Effects of cottonseed oil and ferrous sulfate on the performance and expression of antioxidant enzymes in broilers.

Aiming to reduce feed costs, cottonseed oil (CSO) has been used as an alternative component in diets for broilers. However, this oil contains gossypol, an antinutritional agent that impacts the use of mineral elements, inhibits glucose uptake, and has a direct inhibitory action on intestinal enzymes. Nevertheless, toxic effects of gossypol can be prevented by the addition of iron salts, such as ferrous sulfate (FS), to the diet. This work was conducted to evaluate performance and gene expression of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) in the intestines of 21- and 42-day-old broilers fed 0, 2, 4, or 6% concentrations of CSO, with or without FS. All CSO diets led to weight gain (WG) at 21 D. At 42 D, an increase in WG and a decrease in feed conversion (FCR) in the diets containing FS were observed. In 21-day-old birds supplemented with 4% CSO and FS, an increase in GPx gene expression was observed when compared to the 6% level. Animals (42 day old) supplemented with 6% CSO and FS presented greater expression of SOD gene when compared to 2% CSO and FS. In addition, a higher GPx expression in broilers supplemented with 6% CSO and FS compared to 6% CSO without FS was achieved. In conclusion, including CSO in the diets of broiler favors WG in animals at 21 D of age, independent of the presence or absence of FS; and including 4% CSO and FS in the diet of these animals alters the expression of the GPx gene in the intestine, so it is not necessary to add FS at 21 D. On the other hand, in 42-day-old broilers, the addition of FS is indicated, due to increases WG, decreased FCR and at the 6% CSO level without FS increase in the expression of the SOD and GPx genes.

Dietary L-arginine supplementation increased mammary gland vascularity of lactating sows.

The present study aimed to evaluate the mechanisms modulated by dietary arginine supplementation to sows during lactation regarding antioxidant capacity and vascularization of mammary glands. At 109 days of gestation, animals were transferred to individual farrowing crates equipped with manual feeders and automatic drinker bowls. Environmental temperature and humidity inside the farrowing rooms were registered every 15 min. At farrowing, sows were assigned in a completely randomized design to a control diet (CON) or the CON diet supplemented with 1.0% L-arginine (ARG). A total of three gilts and two sows were fed the CON diet, whereas three gilts and three sows were fed ARG diets. Sows were fed a fixed amount of 6.0 kg/day, subdivided equally in four delivery times (0700, 1000, 1300 and 1600 h) for 21 days. At weaning, sows were slaughtered and mammary tissue samples and blood from the pudendal vein were collected. Data were analyzed considering each sow as an experimental unit. Differences were considered at P<0.05. L-arginine fed sows presented lower messenger RNA (mRNA) expression for prolactin receptor (P=0.002), angiopoietin1 (P=0.03) and receptor tyrosine kinase (P=0.01); higher mRNA expression for prostaglandin synthase 1 (P=0.01); a trend of decrease for glucocorticoid receptor (P=0.06) and IGF receptor 1 (P=0.07); and a trend (P=0.05) for an increased glutathione peroxidase mRNA expression. The angiopoietin2:angiopoietin1 mRNA ratio tended to increase (P=0.07) in ARG fed sows. L-arginine fed sows had greater (P=0.04) volumetric proportion of blood vessels and a trend of enhance (P=0.07) in the number of blood vessels per mm2. These findings show that 1.0% ARG supplementation to sows activates proliferative mechanisms, may improve mammary tissues' angiogenesis and tended to increase mRNA expression of genes that encode antioxidant enzymes in mammary gland of sows.

Acute heat stress and dietary methionine effects on IGF-I, GHR, and UCP mRNA expression in liver and muscle of quails.

This study evaluated the expression of insulin-like growth factor I (IGF-I), growth hormone receptor (GHR), and uncoupling protein (UCP) mRNA in muscle and liver of quails that were in thermal comfort or exposed to heat stress and that were fed diets with or without methionine supplementation. Meat quails were fed a diet that either met the nutritional demands for methionine (MS) or did not meet this demand (methionine-deficient diet, MD). The animals were either kept at a thermal comfort temperature (25°C) or exposed to heat stress (38°C for 24 h starting on the 6th day). RNA was extracted from liver and breast muscle, and cDNA was synthesized and amplified using quantitative reverse transcription-polymerase chain reaction. Animals that were fed the MS diet and remained at the thermal comfort temperature exhibited increased IGF-I mRNA expression in the liver (0.56 AU). The GHR mRNA expression in the liver and muscle was influenced by both the study variables. Animals receiving the MS diet showed higher GHR expression, while increased expression was observed in animals at the thermal comfort temperature. The UCP mRNA expression in the muscle was influenced by both methionine supplementation and heat stress. Higher expression was observed in animals that received the MD diet (2.29 vs 3.77 AU) and in animals kept in thermal comfort. Our results suggest that heat stress negatively affects the expression of growth-related genes and that methionine supplementation is necessary to appropriately maintain the levels of IGF-I, GHR, and UCP transcripts for animal metabolism.

Effect of heat stress and feeding phosphorus levels on pig electron transport chain gene expression.

