广西猪源产肠毒素大肠杆菌毒力因子及多重耐药性分析

doi:10.16431/j.cnki.1671-7236.2019.03.033

摘要/Abstract

摘要：

本研究旨在探究广西某猪场可能存在的病原菌及其致病的主要原因。试验采用细菌分离纯化、形态学鉴定、生长曲线测定、生化分析及药敏试验等进行细菌的分离鉴定，应用PCR技术对菌株16S rRNA、毒力基因及耐药基因进行检测。结果表明，该由病料分离的菌株为无芽孢、有菌毛、两端钝圆的粗短杆状的革兰氏阴性菌，其繁殖能力强、生长迅速，疑似为大肠杆菌。16S rRNA扩增结果显示，出现大小为1 474 bp的条带，测序结果表明其与大肠杆菌的同源性可达99%。该菌株对31种常见抗菌药物表现出极强的耐药性，对庆大霉素、替米考星、四环素、恩诺沙星、青霉素、林可霉素、磺胺甲噁唑等均呈现较高的耐药水平，耐药率高达87.10%（27/32），仅对阿米卡星、黏菌素2种药物敏感。毒力因子检测共鉴定致病岛FyuA、肠毒素STb和LT及黏附素F41和K88共5种毒力因子，检出率为35.70%（5/14）。扩增出β-内酰胺酶类bla_TEM和bla_OXA、四环素类tet（A）、磺胺类Sul2、氨基糖苷类aadA1和aac（3'）-Ⅱa、喹诺酮类GyrA、GyrB和ParC及氯霉素floR共10种耐药基因，检出率为66.70%（10/15），且耐药基因的检出与耐药表型呈正相关。综上，该菌株含多种毒力因子，耐药型复杂，多重耐药性情况严重，需采取相应的预防措施，防止蔓延和传播。

关键词: 产肠毒素; 多重耐药; 大肠杆菌; 毒力因子

Abstract:

This study was aimed to investigate the possible pathogens and the main causes of disease in a pig farm in Guangxi.The bacteria were isolated and identified by bacterial separation and purification,morphological identification,growth curve measurement,biochemical analysis and drug susceptibility test.The strain 16S rRNA,virulence genes and drug resistance genes were detected by PCR.The results showed that the strain isolated from the disease material was a Gram-negative bacterium with no spores,pili,and round and short rods at both ends,which had strong reproductive ability and rapid growth,and was suspected to be Escherichia coli (E.coli).The 16S rRNA amplification results showed that a 1 474 bp band appeared,and the sequencing results showed that the homology of the isolate with E.coli was 99%.The strain showed strong resistance to 31 antibacterial drugs gentamicin,tilmicosin,tetracycline,enrofloxacin,penicillin,lincomycin,sulfamethoxazole,etc.,all showed higher levels of drug resistance,and the drug resistance rate was as high as 87.10% (27/32).It was only sensitive to two drugs,amikacin and colistin.Toxicity factor detection identified a total of five virulence factors,FyuA,enterotoxin STb and LT,adhesin F41 and K88.The detection rate was 35.70% (5/14).It had been amplified β-lactamases bla_TEM and bla_OXA,tetracycline tet(A),sulfonamide Sul2,aminoglycosides aadA1 and aac(3')-Ⅱa,quinolone GyrA,GyrB and ParC,amide alcohol floR 10 drug resistance genes,the detection rate was 66.70% (10/15),and the detection of drug resistance genes was positively correlated with drug resistance phenotype.In summary,the strain presented a variety of complex virulence factors,complex drug-resistance,and serious multi-drug resistance,and corresponding precautions need to be taken to prevent spread and spread.

Key words: enterotoxin production; multi-drug resistance; Escherichia coli; virulence factor

中图分类号:

S852.61

葛晨玲, 石大丽, 胡文, 宋星星, 葛强, 莫玉鹏, 胡传活, 王晓晔. 广西猪源产肠毒素大肠杆菌毒力因子及多重耐药性分析[J]. 《中国畜牧兽医》, 2019, 46(3): 913-923.

GE Chenling, SHI Dali, HU Wen, SONG Xingxing, GE Qiang, MO Yupeng, HU Chuanhuo, WANG Xiaoye. Analysis of Virulence Factors and Multidrug Resistance of Enterotoxin-producing Escherichia coli in Guangxi[J]. , 2019, 46(3): 913-923.

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