Effects of supplemental d-methionine in comparison to l-methionine on nitrogen retention,gut morphology,antioxidant status,and mRNA abundance of amino acid transporters in weanling pigs

An N-balance experiment was conducted to test the hypothesis that d-Methionine (d-Met) has the same bioavailability and efficacy as l-Methionine (l-Met) when fed to weanling pigs. A Met-deficient basal diet containing 0.24% standardized ileal digestible (SID) Met was formulated. Six additional diets were formulated by adding 0.036%, 0.072%, or 0.108% d-Met or l-Met to the basal diet, and these diets, therefore, contained 77%, 87%, or 97% of the requirement for SID Met. Fifty-six barrows (10.53 ± 1.17 kg) were housed in metabolism crates and allotted to the seven diets with eight replicate pigs per diet. Feces and urine were collected quantitatively with 7-d adaptation and 5-d collection periods. Blood and tissue samples from pigs fed the basal diet and pigs fed diets containing 0.108% supplemental Met were collected on the last day. Results indicated that N retention (%) linearly increased (P < 0.01) as supplemental d-Met or l-Met increased in diets. Based on N retention (%) as a response, the linear slope-ratio regression estimated the bioavailability of d-Met relative to l-Met to be 101% (95% confidence interval: 57%–146%). The villus height and crypt depth in the jejunum were not affected by the Met level or Met source. Total antioxidant capacity or thiobarbituric acid reactive substance concentrations in plasma or tissue samples from pigs fed the control diet or diets containing 0.108% supplemental d-Met or l-Met were not different. Abundance of mRNA for some AA transporters analyzed in intestinal mucosa of pigs also did not differ. Therefore, it is concluded that d-Met and l-Met are equally bioavailable for weanling pigs.     

Effects of methionine and guanidinoacetic acid supplementation on performance and energy metabolites in breast muscle of male broiler chickens fed corn-soybean diets

ABSTRACT

1. Guanidinoacetic acid (GAA) is the single endogenous precursor of creatine, which plays a critical role in energy homeostasis of cells. Since GAA is endogenously converted to creatine by methylation, it was hypothesised that the effects of dietary GAA supplementation might determine the methionine (Met) availability in corn-soybean based diets.

2. A total of 540, one-day-old male Ross 308 broilers were allocated to nine dietary treatments with six replicates (10 birds each) in a 3 × 3 factorial arrangement with three graded levels of supplementary Met (+0.4 g/kg per level), whilst cystine was equal across groups, resulting in a low, medium and high level of total sulphur amino acids, and with three levels of GAA (0, 0.6 and 1.2 g/kg). Birds were fed for 42 days.

3. Increasing levels of supplemental Met enhanced performance indices in all rearing periods, although there was no effect on feed conversion ratio in the grower or feed intake in the finisher periods. Final body weight was 8.8% and 14.6% higher in the birds fed medium and high Met diets, respectively, compared to the low Met level. Relative breast weight and protein content in muscle on d 25 linearly increased with higher levels of Met. At low and high Met levels, growth in the finisher phase was negatively affected by supplementing GAA at 1.2 g/kg. It was suggested that disturbances in methylation homeostasis and/or changes in Arg metabolism might explain these findings. At the end of the grower phase, muscle creatine content was higher when feeding GAA at 0.6 and 1.2 g/kg (4464 and 4472, respectively, vs. 4054 mg/kg fresh muscle in the control group).

4. The effects of dietary GAA supplementation were influenced by the dietary Met level only in the finisher period, which indicates the need for proper sulphur amino acid formulation in diets when feeding GAA.     

