新疆维吾尔自治区阿克苏地区某屠宰场羊源大肠杆菌MLST分型、耐药性分析和毒力基因检测

doi:10.12160/j.issn.1672-5190.2024.02.012

畜牧与饲料科学 ›› 2024, Vol. 45 ›› Issue (2): 95-102.doi: 10.12160/j.issn.1672-5190.2024.02.012

新疆维吾尔自治区阿克苏地区某屠宰场羊源大肠杆菌MLST分型、耐药性分析和毒力基因检测

李广西¹, 韩婷婷¹, 吴自豪², 吴静¹

1.塔里木大学动物科学与技术学院/新疆生产建设兵团塔里木动物疫病诊断与防控工程实验室,新疆阿拉尔 843300;
2.塔里木大学生命科学与技术学院,新疆阿拉尔 843300

收稿日期:2024-02-05 出版日期:2024-03-30 发布日期:2024-05-06
通讯作者: 吴静（1978—）,女,副教授,博士研究生,硕士生导师,主要研究方向为分子病原学。
作者简介:李广西（2002—）,男,所学专业为动物医学。
基金资助:
新疆生产建设兵团第九师科技计划项目“牛羊重要细菌性传染病的综合防治与示范”（2021JS001）

Multilocus Sequence Typing （MLST）,Antimicrobial Resistance Analysis and Virulence Gene Detection of Escherichia coli Isolated from Sheep in a Slaughterhouse in Aksu Prefecture, Xinjiang Uygur Autonomous Region

LI Guangxi¹, HAN Tingting¹, WU Zihao², WU Jing¹

1. College of Animal Science and Technology,Tarim University/Engineering Laboratory of Tarim Animal Diseases Diagnosis and Control,Xinjiang Production and Construction Corps,Aral 843300,China;
2. College of Life Science and Technology,Tarim University,Aral 843300,China

Received:2024-02-05 Online:2024-03-30 Published:2024-05-06

摘要/Abstract

摘要： [目的]了解新疆维吾尔自治区阿克苏地区屠宰场羊源大肠杆菌的流行情况以及耐药性和毒力基因分布特征。[方法]从阿克苏地区某屠宰场采集健康羊的淋巴结、胴体拭子、粪便样本共30份,使用选择性培养基分离大肠杆菌,然后采用大肠杆菌特异性phoA基因对其进行分子生物学鉴定;利用PCR扩增及测序技术对大肠杆菌分离株进行多位点序列分型（multilocus sequence typing,MLST）;采用K-B纸片扩散法开展分离菌株对抗菌药物的敏感性试验;应用PCR技术检测分离菌株的耐药基因和毒力基因携带情况;使用96孔板微量肉汤法测定分离菌株的生物被膜形成能力。[结果]从30份样本中分离得到14株大肠杆菌,分离率为46.67%;14株分离株分属于10个MLST型别;对复方新诺明（100%）、青霉素（85.71%）和四环素（50.00%）耐药率较高,9株菌具有多重耐药表型（64.29%）;在14株大肠杆菌中检测到bla_TEM（85.71%）、aph（3′）（71.43%）、tetA（64.29%）、sul1（57.14%）和sul2（57.14%）等12种耐药基因,并且8株菌携带Ⅰ型整合子int-Ⅰ基因（57.14%）;在14株大肠杆菌中检测到fimH（100%）、yijP（92.86%）、mat（92.86%）、sheA（92.86%）、stx1（35.71%）和ibeB（28.57%）等10种毒力基因;10株大肠杆菌具有生物被膜形成能力（71.43%）。[结论]该屠宰场羊源大肠杆菌种内分型丰富,具有较强的耐药性,还可形成生物被膜,并且携带多种耐药基因和毒力基因,对动物性食品安全可能存在潜在威胁。

关键词: 大肠杆菌, MLST分型, 耐药性, 毒力基因, 生物被膜

Abstract: [Objective] The aim of this study was to understand the prevalence as well as antimicrobial resistance and virulence gene distribution of Escherichia coli in sheep in a slaughterhouse in Aksu Prefecture, Xinjiang Uygur Autonomous Region. [Method] A total of 30 samples of lymph node, carcass swabs and feces were collected from the healthy sheep in a slaughterhouse in Aksu Prefecture. Escherichia coli was isolated using selective culture medium, and subsequently molecularly identified using Escherichia coli specific phoA gene. Multilocus sequence typing （MLST） of the isolates was performed by PCR amplification and sequencing. K-B disk diffusion method was used for antimicrobial susceptibility test of the isolates. The presence of the antimicrobial resistance and virulence genes of the isolates were detected by PCR assay. The 96-well plate micro-broth method was used to determine the biofilm formation ability of the isolates. [Result] A total of 14 strains of Escherichia coli were isolated from the 30 samples, with a separation rate of 46.67%. The 14 isolates belonged to 10 MLST types. High resistance rates to sulfamethoxazole-trimethoprim （100%）, penicillin （85.71%） and tetracycline （50.00%） were observed among the isolates. Nine isolates （64.29%） expressed multi-drug resistance phenotype. Twelve antimicrobial resistance genes, including bla_TEM （85.71%）, aph （3′）（71.43%）, tetA （64.29%）, sul1 （57.14%） and sul2 （57.14%） were detected in the 14 isolates, and 8 isolates （57.14%） harboured the typeⅠintegron int-Ⅰgene. Ten virulence genes, including fimH （100%）, yijP （92.86%）, mat （92.86%）, sheA （92.86%）, stx1 （35.71%） and ibeB （28.57%） were detected in the 14 isolates. Ten isolates （71.43%） had the ability to form biofilms. [Conclusion] The sheep derived Escherichia coli strains in the slaughterhouse had abundant intraspecific subtypes, increased antimicrobial resistance and the ability to form biofilms. They also carried multiple antimicrobial resistance and virulence genes, and thus posed a potential threat to animal food safety.

Key words: Escherichia coli, MLST, antimicrobial resistance, virulence gene, biofilm

中图分类号:

S852.612

李广西, 韩婷婷, 吴自豪, 吴静. 新疆维吾尔自治区阿克苏地区某屠宰场羊源大肠杆菌MLST分型、耐药性分析和毒力基因检测[J]. 畜牧与饲料科学, 2024, 45(2): 95-102.

LI Guangxi, HAN Tingting, WU Zihao, WU Jing. Multilocus Sequence Typing （MLST）,Antimicrobial Resistance Analysis and Virulence Gene Detection of Escherichia coli Isolated from Sheep in a Slaughterhouse in Aksu Prefecture, Xinjiang Uygur Autonomous Region[J]. Animal Husbandry and Feed Science, 2024, 45(2): 95-102.

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新疆维吾尔自治区阿克苏地区某屠宰场羊源大肠杆菌MLST分型、耐药性分析和毒力基因检测

Multilocus Sequence Typing （MLST）,Antimicrobial Resistance Analysis and Virulence Gene Detection of Escherichia coli Isolated from Sheep in a Slaughterhouse in Aksu Prefecture, Xinjiang Uygur Autonomous Region

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