牛源肠球菌的分离鉴定及毒力因子基因的检测  

Isolation and identification of bovine Enterococci and detection of virulence factor genes

作　　者：候书宝 阚威[1] 武小虎[1] 张晶晶[1] 张勇[1] 赵兴绪[1] 董翔宸 王华[1] 

机构地区：[1]甘肃农业大学动物医学院,甘肃兰州730070

出　　处：《中国兽医学报》2015年第3期383-387,393,共6页Chinese Journal of Veterinary Science

基　　金："十二五"农村领域国家科技计划资助项目(2011AA10A210);兰州市科技局科技三项资助项目(033143)

摘　　要：为探究牛源性肠球菌中6种毒力因子基因(ccf、asa1、ace、esp、cylA、gelE)的分布情况,采集了不同地区382份疑似隐形乳腺炎的奶样,常规分离纯化细菌,采用16SrRNA和16S^23SrRNA方法相结合的方法,共鉴定出了68株肠球菌并检测了其溶血特性和上述6种毒力基因的存在情况。结果显示,40株(58.82%)表现为α-溶血,10株(14.71%)为β-溶血,18株(26.47%)为γ-不溶血。68株(100%)携带asa1基因,49株(72.06%)携带gelE基因,43株(63.24%)携带ccf基因,12株(17.65%)携带ace基因,10株(14.71%)携带esp基因,6株(8.82%)携带cylA基因。54.41%的分离菌株同时携带gelE、ccf、asa1毒力基因,而青海民和分离株的esp与ace,gelE与ccf同时出现,有一定规律性;但溶血性基因cylA与14株溶血性肠球菌没有相关性。研究牛源肠球菌的溶血特性与毒力基因携带情况,对肠球菌性奶牛乳腺炎的防控具有指导意义,同时也为动物与人之间病原互相传播的研究提供试验依据。Enterococci are widely distributed in the environment,human and animal digestive tract.In recent years,reports on the serious infections of Enterococci have been significantly increased.The conventional bacterial isolates method,16 SrRNA and 16S-23 SrRNA approaches were used to isolate and identify Enterococci from in different regions suffering from the 382 suspected negative mastitis milk samples for the purpose of exploring the case of normal bovine Enterococci flora carrying 6kinds of virulence factor genes(ccf,asa1,ace,esp,cylA,gelE)68 Enterococcus were isolated.Meanwhile,we detected hemolytic characteristics of the 68 isolates and 6kinds of virulence factor genes of Enterococci.Of which 40(58.82%)strains wereα-hemolysin,10(14.71%)strains wereβ-hemolytic and 18(26.47%)wereγ-hemolysin.The detection numbers of asa1,gelE,ccf,ace,esp,cylA genes were 68(100%),49(72.06%),43(63.24%),12(17.65%),10(14.71%),6(8.82%)respectively.It indicated that three of the six genes investigated(ccf,asa1,and gelE)were simultaneously present in 54.41% isolates.E.faecalis isolates in Minhe of Qinghai,harboured simultaneously the esp and ace,gelE and ccf.However,the hemolytic cylA gene were no correlation with 14 hemolytic Enterococci.The study on bovine Enterococcus virulence genes serves as a guidance for preventing and controling Enterococci mastitis,while providing a theoretical basis for the research on the transmission of pathogens between animals and humans.

关 键 词：牛源性肠球菌 分离鉴定 毒力因子基因 

分 类 号：R446.5[医药卫生—诊断学]