羊源D型产气荚膜梭菌分离鉴定及耐药性与致病性分析  

Isolation,identification,drug resistance and pathogenicity analysis of Clostridium perfringens type D from sheep

作　　者：赵学亮 王晨骁 王斌[1] 冯航 叶超 党如意[1] 王娟[1] 杨增岐[1] ZHAO Xueliang;WANG Chenxiao;WANG Bin;FENG Hang;YE Chao;DANG Ruyi;WANG Juan;YANG Zengqi(College of Veterinary Medicine,Northxvest A&F University,Yangling,Shaanxi 712100,China)

机构地区：[1]西北农林科技大学动物医学院,陕西杨凌712100

出　　处：《中国兽医学报》2021年第7期1290-1294,共5页Chinese Journal of Veterinary Science

基　　金：国家绒毛用羊产业技术体系资助项目(CARS-39-14)。

摘　　要：为鉴定2020年9月乌审旗放牧绵羊陆续发生死亡的原因,本试验采集样品进行细菌分离培养、生化鉴定、16S rRNA序列进化树比较分析、毒素基因PCR鉴定、小鼠致病性及药敏检测。结果显示,分离株为D型产气荚膜梭菌,其α、ε毒素基因及16S rRNA高度保守,与参考序列之间核苷酸同源性99.49%;进化树分析显示该菌与序列号为MT613499.1产气荚膜梭菌遗传距离最近;耐药性结果显示分离菌对阿莫西林、氨苄青霉素、头孢噻呋钠、链霉素、庆大霉素、新霉素、替米考星、磺胺异恶唑等完全耐药,对多西环素、金霉素、土霉素、泰妙菌素、氟苯尼考表现为极敏;小鼠在接种分离株后8 h全部死亡,表明其毒力较强。试验结果可为羊源D型产气荚膜梭菌合理用药、减少多重耐药菌株的出现提供依据。The aim of this study is to identify the cause of the successive deaths of grazing sheep in Wushen city of Inner Mongolia in September 2020.Samples were collected for bacterial isolation and culture, biochemical identification, 16S rRNA sequence phylogenetic tree comparative analysis, PCR identification of toxin genes, mouse pathogenicity and drug sensitivity testing.The results showed that the isolated strain was Clostridium perfringens type D,and its α and ε toxin genes and 16 S rRNA were highly conserved, and were 90% homologous to the reference sequence.The phylogenetic tree analysis showed that the bacterium has the closest genetic distance to Clostridium perfringens with sequence number of MT613499.1;the drug resistance results showed that the isolate was resistant to amoxicillin, ampicillin, ceftiofur sodium, streptomycin, gentamicin, neomycin, tilmicosin and sulfisoxazole, and it was completely resistant to doxycycline, chlortetracycline, oxytetracycline, tiamulin, florfenicol.All mice died 8 h after inoculation with the isolate, indicating that its virulence was strong.The test results can provide a basis for rational drug use and reducing the occurrence of multi-drug resistant strains of Clostridium perfringens.

关 键 词：D型产气荚膜梭菌 毒素分型 分离鉴定 药敏试验 

分 类 号：S852.61[农业科学—基础兽医学]