The Effect of Season on Spermatozoa Motility,Plasma Membrane and Acrosome Integrity in Fresh and Frozen–Thawed Semen from Xinong Saanen Bucks

The aim of this study was to evaluate whether the season of ejaculate collection influences seminal quality parameters of pre‐ and post‐freeze–thawing in Xinong Saanen bucks. Ejaculates were collected from eight bucks throughout the four seasons (spring, summer, autumn and winter) in a 12 months’ time period, identified in the Northern Hemisphere. Semen samples were evaluated by the combinations of conventional and Computer‐Assisted Sperm Analysis (CASA) when fresh and after frozen–thawed, respectively. The results clearly demonstrated that season of ejaculate collection influenced (p < 0.05) fresh semen quality. Highest semen quality was observed during autumn. On the contrary, undesirable indices (significantly lower, p < 0.05) were observed in winter as compared with the other remaining seasons. CASA has clearly shown the influences of seasonal variations on semen motility parameters. Furthermore, season of ejaculate collection was also found to influence sperm freezability. Semen characteristics after frozen–thawed followed a similar pattern with that of fresh ejaculate except in spring. The results revealed that sperm quality was higher (p < 0.01) in summer and autumn than in spring and winter. In conclusion, seasonal variation influences semen quality in Xinong Saanen bucks. In addition to summer and autumn, fresh ejaculates in spring can also be successfully used for AI. Sperm from ejaculates collected during summer and autumn are more suitable for cryopreservation. Hence, it is possible to increase the efficiency of goat breeding by manipulating the seasonal variations of semen quality for immediate AI and/or cryopreservation.     

加拿大双低菜粕对奶牛产奶性能的影响

试验主要研究了加拿大双低菜粕对奶牛产奶性能的影响,为加拿大双低油菜籽粕在奶牛生产中的科学应用提供理论依据和实践指导。试验采用单因素完全随机设计,选择32头荷斯坦奶牛,按年龄、体重、胎次、泌乳期与产奶量随机分组。试验包括2部分。第一部分,日粮粗蛋白含量为14.5%,将16头奶牛随机分为对照组与双低菜粕组,每组8头牛,对照组不饲喂双低菜粕,双低菜粕组在精饲料中添加10%的双低菜粕,替代精饲料中全部的豆粕和棉粕。第二部分,日粮粗蛋白含量为12.5%,将16头奶牛随机分为对照组与双低菜粕组,每组8头牛,对照组不饲喂双低菜粕,双低菜粕组在精饲料中添加5%的双低菜粕,替代精饲料中全部的豆粕和棉粕。试验期75 d,其中预饲期15 d,正饲期60 d。结果表明:蛋白含量为14.5%的双低菜粕组可以提高产奶量,在整个试验期内,平均每头奶牛每天多产奶0.76 kg,对乳成分没有显著影响(P0.05),与对照组相比,双低菜粕组的平均乳脂量、乳蛋白量显著增加(P0.05)。在奶牛日粮中蛋白含量为12.5%时,双低菜粕组对奶牛产奶性能没有显著的影响(P0.05)。因此合理使用加拿大双低菜粕提高奶牛产奶性能,从而提高经济效益。     

猪霍乱沙门氏菌LAMP检测方法的建立与应用

利用猪霍乱沙门氏菌lacZ基因设计特异性引物,通过优化各种反应条件,建立了检测猪霍乱沙门氏菌的环介导等温扩增（LAMP）快速检测方法。此方法特异性好,灵敏度高,将细菌DNA稀释到10-8仍可检出,是PCR方法的1000倍。本研究建立的LAMP方法操作简便、灵敏度高、特异性强,为临床检测猪霍乱沙门氏菌和预防人猪霍乱沙门氏菌病提供了技术保障。     

5株水貂阿留申病毒的分离与鉴定

为给水貂阿留申病疫苗的研制奠定基础,对疑似感染阿留申病死亡的水貂内脏处理后,接种猫肾传代细胞（CRFK）分离病毒,并对分离的毒株进行PCR鉴定及动物回归试验。与此同时,首次应用免疫过氧化物酶单层细胞实验（IPMA）对所分离的水貂阿留申病毒进行TCID50的测定。结果成功分离并鉴定出5株水貂阿留申病毒株,分别命名为ADV-DL124、ADV-DL125、ADV-ZJ3、ADV-QD2、ADV-QD3,各分离株的TCID50分别为105.7TCID50/ml、105.0TCID50/ml、104.6 TCID50/mL、105.2 TCID50/mL、104.1 TCID50/mL。动物回归实验显示,所分离的病毒对水貂具有致病性,为水貂阿留申病毒强毒株。     

