固体脂质纳米增强抗菌药物对胞内金黄色葡萄球菌活性的研究进展

金黄色葡萄球菌导致的乳房炎是危害奶牛养殖业最严重的疾病之一。抗菌药在胞内极易排出或失活,导致奶牛金黄色葡萄球菌乳房炎治疗效果差且容易复发。纳米粒子具有在巨噬细胞和上皮细胞等多种细胞中积累的能力,可提高药物在胞内的浓度和胞内停留时间,这使得它们可用于治疗细胞内感染。本文对固体脂质纳米抗金黄色葡萄球菌感染的研究进展进行综述,以指导和推动固体脂质纳米在治疗金黄色葡萄球菌乳腺炎方面的应用。     

香蕉茎叶粉固态发酵条件优化及鹅对其养分利用率的研究

本试验旨在研究应用固态发酵技术改善香蕉茎叶粉养分组成的工艺参数及鹅对其养分的利用率。选择米曲霉和产朊假丝酵母作为发酵菌种,逐步探究5个因素(单菌种发酵、硫酸铵添加量、米曲霉和产朊假丝酵母接种比例、混合菌液接种量)对发酵后香蕉茎叶粉蛋白质含量的影响。利用正交设计筛选最优发酵温度、底物水分和发酵时间的组合。结果显示:米曲霉和产朊假丝酵母单独发酵均能显著或极显著提高发酵后香蕉茎叶粉的粗蛋白质含量(P0.05或P0.01)。添加硫酸铵能显著或极显著提高发酵后香蕉茎叶粉的真蛋白质含量(P0.05或P0.01),其中添加2%硫酸铵组的真蛋白质含量最高。米曲霉∶产朊假丝酵母接种比例为2∶1的处理发酵后香蕉茎叶粉的蛋白质净增加量显著或极显著高于比例为1∶1、1∶3和3∶2的处理(P0.05或P0.01)。正交试验结果显示,以4%的接种量、接种比例为2∶1(米曲霉∶产朊假丝酵母)、2%的硫酸铵添加量,在基质水分为50%,30℃的环境下发酵4 d效果最佳。经过该工艺发酵后的香蕉茎叶粉粗蛋白质含量提高了33.82%,氨基酸分析结果显示,除赖氨酸和精氨酸外其余的15种氨基酸含量均有不同程度的提高。马冈鹅的代谢试验结果显示,发酵后香蕉茎叶粉中的粗蛋白质利用率提高了52.66%,极显著高于发酵前(P0.01);此外,代谢能和能量利用率也都略有提高(P0.05)。由此可见,经过该发酵工艺发酵后的香蕉茎叶粉营养价值不仅得到了改善,也促进了鹅对其养分的消化吸收。     

冰鲜杂鱼和人工配合饲料对大口黑鲈肌肉品质及健康状况影响的评价

本试验通过测定血清生化指标和肌肉营养成分,评价冰鲜杂鱼和人工配合饲料对大口黑鲈肌肉品质及健康状况的影响。养殖过程中冰鲜组大口黑鲈全程投喂冰鲜杂鱼,饲料组大口黑鲈全程投喂大口黑鲈专用人工配合饲料。在商品鱼上市前,冰鲜组和饲料组各随机采集6尾用于测定血清生化指标,再各随机采集6尾用于测定肌肉营养成分。结果显示:饲料组大口黑鲈血清中谷草转氨酶(AST)活性显著高于冰鲜组(0.01≤P0.05),血清中碱性磷酸酶(ALP)活性极显著高于冰鲜组(P0.01),血清中总蛋白(TP)和白蛋白(ALB)含量极显著低于冰鲜组(P0.01)。冰鲜组大口黑鲈肌肉中总氨基酸(TAA)含量显著高于饲料组(0.01≤P0.05),且肌肉中各种氨基酸的氨基酸评分(AAS)和化学评分(CS)以及必需氨基酸指数(EAAI)均高于饲料组。饲料组大口黑鲈肌肉中亚油酸(C18∶2)含量极显著高于冰鲜组(P0.01)。结果表明,本试验条件下,冰鲜组大口黑鲈肌肉营养品质特别是蛋白质品质及氨基酸组成要优于饲料组,且健康状况也优于饲料组,但饲料组大口黑鲈肌肉中亚油酸含量较冰鲜组极显著升高。因此,大口黑鲈的人工配合饲料配方还需要进一步的优化,来满足消费者对大口黑鲈的营养需求。     

金石翁芍散对仔猪大肠杆菌病治疗试验

为了评价金石翁芍散对仔猪大肠杆菌病的临床疗效,设立金石翁芍散高、中、低3个剂量组及药物对照组,通过混饲给药方式治疗仔猪大肠杆菌病,进行临床治疗效果比较。结果表明,金石翁芍散对仔猪大肠杆菌病有较好的治疗效果,高剂量组治愈率为91.6%,总有效率为94.4%;中剂量治愈率为88.9%,总有效率为94.4%;低剂量组治愈率为72.2%,总有效率为77.8%。高剂量组、中剂量组在降低死亡率、提高治愈率等方面均优于恩诺沙星可溶性粉（治愈率为72.2%,总有效率为77.8%）。低剂量组的治疗效果低于高、中剂量组;中、高剂量组死亡率、治愈率和总有效率均无显著性差异。     

基于单时相MODIS数据的草地退化遥感监测研究

以草地NPP减少的相对百分数（由实际NPP和潜在NPP计算）为主要监测指标，提出了一个基于单时相MODIS数据的草地退化遥感监测方法。介绍了利用MODIS数据反演实际NPP的方法，并提出了一个根据实际NPP和草地类型分布图确定潜在NPP的方法。对2003年中国北方草地的监测结果表明，中国北方草地退化严重，退化程度的空间差异性较大，草地退化程度与草地类型密切相关。     