Bioavailability of the calcium salt of dl-methionine hydroxy analog compared with dl-methionine for nitrogen retention and the preference of nursery pigs for diets based on the 2 forms of methionine

Experiments were conducted to determine the relative bioavailability (RBV) of the calcium salt of the hydroxy analog of dl-methionine (MHA-Ca, 84%) to dl-methionine (dl-Met, 99%) as Met sources fed to pigs. In experiment 1, 42 crossbred barrows (initial BW of 15.0 ± 0.7 kg) were allotted to 7 treatments in an N-balance study. The basal diet (BD) was formulated to contain 15.4% CP and 0.22% Met (70% of requirement). Diets included (1) BD, (2) BD + 0.025% dl-Met, (3) BD + 0.050% dl-Met, (4) BD + 0.075% dl-Met, (5) BD + 0.038% MHA-Ca, (6) BD + 0.077% MHA-Ca, and (7) BD + 0.115% MHA-Ca. An increase in dietary inclusion rates of both Met sources linearly increased (P < 0.01) N retained (g/d) and N retention (% of intake). Using linear slope-ratio regression, the RBV value of MHA-Ca to dl-Met for N retained (g/d) was 63.0% on a product-to-product basis (75.0% on an equimolar basis). In experiment 2, 40 crossbred barrows (initial BW of 15.5 ± 1.5 kg) were allotted to 5 treatments in another N-balance study. The BD was formulated to contain 17.0% CP and 0.22% Met (70% of requirement). Diets included (1) BD, (2) BD + 0.030% dl-Met, (3) BD + 0.060% dl-Met, (4) BD + 0.046% MHA-Ca, and (5) BD + 0.092% MHA-Ca. Increasing levels of dl-Met or MHA-Ca increased N retained (g/d) and N retention (% of intake) linearly (P < 0.001) and quadratically (P < 0.05). Using linear slope-ratio regression, a product-to-product RBV value of MHA-Ca to dl-Met was 68.4% (81.4% on an equimolar basis) for N retained (g/d). In experiment 3, 276 pigs (12 barrow and 11 gilt replicates; initial BW of 7.09 ± 1.1 kg) were used in 3 diet preference studies. Pigs were randomly allotted to 1 of 3 treatment comparisons of feed choice: (1) BD (0.23% Met) or BD + 0.07% dl-Met; (2) BD or BD + 0.0825% MHA-Ca, and (3) BD + 0.07% dl-Met or BD + 0.0825% MHA-Ca. Pigs consumed a higher percentage (55 vs. 45%; P = 0.008) of their total feed intake from the diet supplemented with 0.07% dl-Met in Comparison 1, but a lower percentage (45 vs. 55%; P = 0.003) of their total feed intake from the diet supplemented with 0.0825% MHA-Ca in Comparison 2. There was no diet preference for dl-Met or MHA-Ca in Comparison 3. The observed Met source preference differences occurred in the barrow replicates but not in the gilt replicates. These results demonstrated the mean RBV of MHA-Ca to dl-Met of 65.7% on a product-to-product (wt/wt) basis or 78.2% on an equimolar basis and that a preference for Met sources was observed in barrows but not in gilts.     

Methionine as a methyl group donor in growing cattle

Holstein steers were used in two 5 x 5 Latin square experiments to evaluate the sparing of methionine by alternative sources of methyl groups (betaine and choline). Steers were housed in metabolism crates and limit-fed a soybean hull-based diet high in rumen degradable protein. To increase energy supply, ruminal infusions of volatile fatty acids and abomasal infusions of glucose were provided. An amino acid mixture, limiting in methionine, was infused abomasally to ensure that nonsulfur amino acids did not limit protein synthesis. Treatments for Exp. 1 were abomasal infusion of 1) water, 2) 2 g/d L-methionine, 3) 1.7 g/d L-cysteine, 4) 1.6 g/d betaine, and 5) 1.7 g/d L-cysteine + 1.6 g/d betaine. Treatments for Exp. 2 were abomasal infusion of 1) water, 2) 2 g/d L-methionine, 3) 8 g/d betaine, 4) 16 g/d betaine, and 5) 8 g/d choline. In both experiments, nitrogen retention increased in response to methionine (P < 0.05), demonstrating a deficiency of sulfur amino acids. Responses to cysteine, betaine, and choline were all small and not significant. The lack of response to cysteine indicates that the response to methionine was not due to transsulfuration to cysteine or that cysteine supply did not alter the flux of methionine through transsulfuration. The lack of response to betaine suggests that the steers' needs for methyl groups were met by the dietary conditions or that betaine was relatively inefficient in increasing the remethylation of homocysteine to methionine and, thereby, reducing the synthesis of cysteine from homocysteine. Under our experimental conditions, responses to methionine were likely due to a correction of a deficiency of methionine per se rather than of methyl group donors.     

