Pyridoxine regulates hair follicle development via the PI3K/Akt,Wnt and Notch signalling pathways in rex rabbits

This study was conducted to evaluate the effect of pyridoxine on the development of hair follicles in Rex rabbits and the underlying molecular mechanism. Two hundred 3-month-old Rex rabbits were randomly divided into 5 groups and fed diets supplemented with 0, 5, 10, 20, or 40 mg/kg pyridoxine. The hair follicle density on the dorsal skin and the gene and protein expression levels of components of the phosphoinositide 3-kinase (PI3K)/protein kinase B (PKB or Akt), Wnt, Notch and bone morphogenetic protein (BMP) signalling pathways were measured. In addition, free hair follicles were isolated from Rex rabbits and cultured with pyridoxine in vitro to measure hair shaft growth. Furthermore, dermal papilla cells (DPC) were isolated from the skin of Rex rabbits and cultured with pyridoxine in vitro to measure the gene and protein expression levels of components of the PI3K/Akt, Wnt, Notch and BMP signalling pathways. The results showed that the addition of dietary pyridoxine significantly increased the total follicle density, secondary follicle density, and secondary-to-primary ratio (S/P, P < 0.05), that the growth ratio of hair stems was promoted by pyridoxine in basic culture medium, and that the growth length of tentacle hair follicles cultured in the pyridoxine group was longer than that in the control group (P < 0.05). In addition, pyridoxine changed the DPC cycle progression and promoted cell proliferation, and appropriate concentrations of pyridoxine (10 and 20 μmol/L) significantly inhibited cell apoptosis (P < 0.05). Pyridoxine significantly affected the gene expression of components of the PI3K/Akt, Wnt and Notch signalling pathways in the skin and DPC of Rex rabbits (P < 0.05), increased the levels of phosphorylated catenin beta 1 (CTNNB1) and Akt, and decreased the level of phosphorylated glycogen synthase kinase 3 beta (GSK-3β) (P < 0.05). Therefore, the molecular mechanism by which pyridoxine promotes hair follicle density in Rex rabbits probably occurs through activation of the PI3K/Akt, Wnt and Notch signalling pathways, prolonging hair follicle growth and delaying the onset of telogen.     

Comparative assessment of growth performance of three different indigenous goat breeds exposed to summer heat stress

A study was conducted to assess comparatively the growth performance of three different indigenous goat breeds during exposure to summer heat stress. The primary objective of the study was to observe the heat stress impact on the growth performance based on the body weight changes, allometric measurements, growth hormone (GH) concentration and peripheral blood mononuclear cell (PBMC) Insulin‐like growth factor‐1 (IGF‐1) mRNA expression pattern during the summer season in comparison with the local breed (Osmanabadi). Thirty‐six ten‐month‐ to one‐year‐old female goats of Osmanabadi, Malabari and Salem Black breeds were randomly divided into six groups, OC (n = 6; Osmanabadi control), OHS (n = 6; Osmanabadi heat stress), MC (n = 6; Malabari control), MHS (n = 6; Malabari heat stress), SBC (n = 6; Salem Black control) and SBHS (n = 6; Salem Black heat stress). Body weight was recorded at weekly intervals, whereas other growth and allometric measurements and blood collection were carried out at fortnightly intervals. Breed factor significantly (p < .05) influenced only few growth variables such as body weight, body mass index (BMI) and body condition score (BCS). However, heat stress treatment significantly (p < .05) reduced all growth parameters expect BMI. Further, the heat stress significantly (p < .01) increased plasma GH concentration in goats with significantly higher (p < .05) concentration recorded in OHS. Among the stress groups, the lower (p < .05) PBMC IGF‐1 mRNA expression was recorded in OHS, while the higher (p < .05) expression was observed in SBHS indicating the extreme adaptive capability of Salem Black breed. Thus, the results indicated that the Salem Black breed performed much better compared to both Osmanabadi and Malabari breeds indicating the superior ability of this breed to adapt to heat stress challenges. The results also indicated that plasma GH and IGF‐1 gene may act as ideal biomarkers for assessing the heat stress impact on growth performance in indigenous goats.     

