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.     

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 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.     

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.     

Effect of gut stress induced by oxidized wheat gluten on the growth performance,gut morphology and oxidative states of broilers

This study was to investigate the effect of oxidized wheat gluten (OG) on growth performance, gut morphology and its oxidative states of broilers. One hundred and eighty‐day‐old male broilers (10 chicks/pen) were randomly allocated into three dietary treatments: control diet (CON), diet with 8% wheat gluten (WG) and diet with 8% OG with six pens/treatment. Body weight (BW) (21 and 35 days) and average daily gain (ADG) (1–21 days and 22–35 days) decreased (p < .05) and feed conversion ratio (FCR) (1–21 days and 22–35 days) increased (p < .05) in OG treatment. Feed intake (FI) decreased (p < .05) in WG and OG treatments during 22–35 days. However, FI was not influenced by dietary treatments during 1–21 days (p > .05). The OG‐fed broilers had a lower faecal pH value (p < .05) and higher faecal moisture content (p < 05) at 14, 21, 28 and 35 days. Villus height, crypt depth and V/C value were not different (p > .05) among treatments at 21 and 35 days. Lipid peroxidation (LPO) (21 and 35 days) and malondialdehyde (MDA) (35 days) content in crop of OG treatment increased (p < .05). Oxidized glutathione (GSSG) (21 days), LPO (21 and 35 days) and MDA (21 and 35 days) content in ileum of OG treatment increased (p < .05). The reduced glutathione/oxidized glutathione (GSH/GSSG) (21 days) and (GSH) (35 days) in ileum of OG treatment decreased (p < .05). The present findings indicate that OG might be a stressor for broiler gut, which could induce oxidative stress both in crop and in ileum, and the diarrhoea as well. The growth performance of broiler was consequently depressed.     

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.     

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.     

Aspartame supplementation in starter accelerates small intestinal epithelial cell cycle and stimulates secretion of glucagon‐like peptide‐2 in pre‐weaned lambs

The objective of this study was to test the hypothesis that aspartame supplementation in starter diet accelerates small intestinal cell cycle by stimulating secretion and expression of glucagon‐like peptide ?2 (GLP‐2) in pre‐weaned lambs using animal and cell culture experiments. In vivo, twelve 14‐day‐old lambs were selected and allocated randomly to two groups; one was treated with plain starter diet (Con, n = 6) and the other was treated with starter supplemented with 200 mg of aspartame/kg starter (APM, n = 6). Results showed that the lambs received APM treatment for 35 d had higher (p < .05) GLP‐2 concentration in the plasma and greater jejunum weight/live body weight (BW) and jejunal crypt depth. Furthermore, APM treatment significantly upregulated (p < .05) the mRNA expression of cyclin D1 in duodenum; and cyclin A2, cyclin D1, cyclin‐dependent kinases 6 (CDK6) in jejunum; and cyclin A2, cyclin D1, CDK4 in ileum. Moreover, APM treatment increased (p < .05) the mRNA expression of glucagon (GCG), insulin‐like growth factor 1 (IGF‐1) in the jejunum and ileum and mRNA expression of GLP‐2 receptor (GLP‐2R) in the jejunum. In vitro, when jejunal cells were treated with GLP‐2 for 2 hr, the 3‐(4,5‐dimethyl‐2‐thiazolyl)‐2,5‐diphenyl‐2‐H‐tetrazolium bromide (MTT) OD, IGF‐1 concentration, and the mRNA expression of IGF‐1, cyclin D1 and CDK6 were increased (p < .05). Furthermore, IGF‐1 receptor (IGF‐1R) inhibitor decreased (p < .05) the mRNA expression of IGF‐1, cyclin A2, cyclin D1 and CDK6 in GLP‐2 treatment jejunal cells. These results suggest that aspartame supplementation in starter accelerates small intestinal cell cycle that may, in part, be related to stimulate secretion and expression of GLP‐2 in pre‐weaning lambs. Furthermore, GLP‐2 can indirectly promote the proliferation of jejunal cells mainly through the IGF‐1 pathway. These findings provide new insights into nutritional interventions that promote the development of small intestines in young ruminants.     

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.     

