Isolation,Identification and Genotyping of Pasteurella multocida from Fowl Cholera

In order to determine the suspected avian cholera pathogen and geneotypes, bacteria isolation technology was used to isolate and culture pathogenic bacteria, the isolated bacteria were identified by traditional methods and molecular biological methods. Geneotyping of the isolated bacteria were determined using PCR technology and gene sequence analysis. The results demonstrated that the isolated bacteria had typical culture characteristics of Pasteurella multocida (Pm),and the morphology of the colony, the characteristics of cell staining, the physiological and biochemical characteristics were consist with Pm; 457 bp fragment was amplified by PCR. Geneotyping results showed that only A primer amplified the target gene fragment of 1 050 bp, sequence analysis also showed that the isolated bacteria shared 97.6% to 100.0% sequence homology with reference strains, and phylogenetic tree analysis showed that the isolated bacteria and A type Pm were in a branch.The experimental results indicated that the isolated bacteria was identified as capsular serotype A of Pm, the results provided reference for the prevention and control of fowl cholera.     

不同剂量猪繁殖与呼吸综合征病毒ORF5基因疫苗肌肉注射诱导仔猪细胞免疫的研究

将猪繁殖与呼吸综合征病毒(PRRSV)ORF5基因疫苗(pcDNA-PRRSV-SC2-ORF5)分别以200 ug,100 ug和50 ug 3种剂量肌肉注射免疫21日龄仔猪,同时设空载体对照和生理盐水空白对照.于免疫后7 d、15d、30d、45d、60d和70d采用流式细胞仪(FACS)和淋巴细胞增殖试验(MTT法)分别对免疫猪外周血中CIM+、CD8+T淋巴细胞数和淋巴细胞的转化功能进行检测.结果表明3个不同剂量的pcDNA-PRRSV-SC2-ORF5基因疫苗免疫仔猪后均能诱导产生良好的细胞免疫应答,实验组与对照组比较差异极显著(p<0.01)或显著(p<0.05).大剂量基因疫苗免疫仔猪的外周血T淋巴细胞的增殖能力及CD4+、CD8+百分含量高于中剂量和小剂量基因疫苗免疫的仔猪,表现出一定的量效关系.     

鸡胚蛋重比作为评价传染性支气管炎病毒致侏儒化指标的研究

在鸡胚上接种鸡传染性支气管病毒（M41、H120、HN99株）后6 d内,每天称量胚体和各自种蛋的重量,计算胚体/种蛋重量的比值（EE值）。EE值与未接种病毒的对照组鸡胚相比,计算出最小感染量,EE指数通过接种过IBV的鸡胚EE值和对照组EE值相比计算出来。结果表明,在接种M41株和HN99株后第4天EE值与对照组差异显著,在接种H120株后第5天EE值与对照组差异显著,可初步判定是被传染性支气管炎病毒感染,这种方法可更快更有效的观察胚体损伤程度,尤其适用于需经传代才能看到矮小化现象的野毒株。     

海南原鸡PDCD10的克隆、表达及其蛋白的生物信息学分析

通过克隆、表达海南原鸡程序性细胞死亡分子10(Programmed cell death10,PDCD10)基因,对其蛋白进行生物信息学分析。采用RT-PCR方法克隆得到海南原鸡PDCD10基因CDS区,将扩增产物与原核表达载体pET42a连接,构建pET42a-PDCD10重组质粒,经IPTG诱导表达后,进行SDS-PAGE和Western blot分析;运用生物信息学软件对所获得基因的核苷酸序列进行分析,并预测其编码蛋白的理化性质及二级结构等。克隆获得了PDCD10639bp的CDS区核苷酸序列,融合蛋白分子量约为57ku,生物信息学分析发现,PDCD10CDS区包括1个639bp的开放读码框,编码212个氨基酸。该蛋白不含信号肽,无跨膜区,二级结构中存在大量α-螺旋。研究结果为进一步开展海南原鸡PDCD10的功能研究奠定了坚实的基础。     

脱氧雪腐镰刀菌烯醇对猪毒害作用及毒理机制研究进展

脱氧雪腐镰刀菌烯醇(deoxynivalenol, DON)是一种常见的污染粮食、饲料和食品的霉菌毒素,严重影响人和牲畜的健康。猪对DON非常敏感,DON可造成猪拒食、生长抑制、呕吐以及器官损伤、细胞毒性、免疫毒性等毒害作用。本文主要综述了DON对猪的毒害作用、毒理机制以及解毒剂开发等方面的研究进展,尤其关注DON毒理作用的遗传调控研究,旨在为从遗传本质提高猪对脱氧雪腐镰刀菌烯醇的抵抗力提供更全面的参考。     

酸枣仁提取物的生物学作用及其在畜牧生产中的应用

酸枣仁提取物是镇静安神的传统中药,其发挥生物学功能的主要成分是其中的皂苷和黄酮类。该提取物逐步受到中兽医的关注,在畜牧生产中有着广泛的开发利用前景。作者主要综述酸枣仁提取物对中枢神经系统、免疫系统及其在畜牧生产中应用的影响,为其在畜牧生产中的开发应用提供依据。     