Effects of dietary supplementation with betaine on muscle growth,muscle amino acid contents and meat quality in Cherry Valley ducks

The effects of dietary betaine supplementation on growth performance, carcass characteristics, muscle amino acid contents, meat quality, antioxidant capacity, myogenic gene expression and mechanistic target of rapamycin (mTOR) signalling pathway in Cherry Valley ducks were evaluated. A total of 720 1‐day‐old Cherry Valley ducks were randomly distributed into four groups with six replicates of 30 birds for a 42‐day feeding trial. Ducks were fed a basal diet supplemented with 0 (control), 250, 500 or 1,000 mg/kg betaine, respectively. Growth performance was not affected by betaine. Incremental levels of betaine linearly (p < 0.05) increased the breast muscle yield and linearly (p < 0.05) decreased the subcutaneous fat thickness and the abdominal fat yield. The contents of methionine, serine, glycine, glutamate and total non‐essential amino acid in breast muscle were linearly (p < 0.05) increased by betaine supplementation. With increasing betaine levels, the drip loss and the content of malondialdehyde (MDA) were linearly (p < 0.05) decreased, and the redness of meat (linear p < 0.05), the activities of catalase (CAT) (linear p < 0.05) and total superoxide dismutase (T‐SOD) (linear p < 0.05, quadratic p < 0.05) were increased. Moreover, the myogenic differentiation factor 1 (MyoD1) mRNA expression and the mTOR mRNA expression and protein phosporylation were linearly (p < 0.05) up‐regulated, and the myostatin (MSTN) mRNA expression was linearly (p < 0.05) down‐regulated by betaine supplementation. Overall, this study indicated that betaine supplementation did not affect the growth performance of Cherry Valley ducks, but could linearly increase some amino acid contents in breast muscle, especially glycine, and increase muscle antioxidant activity to improve meat quality. Moreover, betaine supplementation could improve the breast muscle yield by increasing MyoD1 mRNA expression, decreasing MSTN mRNA expression and regulating mTOR signalling pathway.     

Absence of Sperm Factors as in the Parthenogenesis Does Not Interfere on Bovine Embryo Sensitiveness to Heat Shock at Pre‐Implantation Stage

Oocyte has been considered the major contributor for embryo thermo‐tolerance. However, it was shown that sperm factors can be transferred to the oocyte during fertilization, raising the question of whether the absence of such factors could interfere on embryo thermo‐tolerance. In this study, we used parthenogenesis to generate bovine embryos without spermatozoa in order to test whether the absence of sperm factors could influence their thermo‐sensitiveness at early stages. In vitro fertilized (IVF) and parthenogenetic (PA) embryos at 44 h post‐insemination/chemical activation were exposed to 38.5°C (control) or 41°C (heat shock) for 12 h and then developed for 48 h and up to blastocyst stage. Apoptosis index and expression of PRDX1, GLUT1, GLUT5 and IGF1r genes in blastocysts derived from heat‐shocked embryos were also evaluated. The heat shock decreased the blastocyst rate at day seven (p < 0.05) for IVF embryos and at day eight (p < 0.01) for both IVF and PA embryos. Total cell number was not affected by heat shock in IVF and PA blastocysts, but there was an increased proportion (p < 0.05) of apoptotic cells in heat‐shocked embryos when compared to controls. There was no interaction (p > 0.05) between method of activation (IVF and PA) and temperature (38.5°C or 41.5°C) for all developmental parameters evaluated. Expression of GLUT1 gene was downregulated (p < 0.05) by heat shock in both IVF and PA blastocyst whereas expression of GLUT5 and IGF1r genes was downregulated (p < 0.05) by heat shock in PA blastocysts. Those data show that the heat shock affects negatively the embryo development towards blastocysts stage, increases the apoptotic index and disturbed the expression of some genes in both IVF and PA embryos, indicating that the presence or absence of sperm factors does not influence the sensitivity of the bovine embryo to heat shock.     

Methionine promotes the development of hair follicles via the Wnt/β-catenin signalling pathway in Rex rabbits

To investigate the effect and molecular mechanism of methionine (Met) on the growth of hair follicles (HFs) in Rex rabbits. A total of 200 weaning Rex rabbits were divided into four groups and fed varying levels of Met-supplemented diets. We measured the HF density on dorsal skin and the Wnt/β-catenin pathway protein expression level. Meanwhile, whole HFs were isolated from Rex rabbit skins and cultured with Met in vitro to measure hair shaft growth. The relationship between Met and the Wnt/β-catenin signalling pathway was also characterized by using the Wnt/β-catenin signalling inhibitor, XAV-939. The results showed that the addition of dietary Met could significantly increase the HF density on dorsal skin (p < .05) and enhance the protein expression level of Wnt10b (p < .05), β-catenin (p < .05) and DSH (p < .05). Methionine stimulation could also prolong the hair shafts growth in vitro (p < .05). And inhibition of Wnt/β-catenin signalling using XAV-939 could eliminate this phenomenon. In summary, Met can increase the density of HFs on dorsal skin in vitro and prolong the hair shaft growth of HFs in vivo via the Wnt/β-catenin signalling pathway.     

