Age-related oxidative stress and antioxidant capacity in heat-stressed broilers.

We aimed to evaluate the effects of acute heat stress (HS) and age on the redox state in broilers aged 21 and 42 days. We evaluated the expression of genes related to antioxidant capacity, the production of hydrogen peroxide (H2O2), and the activity of antioxidant enzymes in the liver, as well as oxidative stress markers in the liver and plasma. The experiment had a completely randomized factorial design with two thermal environments (thermoneutral and HS, 38°C for 24 h) and two ages (21 and 42 days). Twenty-one-day-old animals exposed to HS showed the highest thioredoxin reductase 1 (TrxR1) (P<0.0001) and glutathione synthetase (GSS) (P<0.0001) gene expression levels. Age influenced the expression of the thioredoxin (Trx) (P=0.0090), superoxide dismutase (SOD) (P=0.0194), glutathione reductase (GSR) (P<0.0001) and glutathione peroxidase 7 (GPx7) (P<0.0001) genes; we observed greater expression in birds at 21 days than at 42 days. Forty-two-day-old HS birds showed the highest H2O2 production (222.31 pmol dichlorofluorescein produced/min×mg mitochondrial protein). We also verified the effects of age and environment on the liver content of Glutathione (GSH) (P<0.0001 and P=0.0039, respectively) and catalase (CAT) enzyme activity (P=0.0007 and P=0.0004, respectively). Higher GSH content and lower CAT activity were observed in animals from the thermoneutral environment compared with the HS environment and in animals at 21 days compared with 42 days. Broilers at 42 days of age had higher plasma creatinine content (0.05 v. 0.01 mg/dl) and higher aspartate aminotransferase activity (546.50 v. 230.67 U/l) than chickens at 21 days of age. Our results suggest that under HS conditions, in which there is higher H2O2 production, 21-day-old broilers have greater antioxidant capacity than 42-day-old animals.

Effects of selenium supplementation on the oxidative state of acute heat stress-exposed quails.

This study aimed to evaluate the effect of heat stress (HS) and selenium supplementation on markers of stress, meat quality and gene expression. For this, meat quails of 42 days of age were fed a diet that either met [0.33 mg/kg, nutritional demand for selenium (SS)] or did not meet [0.11 mg/kg, selenium deficient (SD)] the nutritional demands for selenium during the 7 days of evaluation. In addition, the animals were kept at either a thermal comfort temperature (25 °C) or exposed to HS (38 °C for 24 h). Glutathione synthetase (GSS), glutathione reductase (GSR) and uncoupling protein (UCP) gene expression were influenced by the interaction between temperature and diet. Animals subjected to HS and fed the SS diet exhibited the highest GSS and GSR gene expression. In terms of UCP gene expression, the lowest values were observed in HS animals on the SD diet. Glutathione peroxidase 7 (GPX7) gene expression, body temperature (BT) and creatine kinase (CK) activity were influenced by both selenium supplementation and HS. Aspartate aminotransferase (AST), alanine aminotransferase (ALT) activity and creatinine content all were influenced by the diet/environment interaction. The highest AST activity, ALT activity and creatinine levels were observed in animals that were both on the SD diet and exposed to HS. HS animals also exhibited an increased heterophil/lymphocyte ratio and lower triiodothyronine (T3) hormone levels than birds that remained at the comfortable temperature. Animals subjected to HS and fed with selenium supplemented diet showed better results regarding gene expression and, thus, better results for the activities of enzymes used as stress markers, which could be due to the higher antioxidant capacity provided by the action of the studied genes.

Mitochondrial gene expression in the liver and muscle of high and low feed efficiency Japanese quail layers subjected to different environmental temperatures.

