甘肃地区犊牛腹泻病原调查及产气荚膜梭菌的分离鉴定、耐药性分析

doi:10.16431/j.cnki.1671-7236.2025.08.041

摘要/Abstract

摘要： 【目的】掌握甘肃地区规模化牛场犊牛腹泻感染性病原微生物的流行情况，并了解产气荚膜梭菌(Clostridium perfringens，Cp)对常见抗菌药物的耐药现状。【方法】使用5联犊牛腹泻病原微生物核酸检测试剂盒对兰州、张掖和武威等8个地区10个牛场采集的108份犊牛腹泻粪便样本进行Cp、牛轮状病毒(Bovine rotavirus,BRV)、牛冠状病毒(Bovine coronavirus,BCoV)、产肠毒素型大肠杆菌K99(enterotoxigenic Escherichia coli K99，K99)、隐孢子虫(Cryptosporidium，COWP)检测；取Cp核酸阳性的粪便进行细菌分离培养，并使用染色镜检、生化试验对其进行鉴定。同时，采用PCR扩增对分离菌株进行分型；通过纸片扩散法(K-B法)和PCR法分别对分离株进行药敏试验和耐药基因检测。【结果】采集的108份犊牛腹泻粪便样本中Cp核酸阳性率为78.70%(85/108)，其单独感染阳性率为14.81%(16/108)，总混合感染率为63.89%(69/108)；Cp与BRV、BCoV的混合感染率分别为44.44%(48/108)和24.07%(26/108)。共分离得到13株Cp分离株，且均为A型。药敏试验结果显示，84.62%(11/13)分离株对庆大霉素表现耐药，76.92%(10/13)分离株对卡那霉素表现耐药。耐药基因检测结果显示，84.62%(11/13)分离菌株检出Acc(6')/ph(2″)基因，53.85%(7/13)分离菌株检出ermC基因，Sul1、qnrS、和ermB等7种耐药基因均未检出。【结论】甘肃地区犊牛腹泻混合感染较为严重，主要流行的Cp菌株为A型，多数Cp分离菌株对氨基糖苷类药物耐药严重且携带多种耐药基因。

关键词: 甘肃; 犊牛腹泻; 产气荚膜梭菌; 分离鉴定; 耐药性

Abstract: 【Objective】 This experiment was to investigate the prevalence of infectious pathogenic microorganisms causing diarrhea in calves from large-scale cattle farms in Gansu and to understand the resistance status of Clostridium perfringens (Cp) to common antibacterial drugs. 【Method】 5-combined nucleic acid detection kit for pathogenic microorganisms of calf diarrhea was used to test 108 fecal samples of calf diarrhea collected from 10 cattle farms in 8 regions including Lanzhou,Zhangye and Wuwei，which mainly included Cp,Bovine rotavirus (BRV),Bovine coronavirus (BCoV),enterotoxigenic Escherichia coli K99 (K99) and Cryptosporidium (COWP). Cp nucleic acid positive bovine fecal samples were subject to bacterial isolation and culture,and identified using staining microscopy and biochemical tests.At the same time,PCR amplification was used to type the isolated strains.The drug sensitivity test and drug resistance gene detection of the isolated strains were conducted by disk diffusion method (K-B) and PCR，respectively. 【Result】 Among the 108 fecal samples collected from calves with diarrhea,the positive rate of Cp nucleic acid was 78.70% (85/108),the positive rate of individual infection was 14.81% (16/108),and the total mixed infection rate was 63.89% (69/108).The mixed infection rates of Cp with BRV and BCoV was 44.44% (48/108) and 24.07% (26/108),respectively.A total of 13 Cp were isolated,and all of them were type A.The results of drug sensitivity test showed that 84.62% (11/13) of the isolated strains were resistant to gentamicin,and 76.92% (10/13) were resistant to kanamycin.The results of the resistance gene detection showed that 84.62% (11/13) of the isolated strains were detected with Acc(6')/ph(2″) gene,53.85% (7/13) of the isolated strains were detected with ermC gene,and none of the seven resistance genes such as Sul1,qnrS,and ermB were detected. 【Conclusion】 The mixed infection of calf diarrhea in Gansu region was relatively serious.The main prevalent Cp strain was type A，and most Cp isolated strains were severely resistant to aminoglycoside and carried multiple resistance genes.

Key words: Gansu; calf diarrhea; Clostridium perfringens; isolation and identification; drug resistance

中图分类号:

S852.61^＋6.3

赵雪剑, 陈晓良, 冯晓微, 张康, 马小军, 李建喜. 甘肃地区犊牛腹泻病原调查及产气荚膜梭菌的分离鉴定、耐药性分析[J]. 中国畜牧兽医, 2025, 52(8): 3927-3938.

ZHAO Xuejian, CHEN Xiaoliang, FENG Xiaowei, ZHANG Kang, MA Xiaojun, LI Jianxi. Investigation of the Pathogen of Calf Diarrhea in Gansu Region and Isolation, Identification and Drug Resistance Analysis of Clostridium perfringens[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(8): 3927-3938.

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