Ruminal epithelial insulin‐like growth factor‐binding proteins 2, 3, and 6 are associated with epithelial cell proliferation

The aim of this study was to identify factors that regulate ruminal epithelial insulin‐like growth factor‐binding protein (IGFBP) expression and determine its role in rumen epithelial cell proliferation. Primary bovine rumen epithelial cells (BREC) were incubated with short‐chain fatty acids (SCFAs) at pH 7.4 or 5.6, lactate, lipopolysaccharide (LPS), insulin‐like growth factor‐I (IGF‐I), ‐II (IGF‐II), or recombinant bovine IGFBP2 (rbIGFBP2). The mRNA expression levels of IGFBP in BREC were analyzed using quantitative real‐time polymerase chain reaction (qRT‐PCR). The proliferation rate of BREC was analyzed using a WST‐1 assay. IGFBP2 gene expression tended to be lower with SCFA treatment (p < .1), and IGFBP6 gene expression was significantly lower with SCFA treatment (p < .05). IGFBP3 and IGFBP6 gene expression tended to be higher with d ‐Lactate treatment (p < .1). IGFBP3 gene expression was significantly higher (p < .05) with LPS treatment. BREC treated with IGF‐I grew more rapidly than vehicle control‐treated cells (p < .01); however, recombinant bovine rbIGFBP2 inhibited IGF‐I‐induced proliferation. IGF‐II and/or rbIGFBP2 did not affect BREC proliferation. Taken together, SCFA treatment decreased IGFBP2 and IGFBP6 expression in rumen epithelial cells, and lower expression of these IGFBP might promote rumen epithelial cell proliferation by facilitating IGF‐I.     

Quantitative expression of mRNA encoding BMP/SMAD signalling genes in the ovaries of Booroola carrier and non‐carrier GMM sheep

The GMM sheep is a carrier of Booroola fecundity (FecB) gene, which produces the twins and triplets in one lambing. The homozygous carrier GMM (FecB^BB), non‐carrier GMM and Malpura (FecB⁺⁺) ewes were synchronized by progesterone sponges, and the plasma progesterone concentration was measured by RIA. The results showed that the progesterone concentration did not differ significantly (p > .05) in homozygous carrier GMM (5.74 ± 1.2 ng/ml), non‐carrier GMM (5.42 ± 1.4 ng/ml) and non‐carrier Malpura ewes (5.67 ± 1.5 ng/ml). Further, quantitative expression of BMP factors/receptors and SMAD signalling genes were analysed in the ovaries of sheep by qRT‐PCR. The study showed that the expression of BMP2 was slightly higher (p > .05) in carrier GMM than that of non‐carrier GMM, but it was almost similar to Malpura ewes. Expression of BMP4 and BMP7 was significantly higher (p < .001; p < .05) in carrier GMM than that of non‐carrier GMM and Malpura ewes. Although BMP6 expression was higher (p > .05) in carrier GMM than that of non‐carrier GMM, but lower (p > .05) than the Malpura ewes. Expression of BMP15 (p < .05), GDF5 (p < .01) and GDF9 (p < .05) was significantly higher in carrier GMM than non‐carrier GMM ewes. Surprisingly, BMPR1B expression was significantly higher (p < .001) in non‐carrier GMM and Malpura than the carrier GMM ewes, while TGFβRI did not differ significantly (p > .05) among both GMM genotypes. On the other hand, expression of BMPR1A (p > .05) and BMPRII (p < .05) was higher in carrier GMM than the non‐carrier GMM, but significantly lower (p < .001) than the Malpura ewes. It was interesting to note that the expression of SMAD1 (p > .05), SMAD2 (p < .001), SMAD3 (p < .05), SMAD4 (p < .001), SMAD5 (p < .001) and SMAD8 (p < .001) was lower in the carrier GMM than that of non‐carrier GMM ewes. It is concluded that the FecB mutation alters the expression of BMPR1B and SMAD signalling genes in the ovaries of homozygous carrier GMM ewes.     