Dietary levels of protein and free amino acids affect pancreatic proteases activities,amino acids transporters expression and serum amino acid concentrations in starter pigs

The dietary contents of crude protein and free amino acids (AA) may affect the protein digestion and AA absorption in pigs. Trypsin and chymotrypsin activities, AA serum concentrations and expression of AA transporters in the small intestine of pigs fed a low protein, AA‐supplemented (19.2%, LPAA) or a high protein (28.1%, HP), wheat‐soybean meal diet were measured in two 14‐d trials. The LPAA diet contained free L‐Lys, L‐Thr, DL‐Met, L‐Leu, L‐Ile, L‐Val, L‐His, L‐Trp and L‐Phe. All pigs were fed the same amount of feed (890 and 800 g/d for trial 1 and 2 respectively). In trial 1, samples of mucosa (duodenum, jejunum and ileum) and digesta (duodenum and jejunum) were collected from 14 pigs (17.2 ± 0.4 kg); in trial 2, blood samples were collected from 12 pigs (12.7 ± 0.3 kg). The trypsin and chymotrypsin activities in both intestinal segments were higher in pigs fed the HP diet (p < 0.01). Trypsin activity was higher in jejunum than in duodenum regardless the dietary treatment (p < 0.05). Pigs fed the LPAA diet expressed more b^0,+AT in duodenum, B⁰AT1 in ileum (p < 0.05), and tended to express more y⁺LAT1 in duodenum (p = 0.10). In pigs fed the LPAA diet, the expression of b^0,+AT was higher in duodenum than in jejunum and ileum (p < 0.01), but no difference was observed in pigs fed the HP diet. Ileum had the lowest b^0,+AT expression regardless the diet. The serum concentrations of Lys, Thr and Met were higher in LPAA pigs while serum Arg was higher in HP pigs (p < 0.05). Serum concentrations of AA appear to reflect the AA absorption. In conclusion, these data indicate that the dietary protein contents affect the extent of protein digestion and that supplemental free AA may influence the intestinal site of AA release and absorption, which may impact their availability for growth of young pigs.     

A deficient or an excess of dietary threonine level affects intestinal mucosal integrity and barrier function in broiler chickens

The aim of this study was to investigate the effects of deficient or excess of dietary threonine (Thr) levels on intestinal integrity and barrier function of broilers. A total of 432 1‐day‐old commercial broilers (Arbor Acre) were assigned to four experiment groups consisting of six replicates of 18 birds. The treatments were designed as follows: 85%, 100%, 125% and 150% of NRC (Nutrient requirements of poultry (9th edn). Washington, DC: The National Academies Press, 1994) recommendations. The results indicated that expressions of jejunal and ileal secretory immunoglobulin A (sIgA) mRNA were increased linearly or quadratically by increasing Thr (p < .05), and the highest sIgA mRNA abundance was obtained in 125% Thr level. Likewise, the intestinal sIgA content showed similar increasing trend with the intestinal sIgA gene expression in this instance. The high level of Thr inclusion upregulated mucin 2 (MUC2) mRNA expression in the jejunum and ileum (p < .05). In addition, on day 21, the expression levels of jejunal zonula occludens‐2 (ZO‐2) and ileal zonula occludens‐1 (ZO‐1) decreased then increased with increasing Thr level (p < .05), whereas, the mRNA expressions of occludin in the jejunum and ileum had no significant difference amongst groups (p >.05). On day 42, Thr treatments did not affect the mRNA abundance of measured genes in the jejunum and ileum (p > .05). These findings suggested that Thr might be a nutrient immunomodulator that affects intestinal barrier function, moreover, 125% of the NRC (1994) recommendations Thr level was optimum.     

The effects of dietary supplementation with porous zinc oxide on growth performance,intestinal microbiota,morphology, and permeability in weaned piglets

