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越南生肉和贝类的安全性:大肠杆菌分离株抗生素耐药性和毒力基因分析

Safety of raw meat and shellfish in Vietnam: an analysis of Escherichia coli isolations for antibiotic resistance and virulence genes.

作者信息

Van Thi Thu Hao, Chin James, Chapman Toni, Tran Linh Thuoc, Coloe Peter J

机构信息

Biotechnology and Environmental Biology, School of Applied Sciences, RMIT University, Bundoora West Campus, Bundoora, Melbourne, VIC 3083, Australia.

出版信息

Int J Food Microbiol. 2008 Jun 10;124(3):217-23. doi: 10.1016/j.ijfoodmicro.2008.03.029. Epub 2008 Mar 31.

Abstract

This study was conducted to examine a current baseline profile of antimicrobial resistance and virulence of Escherichia coli isolated from foods commonly sold in the market place in Vietnam. E. coli were isolated from 180 samples of raw meat, poultry and shellfish and also isolated from 43 chicken faeces samples. Ninety-nine E. coli isolates recovered from all sources were selected for the investigation of their susceptibility to 15 antimicrobial agents by the disk diffusion method. Eighty-four percent of the isolates were resistant to one or more antibiotics, and multi-resistance, defined as resistance to at least 3 different classes of antibiotics, was detected in all sources. The rates of multi-resistance were up to 89.5% in chicken, 95% in chicken faeces and 75% in pork isolates. Resistance was most frequently observed to tetracycline (77.8%), sulfafurazole (60.6%), ampicillin (50.5%), amoxicillin (50.5%), trimethoprim (51.5%), chloramphenicol (43.4%), streptomycin (39.4%), nalidixic acid (34.3%) and gentamicin (24.2%). In addition, the isolates also displayed resistance to fluoroquinolones (ciprofloxacin 16.2%, norfloxacin 17.2%, and enrofloxacin 21.2%), with chicken isolates showing the highest rates of resistance to these antibiotics (52.6-63.2%). Thirty-eight multi-resistant isolates were selected for further the examination of antibiotic resistance genes and were also evaluated for virulence gene profiles by multiplex and uniplex polymerase chain reaction. The beta-lactam TEM gene and tetracycline resistance tetA, tetB genes were frequently detected in the tested isolates (84.2% and 89.5% respectively). Genes which are responsible for resistance to streptomycin (aadA) (68.4%), chloramphenicol (cmlA) (42.1%), sulfonamides (sulI) (39.5%), trimethoprim (dhfrV) (26.3%) and kanamycin (aphA-1) (23.7%) were also widely distributed. Plasmid-mediated ampC genes were detected in E. coli isolates from chicken and pork. The isolates were tested for the presence of 58 virulence genes for adhesins, toxins, capsule synthesis, siderophores, invasins and others from different E. coli pathotypes. All of the tested isolates contained at least one virulence gene and there were 16 genes detected. Virulence genes detected were fimH (92.1%), bmaE (84.2%), TSPE4.C2 (42.1%), aidA AIDA-I (orfB) (31.6%), east1 (26.3%), traT (23.7%), and others including fyuA, iutA, chuA, yjaA, iss, iroN(E. coli), ibeA, aah (orfA), iha and papG allele III (10.5-2.6%). Typical toxin genes produced by enterohemorrhagic and enterotoxigenic E. coli pathotypes (a heat-stable toxin (ST), heat-labile toxin (LT) and Shiga toxin stx1, stx2) were not detected in any of these 38 isolates. The study has revealed that E. coli in raw foods is a significant reservoir of resistance and virulence genes.

摘要

本研究旨在检测从越南市场上常见食品中分离出的大肠杆菌的抗菌药物耐药性和毒力的当前基线概况。从180份生肉、家禽和贝类样本以及43份鸡粪便样本中分离出大肠杆菌。从所有来源回收的99株大肠杆菌分离株通过纸片扩散法被选择用于检测它们对15种抗菌药物的敏感性。84%的分离株对一种或多种抗生素耐药,并且在所有来源中均检测到多重耐药,即对至少3种不同类别的抗生素耐药。鸡的多重耐药率高达89.5%,鸡粪便中为95%,猪肉分离株中为75%。最常观察到对四环素(77.8%)、磺胺异恶唑(60.6%)、氨苄西林(50.5%)、阿莫西林(50.5%)、甲氧苄啶(51.5%)、氯霉素(43.4%)、链霉素(39.4%)、萘啶酸(34.3%)和庆大霉素(24.2%)耐药。此外,分离株还对氟喹诺酮类药物(环丙沙星16.2%、诺氟沙星17.2%、恩诺沙星21.2%)耐药,鸡分离株对这些抗生素的耐药率最高(52.6 - 63.2%)。选择38株多重耐药分离株进一步检测抗生素耐药基因,并通过多重和单重聚合酶链反应评估毒力基因概况。在测试的分离株中经常检测到β-内酰胺TEM基因和四环素耐药tetA、tetB基因(分别为84.2%和89.5%)。对链霉素(aadA)(68.4%)、氯霉素(cmlA)(42.1%)、磺胺类药物(sulI)(39.5%)、甲氧苄啶(dhfrV)(26.3%)和卡那霉素(aphA-1)(23.7%)耐药的基因也广泛分布。在来自鸡和猪肉的大肠杆菌分离株中检测到质粒介导的ampC基因。对这些分离株检测了来自不同大肠杆菌致病型的58种黏附素、毒素、荚膜合成、铁载体、侵袭素等毒力基因的存在情况。所有测试分离株至少含有一个毒力基因,共检测到16个基因。检测到的毒力基因有fimH(92.1%)、bmaE(84.2%)、TSPE4.C2(42.1%)、aidA AIDA-I(orfB)(31.6%)、east1(26.3%)、traT(23.7%),以及其他包括fyuA、iutA、chuA、yjaA、iss、iroN(大肠杆菌)、ibeA、aah(orfA)、iha和papG等位基因III(10.5 - 2.6%)。在这38株分离株中均未检测到肠出血性和产肠毒素大肠杆菌致病型产生的典型毒素基因(热稳定毒素(ST)、热不稳定毒素(LT)和志贺毒素stx1、stx2)。该研究表明,生食中的大肠杆菌是耐药基因和毒力基因的重要储存库。

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