Lactoferrin concentration and expression in New Zealand cows milked once or twice a day

This study evaluated the concentration and expression of lactoferrin (LF) in cows selected for once a day (OAD) milking compared to twice a day (TAD) milking. Milk samples were collected from the Massey University TAD and OAD herds. Milk traits and expression of LF and insulin‐like growth factor 1 (IGF‐1) were analyzed with a general linear model that included the fixed effects of milking frequency, lactation number, interaction between milking frequency and lactation number, and as covariates proportion of F, heterosis F × J and deviation from the herd median calving date. Cows milked OAD produced milk with higher (p < .01) concentrations of protein and lactose than TAD milked cows. Compared to TAD cows, cows milked OAD had higher expression of the LF gene (1.40 vs. 1.29 folds, p = .03) and the IGF‐1 gene (1.69 vs. 1.48 folds, p = .007). The correlation between the expression of LF gene and the concentration of LF in milk was strong (r = .66 p < .001), but the correlation between the expression of the IGF‐1 gene and LF concentration was stronger (r = .94, p < .001). These results suggest that milking frequency affects the milk composition and expression of milk composition genes at early lactation.     

Insulin‐Like Growth Factor‐1 Regulates the Expression of Luteinizing Hormone Receptor and Steroid Production in Bovine Granulosa Cells

Luteinizing hormone LH plays important roles in follicular maturation and ovulation. The effects of LH are mediated by LH receptor (LHR) in the ovary. However, the factors that regulate the expression of LHR in bovine granulosa cells (GCs) are not well known. Insulin‐like growth factor‐1 (IGF‐1) is known to play a key role in the acquisition and maintenance of functional dominance. To better understand the roles of LHR expression and IGF‐1, we conducted three experiments to determine (i) mRNA expression of LHR in the GCs of developing follicles, (ii) the effects of IGF‐1 on LHR mRNA expression in cultured GCs and (iii) the effects of IGF‐1 on estradiol (E2), progesterone (P4) and androstenedione (A4) production by non‐luteinized GCs. In experiment 1, small follicles (<6 mm Ø) expressed lower levels of LHR than mid‐sized follicles (6–8 mm Ø) and large follicles (≥9 mm Ø) expressed the highest levels of LHR mRNA (p < 0.05). In experiment 2, IGF‐1 (1 and 100 ng/ml) increased (p < 0.05) the expression of LHR mRNA in GCs from small and large follicles. In experiment 3, IGF‐1 (0.1–100 ng/ml) increased A4 and E2 in GCs from both small and large follicles but increased P4 only in large follicles. IGF‐1 in combination with LH (0.1 and 1 ng/ml) increased P4 and A4 in large follicles, and increased E2 and A4 in GCs of small follicles. These findings strongly support the concept that IGF‐1 upregulates LHR mRNA expression as well as A4 and E2 production in GCs and that IGF‐1 is required for determining which follicle becomes dominant and acquires ovulatory capacity.     

The preliminary study on the effects of growth hormone and insulin‐like growth factor‐I on κ‐casein synthesis in bovine mammary epithelial cells in vitro

The effects of growth hormone (GH) and insulin‐like growth factor‐I (IGF‐I) on protein synthesis and gene expression of κ‐casein in bovine mammary epithelial cell in vitro were studied. The treatments were designed as follows: the growth medium without serum was set as the control group, while the treatments were medium supplemented with GH (100 ng/ml), IGF‐I (100 ng/ml), and GH (100 ng/ml) + IGF‐I (100 ng/ml). The quantity of κ‐casein protein was measured by ELISA, and the κ‐casein gene (CSN3) expression was examined by real‐time quantitative PCR (RT‐qPCR). Compared with the control group, all the experimental groups had greater (p < 0.05) expression of CSN3. The concentration of κ‐casein followed a similar response as CSN3, but the difference between the treatments and the control was not statistically significant (p > 0.05). Furthermore, no synergistic effect of GH and IGF‐I was observed for both the κ‐casein concentration and CSN3 expression. It is therefore concluded that GH or IGF‐I can independently promote the expression of CSN3 in bovine mammary epithelial cells in vitro.     