The purpose of this study was to evaluate the effect of temperature and different levels of available phosphorus (aP) on the expression of nine genes encoding electron transport chain proteins in the Longissimus dorsi (LD) muscle of pigs. Two trials were carried out using 48 high-lean growth pigs from two different growth phases: from 15 to 30 kg (phase 1) and from 30 to 60 kg (phase 2). Pigs from growth phase 1 were fed with three different levels of dietary aP (0.107%, 0.321% or 0.535%) and submitted either to a thermoneutral (24°C and RH at 76%) or to a heat stress (34°C and RH at 70%) environment. Pigs from growth phase 2 were fed with three different levels of dietary aP (0.116%, 0.306% or 0.496%) and submitted either to a thermoneutral (22ºC and RH at 77%) or to a heat stress (32ºC and RH at 73%) environment. Heat stress decreased (P<0.001) average daily feed intake at both growth phases. At 24°C, pigs in phase 1 fed the 0.321% aP diet had greater average daily gain and feed conversion (P<0.05) than those fed the 0.107% or 0.535% while, at 34°C pigs fed the 0.535% aP had the best performance (P<0.05). Pigs from phase 2 fed the 0.306% aP had best performance in both thermal environments. Gene expression profile was analyzed by quantitative real-time polymerase chain reaction. Irrespective of growing phase, the expression of six genes was lower (P<0.05) at high temperature than at thermoneutrality. The lower expression of these genes under high temperatures evidences the effects of heat stress by decreasing oxidative metabolism, through adaptive physiological mechanisms in order to reduce heat production. In pigs from phase 1, six genes were differentially expressed across aP levels (P<0.05) in the thermoneutral and one gene in the heat stress. In pigs from phase 2, two genes were differentially expressed across aP levels (P<0.05) in both thermal environments. These data revealed strong evidence that phosphorus and thermal environments are key factors to regulate oxidative phosphorylation with direct implications on animal performance.

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.

The effect of glycerol on mRNA expression of growth hormone, insulin-like growth factor, and mitochondrial breast muscle genes of Japanese quail.

1. A study was carried out to evaluate the expression of growth hormone (GH), insulin-like growth factor I (IGF-I), mitochondrial adenine nucleotide translocase (ANT), cytochrome oxidase III (COX III) and avian uncoupling protein (avUCP) genes in 14-d-old Japanese quail that were fed different levels of glycerol (0%, 4%, and 12% dietary glycerol) which replaced maize as an energy source. 2. Total RNA was extracted from the breast muscle, and cDNA was amplified using real-time PCR with primers specific to the examined genes. 3. Quail fed the diet with 12% glycerol supplementation presented higher growth hormone (GH) mRNA expression than did those fed 0% glycerol. Supplementation with 12% glycerol negatively influenced IGF-I mRNA expression and reduced ANT mRNA expression in comparison with the treatment with no glycerol. COX III mRNA expression in the pectoralis superficialis muscle was decreased by 26% in quail fed 12% glycerol compared with those fed 0 and 4% glycerol. 4. There was no difference in UCP mRNA expression between quail fed 0 and 4% glycerol; however, UCP expression was reduced (73%) in birds fed the 12% glycerol diet compared with the level in those fed the 4% glycerol diet. 5. The inclusion of 4% glycerol in the diet produced results similar to those of the diet with no glycerol. Based on quail performance and the expression of the GH, IGF-I, ANT, COX III and UCP genes, 4% glycerol can be used in quail feeding without any harmful effects.

Phosphorus requirements for 60- to 100-kg pigs selected for high lean deposition under different thermal environments.

Two experiments were conducted to determine the requirement of available P (aP) of pigs selected for high lean deposition during summer (maximum and minimum temperatures of 29.8 ± 3.6 and 21.3 ± 1.1°C, respectively; Exp. 1) and winter (maximum and minimum temperatures of 24.7 ± 2.1 and 9.1 ± 1.6°C, respectively; Exp. 2). Pigs (66.5 ± 2.8 kg, Exp. 1; and 61.1 ± 0.6 kg, Exp. 2) were randomly allotted to 5 dietary treatments. Exp. 1 had 5 pens/treatment and Exp. 2 had 6 pens/treatment, and all pens had 2 pigs. Treatments were composed of a corn-soybean meal basal diet and 4 diets with additional dicalcium phosphate to obtain 5 aP concentrations (0.122, 0.187, 0.252, 0.317, and 0.382% in Exp. 1, and 0.135, 0.200, 0.265, 0.330, and 0.395% in Exp. 2). Pigs were allowed ad libitum access to their respective diets for 30 d. In Exp. 1, increasing aP improved ADFI and ADG (quadratic, P < 0.05), with maximum responses obtained at 0.226 and 0.256% of aP, respectively. Increasing aP improved (P < 0.05) G:F. A broken-line analysis indicated that G:F was maximized (P < 0.05) at 0.295% of aP or 9.04 g of aP/d. Increasing aP also increased (quadratic, P < 0.05) P and Ca (g/kg) in the metacarpals with maximum responses obtained at 0.314 and 0.272% of aP, respectively. The percentage of ash in the metacarpals continued to increase (linear, P < 0.05) with increasing aP. In Exp. 2, ADG and G:F improved (linear and quadratic, P < 0.05) as aP increased in the diet. A broken-line analysis indicated that G:F was maximized (P < 0.05) at 0.316% of aP or 10.58 g of aP per day. However, increasing aP had no effect on ADFI. Percentages of ash, P, and Ca in the metacarpals increased (linear, P < 0.05) with increasing aP. In conclusion, results of this experiment indicated that finishing pigs (60 to 100 kg) kept in a thermoneutral environment require 0.316% aP in the diet (10.58 g/d) to maximize G:F, but the requirement is only 0.295% aP (9.04 g/d) when pigs are under heat stress.