1株羊源D型产气荚膜梭菌的分离鉴定及耐药性分析

doi:10.16431/j.cnki.1671-7236.2025.03.036

摘要/Abstract

摘要： 【目的】D型产气荚膜梭菌是一种革兰阳性厌氧菌，可引起山羊、绵羊和牛等动物发生肠毒血症，对反刍动物的健康造成巨大的影响。本研究旨在了解内蒙古乌审旗地区D型产气荚膜梭菌耐药情况，为该地区由D型产气荚膜梭菌引起的疾病的治疗提供科学依据。【方法】通过培养特性及革兰染色镜检对分离菌进行初步鉴定，通过16S rRNA基因序列比对和毒素基因PCR扩增对可疑菌株进一步鉴定，并采用K-B纸片法进行药敏试验，PCR检测分离菌耐药基因。【结果】分离菌在TSC培养基中长出黑色的圆形菌落，革兰染色镜检可见菌体粗短，成单个或双个排列的革兰阳性直杆菌，形态学及镜检结果符合产气荚膜梭菌特点。16S rRNA基因序列比对结果显示，分离菌与NCBI数据库中已公布的产气荚膜梭菌16S rRNA基因序列相似性均＞99%，鉴定该菌株为产气荚膜梭菌。毒素基因PCR检测扩增出cpa和etx基因，表明该菌株为D型产气荚膜梭菌。药敏试验结果显示，分离菌对头孢曲松、头孢呋新、庆大霉素等16种抗菌药耐药。耐药基因检测结果显示，分离菌检出bla_CTX-M、bla_SHV、qnrA和aac(6')-Ⅰb-cr 4种耐药基因，未检出bla_TEM和qnrS 2种耐药基因。【结论】本试验成功从绵羊体内分离出1株D型产气荚膜梭菌，该菌株存在多重耐药，该研究结果为D型产气荚膜梭菌感染绵羊的诊断、流行病学调查和治疗提供了参考依据。

关键词: 绵羊; D型产气荚膜梭菌; 分离鉴定; 耐药性

Abstract: 【Objective】 Clostridium perfringens type D was a Gram-positive anaerobic bacterium,which could cause enterotoxaemia in animals such as goats,sheep and cattle,which had a great impact on the health of ruminants.The aim of this study was to understand the drug resistance of Clostridium perfringens type D in Wushen county, Inner Mongolia, and provide a scientific basis for the treatment of diseases caused by Clostridium perfringens type D in this region.【Method】 The isolated strain was initially identified by culture characteristics and Gram staining microscopy.The suspected strain was further identified by 16S rRNA sequence comparison and PCR amplification of toxin gene.And the drug sensitivity test was carried out by K-B paper slide method,and PCR was used to detect the drug resistance gene of the isolated strain.【Result】 The isolated strain grew black round colonies in TSC medium,and the Gram staining microscopy showed that the bacterium was thick and short,and became a single or double arrangement of Gram-positive rectobacteria,and the morphological and microscopic results were consistent with the characteristics of Clostridium perfringens.The results of 16S rRNA sequence alignment showed that the similarity of the isolated strain and 16S rRNA gene sequences of Clostridium perfringens published in NCBI database was more than 99%,and the strain was identified as Clostridium perfringens.The PCR detection results of toxin genes showed that cpa and etx genes were amplified,indicating that the strain was Clostridium perfringens type D.The results of drug sensitivity tests showed that the isolated strain was resistant to 16 antimicrobials including ceftriaxone,cefuroxime and gentamicin.The results of resistance gene detection showed that 4 resistance genes,bla_CTX-M,bla_SHV,qnrA and aac(6')-Ⅰb-cr,were detected,and 2 resistance genes,bla_TEM and qnrS,were not detected.【Conclusion】 In this experiment,a strain of Clostridium perfringens type D,which was multi-drug resistant,was successfully isolated from sheep.The results of this study provided a reference basis for the diagnosis,epidemiological investigation,and treatment of Clostridium perfringens type D infected sheep.

Key words: sheep; Clostridium perfringens type D; isolation and identification; drug resistance

中图分类号:

S852.61

李娜, 刘重阳, 张靖靖, 玛丽雅其其格, 珠娜, 陆斌, 海鹰. 1株羊源D型产气荚膜梭菌的分离鉴定及耐药性分析[J]. 中国畜牧兽医, 2025, 52(3): 1352-1359.

LI Na, LIU Chongyang, ZHANG Jingjing, MALIYA Qiqige, ZHU Na, LU Bin, HAI Ying. Isolation,Identification and Drug Resistance Analysis of a Sheep-derived Strain of Clostridium perfringens Type D[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(3): 1352-1359.

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