Effect of dietary nutmeg oil on heat‐stress tolerance‐related parameters in Korean native chicken reared under hot temperature

This study investigated the effect of dietary nutmeg oil (NO) on growth performance, blood parameters, lipid peroxidation and heat shock protein (HSP) 70 expression in Korean native chicken (KNC) reared under hot temperature. We allocated 273 meat‐type KNCs (Hanhyup‐3, 4‐week‐old, body weight [BW] = 539.93 ± 1.75 g) to the following three treatments with seven replicate pens (13 birds/pen) per treatment. Three treatment diets were as follows: (a) Control, basal diet without NO supplementation; (b) NO 250; and (c) NO 500, basal diet supplemented with 250 and 500 ppm NO respectively. Diets and water were provided ad libitum throughout the 6‐week feeding trial. During overall period (0–6 weeks), no differences (p > 0.05) were observed in BW gain (BWG), feed intake (FI) and feed conversion rate (FCR) among treatments. However, the FI at 0–3 weeks decreased (p < 0.05) quadratically with increasing NO levels. Most blood parameters did not differ (p > 0.05) among treatments, although the monocyte level of the NO 500 group was considerably lower (p > 0.05) than that of the other groups. Furthermore, dietary NO did not affect serum triglyceride, cholesterol, total protein, albumin, calcium, phosphorus and alanine aminotransferase (ALT) levels (p > 0.05); however, it linearly decreased serum aspartate aminotransferase (AST) level (p < 0.05). Additionally, serum malondialdehyde (MDA) concentration decreased (p < 0.05) and heart MDA concentration was lower (p = 0.08) with increasing dietary NO supplementation. After a 3‐hr heat (35°C) challenge, the rectal temperature (RT) reduced (p < 0.05) linearly with increasing NO levels. Dietary NO did not affect liver HSP70 (p > 0.05) gene expression. In conclusion, NO potentially enhanced the ability of chickens to alleviate heat stress. Furthermore, our findings suggest that lipid oxidation inhibition by dietary NO likely mediated the enhanced heat‐stress tolerance of the chickens.     

Influence of growth differentiation factor 9 and bone morphogenetic protein 15 on in vitro maturation of canine oocytes

Growth differentiation factor 9 (GDF‐9) and bone morphogenetic protein 15 (BMP‐15) have pivotal roles in oocyte development in many species, therefore the aim was to investigate these factors during in vitro maturation (IVM) of canine oocytes. Canine cumulus oocytes complexes (COCs) were cultured in six groups for 72 hr in a supplemented TCM199‐Hepes medium as (a) Control group; (b) GDF‐9 antibody (Ab); (c) BMP‐15 Ab; (d) recombinant human (rh) GDF‐9; (e) rh BMP‐15 or (f) rh BMP‐15 and GDF‐9. Data were evaluated by ANOVA. The Abs against GDF‐9 or BMP‐15 had a negative impact on meiotic development. Higher (p < 0.05) number of oocytes was arrested at GVBD stage when they were incubated with either GDF‐9 Ab (64.4 ± 2.1%) or BMP‐15 Ab (67.2%± 4.9%) in comparison to those in control group (32.4 ± 7.8%). In contrast, more (p < 0.05) oocytes in control group reached MI (37.4 ± 1.3%) and MII stages (10.2 ± 2.1%) comparing to those groups with GDF‐9 Ab (23.1 ± 4.7% MI; 0.0% MII) or BMP‐15 Ab (16.4 ± 2.4%MI; 5.9% ± 2.1 MII). Higher rates (p < 0.05) of oocytes in control group stayed still arrested at GV (19.9 ± 8.6%) in comparison to those cultured with either rhGDF‐9 (3.7 ± 0.4%) or rhBMP‐15 (10.9 ± 0.7%). However, there were no differences in MII rates between oocytes cultured with GDF‐9 (14.7 ± 3.1) and BMP‐15 (7.8 ± 2.5) separately. But, more oocytes (p < 0.05) reached the MII stage (20.5 ± 3.8%) compared to those exposed to each protein separately and to the control group. These results suggest that these proteins likely contribute to the meiotic development in dogs.     

Methyl donors dietary supplementation to gestating sows diet improves the growth rate of offspring and is associating with changes in expression and DNA methylation of insulin‐like growth factor‐1 gene