The objective of this experiment was to evaluate the effects of dietary supplementation with porous zinc oxide (HiZox) on growth performance, intestinal microbiota, morphology, and permeability in weaned piglets. A total of 128 weaned piglets [(Landrace × Yorkshire) × Duroc] with an average body weight (BW) of (6.55 ± 0.25 kg; 21 d of age) were randomly assigned to four dietary treatments: (1) a corn‐soybean basal diet; (2) basal diet + 3,000 mg/kg conventional ZnO; (3) basal diet + 200 mg/kg HiZox; (4) basal diet + 500 mg/kg HiZox. The experiments lasted for 28 days. Incremental HiZox in the diet increased ADG (linear p = 0.015; quadratic p = 0.043) and ADFI (linear p = 0.027; quadratic p = 0.038), and the diarrhea index decreased linearly and quadratically (p < 0.01) as HiZox supplemented increased. Furthermore, supplementation with HiZox increased the amounts of Lactobacillus spp. (p < 0.05) in the ileum and cecum in comparison with that of control treatment or 3,000 mg/kg ZnO treatment, while decreased the populations of Escherichia coli, Clostridium coccoides, and Clostridium. leptum subgroup (p < 0.05) in the ileum and cecum relative to those in control treatment. The addition of HiZox increased the villus height and villus‐to‐crypt ratio (VC) of duodenum, jejunum, and ileum (p < 0.05), while decreased the crypt depth of jejunum (p < 0.05) and tended to reduce the crypt depth of duodenum (p < 0.10) compared with the control treatment. Piglets fed with 500 mg/kg HiZox had lower serum D‐lactate and diamine oxidase (DAO) than those fed with basal control diet or 3,000 mg/kg ZnO diet (p < 0.01). The results suggested that supplementation with HiZox modulated intestinal microbial composition and improved intestinal morphology, which may exert protective effects on the integrity of the mucosal barrier function of weaned piglets, was as efficacious as pharmaceutical doses of ZnO in enhancing growth performance, indicating that the HiZox may be a promising alternative to pharmaceutical doses of ZnO.     

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%.     

Indices of oxidative stress in blood and pulmonary epithelium lining fluid in horses suffering from recurrent airway obstruction.

To test the hypothesis that reactive oxygen species could be associated to the lower airway disorders occurring in horses suffering from recurrent airway obstruction (RAO), indices of oxidative stress were studied in blood and pulmonary epithelium lining fluid in 5 RAO horses either in clinical remission or 24 h after the onset of a crisis of bronchospasm and in 5 healthy horses. Venous blood and bronchoalveolar lavage fluid (BALF) samples were collected and analysed for reduced glutathione (GSH), oxidised glutathione (GSSG), total glutathione (TGSH), glutathione redox ratio (GRR) in blood haemolysate and pulmonary epithelium lining fluid (PELF). The haemolysate concentrations of GSH, GSSG, TGSH and GRR were similar in the 3 groups. The PELF glutathione status was significantly different in the RAO horses in acute crisis compared to healthy horses, indicating the occurrence of an oxidative stress. When RAO horses were in crisis their GSH and TGSH remained unchanged but their GSSG and GRR were significantly increased compared to the remission. These results support the hypothesis that oxidative stress is associated with lower airway disorders occurring in horses suffering from RAO.     

Bovine colostrum enriched with lyophilized bovine colostrum stimulates intestinal epithelium renewal of Holstein calves in the first days of life

Consumption of a second meal of colostrum with high quality could contribute to the intestinal epithelium development, especially if there is poor supply of colostrum just after birth. The effect of a second colostrum meal was evaluated on histomorphometry of the intestinal mucosa of newborn Holstein calves fed with high‐ and low‐quality first colostrum. Seventy‐two calves were fed with a first colostrum meal with high (HFM, close to 100 mg/ml) or low (LFM, close to 30 mg/ml) IgG concentration. At 12 hr of life, three treatments of second colostrum feeding were applied to the calves either fed high or low first colostrum: calves fed with low (LOW—close to 30 mg/ml) or high (HIGH—close to 100 mg/ml) IgG concentration; and colostrum enriched with lyophilized bovine colostrum with high IgG concentration (ENRICHED—higher than 120 mg/ml), resulting in six groups. Intestinal samples were collected after 24 and 72 hr of life. In the distal jejunum and ileum, LOW showed higher villus height than ENRICHED (p < .05). In the distal jejunum, greater villus perimeter was observed in the LOW compared to ENRICHED at 24 hr (p < .05). In ileum, LFM showed higher villus perimeter compared to HFM (p < .05). LOW showed the highest villus height‐to‐crypt depth ratio in the medium and distal jejunum and ileum, p < .05. ENRICHED and HFM showed decreased muscle layer thickness in the proximal and distal jejunum respectively (p < .05). The results reveal that the high concentration of total solids, crude protein, IgG and IGF‐I of colostrum with high quality worsened the absorptive area, but may have stimulated the activity of cell division in intestinal crypts. Considering the present results, bovine colostrum enriched with lyophilized bovine colostrum stimulates intestinal epithelium renewal of Holstein calves in the first days of life.     