We evaluated the adenine nucleotide translocator (ANT) and cytochrome oxidase subunit III (COX III) mRNA expressions in the muscle and liver of Japanese quails presenting high and low feed efficiency (FE), and subjected them to three different environmental temperatures: comfort, heat stress (38°C), and cold stress (10°C). ANT mRNA expression was lower in the liver of heat-stressed animals. In the muscle, higher ANT and COX III mRNA expressions were observed in high-FE and cold-stressed animals. In the liver, much higher expression of COX III mRNA was observed in cold-stressed animals. These results suggest a possible correlation between the genes involved in energy production by the mitochondria and FE phenotypes, and that environmental temperature can affect the ANT and COX III mRNA expressions. Japanese quails presenting different FE levels respond differently to environmental stimuli.

Effects of methionine supplementation on the redox state of acute heat stress-exposed quails.

The aims of the present study were to evaluate the possible effects of heat stress (HS) on H2O2 production and to evaluate whether methionine supplementation (MS) could mitigate the deleterious effects on cell metabolism and the redox state induced by oxidative stress. Meat quails (Coturnix coturnix coturnix) were fed a diet that either met the nutritional demands for methionine or did not meet this demand (methionine deficient [MD] diet) for 7 d. The animals were either kept at a thermal comfort temperature (25°C) or exposed to HS (38°C for 24 h, starting on the sixth day). Heat stress induced decreased food intake (P = 0.0140), decreased daily weight gain (P < 0.0001), and increased water intake (P = 0.0211). A higher rate of H2O2 production was observed in HS animals (0.0802 vs. 0.0692 nmol of reactive oxygen species [ROS] produced per minute per milligram of protein; P = 0.0042) and in animals fed with the MD diet (0.0808 vs. 0.0686 nmol of ROS produced per minute per milligram of protein; P = 0.0020). We observed effects of the interaction between diet and the environment on the activities of glutathione peroxidase (GP-x) and catalase (P = 0.0392 and P < 0.0001, respectively). Heat stress induced higher levels of GP-x activity in animals on the MS diet and higher catalase activity in animals on the MD diet. Glutathione (GSH) levels were higher in animals on the MS diet (P = 0.0273) and in animals that were kept in thermal comfort (P = 0.0018). The thiobarbituric acid reactive substances level was higher in HS animals fed with the MD diet (P = 0.0386). Significant effects of the interaction between supplementation and environment were observed on uric acid concentration levels, which were higher in HS animals fed the MS diet (P = 0.008), and on creatine kinase activity levels, which were lower in HS animals fed the MD diet (1,620.33 units/L; P = 0.0442). Our results suggest that under HS conditions, in which H2O2 production is increased, MS was able to mitigate ROS-induced damage, possibly by increasing the activities of antioxidant elements such as GSH, GPx activity, and uric acid concentration, which were present in higher levels in animals that were subjected to HS and fed the MS diet.

Valine needs in starting and growing Cobb (500) broilers.

Two independent experiments were conducted with male Cobb × Cobb 500 broilers to determine the optimal valine-to-digestible-lysine ratio for broiler development. We conducted a randomized block experiment with 7 treatments, each with 8 replicates of 25 starter birds (8 to 21 d of age) and 20 finisher (30 to 43 d of age) birds. To prevent any excess of digestible lysine, 93% of the recommended level of digestible lysine was used to evaluate the valine-to-lysine ratio. The utilized levels of dietary digestible lysine were 10.7 and 9.40 g/kg for the starting and growing phases, respectively. A control diet with 100% of the recommended level of lysine and an adequate valine-to-lysine ratio was also used. The feed intake, weight gain, feed conversion ratio, and carcass parameters were evaluated. The treatments had no significant effect on the feed intakes or carcass parameters in the starter and finisher phases. However, during both of the studied phases, we observed a quadratic effect on weight gain and the feed conversion ratio. The broilers of both phases that were fed test diets with the lower valine-to-lysine (Val/Lys) ratio had poorer performance compared with those broilers fed control diets. However, when higher Val/Lys ratios were used for the starting and growing broilers that were fed test diets, the 2 groups had similar performance. During the starting phase, in broilers that were fed a higher Val/Lys ratio, weight gain, and the feed conversion ratio improved by 5.5% compared with broilers fed the basal diets. The broilers in the growing phase also had improved performance (by 7 to 8%) when the test diets had higher Val/Lys ratios. Based on the analysis of the starter phase data, we concluded that the optimal digestible Val/Lys ratio for Cobb × Cobb 500 broilers is 77%, whereas for birds in the finisher phase (30 to 43 d of age), a digestible Val/Lys ratio of 76% is suggested.