The study aimed to investigate the effects of maternal dietary methyl donors on the performance of sows and their offspring, and the associated hepatic insulin‐like growth factor‐1 (IGF‐1) expression of the offspring. A total of 24 multiparous sows were randomly fed the control (CON) or the CON diet supplemented with methyl donors (MD) at 3 g/kg betaine, 15 mg/kg folic acid, 400 mg/kg choline and 150 μg/kg VB₁₂, from mating until delivery. After farrowing, sows were fed a common lactation diet through a 28‐days lactation period and six litters per treatment were selected to be fed until at approximately 110 kg BW. Maternal MD supplementation resulted in greater birthweight (p < 0.05) and increased the piglet weights (p < 0.01) and litter weights (p < 0.05) at the age of day 28, compared with that in CON group. The offspring pigs in the MD group had greater ADG (p < 0.05) and tended to lower F:G ratio (p = 0.07) compared with that of CON group from day 28 to 180 of age. The offspring pigs from MD group had greater serum IGF‐1 concentrations and expressions of hepatic IGF‐1 gene and muscular IGF‐1 receptor (IGF‐1r) protein at birth (p < 0.05), and greater hepatic IGF‐1 protein (p = 0.03) and muscular IGF‐1r gene expressions (p < 0.05) at slaughter, than that from the CON group. Moreover, the methylation at the promoter of IGF‐1 gene in the liver of newborn piglets and finishing pigs was greater in the MD group than that of the CON group (p < 0.05). In conclusion, maternal MD supplementation throughout gestation could enhance the birthweight and postnatal growth rate of offspring, associated with an increased expression of the IGF‐1 gene and IGF‐1r, as well as the altered DNA methylation of IGF‐1 gene promotor.     

Effect of Physiologically Relevant Heat Shock on Development,Apoptosis and Expression of Some Genes in Buffalo (Bubalus bubalis) Embryos Produced In Vitro

For investigating the effects of physiologically relevant heat shock, buffalo oocytes/embryos were cultured at 38.5°C (control) or were exposed to 39.5°C (Group II) or 40.5°C (Group III) for 2 h once every day throughout in vitro maturation (IVM), fertilization (IVF) and culture (IVC). Percentage of oocytes that developed to 8‐cell, 16‐cell or blastocyst stage was lower (p < 0.05) and the number of apoptotic nuclei was higher (p < 0.05) for Group III > Group II > controls. At both 8–16‐cell and blastocyst stages, relative mRNA abundance of stress‐related genes HSP 70.1 and HSP 70.2 and pro‐apoptotic genes CASPASE‐3, BID and BAX was higher (p < 0.05) in Groups III and II than that in controls with the exception of stress‐related gene HSF1. Expression level of anti‐apoptotic genes BCL‐XL and MCL‐1 was also higher (p < 0.05) in Groups III and II than that in controls at both 8–16‐cell and blastocyst stages. Among the genes related to embryonic development, at 8–16‐cell stage, the expression level of GDF9 was higher (p < 0.05) in Group III than that in controls, whereas that of GLUT1, ZAR1 and BMP15 was not significantly different among the three groups. At the blastocyst stage, relative mRNA abundance of GLUT1 and GDF9 was higher (p < 0.05) in Group II than that in controls, whereas that of ZAR‐1 and BMP15 was not affected. The results of this study demonstrate that exposure of buffalo oocytes and embryos to elevated temperatures for duration of time that is physiologically relevant severely compromises their developmental competence, increases apoptosis and affects stress‐, apoptosis‐ and development‐related genes.     

Effect of resveratrol on growth performance,rectal temperature and serum parameters of yellow‐feather broilers under heat stress

This study investigated the effect of dietary resveratrol supplementation on growth performance, rectal temperature, and serum parameters of yellow‐feather broilers under heat stress. A total of 480 yellow‐feather broilers (28‐day‐old) were randomly allotted to five groups with six replicates. A thermoneutral group (TN) (24 ± 2°C) received a basal diet and another four heat‐stressed groups (37 ± 2°C for 8 hr/day and 24 ± 2°C for the remaining time) were fed the basal diet or basal diet with 200, 350, and 500 mg/kg resveratrol for 14 consecutive days. The results revealed that resveratrol supplementation improved average daily gain (p = 0.001), and decreased (p < 0.05) rectal temperature from d 3 when compared with heat‐stressed group without resveratrol. In addition, supplementation with resveratrol at 350 or 500 mg/kg lowered (p < 0.05) the contents of corticosterone, adrenocorticotropic hormone, cholesterol, triglycerides, uric acid, malonaldehyde, and activities of aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase, increased (p < 0.05) the levels of triiodothyronine, the ratio of triiodothyronine to thyroxine, total protein, glutathione, and activities of alkaline phosphatase, total superoxide dismutase, catalase, and glutathione peroxidase, though with few fluctuation. In conclusion, supplementation with resveratrol can improve the growth performance by positively regulating serum metabolic parameters and alleviating tissue oxidant damage of broilers under heat stress.     

