畜牧与饲料科学 ›› 2023, Vol. 44 ›› Issue (3): 109-115.doi: 10.12160/j.issn.1672-5190.2023.03.016

• 动物疾病防控 • 上一篇    下一篇

广州市某马术俱乐部马源大肠杆菌的分离鉴定、毒力基因检测及耐药性分析

刘鏊1,马丁允1,陀海欣1,李静1,王超男1,孙铭飞2,林栩慧2,齐萌1   

  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.广东省农业科学院动物卫生研究所/广东省畜禽疫病防治研究重点实验室,广东 广州 510640
  • 收稿日期:2023-03-13 出版日期:2023-05-30 发布日期:2023-07-12
  • 通讯作者: 齐萌(1985—),男,教授,博士,硕士生导师,主要从事人兽共患病病原微生物学研究工作。
  • 作者简介:刘鏊(1999—),男,硕士研究生,主要研究方向为动物群发病防控。
  • 基金资助:
    新疆生产建设兵团重点领域科技攻关计划项目(2020AB025)

Isolation,Identification,Virulence Genes Detection and Antimicrobial Resistance Profile of Horse Origin Escherichia coli Isolates from an Equestrian Club in Guangzhou City

LIU Ao1,MA Dingyun1,TUO Haixin1,LI Jing1,WANG Chaonan1,SUN Mingfei2,LIN Xuhui2,QI Meng1   

  1. 1. College of Animal Science and Technology,Tarim University/Engineering Laboratory of Tarim Animal Disease Diagnosis and Control,Xinjiang Production and Construction Corps,Alar 843300,China
    2. Institute of Animal Health,Guangdong Academy of Agricultural Sciences/Guangdong Provincial Key Laboratory of Livestock Disease Prevention,Guangzhou 510640,China
  • Received:2023-03-13 Online:2023-05-30 Published:2023-07-12

摘要:

[目的]分离鉴定广州市某马术俱乐部赛马粪便中的大肠杆菌,检测其毒力基因,并进行耐药性分析。[方法]收集广州市某马术俱乐部20份成年赛马粪便样本,通过细菌培养、革兰染色镜检和细菌16S rDNA序列PCR法分离鉴定大肠杆菌,并对分离菌株进行大肠杆菌系统发育群鉴定;采用PCR法检测10种毒力基因和11种耐药基因,应用K-B纸片法开展药物敏感性试验。[结果]经形态学观察和PCR检测,鉴定出13株大肠杆菌,分离率为65.00%(13/20);系统进化分群分别属于A群(23.07%,3/13)、B1群(61.54%,8/13)和D群(15.38%,2/13);共检出5种毒力基因,分别为fimC(61.54%,8/13)、fyuA(7.69%,1/13)、irp2(7.69%,1/13)、stx1(15.38%,2/13)和stx2(7.69%,1/13);共检出9种耐药基因,分别为parC(100%,13/13)、gyrA(100%,13/13)、gyrB(92.31%,12/13)、sul2(76.92%,10/13)、cmlA(30.77%,4/13)、aadA(23.07%,3/13)、tetB)(15.38%,2/13)、blaTEM(7.69%,1/13)和blaCTX(7.69%,1/13);13株大肠杆菌分离株对头孢西丁、庆大霉素、环丙沙星敏感,对四环素的耐药率为38.46%,对头孢噻呋、阿米卡星、氨苄西林的耐药率均为30.77%,对氯霉素、甲氧苄啶/磺胺异噁唑和亚胺培南也呈现出不同程度的耐药性。[结论]广州市某马术俱乐部赛马源大肠杆菌种群具有多样性,携带多种毒力基因和耐药基因,呈现多重耐药性,提示临床上应加强马大肠杆菌病的防治。

关键词: 大肠杆菌, 系统进化群, 毒力基因, 耐药基因,

Abstract:

[Objective] This study was conducted to isolate and identify Escherichia coli strains from the fecal samples of the racing horses in an equestrian club in Guangzhou City, and to characterize the virulence genes and antimicrobial resistance profile of the isolates. [Method] A total of 20 fecal samples were collected from the adult racing horses in an equestrian club in Guangzhou City. The bacterial culture technology, Gram staining and microscopy examination, and 16S rDNA PCR amplification and sequencing method were employed to isolate and identify the Escherichia coli strains. Subsequently, the Escherichia coli phylogenetic group identification was performed on the isolates, and the presence of 10 virulence genes and 11 antimicrobial resistance genes in the isolates was detected using PCR assay. In addition, the antimicrobial susceptibility test of the isolates was carried out using K-B disk method. [Result] A total of 13 Escherichia coli strains were isolated and identified by morphological observation and PCR detection, with an isolation rate of 65.00% (13/20). The phylogenetic group identification demonstrated that the 13 Escherichia coli isolates were classified as group A (23.07%, 3/13), group B1 (61.54%, 8/13), and group D (15.38%, 2/13), respectively. In the isolates, 5 virulence genes including fimC (61.54%, 8/13), fyuA (7.69%, 1/13), irp2 (7.69%, 1/13), stx1 (15.38%, 2/13), and stx2 (7.69%, 1/13) were detected, and 9 antimicrobial resistance genes including parC (100%, 13/13), gyrA (100%, 13/13), gyrB (92.31%,12/13), sul2 (76.92%, 10/13), cmlA (30.77%, 4/13), aadA (23.07%, 3/13), tet (B) (15.38%, 2/13), blaTEM (7.69%, 1/13), and blaCTX (7.69%, 1/13) were detected. Among the 13 Escherichia coli isolates, 38.46% of them were resistant to tetracycline, and 30.77% of them were resistant to ceftiofur, amikacin, and ampicillin. Different levels of resistance to chloramphenicol, trimethoprim/sulfafurazole, and imipenem was also observed. All of the isolates were susceptible to cefoxitin, gentamicin, and ciprofloxacin. [Conclusion] The community of the horse origin Escherichia coli strains isolated from the racing horses in this equestrian club was diversified. The isolates harbored multiple virulence genes and antimicrobial resistance genes, and exhibited multi-drug resistance. Our results suggest that the prevention and treatment of horse colibacillosis should be strengthened in clinical practice.

Key words: Escherichia coli, phylogenetic group, virulence gene, antimicrobial resistance gene, horse

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