy–lysosomal pathway of protein degradation, the GCN2/eIF2a pathway involved in the inhibition of protein synthesis and in the myostatin–Smad2/3 pathway involved in myogenesis were not affected by Met source. Feeding diets with suboptimum Met + Cys concentrations, however, decreased expression of GHR and IGF1 in liver and muscle and increased that of MURF1 involved in the UPP in the broiler's muscle at day 10 and 21, while that of FOXO and atrogin‐1 and FOXO phosphorylation remained unaffected. Additionally, suboptimum dietary Met concentrations increased expression of the autophagy‐related genes ATG5 and BECN1 at day 35. Met supplementation neither affected gene expression nor phosphorylation of proteins involved in the GNC2/eIF2a and mTOR pathways. These data indicate that protein synthesis was not affected on the molecular level, while protein degradation was marginally affected by dietary Met dosage.     

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.     

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.     

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.