Exogenous DNA Uptake of Boar Spermatozoa by a Magnetic Nanoparticle Vector System

The sperm‐mediated gene transfer method is applicable to transgenesis in many species that use spermatozoa for reproduction recently, which has been shown various results. In the current study, we show that transgenic porcine embryos can be efficiently produced by employing a simple transfection method that uses magnetic nanoparticles (MNPs). The complexes formed between plasmid DNA and MNPs were bounded on ejaculated boar spermatozoa at a higher efficiency compared to methods using DNA alone or lipofection. Using confocal microscopy, rhodamine fluorophore‐labelled MNPs were detected on external surfaces of the spermatozoa membrane, which were bounded on zona pellucida of in vitro maturated oocyte during in vitro fertilization. Electron microscopy revealed that clusters of MNPs were detected in inside of plasma membrane and nucleus of the spermatozoa head. Additionally, we found that magnetofected boar spermatozoa could be fertilized with oocytes in vitro and that the resulting gene of green fluorescent protein was detected in fertilized eggs by genomic PCR analysis. Taken together, these results suggest that MNPs can be used to efficiently introduce a transgene into embryo via spermatozoa.     

Use of comparative proteomics to identify the effects of creatine pyruvate on lipid and protein metabolism in broiler chickens

Four hundred male chickens were selected to study the effects of pyruvate (Pyr), creatine pyruvate (CrPyr) and creatine (Cr) on the expression of hepatic mitochondrial and cytoplasm proteins associated with lipid and protein metabolism. Mitochondrial purification was accomplished using the two-step differential centrifugation and density gradient method, and the activities of organelle-specific marker enzymes were determined to assess the purity of the mitochondria. Proteins were extracted and fractionated by two-dimensional electrophoresis and the differential protein spots were assessed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry. CrPyr reduced fatty acid accumulation by down-regulating adipose differentiation-related protein, inhibited ATP synthase expression, and reduced cholesteryl ester transfer protein (CETP) expression, thus reducing the levels of high density lipoprotein and triglycerol (TG) levels (thereby lowering fat and cholesterol deposition). CrPyr increased the expression of eukaryotic translation initiation factor (eIF) 2B, calreticulin (CRT) and eIF3a, thus promoting protein synthesis. CrPyr up-regulated the expression of fatty acid-binding proteins, CETP and apolipoprotein A-IV in cytoplasmic extracts, and these proteins accelerated the decomposition of fatty acids and TG, thus reducing fat deposition. In conclusion, CrPyr plays an important role in lipolysis and protein synthesis, and this effect was more pronounced than was the effect of Pyr and Cr.     

安徽省2015年规模猪场高致病性蓝耳病定点监测与分析

[目的]掌握安徽省规模猪场高致病性蓝耳病的感染状况和免疫抗体水平。[方法]采用抗体阻断ELISA、荧光RT-PCR方法检测了24个猪场的588份血清样本和28个场点的790份组织样品。[结果]高致病性蓝耳病个体抗体免疫合格率为78.1%,群体免疫合格率为75.0%;中型猪场的平均免疫抗体合格率为87.1%、小型猪场为63.9%。高致病性蓝耳病病毒核酸个体阳性率为28.7%,群体阳性率为75.0%;中型猪场、小型猪场和生猪屠宰场的高致病性蓝耳病病毒核酸个体阳性率分别为41.7%、19.5%和6%。[结论]2015年安徽省规模猪场的高致病性蓝耳病抗体免疫合格率较高,但存在带毒现象,建议加强疫病的免疫和综合防控。     

鲜茧生丝与干茧生丝理化性状的比较研究

为了明确鲜茧生丝与干茧生丝的理化性能,采用六个庄口的蚕茧分别缫制的鲜茧生丝和干茧生丝,进行生丝样品的器械检测.结果表明,来源不同庄口的鲜茧生丝与干茧生丝,其清洁、洁净、断裂强度、断裂伸长率、切断、丝胶溶失率等性能无明显差异,而干茧生丝的抱合性能优于鲜茧生丝.鲜茧生丝与干茧生丝的热学性能和丝蛋白结构也无明显差异.     

Cytokine gene expression in ileal tissues of cattle infected with Mycobacterium paratuberculosis

Cytokine gene expression in ileal tissues of cattle infected with Mycobacterium paratuberculosis was evaluated. The effects of infection with M. paratuberculosis on cytokine production may influence immune regulation at the site of colonization, resulting in the chronic inflammatory state associated with the latter stages of this disease. Ileal samples were obtained at necropsy from noninfected control cows (n=8) and clinically infected cows (n=7) and processed for immunohistochemistry and in situ hybridization. Cows infected with M. paratuberculosis were in the latter stages of disease with clinical signs such as weight loss, watery diarrhea, and inappetence. Among cytokines we studied, interleukin-1alpha (IL-1alpha), IL-1beta, IL-6, and interferon-gamma (IFN-gamma) were expressed significantly more in infected animals than in noninfected control animals. The expression of tumor necrosis factor-alpha (TNF-alpha), however, was not different between the two groups of cattle. In addition, immunohistochemical staining demonstrated that the number of resident macrophages in the ileum of infected animals was three times greater than that of noninfected cows. In contrast to this, ileal tissues from noninfected control animals contained 1.5 times more neutrophils than the ileal tissues from cows infected with M. paratuberculosis. These data demonstrate that localized ileal cytokine production is different between cows chronically infected with M. paratuberculosis and noninfected control cows.     

纳罗克非洲狗尾草种子产量因子与产量的通径分析

采用多因素裂区试验设计,通过大样本相关、通径和逐步回归分析后,结果表明,纳罗克非洲狗尾草Setaria sphacelata cv. Narok的6个种子产量因子对种子产量的直接贡献大小排序为：x4（单位长度小花数）>x6（千粒重）>x1（花序长）>x2（单位面积生殖枝数）>x3（单位长度小穗数）>x5（单位长度种子数）,说明提高单位长度小花数、千粒重和花序长是最有效提高纳罗克非洲狗尾草种子产量的途径,其次提高单位长度小穗数、单位长度小花数和单位面积生殖枝数。     

家蚕母种继代蛾区选择方法对种性维持的影响

为了有效保持生产实用家蚕品种的优良性状,在家蚕母种继代繁育中采用5种选择方法选择继代蛾区,探讨种性维持效果。分析8年12代及F1代的成绩表明:所有性状以保持种性为主,采用选留平均数上下1个标准差范围内蛾区的稳定化选择和以茧丝质指标在平均数-0.5~1.0标准差范围内选择,并结合健康性指标选留高值的综合选择,有利于家蚕母种的种性维持。     

Molecular epidemiology of Newcastle disease viruses in Vietnam

Newcastle disease virus (NDV) causes significant economic losses to the poultry industry in Southeast Asia. In the present study, 12 field isolates of NDV were recovered from dead village chickens in Vietnam between 2007 and 2012, and were characterized. All the field isolates were classified as velogenic. Based on the sequence analysis of the F variable region, two distinct genetic groups (Vietnam genetic groups G1 and G2) were recognized. Phylogenetic analysis revealed that all the 12 field isolates fell into the class II genotype VII cluster. Ten of the field isolates, classified as Vietnam genetic group G1, were closely related to VIIh viruses that had been isolated from Indonesia, Malaysia, and Cambodia since the mid-2000s, while the other two field isolates, of Vietnam genetic group G2, clustered with VIId viruses, which were predominantly circulating in China and Far East Asia. Our results indicate that genotype VII viruses, especially VIIh viruses, are predominantly responsible for the recent epizootic of the disease in Vietnam.