Methionine alleviates aflatoxinb1-induced broiler chicks embryotoxicity through inhibition of caspase-dependent apoptosis and enhancement of cellular antioxidant status.

Practical methods for preventing embryotoxicity in chickens that are caused by aflatoxin-B1 (AFB1) are currently rare. Binding absorbers are commonly used in feeding stuff to reduce laying hens' exposure to off-contaminated diets, thus reducing residue exposure to fertilized eggs. Nonetheless, several adsorbents have been shown to affect the use of nutrients and the absorption of minerals in poultry. Thus, seeking an effective strategy to counter or control embryotoxicity in broiler chicks caused by AFB1 is a problem. A total of 180 embryonated eggs were injected with 36 ng AFB1 with or without 5.90 mg L-methionine (Met) 30 embryonated eggs each, followed by incubation in an incubator until hatching time. The in ovo injection of Met significantly reduced toxicity caused by AFB1 in broiler embryos by enhancing the liver and kidney functions, lipid profiles, and alleviated oxidative stress during the incubation period. Furthermore, the relative gene expressions (SSTR5, TSH-ß, Bcl-2, GSH-Px, GST-a, and SOD in the liver) were up-regulated with in ovo injection of AFB1+Met compared to AFB1 alone. Moreover, there was a dowin-regulated trend in Bax, Caspases-3, Caspases-7, Caspases-9, CYP1A1, CYP2H1, and P53 gene expression with in ovo injection of AFB1+Met compared to AFB1 alone. The in ovo injection of Met led to less apoptotic cells in liver tissues. Such results might be necessary for the poultry industry as it is focused on managing the embryotoxicity of AFB1, which affecting poultry production and welfare. Results from this study demonstrated that in ovo Met injection could alleviate AF-induced toxicity in chicken embryos.

Effects of iron glycine chelate on laying performance, antioxidant activities, serum biochemical indices, iron concentrations and transferrin mRNA expression in laying hens.

This study was conducted to evaluate the effects of iron glycine chelate (Fe-Gly) on laying performance, antioxidant enzyme activities, serum biochemical indices and iron concentrations in laying hens. A total of 810 laying hens (Hy-Line Variety White, 26 weeks old) were randomly assigned to six groups with five replicates of 27 layers. Hens in the control group received diet supplemented with 60 mg Fe/kg as FeSO4 , while hens in other five groups received the diet supplemented with 0, 20, 40, 60 and 80 mg Fe/kg from Fe-Gly respectively. The results showed that dietary Fe-Gly treatments significantly influenced (p < 0.01) the laying rate and egg weight of layers, compared with the control group. Concerning to CuZn-superoxide dismutase (CuZn-SOD) and total superoxide dismutase (T-SOD) activity, Fe-Gly groups (60, 80 mg Fe/kg) were promoted significantly (p < 0.01) compared with 0 mg Fe/kg group. The concentrations of Fe in serum, liver, kidney, spleen and ovary were increased significantly with the level dietary Fe-Gly raised where Fe-Gly groups (60, 80 mg Fe/kg) had observably higher Fe concentration than the control (p < 0.01) in serum, kidney and spleen. There was a trend that transferrin mRNA expression was decreased with the increase of Fe as Fe-Gly in diets, and compared with the control, the expression was lower in the group fed diet with 60 mg/kg Fe as Fe-Gly. In conclusion, Fe-Gly (60 mg Fe/kg) had improved laying rate, egg weight, SOD enzyme activity, Fe absorption and protein synthesis in body and promoted iron metabolism in laying hens. Moreover, Fe-Gly (40 mg/kg Fe) had the similar effect with control group. It revealed that FeSO4 could be substituted by lower concentration of Fe-Gly and Fe-Gly may be superior to FeSO4 for iron fortification to laying hens.

Differential expression of fatty acid transporters and fatty acid synthesis-related genes in crop tissues of male and female pigeons (Columba livia domestica) during incubation and chick rearing.

1. The growth performance of squabs reared solely by male or female parent pigeons was measured, and the changes of lipid content of crop milk and the expression profiles of genes potentially involved in lipid accumulation by crop tissues of parent pigeons were evaluated during incubation and chick rearing. 2. Squabs increased in body weight during 25 d of rearing, whereas both male and female pigeons lost weight after finishing rearing chicks, and the weight loss of male pigeons was significantly greater than that of female parent pigeons. Lipid content of crop milk from both parent pigeons gradually decreased to the crude fat level in the formulated diet after 10 d (R10) of chick rearing. 3. The gene expression of fatty acid translocase (FAT/CD36), fatty acid-binding protein 5 (EFABP) and acyl-CoA-binding protein (ACBP) in male pigeon crop tissue were the greatest at 17 d (I17) of incubation. In female pigeons, FAT/CD36 expression was the highest at I14, and both EFABP and ACBP expression peaked at I14 and R7. The expression of acetyl-CoA carboxylase and fatty acid synthase in male pigeons reached the maximum level at R1, while they peaked at I14 and I17, respectively in female pigeons. The gene expression of peroxisome proliferators-activated receptor-gamma (PPARγ) was the greatest at I17 in the male, while it was at I14 in the female. However, no regular changing pattern was found in PPARα gene expression in male pigeons. 4. These results indicated that male and female pigeons may make different contributions in rearing squabs. The gene expression study suggested that fatty acids used in lipid biosynthesis of crop milk probably originated from both exogenous supply and de novo synthesis. The sex of the parent pigeon affected the lipid content of crop milk and the expression profiles of genes involved in fatty acid transportation and lipogenesis.

