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利用全基因组测序对源自奥利亚罗非鱼养殖场的致病性大肠杆菌菌株进行特性分析。

Characterization of a Pathogenic Escherichia coli Strain Derived from Oreochromis spp. Farms Using Whole-Genome Sequencing.

作者信息

Magaña-Lizárraga José Antonio, Gómez-Gil Bruno, Enciso-Ibarra Julissa, Báez-Flores María Elena

机构信息

Unidad de Investigaciones en Salud Pública "Dra. Kaethe Willms", Facultad de Ciencias Químico Biológicas, Universidad Autónoma de Sinaloa, Ciudad Universitaria.

Centro de Investigación en Alimentación y Desarrollo, A. C. (CIAD), Unidad Mazatlán en Acuicultura y Manejo Ambiental.

出版信息

J Vis Exp. 2022 Dec 23(190). doi: 10.3791/64404.

Abstract

Aquaculture is one of the fastest-growing food-producing sectors worldwide and tilapia (Oreochromis spp.) farming constitutes the major freshwater fish variety cultured. Because aquaculture practices are susceptible to microbial contamination derived from anthropogenic sources, extensive antibiotic usage is needed, leading to aquaculture systems becoming an important source of antibiotic-resistant and pathogenic bacteria of clinical relevance such as Escherichia coli (E. coli). Here, the antimicrobial resistance, virulence, and mobilome features of a pathogenic E. coli strain, recovered from inland farmed Oreochromis spp., were elucidated through whole-genome sequencing (WGS) and in silico analysis. Antimicrobial susceptibility testing (AST) and WGS were performed. Furthermore, phylogenetic group, serotype, multilocus sequence typing (MLST), acquired antimicrobial resistance, virulence, plasmid, and prophage content were determined using diverse available web tools. The E. coli isolate only exhibited intermediate susceptibility to ampicillin and was characterized as ONT:H21-B1-ST40 strain by WGS-based typing. Although only a single antimicrobial resistance-related gene was detected [mdf(A)], several virulence-associated genes (VAGs) from the atypical enteropathogenic E. coli (aEPEC) pathotype were identified. Additionally, the cargo of plasmid replicons from large plasmid groups and 18 prophage-associated regions were detected. In conclusion, the WGS characterization of an aEPEC isolate, recovered from a fish farm in Sinaloa, Mexico, allows insights into its pathogenic potential and the possible human health risk of consuming raw aquacultural products. It is necessary to exploit next-generation sequencing (NGS) techniques for studying environmental microorganisms and to adopt a one health framework to learn how health issues originate.

摘要

水产养殖是全球增长最快的粮食生产部门之一,罗非鱼(Oreochromis spp.)养殖是养殖的主要淡水鱼品种。由于水产养殖活动易受人为来源的微生物污染影响,因此需要大量使用抗生素,这导致水产养殖系统成为具有临床相关性的抗生素耐药菌和病原菌(如大肠杆菌)的重要来源。在此,通过全基因组测序(WGS)和计算机分析,阐明了从内陆养殖的罗非鱼中分离出的一株致病性大肠杆菌菌株的抗菌耐药性、毒力和可移动基因组特征。进行了抗菌药敏试验(AST)和WGS。此外还使用了各种可用的网络工具来确定系统发育群、血清型、多位点序列分型(MLST)、获得性抗菌耐药性、毒力、质粒和前噬菌体含量。该大肠杆菌分离株仅对氨苄青霉素表现出中度敏感性,通过基于WGS的分型被鉴定为ONT:H21-B1-ST40菌株。尽管仅检测到一个与抗菌耐药性相关的基因[mdf(A)],但从非典型肠致病性大肠杆菌(aEPEC)致病型中鉴定出了几个与毒力相关的基因(VAGs)。此外,还检测到了来自大质粒组的质粒复制子和18个前噬菌体相关区域。总之,对从墨西哥锡那罗亚州一个养鱼场分离出的aEPEC分离株进行WGS表征,有助于深入了解其致病潜力以及食用生鲜水产养殖产品可能对人类健康造成的风险。有必要利用下一代测序(NGS)技术来研究环境微生物,并采用“同一健康”框架来了解健康问题的起源。

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