巴基斯坦商业鸡肉源大肠杆菌中介导黏菌素（mcr-1）和 ESBLs 基因协同耐药的分子机制。
Molecular mechanism of antimicrobial co-resistance Colistin (mcr-1) and ESBLs genes among Escherichia coli isolates from commercial chickens in Pakistan.
机构信息
Government College University, Department of Microbiology, Faisalabad, Pakistan.
University of Agriculture, Institute of Microbiology, Faisalabad, Pakistan.
出版信息
Braz J Biol. 2023 Jan 30;84:e267494. doi: 10.1590/1519-6984.267494. eCollection 2023.
Emergence of plasmid mediated colistin and extended spectrum β-lactamases (ESBL) resistant genes has been impacted the efficacy of colistin and β-lactams drugs like 3rd, 4th generation cephalosporin. Current study was aimed to investigate antimicrobial resistance genes (ARGs) among Escherichia coli isolates from meat producing commercial broilers in Pakistan. Two hundred (n=200) fecal samples were collected during January-2018 to August-2019. For isolation of E. coli, pink colonies on MacConkey agar were transferred to EMB agar. Metallic sheen color colonies were tested biochemically using API-20E kit. The molecular identification of E. coli (n=153) was targeted by amplification of uid gene through polymerase chain reaction (PCR) and different ARGs i.e. gentamicin, streptomycin, tetracycline, colistin, β-lactams drugs, quinolone and ampicillin followed by sequence analysis. Genotypically, followed by phenotypically of resistant ARGs of isolated PCR-confirmed E. coli (153) shoed resistant against gentamicin (aac(3)-IV), streptomycin (aadA1), tetracycline (tetA), colistine (mcr-1), ampicillin (bla-TEM) and bla-CTX-M were 86%, 88%, 86%, 88%, 83% & 77% respectively. 33/38 (86%) of the isolate was positive for quinolone resistance. Colistine (mcr-1), ESBLs (bla-TEM) and (bla-CTX-M) resistance genes were 88%, 83% and 77% respectively. About 33 isolated E. coli harbored the both mcr-1 and ESBLs genes. All of E. coli isolates were found sensitive to ceftriaxone (CTX-30) and imipenem (IMP-10). The Isolated E. coli showed single or multi clade decadency. The E. coli and ARGs sequences showed single or multi clade decadency. This is first comprehensive study from Pakistan that described the molecular evidences of ARGs and their co-existence in single isolates originated from commercial poultry. Commercial chicken (Broilers) can act as melting pot of antibiotic resistance genes for human being. It is alarming situation for surveillance of antibiotic resistance program because of more regulated prescription of antimicrobial agents in Pakistan.
质粒介导的粘菌素和超广谱β-内酰胺酶（ESBL）耐药基因的出现，已经影响了粘菌素和β-内酰胺类药物（如第三代、第四代头孢菌素）的疗效。本研究旨在调查巴基斯坦商业肉鸡生产中大肠杆菌分离株的抗菌药物耐药基因（ARGs）。2018 年 1 月至 2019 年 8 月期间，共采集了 200 份（n=200）粪便样本。为了分离大肠杆菌，在麦康凯琼脂上的粉红色菌落被转移到 EMB 琼脂上。采用 API-20E 试剂盒对具有金属光泽的菌落进行生化测试。通过聚合酶链反应（PCR）扩增 uid 基因，对 153 株大肠杆菌进行分子鉴定，然后对不同的 ARGs 进行检测，包括庆大霉素、链霉素、四环素、粘菌素、β-内酰胺类药物、喹诺酮类和氨苄西林，随后进行序列分析。表型和基因型分析显示，153 株经 PCR 确认的大肠杆菌分离株对庆大霉素（aac(3)-IV）、链霉素（aadA1）、四环素（tetA）、粘菌素（mcr-1）、氨苄西林（bla-TEM）和 bla-CTX-M 的耐药率分别为 86%、88%、86%、88%、83%和 77%。38 株中的 33 株（86%）对喹诺酮类药物具有耐药性。粘菌素（mcr-1）、ESBLs（bla-TEM）和（bla-CTX-M）耐药基因的耐药率分别为 88%、83%和 77%。约 33 株分离的大肠杆菌同时携带 mcr-1 和 ESBLs 基因。所有大肠杆菌分离株对头孢曲松（CTX-30）和亚胺培南（IMP-10）均敏感。分离的大肠杆菌显示出单或多分支衰退。大肠杆菌和 ARGs 序列显示出单或多分支衰退。这是巴基斯坦首次全面描述 ARGs 及其在单一分离株中的共存的研究，这些分离株来自商业家禽。商业鸡（肉鸡）可能成为人类抗生素耐药基因的“熔炉”。由于巴基斯坦对抗菌药物的使用更加规范，这对监测抗生素耐药性计划来说是一个令人担忧的情况。
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