Effects of lycopene supplementation in both maternal and offspring diets on growth performance,antioxidant capacity and biochemical parameters in chicks

This study investigated the effects of different supplementation ways of lycopene during pre‐hatch (from the diet of hens) and post‐hatch (from the diet of progeny) on production performance, antioxidant capacity and biochemical parameters in chicks. In total, 360 hens were fed diets supplemented with 0 (control group) or 40 mg lycopene/kg diet. From 28 to 34 days after the start of supplementation (30 weeks old), 650 qualified eggs were collected to artificial incubation. In this trial, 2 × 2 factorial designs were used. Male chicks hatched from hens fed with 0 or 40 mg lycopene/kg diet were fed a diet containing either 0 or 40 mg lycopene/kg diet. The results showed that, relative to control, in ovo‐deposited lycopene significantly increased chick birth body weight, improved liver total antioxidant capacity (T‐AOC), glutathione peroxidase (GSH‐Px) and glutathione to oxidized glutathione ratio (GSH: GSSG), and significantly declined liver malondialdehyde (MDA) level and increased liver lycopene content during 0–14 days after hatching. On days 14 after hatching, dietary lycopene in diet began to take over gradually. Both supplementation ways of lycopene increased immune organ index, serum high‐density lipoprotein (HDL) cholesterol, villus length and villus/crypt in duodenum, jejunum and ileum. Data in this study suggested lycopene supplementation could improve antioxidant capacity and immune function, and regulate lipid metabolism in chicks.     

Evidence that oxidative stress is higher in replacement gilts than in multiparous sows

The recent success obtained in term of increasing the litter size of sows has not correlated with a reduction of replacement rate. There is thus an increased economic demand for gilts with optimal reproductive potential and longevity. Unfortunately, replacement gilts are known to be more susceptible to diseases and less productive than multiparous sows. Interestingly, reproductive performance, resistance to diseases and longevity could all be largely affected by oxidative stress. To investigate whether oxidative stress conditions could account for the poor longevity of gilts, three distinct groups of conventional Yorkshire × Landrace sows were formed based on their similar age and parity (gilts, second parity sows as well as fourth to fifth parity sows). All animals were slaughtered during the post‐ovulatory period, and blood as well as tissue samples were collected. Biomarkers of oxidative damage to proteins (carbonyls) and DNA (8‐OHdG) were analysed in samples. Specific mRNA expression of major antioxidants such as glutathione peroxidases 1, 3 and 4 (GPx1, GPx3, GPx4) as well as superoxide dismutases 1 and 2 (Sod1, Sod2) were monitored in liver and kidney samples by quantitative RT‐PCR. Specific enzymatic activities of both GPx and SOD were measured by spectrophotometric assays. The plasma concentration of protein carbonyls was significantly different between the three groups with the highest concentration being observed in gilts (p ≤ 0.001). The mRNA expression levels of GPx1 and GPx4 were also significantly increased in the liver of gilts when compared to multiparous sows (p ≤ 0.05). SOD2 enzymatic activity was found to be higher in the liver of gilts than multiparous sows (p ≤ 0.05). Taken together, these results indicate that replacement gilts sustain significantly higher oxidative conditions than multiparous sows. Current findings may contribute to the design of nutritional regimens that will increase the productivity of gilts by counteracting oxidative stress.     

Transport stress induces pig jejunum tissue oxidative damage and results in autophagy/mitophagy activation

Pig transportation is associated with intestinal oxidative stress and results in destruction of intestinal integrity. Autophagy has been contributed to maintain cell homeostasis under stresses. The purpose of this study was to evaluate the effects of transport stress on morphology, intestinal mucosal barrier and autophagy/mitophagy levels in pig jejunum. A total of 16 finishing pigs were randomly divided into two groups. The control group was directly transported to the slaughterhouse and rested for 24 hr. The experimental groups were transported for 5 hr and slaughtered immediately. The results showed that transportation induced obvious stress responses with morphological and histological damage in jejunum accompanying with an elevated level of malondialdehyde (MDA; p < .05), endotoxin (LPS; p < .05), lactic dehydrogenase (LDH; p < .05) and a decreased level of serum superoxide dismutase (SOD; p < .05). Also, hemeoxy genase 1 (HO‐1; p < .01) as well as tight junction protein (claudin‐1 [p < .001], occludin [p < .05] and zonula occludens 1 [ZO‐1; p < 0.05]) levels were attenuated in jejunum tissue, and NADPH oxidase 1 (NOX1; p < .01) mRNA expression was up‐regulated. Further research indicated that transport stress could induce autophagy through increasing microtubule‐associated protein light chain 3 (LC3; p < .05) and autophagy‐related gene 5 (ATG5; p < .01) levels and suppressing p62 expression. Additionally, transport stress increased the protein levels of PTEN‐induced putative kinase 1 (PINK1; p < .05) and Parkin (p < .05) which was associated with mitophagy. In conclusions, transport stress could induce the destruction of intestinal integrity and involve in the intestinal mucosal barrier oxidative damage, and also contribute to activation of autophagy/mitophagy.     

