伴侣动物源肺炎克雷伯氏菌的分离鉴定及毒力和耐药性分析

doi:10.16431/j.cnki.1671-7236.2020.05.033

摘要/Abstract

摘要： 为探究南宁伴侣动物源肺炎克雷伯氏菌的毒力和耐药情况，本研究采集犬、猫粪便拭子，通过分离培养、形态学观察、药敏试验及PCR扩增16S rRNA、khe基因、毒力基因及耐药基因等方法对细菌特性进行分析。结果显示，分离的菌株中有4株能在麦康凯培养基上形成液状菌落，轻挑拉丝且镜检为短杆状革兰氏阴性菌，疑为肺炎克雷伯氏菌。16S rRNA测序结果显示，分离菌与肺炎克雷伯氏菌同源性达99%，同时肺炎克雷伯氏菌特异性基因（khe）阳性。其中，分离株GXKP-C14、GXKP-D15对大部分临床常用抗菌药物敏感，分离株GXKP-D1则表现出高水平多重耐药，分离株GXKP-D4耐药性稍低于GXKP-D1，对临床常用药氨苄西林、哌拉西林、头孢呋辛、四环素、多西环素、呋喃妥因、复方新诺明表现耐药，对头孢吡肟、美罗培南、亚胺培南、阿米卡星、庆大霉素、链霉素、多黏菌素等敏感。部分菌株携带tet（A）、QnrS、bla_SHV、sul2、mcr-1等耐药基因和WabG毒力基因。本研究结果为犬、猫源肺炎克雷伯氏菌病的检测、诊断及治疗提供了试验依据。同时，黏菌素耐药基因mcr-1的检出将为多黏菌素的耐药性控制和合理使用提供参考。

关键词: 肺炎克雷伯氏菌; 伴侣动物; 多重耐药; 毒力

Abstract: In order to investigate the virulence and drug resistance of Klebsiella pneumoniae from Nanning companion animals,the fecal swabs of dogs and cats were collected,and the bacterial characteristics were analyzed by isolation and culture,morphological observation,drug sensitivity test and PCR amplification of 16S rRNA,khe gene,virulence genes and drug resistance genes.The results showed that four of the isolated strains could form a liquid colony on MacConkey medium,and they were short rod-shaped Gram-negative bacteria,suspected to be Klebsiella pneumoniae.The results of 16S rRNA sequencing showed that the homology between the isolated strain and Klebsiella pneumoniae was 99%,and Klebsiella pneumoniae specific gene (khe) was positive.Among them,GXKP-C14 and GXKP-D15 were sensitive to most commonly used antibiotics,GXKP-D1 showed high level of multi-drug resistance,followed by GXKP-D4 which showed ampicillin,piperacillin,cefuroxime,tetracycline,doxycycline,nitrofurantoin and sulfamethoxazole resistance,but sensitive to cefepime,meropenem,imipenem,amikacin,gentamicin,streptomycin and colistin.Some strains carried drug-resistant genes such as tet(A),QnrS,bla_SHV,sul2,mcr-1 and virulence gene WabG.The results of this study provided experimental basis for the detection,diagnosis and treatment of Klebsiella pneumoniae in dogs and cats.At the same time,the detection of colistin resistance gene mcr-1 would provide a reference for the control and rational use of polymyxin resistance.

Key words: Klebsiella pneumoniae; companion animals; multiple drug resistance; virulence

中图分类号:

S852.61

张聪, 肖亦辰, 陈怀君, 袁敬知, 韦德源, 王晓晔. 伴侣动物源肺炎克雷伯氏菌的分离鉴定及毒力和耐药性分析[J]. 中国畜牧兽医, 2020, 47(5): 1583-1592.

ZHANG Cong, XIAO Yichen, CHEN Huaijun, YUAN Jingzhi, WEI Deyuan, WANG Xiaoye. Isolation,Identification and Analysis of Virulence and Drug Resistance of Klebsiella pneumoniae from Companion Animals[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(5): 1583-1592.

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