Effect of arginine supplementation on the morphology and function of intestinal epithelia and serum concentrations of amino acids in pigs exposed to heat stress

The exposure of pigs to heat stress (HS) appears to damage their intestinal epithelia, affecting the absorption of amino acids (AAs). Arg is involved in the restoration of intestinal epithelial cells but HS reduces Arg intake. The effect of dietary supplementation with Arg on the morphology of intestinal epithelia, AA transporter gene expression, and serum concentration (SC) of free AAs in HS pigs was analyzed. Twenty pigs (25.3 ± 2.4 kg body weight) were randomly assigned to two dietary treatments: Control (0.81% Arg), wheat–soybean meal diet supplemented with l-Lys, l-Thr, dl-Met, and l-Trp, and the experimental diet where 0.16% free l-Arg was supplemented to a similar Control diet (+Arg). All pigs were individually housed and exposed to HS, fed ad libitum with full access to water. The ambient temperature (AT), recorded at 15-min intervals during the 21-d trial, ranged on average from 29.6 to 39.4 °C within the same day. Blood samples were collected on day 18 at 1600 hours (AT peak); serum was separated by centrifugation. At the end of the trial, five pigs per treatment were sacrificed to collect samples of mucosa scratched from each small intestine segment. The expression of AA transporters in intestinal mucosa and the SC of AAs were analyzed. Villi height (VH) was higher (P < 0.01) in the duodenum, jejunum, and ileum but the crypt depth did not differ between the Control and the +Arg pigs. Supplementation of l-Arg increased the mRNA coding for the synthesis of the cationic AA transporter b^0,+ (P < 0.01) and the neutral AA transporter B⁰ (P < 0.05) in the duodenum by approximately 5-fold and 3-fold, respectively, but no effect on mRNA abundance was observed in the jejunum and ileum. The supplementation of l-Arg increased serum Arg, His, Met, Thr, Trp, and urea (P < 0.05) and also tended to increase Val (P < 0.10) but did not affect Ile, Lys, Leu, and Phe. These results indicate that supplementing 0.16% l-Arg to the Control diet may help to improve the function of the small intestine epithelium, by increasing the VH, the abundance of AA transporters, and the SC of most indispensable AAs in pigs exposed to HS conditions. However, the lack of effect of supplemental Arg on both Lys SC and weight gain of pigs suggests that increasing the Lys content in the +Arg diet might be needed to improve the performance of HS pigs.     

Bioavailability of the dl-methionine and the calcium salt of dl-methionine hydroxy analog compared with l-methionine for nitrogen retention in starter pigs

Two nitrogen balance studies were conducted to evaluate the relative bioavailability values (RBV) of dl-methionine (dl-Met) and dl-methionine hydroxy analog calcium salt (MHA-Ca) to l-methionine (l-Met) as Met sources fed to pigs. In experiment 1, 42 pigs were assigned to 7 treatments feeding with basal diet (BD) formulated to be deficient in Met (0.22% standardized ileal digestible basis) but adequate in other amino acids. Diets included (1) BD, (2) BD + 0.025% dl-Met, (3) BD + 0.050% dl-Met, (4) BD + 0.075% dl-Met, (5) BD + 0.025% l-Met, (6) BD + 0.050% l-Met, and (7) BD + 0.075% l-Met. Increasing levels of l-Met and dl-Met enhanced N retained (g/d) and N retention (% of intake) linearly (P < 0.01). Using a linear slope ratio procedure, a product-to-product RBV of dl-Met compared with l-Met was 94% (95% confidence limits: 65% to 123%) based on N retained expressed as g/d and 99% (95% confidence limits: 70% to 128%) for N retention expressed as % of intake. In experiment 2, 42 pigs were allotted to 7 treatments in another N-balance trial. Diets included (1) BD, (2) BD + 0.025% l-Met, (3) BD + 0.050% l-Met, (4) BD + 0.075% l-Met, (5) BD + 0.030% MHA-Ca, (6) BD + 0.060% MHA-Ca, and (7) BD + 0.089% MHA-Ca. An increase in dietary inclusion rates of l-Met increased (P < 0.01) N retained (g/d) linearly while increasing levels of MHA-Ca had no effects (P > 0.05) on N retained (g/d) and N retention (% of intake). Using linear slope-ratio regression, the RBV of MHA-Ca compared with l-Met was 70% (95% confidence limits: 59% to 81%) on a product-to-product basis or 83% on equimolar basis based on N retained expressed as g/d. Overall, the mean RBV of dl-Met to l-Met of 97% (95% confidence limits cover 100%) indicated that dl-Met and l-Met are equally bioavailable as Met sources in pigs. Compared with l-Met, the RBV of MHA-Ca was lower at 70% (95% confidence limits: 59% to 81%) on a product-to-product basis or 83% on equimolar basis in starter pigs.     