应用荧光定量RT-PCR检测J亚群禽白血病病毒

针对J亚群禽白血病病毒（ALV-J）基因序列保守区域设计一对特异性引物和一条特异性探针,通过构建重组阳性标准质粒作为阳性标准品,建立了检测ALV-J核酸的荧光定量PCR方法。优化反应体系和条件后进行特异性、敏感性、重复性试验。结果显示,该检测方法特异性强,与其它禽源病毒如A亚群禽白血病病毒（ALV-A）、B亚群禽白血病病毒（ALV-B）、新城疫病毒（ NDV）、禽流感病毒（AIV）、鸡传染性贫血病毒（CIAV）和马立克病病毒（MDV）均不发生交叉反应;该方法可检测到3.2×102拷贝/μL的病毒核酸,与常规RT-PCR相比,敏感性高100倍;重复性试验的变异系数小于2%。研究结果表明,建立的Real-time RT-PCR 检测方法特异性强、灵敏度高、可重复性好,可用于ALV-J的定量检测。     

鱼类神经坏死病毒纳米探针的制备与特征分析

为了快速检测鱼类神经坏死病毒,本研究设计了病毒特异性的发夹型探针,经硫醇修饰后与纳米银溶胶结合,制备了鱼类神经坏死病毒纳米探针（Ag-NNV）,并使用透射电镜、多功能酶标仪、表面增强拉曼光谱仪等对纳米探针Ag-NNV的表征及发夹型探针在纳米银粒子表面的覆盖率进行了测定和分析。结果表明,裸露的纳米银粒子为球形,平均直径50 nm;制备的纳米探针颗粒为球形,平均直径90 nm。纳米银粒子和纳米探针在溶液中均具有良好的分散性。平均每个纳米银粒子表面结合发夹型探针约100条,单位表面积的探针覆盖量为0.56 pmol/cm2。该纳米探针制备简便,适合用于表面增强拉曼散射法对鱼类神经坏死病毒进行快速检测。     

蛋白原料发酵后营养价值评定研究

本试验把蛋白原料膨化豆粕、棉籽粕、菜籽粕经过啤酒酵母菌、凝结芽孢杆菌以及德氏乳酸杆菌发酵处理后,得到相应的发酵蛋白原料产品指标符合国家饲料卫生标准,小肽含量及其他营养成分得到改善,抗营养因子含量基本消除和降低,显著提高了酵母菌、乳酸杆菌、芽孢杆菌和总菌数,大大提高了蛋白原料的附加值,为节约蛋白饲料,有效提高蛋白原料的综合利用价值开辟了一条新的途径.     

No effect of exogenous melatonin on development of cryopreserved metaphase II oocytes in mouse

Background

This study was conducted to investigate effect of exogenous melatonin on the development of mouse mature oocytes after cryopreservation.

Results

First, mouse metaphase II (MII) oocytes were vitrified in the open-pulled straws (OPS). After warming, they were cultured for 1 h in M₂ medium containing melatonin at different concentrations (0, 10⁻⁹, 10⁻⁷, 10⁻⁵, 10⁻³ mol/L). Then the oocytes were used to detect reactive oxygen species (ROS) and glutathione (GSH) levels (fluorescence microscopy), and the developmental potential after parthenogenetic activation. The experimental results showed that the ROS level and cleavage rate in 10⁻³ mol/L melatonin group was significantly lower than that in melatonin-free group (control). The GSH levels and blastocyst rates in all melatonin-treated groups were similar to that in control. Based on the above results, we detected the expression of gene Hsp90aa1, Hsf1, Hspa1b, Nrf2 and Bcl-x1 with qRT-PCR in oocytes treated with 10⁻⁷, or 10⁻³ mol/L melatonin and untreated control. After warming and culture for 1 h, the oocytes showed higher Hsp90aa1 expression in 10⁻⁷ mol/L melatonin-treated group than in the control (P < 0.05); the Hsf1, Hsp90aa1 and Bcl-x1 expression were significantly decreased in 10⁻³ mol/L melatonin-treated group when compared to the control. Based on the above results and previous research, we detected the development of vitrified-warmed oocytes treated with either 10⁻⁷ or 0 mol/L melatonin by in vitro fertilization. No difference was observed between them.

Conclusions

Our results indicate that the supplementation of melatonin (10⁻⁹ to 10⁻³ mol/L) in culture medium and incubation for 1 h did not improve the subsequent developmental potential of vitrified-warmed mouse MII oocytes, even if there were alteration in gene expression.     

银杏大蚕蝗白僵菌的分离、培养及应用试验

从银杏大蚕蛾僵死虫（蛹）尸上分离的白僵菌B21.1，经过室内扩大培养，生产出含孢量185亿孢子／g菌剂，以其2亿孢子／ml菌液对银杏大蚕蛾幼虫室内毒力测定，表现出很高的致病能力：4天死亡率近半，10天校正死亡率达到98.51%；林间喷雾防治,10天左右致病死亡近50％，21天校正死亡率为84.36％；与其它菌株（剂）试验比较，其致病死亡率和杀虫效果均明显高于其他各供试菌株，表现了较强的专化性。     

我国林业定位的研究

随着我国经济的发展，人民生活水平的提高，以及自然生态问题的日趋严重，林业被赋予了更新的使命。对林业的正确定位正是保证林业合理发挥其作用的重要前提。本文从市场定位学的思想出发，结合我国目前林业发展的实际情况，对林业定位的历史过程、林业定位所要考虑的因素进行了简单的回顾和分析，并对如何定位进行了探讨。