Dietary methionine effects on IGF-I and GHR mRNA expression in broilers.

This study aimed to evaluate liver and breast muscle insulin-like growth factor I (IGF-I) and growth hormone receptor (GHR) gene expression between broilers fed different methionine levels and sources. Broiler chicks were 22 to 42 days old, distributed in 5 treatments (control diet, DL1 - 0.08% DL-methionine, DL2 - 0.24% DL-methionine, MHA-FA1 - 0.11% methionine hydroxy analogue-free acid, and MHA-FA2 - 0.33% methionine hydroxy analogue-free acid). The broilers were euthanized by cervical dislocation. RNA was extracted from liver and breast muscle, followed by cDNA synthesis and amplification using qRT-PCR. DL2 methionine supplementation provided best animal performance results. GHR and IGF-I gene expression in the muscle tissue was not affected by methionine supplementation. IGF-I gene expression in the liver was higher in animals fed methionine supplementation than in animals fed control diet. IGF-I mRNA levels in broilers fed DL2 were greater than DL1 (1.56 vs 0.97 AU) and greater than MAH-FA1 and MAH-FA2. Broilers fed DL2 increased significantly GHR gene expression in the liver than animals fed the control diet. Addition of methionine improved animal performance by stimulating synthesis and release of growth factor.

Expression of growth genes in response to glycerol use in Japanese quail diets.

Glycerol can be used as a substitute for corn for feeding poultry, but there are concerns about how it may affect growth performance and health of the birds. We evaluated the expression of mRNA of growth hormone (GH) and insulin-like growth factor I (IGF-I) in 35-day-old Japanese quails fed different glycerol levels (0, 4, and 8% dietary glycerol instead of corn). Total RNA was extracted from the breast muscle and cDNA was amplified with the use of specific primers for these genes using real-time PCR. Quails fed the diet with 8% glycerol supplementation had significantly lower GH mRNA and IGF-I mRNA expression than those fed no glycerol or 4% glycerol. No significant effect of the treatments was found on quail weight gain or feed intake. Feed conversion ratio was influenced by dietary glycerol levels: the group fed 8% glycerol displayed the worst feed conversion ratio (2.54) compared with that of quail fed the control diet (2.35) or 4% glycerol (2.36). Considering quail performance and the expression of the genes GH and IGF-I, a level of 4% glycerol can be used in quail feeding without any harmful effects.

Age-related changes in mitochondrial UCP, ANT and COX III gene expression in the breast muscle of quails (Coturnix coturnix japonica).

Efficient conversion of food into body mass has been associated with altered gene expression of some proteins of the electron transport chain. We evaluated the effect of age on mRNA expression of uncoupling protein, adenine nucleotide translocase and cytochrome c oxidase complex III in quails. One-day-old broiler quails were sacrificed after 7, 14, 21, or 28 days. Six quails of each age were slaughtered by cervical dislocation. Total RNA was extracted from the breast muscle and reverse transcribed into cDNA. Real-time PCR analysis was performed using specific primers for the genes. As the quails aged, there was reduced expression of all three genes. The greatest reduction when comparing 7- and 28-day-old birds was observed in the uncoupling protein mRNA levels (67%), followed by adenine nucleotide translocase (56%) and cytochrome c oxidase complex III (30%). We conclude that expression of some mitochondrial genes of the electron transport chain changes with age in quails.