Effects of energy source on methionine utilization by growing steers

We evaluated the effects of different supplemental energy sources on Met use in growing steers. Ruminally cannulated Holstein steers were used in two 6 x 6 Latin squares, and data were pooled for analyses. In Exp. 1, steers (148 kg) were fed 2.3 kg of DM/d of a diet based on soybean hulls. Treatments (2 x 3 factorial) were abomasal infusion of 0 or 3 g of l-Met/d, and supplementation with no energy or with glucose (360 g/d) or fat (150 g/d) continuously infused into the abomasum. In Exp. 2, steers (190 kg) received 2.6 kg of dietary DM/d and were provided (2 x 3 factorial) with 0 or 3 g of l-Met/d, and with no supplemental energy or with acetate (385 g/d) or propionate (270 g/ d) continuously infused into the rumen. In both experiments, the energy sources supplied 1.3 Mcal of GE/d, and all steers received basal infusions of 400 g of acetate/d into the rumen and a mixture (125 g/d) of all essential AA except Met into the abomasum. Nitrogen balance (18.8 vs. 23.5 g/d; P < 0.01) and whole-body protein synthesis (2.1 vs. 2.3 kg/d; P < 0.07) were increased by Met supplementation, indicating that protein deposition was limited by Met. Supplemental energy reduced (P < 0.01) urinary N excretion and increased (P < 0.01) N retention without differences among energy sources. Increases in N retention in response to Met were numerically greater when energy was supplemented. Efficiency of supplemental Met use was 11% when no energy was supplemented but averaged 21% when 1.3 Mcal of GE/d was provided. Whole-body protein synthesis and degradation were not affected by energy supplementation. Serum insulin concentrations were increased by glucose and propionate supplementation. Serum IGF-I concentrations were increased by supplementation with Met or glucogenic sources of energy. In growing steers, N retention was increased by energy supplementation even though protein deposition was limited by Met, suggesting that energy supplementation improves the efficiency of AA use. These responses were independent of the source of energy.     

Effect of gestation dietary methionine-to-lysine ratio on methionine metabolism and antioxidant ability of high-prolific sows