Supplemental lipoic acid relieves post‐weaning diarrhoea by decreasing intestinal permeability in rats

Lipoic acid (LA) is a naturally existing substance which widely distributed in the cellular membranes and cytosol of animal cells. Its intracellular functions include quenching of free radicals and repairing oxidized proteins. The purpose of this study was to evaluate the effects of LA on post‐weaning diarrhoea using a rat model. Sixty weaned rats were fed either a basal diet or a LA‐supplemented diet, or a zinc oxide (ZnO)‐supplemented diet as a positive control. Rats in the LA and ZnO groups had better performance and reduced incidence of diarrhoea (p < 0.05). Both LA and ZnO treatments enhanced intestinal homeostatic and architecture, significantly decreased urinary lactulose to mannitol ratios (p < 0.05) and increased the expression of the intestinal mucosal tight junction proteins occludin (OCLN) and zonula occludens protein‐1 (ZO‐1) (p < 0.05). LA significantly increased the activities of antioxidant enzymes, and reduced glutathione while decreasing the levels of oxidative glutathione and malondialdehyde in the intestinal mucosa (p < 0.05). Furthermore, an in vitro study indicated that supplementation with LA in IEC‐6 intestinal epithelial cells significantly enhanced the expression of OCLN and ZO‐1 under hydrogen peroxide‐induced oxidative stress. Collectively, these results suggest that LA relieves post‐weaning diarrhoea by reducing intestinal permeability and improving antioxidant indices.     

Effects of oxidative stress induced by high dosage of dietary iron ingested on intestinal damage and caecal microbiota in Chinese Yellow broilers

The objective of this trial was to test the effects of oxidative stress induced by a high dosage of dietary iron on intestinal lesion and the microbiological compositions in caecum in Chinese Yellow broilers. A total of 450 1‐day‐old male chicks were randomly allotted into three groups. Supplemental iron (0, 700 and 1,400 mg/kg) was added to the basal diet resulting in three treatments containing 245, 908 and 1,651 mg/kg Fe (measured value) in diet respectively. Each treatment consisted of six replicate pens with 25 birds per pen. Jejunal enterocyte ultrastructure was observed by transmission electron microscopy. The results showed that a high dosage of dietary iron induced oxidative stress in broilers. Dilated endoplasmic reticulum (ER), autophagosome formation of jejunal enterocytes and decreased villi were caused by this oxidative stress. Compared to the control, concentration of the malondialdehyde (MDA) in jejunal mucosa in the 908 and 1,651 mg/kg Fe groups increased by 180% (p < .01) and 155% respectively (p < .01); activity of copper‐zinc superoxide dismutase (Cu/ZnSOD) increased in jejunum (p < .01); and the concentration of plasma reduced glutathione (GSH) decreased by 34.9% (p < .01) in birds fed 1,651 mg/kg Fe. Gene expression of nuclear factor, erythroid‐derived 2‐like 2 (Nrf2) and zonula occludens‐1 (ZO‐1), in the higher dietary Fe groups was enhanced (p < .05). Species of microbial flora in caecum increased caused by oxidative stress. The PCR‐DGGE (denaturing gradient gel electrophoresis) dendrograms revealed different microbiota (65% similarity coefficient) between the control and iron‐supplemented groups (p < .05). These data suggest high dosage of iron supplement in feed diet can induce oxidative stress in Chinese Yellow broilers, and composition of microbiota in the caecum changed. It implied there should be no addition of excess iron when formulating diets in Chinese Yellow broilers.