马源蜡样芽孢杆菌的分离鉴定及毒力基因检测

doi:10.3969/j.issn.1004-1524.2019.02.06

浙江农业学报 ›› 2019, Vol. 31 ›› Issue (2): 216-221.DOI: 10.3969/j.issn.1004-1524.2019.02.06

马源蜡样芽孢杆菌的分离鉴定及毒力基因检测

崔一龙, 石芸, 杨达汉, 尹有勤, 薛江东, 霍晓伟, 马德慧^*

1.内蒙古民族大学动物科学技术学院,内蒙古通辽 028000;
2.内蒙古自治区肉牛疾病防控工程技术研究中心,内蒙古通辽 028042

收稿日期:2018-04-26 出版日期:2019-02-25 发布日期:2019-03-06
通讯作者: 马德慧,E-mail: mdh1962@163.com
作者简介:崔一龙(1995—),男,内蒙古通辽人,硕士研究生,主要从事家畜病理学研究。E-mail: 748709954@qq.com
基金资助:
国家自然科学基金(31460662); 内蒙古自治区自然科学基金(2017MS0319)

Isolation,identification of horse Bacillus cereus and its virulence genes detection

CUI Yilong, SHI Yun, YANG Dahan, YIN Youqin, XUE Jiangdong, HUO Xiaowei, MA Dehui^*

1. College of Animal Science and Technology, Inner Mongolia University for the Nationalities, Tongliao 028000, China;
2. Inner Mongolia Engineering Technology Research Center of Prevention and Control the Beef Cattle Disease, Tongliao 028042, China

Received:2018-04-26 Online:2019-02-25 Published:2019-03-06

摘要/Abstract

摘要： 为鉴定从内蒙古通辽市某马术俱乐部发病及死亡赛马体内分离出的一株病原菌,本研究对分离菌株进行革兰氏染色镜检、动物致病性试验、全自动微生物分析系统检测、16S rRNA基因扩增、系统进化树分析及部分毒力基因PCR检测。结果显示:分离株生化特性符合蜡样芽孢杆菌的特性;其对克林霉素、庆大霉素等药物敏感,对头孢西丁、环丙沙星等药物中度敏感,对氨曲南、头孢唑林等药物耐药;16S rRNA基因序列与GenBank中已登录的蜡样芽孢杆菌序列同源性为100%;系统进化树分析表明,分离株与蜡样芽孢杆菌等亲缘关系最近;毒力基因PCR检测结果显示,8种潜在的毒力基因entFM、cytK、hblA、hblC、hblD、nheA、nheB、nheC均呈阳性。试验结果表明,该分离菌株为具有严重致病性的蜡样芽孢杆菌。

关键词: 蜡样芽孢杆菌, 分离鉴定, 16S rRNA, 毒力基因

Abstract: In order to identify a pathogenic bacterium isolated from the incidence and death racing of an equestrian club in Tongliao city,Inner Mongolia,the isolated strains were subjected to Gram stain microscopy,animal pathogenicity tests,automatic microbiological analysis system detection,16S rRNA gene amplification,phylogenetic tree analysis and partial virulence gene PCR detection. The results showed that the biochemical characteristics of isolates were consistent with the characteristics of Bacillus cereus(B. cereus); it was sensitive to clindamycin, gentamicin and other drugs, moderately sensitive to cefoxitin, ciprofloxacin and other drugs,and resistant to aztreonam,cefazolin and other drugs; 16S rRNA gene sequence was 100% homologous to the registered B. cereus sequence in GenBank; phylogenetic tree analysis showed that the isolated strains were closest to the relatives of B. cereus;the detection results of virulence gene PCR showed that 8 potential virulence genes entFM,cytK,hblA,hblC,hblD,nheA,nheB and nheC were all positive.The results showed that the isolated strain was a serious pathogenic B. cereus.

Key words: Bacillus cereus, isolation and identification, 16S rRNA, virulence genes

中图分类号:

S858.21

崔一龙, 石芸, 杨达汉, 尹有勤, 薛江东, 霍晓伟, 马德慧. 马源蜡样芽孢杆菌的分离鉴定及毒力基因检测[J]. 浙江农业学报, 2019, 31(2): 216-221.

CUI Yilong, SHI Yun, YANG Dahan, YIN Youqin, XUE Jiangdong, HUO Xiaowei, MA Dehui. Isolation,identification of horse Bacillus cereus and its virulence genes detection[J]. , 2019, 31(2): 216-221.

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马源蜡样芽孢杆菌的分离鉴定及毒力基因检测

Isolation,identification of horse Bacillus cereus and its virulence genes detection

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