Bhattarai Rebanta K, Basnet Hom B, Dhakal Ishwari P, Devkota Bhuminand
Department of Veterinary Microbiology and Parasitology, Faculty of Animal Science, Veterinary Science and Fisheries, Agriculture and Forestry University, Nepal.
Department of Medicine and Public Health, Faculty of Animal Science, Veterinary Science and Fisheries, Agriculture and Forestry University, Nepal.
Vet World. 2024 Feb;17(2):480-499. doi: 10.14202/vetworld.2024.480-499. Epub 2024 Feb 29.
Antimicrobials are extensively used in poultry production for growth promotion as well as for the treatment and control of diseases, including avian pathogenic (APEC). Poor selection, overuse, and misuse of antimicrobial agents may promote the emergence and dissemination of antimicrobial resistance (AMR) in APEC. This study aimed to assess antimicrobial susceptibility patterns and detect antibiotic resistance genes (ARGs) in APEC isolated from clinical cases of colibacillosis in commercial broiler, layer, and breeder chickens.
A total of 487 APEC were isolated from 539 across 300 poultry farms in various regions of Nepal. Antimicrobial susceptibility patterns was determined using the Kirby-Bauer disk diffusion and broth microdilution methods. The index of AMR, such as multiple antibiotic resistance (MAR) index, resistance score (R-score), and multidrug resistance (MDR) profile, were determined. Polymerase chain reaction was employed to detect multiple ARGs and correlations between phenotypic and genotypic resistance were analyzed.
The prevalence of APEC was 91% (487/539). All of these isolates were found resistant to at least one antimicrobial agent, and 41.7% of the isolates were resistant against 8-9 different antimicrobials. The antibiogram of APEC isolates overall showed the highest resistance against ampicillin (99.4%), whereas the highest intermediate resistance was observed in enrofloxacin (92%). The MAR index and R-score showed significant differences between broiler and layers, as well as between broiler breeder and layers. The number of isolates that were resistant to at least one agent in three or more antimicrobial categories tested was 446 (91.6%) and were classified as MDR-positive isolates. The ARGs were identified in 439 (90.1%) APEC isolates, including the most detected mobilized colistin resistanc (1) which was detected in the highest (52.6%) isolates. Overall, resistance gene of beta-lactam (), , resistance gene of sulphonamide () and resistance gene of tetracycline () (in broiler), were detected in significantly higher than other tested genes (p < 0.001). When examining the pair-wise correlations, a significant phenotype-phenotype correlation (p < 0.001) was observed between levofloxacin and ciprofloxacin, chloramphenicol and tetracycline with doxycycline. Similarly, a significant phenotype-genotype correlation (p < 0.001) was observed between chloramphenicol and the , and colistin with and resistance gene of quinolone ().
In this study, the current state of APEC AMR in commercial chickens is revealed for the first time in Nepal. We deciphered the complex nature of AMR in APEC populations. This information of molecular surveillance is useful to combat AMR in APEC and to contribute to manage APEC associated diseases and develop policies and guidelines to enhance the commercial chicken production.
抗菌药物在禽类生产中被广泛用于促进生长以及治疗和控制包括禽致病性大肠杆菌(APEC)在内的疾病。抗菌药物的选择不当、过度使用和滥用可能会促使APEC中抗菌药物耐药性(AMR)的出现和传播。本研究旨在评估从商业肉鸡、蛋鸡和种鸡大肠杆菌病临床病例中分离出的APEC的抗菌药物敏感性模式,并检测抗生素耐药基因(ARGs)。
从尼泊尔不同地区300个家禽养殖场的539份样本中总共分离出487株APEC。采用 Kirby-Bauer 纸片扩散法和肉汤微量稀释法测定抗菌药物敏感性模式。确定了AMR指数,如多重抗生素耐药性(MAR)指数、耐药评分(R评分)和多重耐药性(MDR)谱。采用聚合酶链反应检测多种ARGs,并分析表型耐药性和基因型耐药性之间的相关性。
APEC的流行率为91%(487/539)。所有这些分离株均被发现对至少一种抗菌药物耐药,41.7%的分离株对8 - 9种不同的抗菌药物耐药。APEC分离株的抗菌谱总体上显示对氨苄西林的耐药性最高(99.4%),而对恩诺沙星的中介耐药性最高(92%)。MAR指数和R评分在肉鸡和蛋鸡之间以及肉种鸡和蛋鸡之间存在显著差异。在测试的三个或更多抗菌类别中对至少一种药物耐药的分离株数量为446株(91.6%),被归类为MDR阳性分离株。在439株(90.1%)APEC分离株中鉴定出ARGs,其中检测到最多的是可移动黏菌素耐药基因(mcr-1),在最高比例(52.6%)的分离株中被检测到。总体而言,β-内酰胺类耐药基因(blaTEM)、磺胺类耐药基因(sul1)和四环素类耐药基因(tetA,在肉鸡中)的检测率显著高于其他测试基因(p < 0.001)。在检查成对相关性时,观察到左氧氟沙星和环丙沙星、氯霉素和四环素与强力霉素之间存在显著的表型 - 表型相关性(p < 0.001)。同样,观察到氯霉素与catA1、黏菌素与mcr-1和喹诺酮类耐药基因(qnrS)之间存在显著的表型 - 基因型相关性(p < 0.001)。
在本研究中,首次在尼泊尔揭示了商业鸡中APEC AMR的现状。我们解读了APEC群体中AMR的复杂性质。这种分子监测信息有助于对抗APEC中的AMR,并有助于管理与APEC相关的疾病,以及制定政策和指南以提高商业鸡生产。