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奶牛场环境中大肠杆菌抗菌药物耐药性的分子特征分析

Molecular characterization of antimicrobial resistance in Escherichia coli from dairy farm environment.

作者信息

Narwal Namita, Amarlapudi Monica Rose, Vinchurkar Ravikant V, Behera Manisha, De Sachinandan, Vishweswaraiah Raghu Hirikyathanahalli, Puniya Anil Kumar, Mallappa Rashmi Hogarehalli, Pradhan Diwas

机构信息

Molecular Biology Unit, Dairy Microbiology Division, ICAR - National Dairy Research Institute, Karnal, Haryana, 132001, India.

Animal Genomics Lab, Animal Biotechnology Centre, National Dairy Research Institute, Karnal, 32001, Haryana, India; Department of Zoology, Hindu College, University of Delhi, Delhi, 110007, India.

出版信息

Int J Hyg Environ Health. 2025 Jul;268:114607. doi: 10.1016/j.ijheh.2025.114607. Epub 2025 Jun 7.

Abstract

The present study was carried out to study the prevalence and genetic mechanisms of antimicrobial resistance in E. coli strains from dairy farms. A total of 60 E. coli strains were initially isolated from 192 dairy farm samples using a selective antibiotic approach and confirmed as E. coli by PCR. Among these, 48 E. coli isolates predominantly from fecal samples were further studied. These isolates were majorly classified in phylogroup A (43.75 %) and B1 (16.66 %) and showed predominant resistance against ampicillin (60.41 %) followed by piperacillin (56.25 %), tetracycline (54.16 %), and other β-lactams such as cefotaxime (47.91 %) and cefuroxime (43.75 %). A significant portion (22.9 %) of the E. coli isolates were multidrug-resistant (MDR) and 50 % were ESBL-positive. Multiple antibiotic resistance (MAR) index ≥0.4 was exhibited by three isolates. Genotypic analysis identified resistance genes associated with β-lactams (bla, 64.58 %; bla, 35.41 %; bla 4.16 %), tetracycline (tetA,37.58 %; tetB, 47.91 %), trimethoprim (dfrA17, 16.66 %), aminoglycosides [aac(6')-Ib-cr, 2.08 %] and fluoroquinolones [qnrB, 25 %; qnrS, 16.66 %; gyrB (S492N), 45.83 %; gyrA (S83L), 45.83 %; gyrA(S87L), 39.58 %; parC (S80I), 14.58 %]. E. coli isolates also showed a high frequency of mobile genetic elements (MGEs) such as IS26 (56.25 %), IncFIB plasmids (52.08 %), and Tn3 transposons (56.25 %). Class 1 integrons harbouring 200 and 800 bp gene cassettes were also detected in 5 E. coli isolates (10.4 %). Overall, this study highlights the high prevalence of diverse AMR genes in cattle-derived E. coli and their strong association with various MGEs. These findings emphasize the need for continuous genomic surveillance to mitigate resistance spread, particularly within and from dairy environments.

摘要

本研究旨在调查奶牛场大肠杆菌菌株中抗菌药物耐药性的流行情况及其遗传机制。最初采用选择性抗生素方法从192份奶牛场样本中分离出60株大肠杆菌菌株,并通过PCR确认为大肠杆菌。其中,对主要来自粪便样本的48株大肠杆菌分离株进行了进一步研究。这些分离株主要分为A群(43.75%)和B1群(16.66%),对氨苄西林的耐药性最为突出(60.41%),其次是哌拉西林(56.25%)、四环素(54.16%)以及其他β-内酰胺类药物,如头孢噻肟(47.91%)和头孢呋辛(43.75%)。相当一部分(22.9%)大肠杆菌分离株具有多重耐药性(MDR),50%为ESBL阳性。三株分离株的多重抗生素耐药(MAR)指数≥0.4。基因型分析确定了与β-内酰胺类(bla,64.58%;bla,35.41%;bla 4.16%)、四环素(tetA,37.58%;tetB,47.91%)、甲氧苄啶(dfrA17,16.66%)、氨基糖苷类[aac(6')-Ib-cr,2.08%]和氟喹诺酮类[qnrB,25%;qnrS,16.66%;gyrB (S492N),45.83%;gyrA (S83L),45.83%;gyrA(S87L),39.58%;parC (S80I),14.58%]相关的耐药基因。大肠杆菌分离株还显示出高频的可移动遗传元件(MGEs),如IS26(56.25%)、IncFIB质粒(52.08%)和Tn3转座子(56.25%)。在5株大肠杆菌分离株(10.4%)中还检测到携带200和800 bp基因盒的1类整合子。总体而言,本研究突出了牛源大肠杆菌中多种抗菌药物耐药基因的高流行率及其与各种MGEs的强关联。这些发现强调了持续进行基因组监测以减轻耐药性传播的必要性,特别是在奶牛场环境内部及其来源方面。

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