The uptake and metabolism of methionine (Met) are critical for epigenetic regulation, redox homeostasis, and embryo development. Our previous study showed that appropriate supplementation of dietary Met promoted the birth weight and placental angiogenesis of high-prolific sows. To further explore the metabolic effect of Met on pregnant sows, we have evaluated the influence of dietary Met level on Met metabolism, and the relationship between metabolites of Met and reproductive performance, antioxidant ability, and placental angiogenesis throughout the gestation of high-prolific sows. Sixty sows (the 3rd parity, Large White) were randomly divided into 5 groups that were fed diets with standardized ileal digestible (SID) methionine-to-lysine (Met:Lys) ratios of 0.27 (control), 0.32, 0.37, 0.42, and 0.47 from the mating day (gestational d 0, G0d) until the farrowing day. HPLC-MS/MS analysis was used to simultaneously evaluate the metabolites related to Met, e.g. S-adenosylmethionine (SAM), S-adenosylhomocysteine (SAH), homocysteine (Hcy), cysteine (Cys), and glutathione (GSH). The concentration of SAM and SAH in plasma had significant fluctuations, especially in late pregnancy. Increasing dietary Met supplementation significantly improved the plasma SAM and methylation potential (SAM-to-SAH ratio) at d 114 of pregnancy (G114d). Moreover, a positive association of the plasma SAM concentration at G114d was observed with the litter weight of born alive (P < 0.05; R² = 0.58). Furthermore, Hcy concentration in plasma was at the lowest level for 0.37 ratio group at G114d. However, it significantly increased during late pregnancy. Moreover, there were negative correlations between plasma Hcy concentration at G114d (P < 0.05) and the placental vascular density in the fold and stroma (P < 0.05). Compared with the control group, the expression of vascular endothelial growth factor-A (VEGF-A) in the placenta tissue of 0.37 ratio group increased significantly (P < 0.05). Collectively, these findings indicate that dietary Met:Lys ratio (0.37 to 0.57) in the pregnant diet dose not influence the antioxidant ability of the high-prolific sows; however, the improvement of fetal development and placental angiogenesis of high-prolific sows by supplementation of Met are closely associated to the key Met-related metabolite of SAM and Hcy, respectively.     

Metabolize methionine and lysine requirements of growing cattle

Two growth studies were conducted to determine the Met and Lys requirements of growing cattle. In each 84-d trial, steer calves were fed individually diets containing 44% sorghum silage, 44% corn cobs, and 12% supplement (DM basis) at an equal percentage of BW. In Trial 1, 95 crossbred steers (251 kg) were supplemented with urea or meat and bone meal (MBM). Incremental amounts of rumen-protected Met were added to MBM to provide 0, .45, .9, 1.35, 3, and 6 g/d metabolizable Met. In Trial 2, 60 steers (210 kg) were supplemented with urea or corn gluten meal (CGM). Incremental amounts of rumen-protected Lys were added to CGM to provide 0, 1, 2, 3, 4, 5, 6, 8, and 10 g/d metabolizable Lys. Supplementation with MBM and CGM increased the supply of metabolizable protein to the animal. Steers fed MBM plus 0 Met gained 49 g/d more than steers fed urea, whereas steers fed CGM plus 0 Lys gained 150 g/d more than steers fed urea. Supplementation of rumen-protected Met and Lys improved ADG in steers fed MBM and CGM, respectively (P < .10). Nonlinear analysis, comparing gain vs supplemental Met and Lys intake, predicted supplemental Met and Lys requirements of 2.9 and .9 g/d, respectively. This amount of additional Met promoted .13 kg/ d gain greater than MBM alone, and this amount of additional Lys promoted .10 kg/d gain greater than the CGM alone. Metabolizable Met and Lys requirements were predicted from Level 1 of NRC (1996) calculated metabolizable protein supply, amino acid analysis of abomasal contents, and the maximum response to supplemental AA. Steers gaining .39 kg/d required 11.6 g/ d Met or 3. 1% of the metabolizable protein requirement, whereas steers gaining .56 kg/d required 22.5 g/d Lys or 5.7% of the metabolizable protein requirement.     

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 ammonia load on methionine utilization by growing steers

Seven ruminally cannulated Holstein steers (194 +/- 16 kg) housed in metabolism crates were used in a 6 x 6 Latin square, with one additional steer, to study effects of ruminal ammonia load on methionine (Met) use. All steers received a diet based on soybean hulls (2.6 kg DM/d), ruminal infusions of 200 g/d of acetate, 200 g/d of propionate, and 50 g/d of butyrate, as well as abomasal infusion of 300 g/d of glucose to provide energy without increasing microbial protein supply, and abomasal infusions of a mixture (248 g/d) of all essential AA except Met. Treatments were arranged as a 3 x 2 factorial and included urea (0, 40, or 80 g/d) infused ruminally to supply metabolic ammonia loads and Met (2 or 5 g/d) infused abomasally. Supplementation with the greater amount of Met decreased (P < 0.05) urinary N excretion from 68.8 to 64.8 g/d and increased (P < 0.05) retained N from 22.0 to 27.5 g/d. Urea infusions linearly increased (P < 0.05) urinary N excretions, plasma urea concentrations, and urinary urea excretions, but retained N was not affected. The efficiency of deposition of supplemental Met, calculated by assuming that Met deposition is 2.0% of protein deposition (6.25 x retained N), ranged between 18 and 27% when steers received 0 or 80 g/d of urea, respectively. There were no (P > or = 0.40) effects of treatments on serum insulin or IGF-I concentrations. In our model, increasing ammonia load did not affect whole-body protein deposition in growing steers when Met was limiting.     