Evaluation of embryonic age and the effects of different proteases on the isolation and primary culture of chicken intestinal epithelial cells in vitro.

The present study evaluates the effects of embryonic age and proteolytic enzymes on the isolation and primary culture of chicken enterocyte and to establish an effective technique for chicken intestinal epithelial cell (IEC) cultivation. Fourteen-day-old, 16-day-old and 18-day-old embryos (average weight: 52.23 ± 0.76 g, 50.86 ± 0.99 g, 48.98 ± 1.03 g) were the source for preparation of enterocyte culture, and trypsin-ethylene diamine tetraacetic acid, collagenase, thermolysin and combination of collagenase and thermolysin were used for digestion medium. Optimal culture protocols were determined by qualitative assays of proliferation. Cells isolated by using 14-day-old embryo and collagenase obtain the best attachment and growth in culture, and the production of continuously growing IEC cultures. Thus, we conclude that the use of collagenase as a dissociating enzyme and 14-day-old embryo as a source can be advantageously applied to the isolation of chicken IEC and this method may be useful for various applications and basic studies of the intestinal tract concerning such objects as physiology, immunology and toxicology.

Effect of expanded cottonseed meal on laying performance, egg quality, concentrations of free gossypol in tissue, serum and egg of laying hens.

Three hundred and sixty Hy-Line Brown hens, 40 week of age, were allocated to five treatments, each of which included four replicates of 18 hens. After an expanded process of cottonseed meal (CSM), free gossypol content in CSM was decreased from 1.24 to 0.40 g/kg. The dietary treatments were corn-soybean meal based diets including 6% CSM and 6%, 8% and 10% expanded cottonseed meal (ECSM). Hens fed 8% ECSM had higher (P<0.05) laying rate and average egg weight than those fed 6% CSM. The albumen height and Haugh unit in the control group, 6% and 8% ECSM groups were superior (P<0.05) to other treatments. Hens fed 6% CSM resulted in severe (P<0.05) egg yolk discoloration. Free gossypol (FG) concentrations in yolk and albumen and tissues of the 6% CSM group were greater (P<0.05) than those in any ECSM treatments. Hens fed 6% CSM and 10% ECSM had the highest (P<0.05) FG concentrations in the liver compared with those in the kidney and muscle, and higher (P<0.05) FG residues in yolk than those in albumen. In conclusion, FG in CSM can be reduced by 68% through an expanded process and ECSM can be available in laying hens at up to 10% of the total diet and an appropriate replacement of soybean meal with ECSM may improve performance in laying hens.

Effect of dietary γ-aminobutyric acid on laying performance, egg quality, immune activity and endocrine hormone in heat-stressed Roman hens.

This study was conducted to evaluate the effect of γ-aminobutyric acid (GABA) on laying performance, egg quality, digestive enzyme activity, hormone level and immune activities in Roman hens under heat stress. Roman hens (320 days old) were fed with 0, 25, 50, 75 and 100 mg/kg GABA, respectively during a 60-day experiment. Compared with control, supplementation of 50 mg/kg GABA improved the laying performance and egg quality by significantly increasing egg production, average egg weight and shell strength (P < 0.05), while decreasing the feed-egg ratio and cholesterol level. Anti-oxidation activity was improved by significantly increasing the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), but decreasing malondialdehyde level in serum (P < 0.05), while significantly increasing the glucose and total protein (TP) level, follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E(2) ), insulin, triiodothyronine (T(3) ) and free triiodothyronine (FT(3) ) levels, and IgG, IgA and complement (C3)activity in serum (P < 0.05). The results indicated that oral GABA improved laying performance and physical condition mainly by modulating hormone secretion, enhancing anti-oxidation and immune activity, and maintaining electrolyte balance. Fifty mg/kg was the optimum level for laying hens under heat stress in the present study.

Chitosan nanoparticles act as an adjuvant to promote both Th1 and Th2 immune responses induced by ovalbumin in mice.