Bone Morphogenetic Protein‐6 (BMP‐6) Stimulates the Antrum Formation by the Regulation of its Signalling Pathway in Caprine Pre‐antral Follicles Cultured In Vitro

BMP‐6 has been found to be important to ovarian cells and oocyte, as well as to uterus. Thus, this study investigated the effect of bone morphogenetic protein (BMP‐6) and recombinant follicle‐stimulating hormone (rFSH) alone or in combination on the in vitro culture (IVC) of isolated caprine secondary follicles (Experiment 1) and the mRNA levels for BMP receptors/Smad signalling pathway (BMPR1A, BMPR2, SMAD1, SMAD4, SMAD5, SMAD6, SMAD7 and SMAD8) in vivo and in vitro using BMP‐6 (Experiment 2). Secondary follicles were cultured in αMEM⁺ alone (control medium) or supplemented with BMP‐6 at 1 or 10 ng/ml and rFSH alone or the combination of both BMP‐6 concentrations and rFSH. The results from Experiment 1 showed that the antrum formation rate was higher in the BMP‐6 at 1 ng/ml (p < 0.05) than in MEM. In Experiment 2, the mRNA expression for BMPR2, SMAD1, SMAD5 and SMAD6 was detected in non‐cultured control and after in vitro culture (MEM and 1 ng/ml BMP‐6); while the expression of SMAD7 and SMAD8 mRNA was only detected after IVC, SMAD4 was only detected in the BMP‐6 at 1 ng/ml treatment. In conclusion, the low BMP‐6 concentration positively influenced antrum formation and ensured normal mRNA expression for BMP receptor and Smads after IVC of caprine secondary follicles.     

The effect of Epigallocatechin‐3‐gallate on small intestinal morphology,antioxidant capacity and anti‐inflammatory effect in heat‐stressed broilers

This study was to investigate the effects of Epigallocatechin‐3‐gallate (EGCG) on intestinal morphology, antioxidant capacity and anti‐inflammatory response in heat‐stressed broiler. A total of 192 2‐week‐old Arbour Acres broilers chickens were divided into four groups with six replicates per group and eight chickens per replicate: one thermoneutral control group (28°C, group TN), which was fed the basal diet; and three cyclic high‐temperature groups (35°C from 7:00 to 19:00 hr; 28°C from 19:00 hr to 7:00 hr, heat stress group), which were fed the basal diet supplementation with EGCG 0 mg/kg (group HS0), 300 mg/kg (group HS300) and 600 mg/kg (group HS600). The gut morphology and intestinal mucosal oxidative stress indicators, as well as intestinal barrier‐related gene expression, were analysed. The results showed that compared with group TN, heat stress reduced the villus height (VH), activities of glutathione peroxidase (GSH‐Px), superoxide dismutase (SOD)and catalase (CAT), increased the crypt depth (CD) and malondialdehyde (MDA)content at 21, 28 and 35 days (p < 0.05). After the heat‐stressed broilers were supplemented with EGCG, VH, VH/CD (V/C), and the activities of GSH‐Px, SOD and CAT were increased, and CD and MDA content were reduced compared with those in group HS0 without EGCG supplementation at 21, 28 and 35 days (p < 0.05). The EGCG supplementation promoted the gene expression of nuclear factor‐erythroid 2‐related factor 2 (Nrf2), Claudin‐1, Mucin 2 (Muc2) and alleviated the nuclear factor‐kappa B (NF‐κB) and lipopolysaccharide‐induced tumour necrosis factor (LITAF) gene expression compared with group HS0 (p < 0.05). Moreover, intestinal morphology was strongly correlated with antioxidant ability and inflammatory response. In conclusion, EGCG alleviated the gut oxidative injury of heat‐stressed broilers by enhancing antioxidant capacity and inhibiting inflammatory response.     