Isolation and Identification of Variant HeNZK-2014 of Pseudorabies Virus and Sequence Analysis of gE Gene

In order to learn the situation of pig pseudorabies virus (PRV) variant in this study, tissue samples such as lymph nodes which were collected from clinical pigs with suspected PRV infection were identified by PCR. PRV positive sample were inoculated on PK-15 cells after grinding and degerming, with further experiment including virus isolation, plague purification, PCR and IFA identification, TCID₅₀ confirmed by Reed-Muench method, inoculation test and observation of clinical symptoms in mice. The gE gene of purified PRV and brain tissue samples of dead mice were identified by sequencing analysis. The results showed that the virus grown on PK-15 cells could produce typical cytopathic effect (CPE) after 24 h; After three rounds of plaque purification,the isolate was PRV positive identified by PCR and IFA, and nominated as HeNZK-2014; The TCID₅₀ of the isolate was 10^-9.77/0.1 mL; The virus in 1×10⁸ TCID₅₀ inoculation was able to cause itching, tearing, death in infected mice, and PRV could be detected in tissues of dead mice; The molecular genetic variation analysis of gE gene by PCR amplification and clone sequencing indicated that the gE gene from brain tissue of infected mice shared 100.0% homology with HeNZK-2014, and located in a relatively independent branch with newly pandemic isolates in recent years after 2011, but was far from the classical strains before 2011, and both had two insertion of aspartic acid (D) at sites of 48 and 496 amino acids, which were considered to be the typical characteristics of PRV variants. This study successfully isolated a PRV variant, which laid a foundation for further research on vaccine development, prevention and control against PRV variants.     

奶牛乳房炎金黄色葡萄球菌8种毒力基因的PCR检测

为研究奶牛乳房炎金黄色葡萄球菌(Staphylococcus aureus,SA)主要毒力因子的分布情况,本试验利用PCR对70株临床型乳房炎、55株隐性乳房炎金黄色葡萄球菌的8种主要毒力基因进行检测。结果显示,nuc、ClfA、TSST-1、PVL、Hla、Hlb、FnBPA和FnBPB 8种毒力基因在临床型乳房炎SA菌株中的检出率分别为:100.0%(70株)、100.0%(70株)、0(0株)、5.7%(4株)、100.0%(70株)、11.4%(8株)、97.1%(68株)和100.0%(70株);在隐性乳房炎SA菌株中的检出率分别为:100.0%(55株)、100.0%(55株)、0(0株)、70.9%(39株)、98.2%(54株)、9.1%(5株)、100.0%(55株)和100.0%(55株)。结果表明,nuc、ClfA、Hla、Hlb和FnBPA 5种毒力基因是引起奶牛乳房炎的SA的最主要毒力基因;PVL基因可能是引起隐性乳房炎的重要致病基因。     

广西10个地方优质品种鸡γ-干扰素基因的克隆与序列分析

根据基因库中鸡γ-干扰素的基因序列设计了1对特异性引物，应用反转录-聚合酶链式反应（RT-PCR）技术，从ConA诱导培养的广西10个地方优质品种鸡外周血淋巴细胞RNA中扩增了γ-干扰素基因，结果，均得到了大小为520 bp的特异性片段。将扩增产物纯化并克隆到pMD18-T载体上，获得重组质粒，经PCR和EcoRⅠ＋SalⅠ双酶切鉴定后测序。序列分析结果表明，广西10个地方优质品种鸡γ-干扰席基因均编码145个氨基酸的成熟蛋白，分子质量约为16．8ku，与哺乳动物和其他品种鸡的核苷酸序列同源性分别为38．8％～39．0％和99．2％～100％，氨基酸序列同源性分别为23．69／6～24．8％和97．6％～99．4％。     

水牛卵母细胞去核方法的比较

水牛卵母细胞体外成熟22h后，分别用盲吸法、点击法和纺锤体成像系统（Spindle—View System）去核。结果，三者的去核率分别为65．1％、81．8％和95．0％，差异极显著（P〈0．01）。以水牛胎儿成纤维细胞为供体，通过核移植构建的重构胚的融合率、分裂率和囊胚发育率，在上述3种方法之间均没有显著差异（P〉0．05）。认为点击法是一种较为简单、实用和有效的去核方法。     

Genetic diversity decreases as population density declines: Implications of temporal variation in mitochondrial haplotype frequencies in a natural population of Tscherskia triton

Although the spatial genetic differentiation that occurs in animal populations has been extensively studied, information on temporal variations in genetic structure and diversity is still lacking, especially for animals with oscillating populations. In the present study, we used the mtDNA D‐loop sequence to assess the temporal genetic variation in samples from six successive years for the greater long‐tailed hamster, Tscherskia triton. Sampling was carried out between 1998 and 2003 in cropland on the North China Plain, China. A total of 108 individuals were analyzed. The temporal samples showed a high level of genetic diversity. Substantial genetic changes in haplotype frequencies over time were detected for the hamster population. Random genetic drift and migration are likely to be the major factors responsible for the observed temporal pattern. The genetic diversity of the hamster population was higher in years with higher population density, and lower in years with lower population density. The result supports our hypothesis that genetic diversity decreases when population density declines in animals whose population oscillates greatly between years. The combined effects of inbreeding and genetic drift caused by reproduction, dispersal and population size might play important roles in the observed changes in genetic structure and diversity between years.