浣熊源粪肠球菌的分离鉴定与耐药性分析

doi:10.16431/j.cnki.1671-7236.2021.03.036

摘要/Abstract

摘要： 为探究从四川某动物园1例病死浣熊肝脏中分离到的1株细菌的特性和耐药程度，本试验采用病原分离、革兰氏染色、生化鉴定、16S rRNA测序、致病性试验、药敏纸片扩散法及耐药基因扩增的方法对该分离株进行了形态学及生化鉴定、致病能力和耐药程度分析。结果显示，分离株在普通LB培养基上呈现表面光滑、圆形凸起、边缘整齐、针尖大小的半透明菌落。经革兰氏染色在油镜下观察到蓝紫色球菌，测序结果与GenBank登录号为KY003107.1粪肠球菌（Enterococcus faecalis）的同源性高达99.5%，形态学与生化鉴定特性与粪肠球菌特性相符。致病性试验表明，该菌可导致肝细胞肿胀、变性，具有一定的致病能力；药敏纸片和耐药基因扩增试验表明，该菌对14种常见的抗生素头孢西丁、头孢哌酮、头孢曲松、头孢吡肟、头孢噻肟、氨苄西林、卡那霉素、链霉素、奈替米星、丁胺卡那、阿奇霉素、红霉素、多西环素、四环素耐药，仅对万古霉素和庆大霉素中度敏感。四环素耐药基因tetC检测为阳性；tetM、tetA检测为阴性，氨基糖苷类耐药基因aac （6'）/aph（2″）检测为阳性，aph（3'）-Ⅲ、ant（6）-Ⅰ检测为阴性；大环内酯类耐药基因ermB检测为阳性，ermA、ermC检测为阴性；万古霉素耐药基因vanA、vanB和β-内酰胺类耐药基因TEM检测均为阴性。该试验从浣熊肝脏分离的粪肠球菌表现出多重耐药特性并具有一定的致病能力，结果可为动物园临床肠球菌感染的病例提供一定的药物选择参考，制定合理的感染防治方案。

关键词: 粪肠球菌; 生化鉴定; 16S rRNA; 致病性试验; 药敏试验; 耐药基因

Abstract: To investigate the characteristics and antimicrobial resistance of a dominant bacterium isolated from the liver of a dead raccoon in a zoo in Sichuan province.In this study,Gram staining,biochemical identification,16S rRNA sequence determination,pathogenicity test,antimicrobial susceptibility test and resistance gene amplification test were used to conduct morphological,biochemical identification and study on pathogenicity and drug resistance of the isolated strain.The result showed that the isolated strain showed a translucent colony with smooth surface,round protrusion,neat edge and needle tip size on LB medium.Blue-purple cocci could be observed with Gram staining under oil microscope.The isolate was 99.5% similar to Enterococcus faecalis whose GenBank accession No. was KY003107.1,and the morphological and biochemical characteristics were consistent with those of Enterococcus faecalis.The pathogenicity test showed that the bacterium could cause hepatocytes swelling and degeneration,and had certain pathogenicity.Antimicrobial susceptibility test and resistance gene amplification test showed that the strain was resistant to 14 common antibiotics,cefoxitin,cefoperazone,ceftriaxone,cefepime,cefotaxime,ampicillin,kanamycin,streptomycin,netimicin,butanicin,azithromycin,erythromycin,doxycycline and tetracycline,and was only moderately sensitive to vancomycin and gentamicin.Tetracycline resistance gene tetC was positive,tetM and tetA gene were negative.Aminoglycoside resistance gene aac(6')/aph(2″) was positive,aph(3')-Ⅲ and ant(6)-Ⅰ were negative.The ermB of macrolide resistance genes was positive and ermA and ermC were negative.Vancomycin resistance gene vanA,vanB and β-lactam resistance gene TEM were all negative.In this study,the Enterococcus faecalis isolated from raccoons showed high drug resistance and pathogenicity,which could be used as a reference for the selection of drugs for clinical Enterococcus faecalis infection,and a reasonable infection prevention and treatment scheme could be formulated.

Key words: Enterococcus faecalis; biochemical identification; 16S rRNA; pathogenicity test; antimicrobial susceptibility test; resistance genes

中图分类号:

Q93-337

钟锐, 何廷美, 毕波, 杨兴涛, 施小刚, 金森龙, 冯刚, 彭广能. 浣熊源粪肠球菌的分离鉴定与耐药性分析[J]. 中国畜牧兽医, 2021, 48(3): 1112-1120.

ZHONG Rui, HE Tingmei, BI Bo, YANG Xingtao, SHI Xiaogang, JIN Senlong, FENG Gang, PENG Guangneng. Isolation,Identification and Drug Resistance Analysis of Enterococcus faecalis from Raccoon[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(3): 1112-1120.

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