O型口蹄疫病毒VP1 T细胞与B细胞表位基因双拷贝串联表达产物的免疫应答

以一株O型口蹄疫病毒(FMDV)外壳蛋白VP1基因为模板,合成与细胞免疫及体液免疫相关抗原表位肽基因:21-40肽(20AA)和141-160肽(20AA)基因序列,运用基因工程技术构建了含有串联结构21-40(20AA)～141-160(20AA)～21-40(20AA)～141-160(20AA)的2020-2020VP1融合基因表达载体r2020-2020,转化宿主菌BL21(DE3)RIL后诱导表达,表达产物经SDS-PAGE及Western Blot分析显示重组融合蛋白的分子量约为18Ku.动物实验表明,较小剂量的融合蛋白就能诱导豚鼠产生特异性T淋巴细胞增殖反应及抗FMDV中和抗体,证明该融合蛋白可同时激活细胞免疫及体液免疫反应,具有开发成为抗FMDV疫苗的应用价值.     

实验性脾虚证大鼠肝脏和肌肉组织中糖原变化的组织学观察

将45只大白鼠随机分成对照组、脾虚模型组及治疗组,每组15只。以利血平(1mg/kg·d)复制大白鼠脾虚证模型,四君子汤辅助治疗,在实验进行至第7天和第14天分别捕杀各组大白鼠,取肝脏、肌肉组织,用组织化学方法测定在不同组间和不同实验阶段其中糖原的变化。结果,在第7天时,脾虚组和治疗组糖原含量明显低于正常对照组,第14天时,与正常对照组相比较,脾虚组和治疗组糖原含量显著升高,且脾虚组糖原含量较其它两组高,提示肝脏和肌肉组织中糖原的变化可能是脾虚证发生发展的重要指标。     

Evaluation of Joint Toxicity of Enrofloxacin and Three Antimicrobials on MRC-5 Cells

The aim of this study was to evaluate the combined toxicity of enrofloxacin with each of three antimicrobials (ciprofloxacin,florfenicol and sulfamethazine).MRC-5 cells were used as a cell model to simulate the damage of lung cells caused by mixed contamination of antimicrobials.Multiple concentration gradients and mixing ratios were set.CCK-8 method was used to determine the inhibition rate of cell growth caused by four antimicrobials and the inhibition rate of cell growth caused by enofloxacin mixed with three antimicrobials,respectively.Then Chou-Talalay method was used to fit the median effect plot and to calculate the combination index (CI) value.The results showed that the growth inhibition rates of MCR-5 cells caused by four single drugs went up in a step-like manner with the increase of drug concentration in the tested concentration range,among them,the growth inhibitory rate of MCR-5 cells by florfenicol was low (<4.5%).The combined toxicity of the three binary combinations showed a concentration-dependent and mixing-ratio dependence.Mixing enrofloxacin and ciprofloxacin showed synergistic toxicity (CI<1) on MRC-5 cells at high and middle concentration groups,and antagonistic toxicity (CI>1) at the very-low-concentration groups.Mixing ciprofloxacin and florfenicol mainly showed an inhibited toxicity (CI>1).Binary combination enrofloxacin and sulfamethazine might present either a synergistic joint toxicity (CI<1) or an antagonistic joint toxicity (CI>1) as the concentration and mixing ratio changing.This study showed that it was necessary to assess the combined toxicity of antimicrobials in the toxicity evaluation of antimicrobials.Using Chou-Talalay method,the joint toxicity of multiple antimicrobials could be quickly and efficiently determined at cellular level.     

Association between porpoise presence and fish choruses: implications for feeding strategies and ecosystem-based conservation of the East Asian finless porpoise

The associations between feeding activities and environmental variables inform animal feeding tactics that maximize energetic gains by minimizing energy costs while maximizing feeding success. Relevant studies in aquatic animals, particularly marine mammals, are scarce due to difficulties in the observation of feeding behaviors in aquatic environments. This data scarcity concurrently hinders ecosystem-based fishery management in the context of small toothed-cetacean conservation. In the present study, a passive acoustic monitoring station was deployed in an East Asian finless porpoise habitat in Laizhou Bay to investigate potential relationships between East Asian finless porpoises and their prey. The data revealed that porpoises were acoustically present nearly every day during the survey period. Porpoise detection rates differed between spring and autumn in concert with activities of fish choruses. During spring, fish choruses were present throughout the afternoon, and this was the time when porpoise vocalizations were the most frequently detected. During autumn, when fish choruses were absent, porpoise detection rates decreased, and diurnal patterns were not detected. The close association between fish choruses and finless porpoise activities implies an “eavesdropping” feeding strategy to maximize energetic gains, similar to other toothed cetaceans that are known to engage similar feeding strategies. Underwater noise pollution, particularly those masking fish choruses, could interrupt finless porpoises’ feeding success. Fisheries competing soniferous fishes with finless porpoise could impact finless porpoise viability through ecosystem disruption, in addition to fishing gear entanglement.