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.     

Performance and physiological responses of broiler chickens to supplemental guanidinoacetic acid in arginine-deficient diets

1. The present experiment was designed to examine arginine (Arg)-sparing effects of guanidinoacetic acid (GAA) on production performance, intestinal morphology and certain blood parameters in broiler chickens.

2. A total of 300 male broiler chicks (Ross 308) were randomly allotted to 5 dietary treatments in a completely randomised design during 1–15 and 15–35 d rearing periods. Experimental treatments consisted of a basal diet with a reduction in Arg content (CON?; starter: 12.1 g/kg and grower: 11.3 g/kg Arg), a CON? diet supplemented with synthetic Arg equal to the required Arg level (CON+; starter: 13.80 g/kg; grower: 12.37 g/kg) and CON? diet supplemented with three levels of GAA (GAA0.6; GAA1.2; and GAA1.8; g/kg).

3. Supplemental GAA at 0.6 and 1.2 g/kg reduced the adverse effects of the CON? diet on weight gain and feed conversion ratio in the starter period, equal to the effect of CON+ diet. During the entire rearing period, such an effect was only seen for broilers fed CON? diet supplemented with 1.2 and 1.8 g/kg GAA (P < 0.05). Proportional weights of liver and abdominal fat were decreased in birds fed diets containing supplemental Arg and 1.8 g/kg GAA compared to the CON? broilers (P < 0.05). Jejunal villus height and villus height to crypt depth ratio were increased in broilers receiving the CON? diet (P < 0.05). Serum concentration of nitric oxide increased when the CON? diet was supplemented with either Arg or 1.8 g/kg GAA (P < 0.05).

4. Dietary inclusion of GAA had an Arg-sparing effect, whereby 1.2 and 1.8 g/kg of supplemental GAA resulted in greater growth performance during the starter and entire rearing periods, respectively. Supplementation with 1.8 g/kg GAA had significant effects on some carcass and physiological parameters.     

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.     

Effects of energy level on methionine utilization by growing steers

We evaluated the effect of energy supplementation on Met use in growing steers. Six ruminally cannulated Holstein steers (228 +/- 8 kg of BW) were used in a 6 x 6 Latin square and fed 2.8 kg of DM/d of a diet based on soybean hulls. Treatments were abomasal infusion of 2 amounts of Met (0 or 3 g/d) and supplementation with 3 amounts of energy (0, 1.3, or 2.6 Mcal of GE/d) in a 2 x 3 factorial arrangement. The 1.3 Mcal/d treatment was supplied through ruminal infusion of 90 g/d of acetate, 90 g/d of propionate, and 30 g/d of butyrate, and abomasal infusion of 30 g/d of glucose and 30 g/d of fat. The 2.6 Mcal/d treatment supplied twice these amounts. All steers received basal infusions of 400 g/d of acetate into the rumen and a mixture (125 g/d) containing all essential AA except Met into the abomasum. No interactions between Met and energy levels were observed. Nitrogen balance was increased (P < 0.05) by Met supplementation from 23.6 to 27.8 g/d, indicating that protein deposition was limited by Met. Nitrogen retention increased linearly (P < 0.05) from 23.6 to 27.7 g/d with increased energy supply. Increased energy supply also linearly reduced (P < 0.05) urinary N excretion from 44.6 to 39.7 g/d and reduced plasma urea concentrations from 2.8 to 2.1 mM. Total tract apparent OM and NDF digestibilities were reduced linearly (P < 0.05) by energy supplementation, from 78.2 and 78.7% to 74.3 and 74.5%, respectively. Whole-body protein synthesis and degradation were not affected significantly by energy supplementation. Energy supplementation linearly increased (P < 0.05) serum IGF-I from 694 to 818 ng/mL and quadratically increased (P < 0.05) serum insulin (0.38, 0.47, and 0.42 ng/mL for 0, 1.3, and 2.6 Mcal/d, respectively). In growing steers, N retention was improved by energy supplementation, even when Met limited protein deposition, suggesting that energy supplementation affects the efficiency of AA use.     