Effect of probiotic supplementation on growth performance,intestinal morphology,barrier integrity,and inflammatory response in broilers subjected to cyclic heat stress

This study investigated the protective effects of probiotic on heat stress‐induced intestinal injury and inflammatory response in broilers. A total of 180 male broilers were randomly allocated to three treatments with four replicates each from 22 to 42 days of age. The broilers were either raised under thermoneutral (TN) conditions (23 ± 1°C) or subjected to cyclic heat stress (28–35–28°C for 12 hr daily). The broilers kept at TN conditions were fed a basal diet, and those exposed to heat stress were fed basal diets supplemented with or without probiotic at a dose of 1.5 × 10⁸ cfu/kg. Compared with the TN group, heat stress decreased (p < .05) the growth performance, reduced (p < .05) villus height and villus height: crypt depth ratio in intestinal mucosa, increased (p < .05) serum levels of D‐lactic acid on day 28 and endotoxin, TNF‐α and IL‐6 on day 42, and decreased (p < .05) serum IL‐10 content on day 42. Dietary supplementation of probiotic reversed (p < .05) all these changes except for the growth performance in heat‐stressed broilers. In conclusion, dietary inclusion of probiotic could improve intestinal morphology and barrier function, alleviate inflammatory response, but exert no ameliorative effect on growth performance of broilers under cyclic heat stress.     

Effects of chestnut tannins on intestinal morphology,barrier function,pro‐inflammatory cytokine expression,microflora and antioxidant capacity in heat‐stressed broilers

A study was conducted to evaluate the effects of chestnut tannins (CT) on intestinal morphology, barrier function, pro‐inflammatory cytokine expression, microflora and antioxidant capacity in heat‐stressed broilers. Four hundred 28‐day‐old male Ross 308 broilers were randomly assigned into four groups, with 10 replicates per group and 10 broilers per replicate. The broilers in the normal (NOR) group were kept at 22 ± 1°C and fed the basal diet, and each of the other three groups were treated with cyclic heat (33 ± 1°C from 0800 to 1800 and 22 ± 1°C from 1800 to 0800) and fed the basal diet with 0 (HT), 1 (CT1) or 2 (CT2) g of CT/kg of diet. The experiment lasted for 14 days. Compared with the HT group, broilers in the NOR and CT2 groups had higher (p < .05) average daily gain and villus height in the jejunum and lower serum d ‐lactate (p < .001) and diamine oxidase (p < .01) levels. The addition of 2 g CT/kg of diet increased the total antioxidant capacity (p < .001) and superoxide dismutase activities (p < .05) and zonula occludens‐1 mRNA expression level (p < .05) and decreased the malondialdehyde concentration (p < .01) and mRNA expression levels of interleukin‐6 (p < .001) and nuclear factor kappa B (p < .001) in the jejunal mucosa of heat‐stressed broilers. The populations of Escherichia coli and Clostridium in the jejunum (p < .01) and caecum (p < .05) of broilers in the HT group were higher than those in the NOR and CT2 groups. In conclusion, the addition of 2 g CT/kg of diet seemed to be a feasible means of alleviating the negative effects of heat stress on the growth performance and intestinal function of broilers.     

Comparative study of stress response,growth and development of uteri in post‐weaning gilts challenged with zearalenone and estradiol benzoate

The objective of this study was to evaluate the effects of zearalenone (ZEA) and estradiol benzoate (EB) on stress injury and uterine development in post‐weaning gilts. Thirty healthy post‐weaning female gilts (Duroc × Landrace × Large White) aged 28–32 days were randomly allocated to three treatments as follows: (a) basal diet (Control), (b) basal diet plus 1.0 mg/kg purified ZEA (ZEA) and (c) basal diet plus 0.75 ml (1.5 mg) EB per pig at 3‐days intervals by intramuscular injection (EB). The serum estradiol (E₂), the final and the increased vulvar area, uterine index, thickness of the myometrium and endometrium, and protein expression of heat shock protein 70 (HSP70) in ZEA group were higher than those in the control group (p < .05), but lower than those in the EB group (p < .05). The serum luteinizing hormone in ZEA group was lower than that of the control group (p < .05), but higher than that in the EB group (p < .05). Higher serum follicle‐stimulating hormone and progesterone were observed in the ZEA and control groups than those in the EB group (p < .05). The serum glutathione peroxidase activity in the ZEA group was lower than that in the control and EB groups (p < .001), and the malondialdehyde in the ZEA group was higher than that in the control and EB groups (p < .001). Moreover, the relative mRNA and protein expression of growth hormone receptor (GHR) and relative mRNA expression of HSP70 in the ZEA and EB groups were higher than those in the control group (p < .05). In conclusion, both ZEA (1.0 mg/kg) and EB (1.5 mg at 3 days intervals by intramuscular injection) stimulated vulvar swelling and uterine hypertrophy by disordering serum hormones and up‐regulating GHR expression, and induced stress by different mechanisms in this study. Furthermore, the observed up‐regulating HSP70 expression challenged by ZEA or EB may be part of the mechanism to resist stress injury.