Olorunleke Solomon O, Kirchner Miranda, Duggett Nicholas, AbuOun Manal, Okorie-Kanu Onyinye J, Stevens Kim, Card Roderick M, Chah Kennedy Foinkfu, Nwanta John A, Brunton Lucy A, Anjum Muna F
Veterinary Epidemiology, Economics and Public Health Group, Department of Pathobiology and Population Sciences, Royal Veterinary College, London, United Kingdom.
Department of Bacteriology, Animal and Plant Health Agency, Weybridge, United Kingdom.
Front Microbiol. 2022 Jul 22;13:937968. doi: 10.3389/fmicb.2022.937968. eCollection 2022.
The rise in antimicrobial resistance (AMR) in bacteria is reducing therapeutic options for livestock and human health, with a paucity of information globally. To fill this gap, a One-Health approach was taken by sampling livestock on farms ( = 52), abattoir ( = 8), and animal markets ( = 10), and in-contact humans in Southeast Nigeria. Extended spectrum cephalosporin (ESC)-resistant (ESC-R) was selectively cultured from 975 healthy livestock faecal swabs, and hand swabs from in-contact humans. Antimicrobial susceptibility testing (AST) was performed on all ESC-R . For isolates showing a multi-drug resistance (MDR) phenotype ( = 196), quantitative real-time PCR (qPCR) was performed for confirmation of extended-spectrum β-lactamase (ESBL) and carbapenemase genes. Whole-genome sequencing (WGS) was performed on a subset ( = 157) for detailed molecular characterisation. The results showed ESC-R was present in 41.2% of samples, with AST results indicating 48.8% of isolates were phenotypically MDR. qPCR confirmed presence of ESBL genes, with present in all but others in a subset [ (62.8%) and (0.5%)] of isolates; none harboured transferable carbapenemase genes. Multi-locus sequence typing identified 34 Sequence Types (ST) distributed among different sampling levels; ST196 carrying was predominant in chickens. Large numbers of single nucleotide polymorphisms (SNPs) in the core genome of isolates, even within the same clade by phylogenetic analysis, indicated high genetic diversity. AMR genotyping indicated the predominant variant was (87.9%), although , and were present; it was notable that , common in livestock, was absent. Other predominant AMR genes included: 2, S1, B, , A-v2, and A14, with prevalence varying according to host livestock species. A harbouring plasmid from livestock isolates in Ebonyi showed high sequence identity to one from river/sewage water in India, indicating this ESBL plasmid to be globally disseminated, being present beyond the river environment. In conclusion, ESC-R was widespread in livestock and in-contact humans from Southeast Nigeria. WGS data indicated the isolates were genetically highly diverse, probably representing true diversity of wild type ; they were likely to be MDR with several harbouring Surprisingly, human isolates had highest numbers of AMR genes and pigs the least.
细菌中抗菌药物耐药性(AMR)的增加正在减少用于牲畜和人类健康的治疗选择,而全球范围内这方面的信息匮乏。为填补这一空白,我们采用了一种“同一健康”方法,对尼日利亚东南部的农场(n = 52)、屠宰场(n = 8)和动物市场(n = 10)的牲畜以及与之接触的人类进行采样。从975份健康牲畜粪便拭子和与之接触的人类手部拭子中选择性培养出对超广谱头孢菌素(ESC)耐药(ESC-R)的大肠杆菌。对所有ESC-R菌株进行了抗菌药物敏感性测试(AST)。对于表现出多重耐药(MDR)表型的分离株(n = 196),进行了定量实时PCR(qPCR)以确认超广谱β-内酰胺酶(ESBL)和碳青霉烯酶基因。对一个子集(n = 157)进行了全基因组测序(WGS)以进行详细的分子特征分析。结果显示,41.2%的样本中存在ESC-R大肠杆菌,AST结果表明48.8%的分离株在表型上为多重耐药。qPCR证实了ESBL基因的存在,除了一个子集中的其他菌株外,所有菌株中均存在blaCTX-M(62.8%)和blaTEM(0.5%);没有分离株携带可转移的碳青霉烯酶基因。多位点序列分型鉴定出34种序列类型(ST),分布在不同的采样水平;携带blaCTX-M的ST196在鸡中占主导地位。通过系统发育分析,即使在同一进化枝内,分离株核心基因组中也存在大量单核苷酸多态性(SNP),表明遗传多样性高。AMR基因分型表明,主要的blaCTX-M变体是blaCTX-M-15(87.9%),尽管也存在blaCTX-M-1、blaCTX-M-2和blaCTX-M-9;值得注意的是,在牲畜中常见的blaCMY不存在。其他主要的AMR基因包括:blaSHV、blaS1、blaB、blaOXA-1、blaAmpC-v2和blaA14,其流行率因宿主牲畜种类而异。埃邦伊州一份携带来自牲畜分离株质粒的blaCTX-M-15与印度河流/污水中的一个质粒具有高度序列同一性,表明这种ESBL质粒在全球范围内传播,存在于河流环境之外。总之,ESC-R大肠杆菌在尼日利亚东南部的牲畜和与之接触的人类中广泛存在。WGS数据表明,分离株在遗传上高度多样,可能代表了野生型大肠杆菌的真实多样性;它们可能是多重耐药的,有几个携带blaCTX-M。令人惊讶的是,人类分离株中的AMR基因数量最多,而猪的最少。