内蒙古部分地区牛源产气荚膜梭菌的分离鉴定及耐药性分析

doi:10.16431/j.cnki.1671-7236.2025.04.039

摘要/Abstract

摘要： 【目的】探明内蒙古部分地区肉牛源产气荚膜梭菌(Clostridium perfringens, C.perfringens)分离株的流行特性和耐药性，为该地区肉牛产气荚膜梭菌病的防治提供科学依据。【方法】试验采集37头病死肉牛的组织样品，采用分离培养、染色镜检、生化试验等方法进行细菌分离鉴定，PCR检测分离菌的毒素类型及耐药基因，并采用K-B纸片法进行药敏试验。【结果】分离菌在TSC培养基上呈黑色圆形菌落，在TSA培养基中浑浊产气，镜检见短粗的革兰阳性直杆菌。分离菌发酵蔗糖、葡萄糖、麦芽糖、硫化氢，使明胶液化，在含铁牛乳培养基中“爆裂发酵”。从37头患畜组织样品中分离出8株产气荚膜梭菌，分离率为21.62%。毒素基因PCR分型结果显示，分离株均只含有cpa毒素，为A型产气荚膜梭菌。耐药基因PCR检测结果显示，检出多种耐药基因，其中bla_TEM、tetM、ermB和sul2基因检出率最高，均为100%；耐药基因aph(3')-Ⅲ-F、tetB(P)、ermC检出率分别为87.5%、62.5%和62.5%，其余耐药基因未检出。药敏试验结果显示，8株产气荚膜梭菌对氨基糖苷类药物耐药，对丁胺卡那、链霉素耐药率均为100%；对庆大霉素、卡那霉素耐药率均为87.5%；对复方新诺明、四环素、红霉素耐药率分别为100%、75.0%和50.0%；对氨苄西林、头孢唑林、氯霉素、克林霉素均表现为敏感。【结论】内蒙古部分地区牛群中存在产气荚膜梭菌流行，主要以A型为主，对抗生素产生了较严重的耐受情况，且存在多重耐药。试验结果可为产气荚膜梭菌流行病学研究、临床合理用药和科学防控提供科学依据。

关键词: 产气荚膜梭菌; 分离鉴定; 耐药性; 耐药基因

Abstract: 【Objective】 The purpose of this study was to investigate the epidemiological and antibiotic resistance characteristics of Clostridium perfringens isolates from beef cattle in some areas of Inner Mongolia,and provide scientific basis for the prevention and treatment of Clostridium perfringens disease in beef cattle in these areas.【Method】 The tissue samples of 37 dead beef cattle were collected,and the bacteria were isolated and identified by isolation culture,staining microscopy and biochemical test.The toxin types and drug resistance genes of the isolated bacteria were detected by PCR,and the drug susceptibility test was conducted by K-B disk method.【Result】 The isolated bacteria appeared as black circular colonies on TSC medium and produced cloudy gas on TSA medium.Microscopic examination revealed short and thick Gram positive rods.The isolated bacteria fermented sucrose,glucose,maltose and hydrogen sulfide,causing gelatin to liquefy and undergo "burst" fermentation in an iron containing milk culture medium.Eight strains of Clostridium perfringens were isolated from the samples of 37 dead beef cattle,with an isolation rate of 21.62%.The PCR typing results of toxin genes showed that all the isolated strains only had cpa toxin and were type A Clostridium perfringens.The PCR detection results of resistance genes showed that multiple resistance genes were detected,with bla_TEM,tetM,ermB and sul2 genes having the highest detection rate of 100%.The detection rates of resistance genes aph(3')-Ⅲ-F,tetB(P) and ermC were 87.5%,62.5% and 62.5%,respectively,while the remaining resistance genes were not detected.The results of the drug sensitivity test showed that 8 strains of Clostridium perfringens were resistant to aminoglycoside drugs,with resistance rates of 100% for amikacin and streptomycin,and 87.5% for gentamicin and kanamycin.The resistance rates to compound sulfamethoxazole tetracycline and erythromycin were 100%,75.0% and 50.0%,respectively.And they were sensitive to ampicillin,cefazolin,chloramphenicol and clindamycin.【Conclusion】 There was a prevalence of Clostridium perfringens in cattle herds in some areas of Inner Mongolia,mainly type A,which had developed severe tolerance to antibiotics and multiple drug resistance.The experimental results could provide scientific basis for the epidemiological research,rational clinical drug use,and scientific prevention and control of Clostridium perfringens.

Key words: Clostridium perfringens; isolation and identification; drug resistance; drug resistance genes

中图分类号:

S852.61⁺6.3

蔺冰冰, 赵洪哲, 关娜, 乌日古木拉, 其根, 张杨, 温永俊, 王凤雪. 内蒙古部分地区牛源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(4): 1873-1883.

LIN Bingbing, ZHAO Hongzhe, GUAN Na, WU Rigumula, QI Gen, ZHANG Yang, WEN Yongjun, WANG Fengxue. Isolation,Identification and Drug Resistance Analysis of Clostridium perfringens from Cattle in Some Areas of Inner Mongolia[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(4): 1873-1883.

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