爱拔益加肉仔鸡对胍基乙酸的耐受性评价

本试验旨在研究饲粮添加胍基乙酸(GAA)对爱拔益加(AA)肉仔鸡生长性能、血液学指标、脏器指数、组织同型半胱氨酸含量以及组织形态的影响,以系统评价AA肉仔鸡对GAA的耐受性。试验选用540只1日龄AA肉仔鸡公雏,随机分为5个组,每组6个重复,每个重复18只鸡。5组试鸡分别饲喂在基础饲粮中添加0、800、1 600、4 000和8 000 mg/kg GAA的饲粮。试验期42 d,分为前期(1~21日龄)和后期(22~42日龄)2个阶段。结果表明:与对照组相比,饲粮添加800~4 000 mg/kg GAA显著提高了肉仔鸡前期、后期和全期平均日增重(P0.05),显著降低其后期和全期料重比(P0.05),但对各期平均日采食量均无显著影响(P0.05)。饲粮添加8 000 mg/kg GAA组的平均日增重、平均日采食量和料重比与对照组相比差异不显著(P0.05)。饲粮添加800~8 000 mg/kg GAA对21、42日龄肉仔鸡血常规指标、血清生化指标、脏器指数和组织同型半胱氨酸含量均无显著影响(P0.05)。饲粮添加4 000、8 000 mg/kg GAA对肉仔鸡肝脏和肾脏组织形态无不良影响。由此可见,肉仔鸡饲粮中添加GAA对其生长性能、血液学指标、脏器指数、组织同型半胱氨酸含量和组织形态无不良影响,肉仔鸡可耐受8 000 mg/kg GAA。     

胍基乙酸对肉鸡的适用阶段及饲粮蛋氨酸配套强化效果研究

本研究基于生长性能和肉品质,评价胍基乙酸(GAA)对肉鸡的适用阶段和饲粮蛋氨酸(Met)配套强化效果。选取1日龄雄性爱拔益加肉仔鸡1920只,随机分为16组,每组设6个重复,每个重复20只鸡。试验分前期(1~21日龄)和后期(22~42日龄)2个饲养阶段,1组为对照组,饲喂常规饲粮(Met水平前期为0.50%,后期为0.40%)。2~6组仅在前期设置Met或GAA处理,其中,2组只把饲粮Met的水平提高至0.65%;3~6组采用双因素完全随机设计,2个饲粮Met水平分别为0.50%和0.65%,2个GAA添加量分别为300和600 mg/kg。7~11组仅在后期设置Met或GAA处理,其中,7组只把饲粮Met水平提高至0.55%;8~11组采用双因素完全随机设计,2个饲粮Met水平分别为0.40%和0.55%,2个GAA添加量分别为400和800 mg/kg。12~16组在全期(1~42日龄)设置Met或GAA处理,前期和后期饲粮设置分别同2~6组和7~11组。试验期42 d。结果发现:1)与对照组相比,只在前期饲喂高剂量GAA并强化Met供应能够显著提高1~21日龄平均日增重和21日龄体重(P<0.05),显著降低1~21日龄料重比(P<0.05);仅在后期饲喂高剂量GAA能够显著提高22~42日龄平均日增重,显著降低22~42日龄料重比(P<0.05),同时强化Met供应还可显著提高1~42日龄平均日增重及42日龄体重、胸肌指数(P<0.05),显著降低22~42日龄和1~42日龄料重比及42日龄腹脂指数(P<0.05);全期饲喂高剂量GAA能够提高22~42日龄和1~42日龄平均日增重及42日龄体重和胸肌指数(P<0.05),显著降低22~42日龄和1~42日龄料重比及42日龄腹脂指数(P<0.05),同时强化Met供应还可显著提高1~21日龄平均日增重和21日龄体重(P<0.05),显著降低1~21日龄、22~42日龄和1~42日龄料重比(P<0.05)。2)与对照组相比,全期单独强化饲粮Met或联合添加低剂量GAA可以显著降低42日龄胸肌、腿肌亮度(L^∗)值和剪切力(P<0.05),提高红度(a^∗)值和24 h pH(pHu)(P<0.05);单独饲喂GAA对肉品质无显著影响(P>0.05)。由此可见,当全程饲喂高剂量GAA时,无需额外补充Met就可使肉鸡终末体重和胴体质量达到最佳;若要阶段性使用GAA,宜在后期高剂量添加且同时补充Met。     

