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Abstract: In order to study the molecular mechanism involved in meat quality differences especially tenderness variance caused by castration in Qinchuan cattle males, this study investigated the gene expression profile of Longissimus thoracis (LT) muscle and screened differentially expressed genes in LT muscle from both intact male and castrated male Qinchuan cattle at 36months of age utilising Bovine Genome Array. Significance Analysis of Microarrays (SAM) was used to identify the differentially expressed genes, Go (Gene Ontology) and pathway analyses were conducted on which by a free Web-based Molecular Annotation System 2.0 (MAS 2.0). Approximately 11,000 probe sets representing 10,000 genes were detected in LT muscle of 36-month-old Qinchuan cattle. After SAM analysis of the microarray data, 142 genes were shown to be differentially expressed. These genes were predominantly involved in collagen fibril organization and synthesis, regulation of cell growth and development and striated muscle contraction. The significant pathways involved mainly included ECM–receptor (extracellular matrix-receptor) interaction, cell communication and focal adhesion. Quantitative real-time PCR (qRT-PCR) was performed to validate some differentially expressed genes identified by microarray. These patterns of gene expression may contribute to understanding the molecular mechanism of better meat quality of beef derived from castrates than from intact males. [Copyright &y& Elsevier]

The antioxidant systems of superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH), and glutathione peroxidase (GSH–Px) are important in maintaining sperm motility and viability. The purpose of this study was to determine the effects of varyilig doses of trehalose on in vitro semen quality variables and antioxidant activities of frozenthawed bovine semen. The semen samples, diluted with an extender containing trehalose (0, 25, 50, 100, and 200 mM), were evaluated. The extender supplemented with 100 mM trehalose exhibited the greatest percentages of sperm motility, acrosomal membrane integrity, and plasma membrane integrity in comparison with the control group (P < 0.05). No difference was observed for sperm motility between trehalose 50 and 100 mM groups (P > 0.05). Extender supplemented with trehalose did not affect SOD levels. Compared with the other groups, CAT was greater with the supplementation of trehalose at 100 and 200 mM (P < 0.05). The extender supplemented with trehalose had enhanced GSH–Px activity compared with the control group (P < 0.05). However, increasing the doses of trehalose (100, 200 mM) decreased GSH–Px activity, compared with 50 mMtrehalose (P < 0.05). Compared with the other groups, trehalose at the concentration of 25 and 50 mM increased GSH activity (P < 0.05). The application of 200 mM trehalose produced the least amount of GSH activity among all of the groups (P < 0.05). In conclusion, extender supplemented with trehalose reduced the oxidative stress induced by freeze–thaw and improved measures of bovine semen quality. The antioxidant characteristics of trehalose may be related to its effectiveness in membrane cryopreservation. Further studies are required to obtain more concrete results on the determination of lipid peroxidation and antioxidant capacities of trehalose in cryopreserved bovine semen. [ABSTRACT FROM AUTHOR]

Milk fat determines the quality of milk and is also a main targeted trait in dairy cow breeding. Recent studies have revealed important regulatory roles of microRNAs (miRNA) in milk fat synthesis in the mammary gland. However, the role of miRNA in bovine mammary epithelial cells (BMEC) remains largely unknown. In this study, we found that the overexpression of miR-130a significantly decreased cellular triacylglycerol (TAG) levels and suppressed lipid droplet formation, whereas the inhibition of miR-130a resulted in greater lipid droplet formation and TAG accumulation in BMEC. MiR-130a also significantly affected mRNA expression related to milk fat metabolism. Specifically, the overexpression of miR-130a reduced the mRNA expression of PPARG, FABP3, PLIN2, and FATP1, whereas the downregulation of miR-130a increased the mRNA expression of PPARG, C/EBPβ, C/EBPα, PLIN2, FABP3, and FATP1. Furthermore, western blot analysis revealed the protein level of PPARG in miR-130a mimic and inhibitor transfection groups to be consistent with the mRNA expression response. Finally, luciferase reporter assays verified that PPARG was the direct target of miR-130a. This study provides the first experimental evidence that miR-130a directly affects TAG synthesis in BMEC by targeting PPARG, suggesting that miR-130a potentially could be used to improve beneficial milk components in dairy cows. [ABSTRACT FROM AUTHOR]

