奶牛临床型乳房炎Corynebacterium phoceense的分离鉴定及药敏试验  

Isolation,Identification and Drug Sensitivity Test of Corynebacterium phoceense of Clinical Mastitis Pathogens in Bovine

作　　者：姜东凤[1] 唐启伟 肖艳飞 任晓丽 皇甫和平[1] 石冬梅[1] JIANG Dongfeng;XIAO Fanfei;HUANGFU Heping(Henan University of Animal Husbandry and Economy,Zhengzhou,Henan 450011,China;Zhengzhou Xinlicheng Business Management&Consulting Co.,Ltd,Zhengzhou,Henan 450011,China)

机构地区：[1]河南牧业经济学院,河南郑州450011 [2]郑州新里程企业管理咨询有限公司,河南郑州450011

出　　处：《家畜生态学报》2021年第4期63-66,83,共5页Journal of Domestic Animal Ecology

基　　金：河南省畜禽遗传资源保护工程技术研究中心(53000208);河南牧业经济学院科技创新团队(HUAHE2015004);河南牧业经济学院院级重点培育学科建设项目(41000002)。

摘　　要：对太原市某奶牛场临床型乳房炎病牛进行病原菌的分离鉴定及药敏试验,以期了解主要的病原菌及其药物敏感性,为该场乳房炎的防控提供参考依据。本研究采用病原菌分离培养、16S rRNA基因的PCR扩增及序列分析、16联牛乳房炎病原菌核酸qPCR-荧光探针法鉴定了该牧场4头患病牛的9份乳样,并用纸片法对分离菌株进行了药敏试验。结果表明,共分离出8株细菌,包括Corynebacterium phoceense 5株(62.5%)、气球菌1株、链球菌1株和嗜冷杆菌1株,这些菌属对环丙沙星、庆大霉素、阿莫西林拉维酸钾、利福平、多西环素、头孢噻呋均为高度敏感,而对氨曲南、磺胺异恶唑有一定耐药性;同时,通过TaqMan探针实时荧光PCR筛选出2种弱阳性菌属和6种疑似菌属,但是病原菌含量均不高,且大多为环境条件性致病菌。研究结果说明分离培养的方法不能全面地反映引起乳房炎的病原菌感染状况。所以在生产中须借助分子生物学方法进行综合检测分析。该试验也初步判定Corynebacterium phoceense可能是引起乳房炎的主要致病菌之一。Through the isolation and identification of the pathogenic bacteria and drug sensitive test based on the clinical practice in a dairy farm in Taiyuan China,this paper was aimed to find out the main pathogenic bacteria of cow mammitis and its drug sensitivity to take defensive measures against the disease.9 milk samples of 4 diseased cows in the pasture were identified by the isolation and cultivation of pathogenic bacteria,PCR amplification and sequence analysis of 16S rRNA gene,and 16-linked bovine mastitis pathogen nucleic acid PCR-fluorescence probe method.Drug sensitivity tests were carried out on the isolated strains.The results showed that a total of 8 strains of bacteria were isolated,including 5 strains of“Corynebacterium phoceense”(62.5%),1 strain of balloon fungus,1 strain of streptococcus and 1 strain of psychrotrophic bacterium,which were highly sensitive to Amoxicillin potassium,rifampicin,rifampicin,doxycycline,ceftiofur,and had a certain resistance to aztreonam,sulfamethoxazole;meanwhile,2 weakly positive bacteria and 6 suspected bacteria were screened out through the TaqMan real-time probe fluorescent PCR.However,the content of pathogens was not high,and most of them were environmentally conditioned pathogens.Thus the results showed that the pathogenic bacteria of mastitis could not be detected completely by the isolation method solely,it is necessary to detect them by means of molecular biology in the production.The experiment also reached a preliminary conclusion that Corynebacterium phoceense may be one of the main pathogenic bacteria of cow mammitis.

关 键 词：奶牛乳房炎 Corynebacterium phoceense 分离鉴定 药敏试验 16S rRNA基因检测 qPCR鉴定 

分 类 号：S852.61[农业科学—基础兽医学] S858.23[农业科学—兽医学]