The study was conducted to investigate the promoted immune response to ovalbumin in mice by chitosan nanoparticles (CNP) and its toxicity. CNP did not cause any mortality or side effects when mice were administered subcutaneously twice with a dose of 1.5 mg at 7-day intervals. Institute of Cancer Research (ICR) mice were immunized subcutaneously with 25 µg ovalbumin (OVA) alone or with 25 µg OVA dissolved in saline containing Quil A (10 µg), chitosan (CS) (50 µg) or CNP (12.5, 50 or 200 µg) on days 1 and 15. Two weeks after the secondary immunization, serum OVA-specific antibody titers, splenocyte proliferation, natural killer (NK) cell activity, and production and mRNA expression of cytokines from splenocytes were measured. The serum OVA-specific IgG, IgG1, IgG2a, and IgG2b antibody titers and Con A-, LPS-, and OVA-induced splenocyte proliferation were significantly enhanced by CNP (P < 0.05) as compared with OVA and CS groups. CNP also significantly promoted the production of Th1 (IL-2 and IFN-γ) and Th2 (IL-10) cytokines and up-regulated the mRNA expression of IL-2, IFN-γ and IL-10 cytokines in splenocytes from the immunized mice compared with OVA and CS groups. Besides, CNP remarkably increased the killing activities of NK cells activity (P < 0.05). The results suggested that CNP had a strong potential to increase both cellular and humoral immune responses and elicited a balanced Th1/Th2 response, and that CNP may be a safe and efficacious adjuvant candidate suitable for a wide spectrum of prophylactic and therapeutic vaccines.

Chemical composition and microstructure of uroliths associated with the feeding of high-level cottonseed meal diet to sheep.

The chemical composition and microstructure of five urolith samples (4 bladder stones and one kidney stone) associated with the feeding of high level of cottonseed meal (CSM) diet to Chinese merino fine wool sheep (Junken breed, Xinjiang) were examined by optical microscope, X-ray diffraction, X-ray energy dispersive spectrometry (EDS), scanning electron microscopy (SEM), and infrared spectroscopy analysis. The bladder stone samples appeared yellow or white, small powder and loose mass, and as finely granular under the optical microscope. However, the kidney stone samples from a experimental sheep were found as small brown mass, higher hardness, and as a cracklike structure. Oxygen, phosphorus, potassium and magnesium were found as four major elements in these uroliths by X-ray energy dispersive spectrometry (EDS). Potassium magnesium phosphate (MgKPO(4)) and potassium magnesium phosphate hexahydrate (MgKPO(4)·6H(2)O) were major components in the bladder stones, while less magnesium ammonium phosphate hexahydrate (MgNH(4)PO(4)·6H(2)O) examined by X-ray diffraction and infrared spectroscopy analysis. However, the newly found prismatic crystals, which were rich in magnesium and pyrophosphate, were identified as magnesium pyrophosphate (Mg(2)P(2)O(7)) in the kidney stone. The bladder stone samples appeared irregular mass and balls, cracked under SEM with low magnification, while appeared cracked, irregular layer-like, honeycomb-like or tiny balls under high magnification. The kidney stone samples were observed as cone, irregular block or layered crystal structures.

[Research and development of Lipin family.].

Lipin family including at least three members Lipin 1, Lipin 2, and Lipin 3 is a critical regulatory enzyme identified recently, which plays dual roles in lipid metabolisms. Lipin family has physiological effects not only on regulating lipid metabolism, but also on maintaining normal peripheral nervous functions, liver lipoprotein secretion, cell morphous, reproductive functions, and energy homeostasis. Since mutations in Lipin gene express may be associated with AIDS, insulin resistance, obesity, diabetes mellitus, and the other diseases of metabolic syndrome, Lipin may be a new useful target in treatment of above-mentioned clinical-related diseases. In this article, we focused on discovery, construction features, expression, regulatory mechanism, and biological functions of Lipin, as well as its correlation research with clinical-related diseases.

Ghrelin and its biological effects on pigs.

Ghrelin is a 28 amino acid peptide, which produces its marked effects through binding to the endogenous ligand of the growth hormone secretagogue receptor (GHS-R). Based on the contemporary literatures, it was shown that ghrelin was involved in a series of biological functions including regulation of food intake, body weight, gastrointestinal (GI) motility, hormone secretion, glucose release, cardiovascular functions, enzyme release, cell proliferation and reproduction in pigs through binding to GHS-R 1a or unidentified receptors. It was also observed that ghrelin induced adipocyte and hepatocyte proliferation of primary cultured piglet. In this paper, recent research on ghrelin structure, distribution, GHS-R receptor, biological functions and its regulatory mechanisms for pigs are presented.

Is GPR39 the natural receptor of obestatin?

GPR39, an orphan receptor belonging to the family of G protein-coupled receptors, was originally reported to be the receptor of obestatin. However recently, numerous reports have questioned this conclusion. In mammals, GPR39 was reported to be involved in the regulation of gastrointestinal and the metabolic functions. In this article, a latest and brief review on the receptor family, structure, distribution and physiological functions of GPR39 has been reported.