藏猪源粪肠球菌的分离鉴定及耐药基因与毒力基因检测

doi:10.16431/j.cnki.1671-7236.2025.04.041

摘要/Abstract

摘要： 【目的】对采集于西藏自治区林芝市的藏猪样品进行粪肠球菌分离鉴定并明确其耐药基因与毒力基因情况，为藏猪源肠球菌病的病原学诊断提供一定的参考。【方法】对采集的藏猪样品进行细菌分离培养、革兰染色、生化试验、Ef0027特异性基因PCR鉴定、16S rRNA鉴定，并通过PCR方法检测分离株的耐药基因和毒力基因。【结果】分离菌在PSE琼脂培养基上出现棕黑色菌落，疑似粪肠球菌。革兰染色镜检显示，分离菌呈单个、成双的卵圆形球菌。生化鉴定结果显示，分离菌阿拉伯糖、鼠李糖、棉籽糖呈阴性，葡萄糖、山梨醇、蔗糖、乳糖、精氨酸双水解酶、甘露醇呈阳性。PCR扩增结果显示，获得大小约为518 bp的Ef0027基因条带，16S rRNA基因扩增出大小约为1 500 bp的条带，与NCBI中登录的粪肠球菌序列相似性均＞98%。本研究分离鉴定出8株粪肠球菌。8株菌均检出tetB、catⅠ、gyrA、bla_TEM-₁、aac3、SUL-Ⅰ耐药基因，未检出fosA7基因。毒力基因检测结果显示，8株菌均检出Esp、EfaA、GelE和Agg基因，CylA基因检出率为87.5%，未检出Ace基因。【结论】本研究分离出8株藏猪源粪肠球菌，均具有多重耐药性且含有多种耐药基因和毒力基因。本研究结果可为藏猪源粪肠球菌病的病原学诊断提供一定的参考。

关键词: 粪肠球菌; 分离鉴定; 耐药基因; 毒力基因

Abstract: 【Objective】 This study was aimed to isolate and identify Enterococcus faecalis and determine the status of drug resistance and virulence genes in samples collected from Tibetan pigs in Linzhi,Tibet,and provide a certain reference for the pathogenic diagnosis of enterococcosis in Tibetan pigs.【Method】 Bacterial isolation and culture,Gram staining,biochemical experiments,Ef0027 specific gene PCR identification,16S rRNA identification were performed on samples collected from Tibetan pigs,and the drug resistance and virulence genes of the isolated strains were detected by PCR method.【Result】 The isolated bacteria appeared brown black colonies on PSE agar medium,suspected to be Enterococcus faecalis.Gram staining microscopy showed that the isolated bacteria appeared as single or double oval shaped cocci.The biochemical identification results showed that the isolated bacteria were negative for arabinose, rhamnose and raffinose, while they were positive for glucose, sorbitol, sucrose, lactose, arginine dihydrolase and mannitol. PCR amplification results showed that an Ef0027 gene band with a size of approximately 518 bp was obtained.A band with a size of approximately 1 500 bp was amplified from 16S rRNA gene,and the similarity with the sequence of Enterococcus faecalis registered in NCBI was greater than 98%.Eight strains of Enterococcus faecalis were isolated and identified in this study.tetB,catⅠ,gyrA,bla_TEM-1,aac3 and SUL-Ⅰ resistance genes were detected in all 8 strains,while fosA7 gene was not detected.The results of virulence gene detection showed that Esp,EfaA,GelE and Agg genes were detected in all the 8 strains,the detection rate of CylA gene was 87.5%,and no Ace gene was detected.【Conclusion】 In this study,8 strains of Enterococcus faecalis of Tibetan pig origin were isolated,all of which had multiple drug resistance genes and virulence genes.The results of this study could provide some reference for the etiological diagnosis of Enterococcus faecalis disease from Tibetan pigs.

Key words: Enterococcus faecalis; isolation and identification; drug resistance gene; virulence gene

中图分类号:

S852.61⁺1

何宇轩, 次仁卓玛, 王宇, 刘怀志, 杨金鹏, 韦明邦, 商鹏. 藏猪源粪肠球菌的分离鉴定及耐药基因与毒力基因检测[J]. 中国畜牧兽医, 2025, 52(4): 1895-1904.

HE Yuxuan, CIRING Zhuoma, WANG Yu, LIU Huaizhi, YANG Jinpeng, WEI Mingbang, SHANG Peng. Isolation and Identification of Tibetan Pig-derived Enterococcus faecalis and Detection of Drug Resistance and Virulence Genes[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(4): 1895-1904.

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