Growing evidence has revealed that microRNA are central elements in milk fat synthesis in mammary epithelial cells. A negative regulator of adipocyte fat synthesis, miR-27a has been reported to be involved in the regulation of milk fat synthesis in goat mammary epithelial cells; however, the regulatory role of miR-27a in bovine milk fat synthesis remains unclear. In the present study, primary bovine mammary epithelial cells (BMEC) were harvested from mid-lactation cows and cultured in Dulbecco's modified Eagle's medium/F-12 medium with 10% fetal bovine serum, 5 µg/mL of insulin, 1 µg/mL of hydrocortisone, 2 µg/mL of prolactin, 1 µg/mL of progesterone, 100 U/mL of penicillin, and 100 µg/mL of streptomycin. We found that the overexpression of miR-27a significantly suppressed lipid droplet formation and decreased the cellular triacylglycerol (TAG) levels, whereas inhibition of miR-27a resulted in a greater lipid droplet formation and TAG accumulation in BMEC. Meanwhile, overexpression of miR-27a inhibited mRNA expression of peroxisome proliferatoractivated receptor gamma (PPARG), CCAAT/enhancer- binding protein beta (C/EBPβ), perilipin 2 (PLIN2), and fatty acid binding protein 3 (FABP3), whereas miR-27a downregulation increased PPARG, C/EBPβ, FABP3, and CCAAT enhancer binding protein alpha (C/EBPa) mRNA expression. Furthermore, Western blot analysis revealed the protein level of PPARG in miR-27a mimic and inhibitor transfection groups to be consistent with the mRNA expression response. Moreover, luciferase reporter assays verified that PPARG was the direct target of miR-27a. In summary, these results indicate that miR-27a has the ability to control TAG synthesis in BMEC via targeting PPARG, suggesting that miR-27a could potentially be used to improve beneficial milk components in dairy cows. [ABSTRACT FROM AUTHOR]

Isolation of genomic DNA is a prerequisite for assessment of milk quality. As a source of genomic DNA, milk Somatic cells from milking ruminants are practical, animal friendly, and cost-effective sources. Extracting DNA from milk can avoid the stress response caused by blood and tissue sampling of cows. In this study, we optimized a novel DNA extraction method for amplifying long (>1,000 bp) DNA fragments and used it to evaluate the isolation of DNA from small amounts of milk. The techniques used for the separation of milk omatic cell were explored and combined with a sodium dodecyl sulfate (SDS)-phenol method for optimizing DNA extraction from milk. Spectrophotometry was used to determine the concentration and purity of the extracted DNA. Gel electrophoresis and DNA amplification technologies were used for to determine DNA size and quality. The DNA of 112 cows was obtained from milk (samples of 13 ± 1 mL) and the corresponding optical density ratios at 260:280 nm were between 1.65 and 1.75. Concentrations were between 12 and 45 µg/µL and DNA size and quality were acceptable. The pecific PCR amplification of 1,019- and 729-bp bovine DNA fragments was successfully carried out. This novel method can be used as a practical, fast, and economical mean for long genomic DNA extraction from a small amount of milk. [ABSTRACT FROM AUTHOR]

The present study aimed to investigate the effects of vitamin B supplementation on standard bovine semen quality parameters and anti-oxidative enzyme activities. Vitamin B was supplemented at concentrations of 1.25, 2.5, 3.75 and 5.0 mg/ml to bovine semen cryoprotective medium. The results indicated that the motility and straight line velocity, curvilinear velocity, mean coefficient, velocity of the average path values of sperm supplemented with 2.50 mg/ml vitamin B were significantly higher than that of other groups (p < 0.05). No significant difference was observed for linearity index, lateral head displacement values and the percentage of grade A spermatozoa between the extenders containing 2.50 and 3.75 mg/ml vitamin B (p > 0.05). The percentages of acrosome-intact and plasma membrane-intact spermatozoa were significantly improved (p < 0.05) by supplementing with 2.50 mg/ml vitamin B. The results of biochemical assay revealed that vitamin B supplementation did not cause significant changes in superoxide dismutase levels compared with control (p > 0.05). However, the catalase levels were higher in the treatment supplemented with vitamin B at 2.50 mg/ml, when compared with other groups (p < 0.05). The extender supplemented with vitamin B significantly decreased glutathione peroxidase activity compared with the control (p < 0.05). The supplementation of 3.75 mg/ml vitamin B caused the highest value of glutathione reductase activity, compared with other groups (p < 0.05). In conclusion, the extender supplemented with vitamin B could reduce the oxidative stress provoked by freezing-thawing and improve bovine semen quality. Further studies are required to obtain more concrete results on the determination of lipid peroxidation and antioxidant capacities of vitamin B in cryopreserved bovine semen. [ABSTRACT FROM AUTHOR]

Twenty-seven domesticated yellow cattle breeds of China and three introduced cattle breeds were analysed by means of 30 microsatellite markers to determine the level of genetic variation within and among populations as well as the population structure. In all, 480 microsatellite alleles were observed across the 30 breeds with the mean number of alleles per locus of 9.093 for native breeds and 6.885 for the three introduced breeds. Mean F-statistics (0.08) for Chinese native cattle breeds implied that 92% of the total genetic variation was from genetic differentiation within each breed and 8% of the genetic variation existed among breeds. A phylogenetic tree was constructed based on Nei's genetic distances, and three clusters were obtained. According to the tree, the three introduced breeds were distinct from the 27 native breeds. The indigenous cattle breeds were divided into two clusters, one cluster including five humpless breeds and the other cluster containing 22 humped breeds. This study identifies multiple origins of yellow cattle of China from Bos taurus and Bos indicus. Furthermore, population structure analysis implies that there are possibly five independent original domestications for yellow cattle in China. Four of five origins were four different Bos indicus types, mainly in areas of the Chang Jiang, the Zhu Jiang River basin, the Yellow River and the Huai River basin. The other origin was for Bos taurus type of Mongolian descent, mainly located in Northwestern China, the Mongolian plateau and Northeastern China or north of the Great Wall. [ABSTRACT FROM AUTHOR]