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.     

Effects of Intestinal Modification by Antibiotics and Antibacterials on Utilization of Methionine Sources by Broiler Chickens

A study was conducted to determine if the response to different sources of Met was influenced by the presence or absence of antibiotics and antibacterials that might alter intestinal microflora. A Met-deficient diet (0.33% by analysis), based on corn and soybean meal, was fed with or without a mixture providing 200 g/ton of bacitracin methylene disalicylate, 200 g/ton of chlortetracycline, 100 g/ton of penicillin, and 100 g/ton of sulfaquinoxaline. Diets were fortified with DL-Met or the liquid form of 2-hydroxy-4-methylbutanoic acid (HMB) to provide supplemental levels of Met ranging from 0.0 to 0.20% in increments of 0.04%, based on 99% activity for DLDL-Met and 88% activity for HMB. The experimental treatments consisted of a 2 × 2 × 6 factorial arrangement of treatments with 2 diet types (medicated and unmedicated), 2 Met sources (DL-Met and HMB) at 6 levels of supplementation for a total of 24 dietary treatments. Each of these was fed to 6 replicate pens of 5 male chicks from 0 to 21 d, stratified across tiers in the battery.Feeding the medicated diets resulted in a significant reduction in intestinal bacterial as measured by total aerobic plate count of ileal contents, with significant improvements in BW and feed conversion. However, there were no significant interactions between the medications and the response to the 2 sources of Met. There was no difference in BW or feed conversion related to the 2 different sources of Met when fed to provide equimolar amounts of Met activity. Chicks responded to increasing levels of Met, but there was no interaction between source and level of Met for BW or feed conversion. Peak response appeared to occur at approximately 0.08% supplemental (0.41% total) Met, somewhat below current NRC recommendations. It does not appear that a difference in antibiotic content of test diets is responsible for the discrepancy in reported responses to the 2 sources of Met.     

Combined effects of guanidinoacetic acid,coenzyme Q10 and taurine on growth performance,gene expression and ascites mortality in broiler chickens

High levels of guanidinoacetate acid (GAA) deteriorate growth response in broiler chickens. We propose using coenzyme Q₁₀, an antioxidant, and taurine (TAU), a methyl donor, to cope with the situation when high level of GAA included in diet. GAA was supplemented at 0 (control), 0.75, 1.5 and 2.25 g/kg in isoenergetic and isonitrogenous diets and fed to broilers (Cobb 500) from 1 to 40 days post‐hatch. Three additional diets were prepared by adding CoQ₁₀ (40 mg/kg), TAU (40 mg/kg) or their combination (both CoQ₁₀ and TAU at 40 mg/kg) to the 2.25 g/kg GAA group. The experimental design used was a completely randomized design. While weight gain (p = 0.038) and feed conversion ratio (p = 0.024) improved when GAA added at 1.5 g/kg, higher supplementation (2.25 g/kg) deteriorated these responses. These responses, however, were significantly restored by using CoQ₁₀, TAU or their combination. Abdominal fat deposition was significantly decreased when TAU added to broiler diets by virtue of upregulating peroxisome proliferator‐activated receptor alpha. Supplementing broiler diets with CoQ₁₀ and TAU or their combination significantly decrease ascites mortality. In conclusion, CoQ₁₀ and TAU have shown beneficial effects when high level of GAA included in broiler diets.