Abstract

Enterohaemorrhagic Escherichia coli (EHEC) is an important human pathogen worldwide. Although serotype O157 is currently the most dominant and important EHEC strain, serotypes O26, O111, O91, O103 and O121 are also recognized as serious pathogens that affect public health. EHEC outbreaks often occur in nurseries and elderly care facilities. In 2012, a nursery outbreak of EHEC O121 occurred during which the bacterium acquired a plasmid-borne extended-spectrum β-lactamase (ESBL) gene. ESBL-producing E. coli O86 was concurrently isolated from one of the EHEC patients. Therefore, we investigated the isolates by whole-genome sequence (WGS) analysis to elucidate the transmission dynamics of the EHEC strains and the ESBL plasmid. According to WGS-based phylogeny, all 17 EHEC O121 isolates were clonal, while E. coli O86 was genetically distant from the EHEC O121 isolates. The complete sequence of an ESBL plasmid encoding the CTX-M-55 β-lactamase was determined using S1-PFGE bands, and subsequent mapping of the WGS reads confirmed that the plasmid sequences from EHEC O121 and E. coli O86 were identical. Furthermore, conjugation experiments showed that the plasmid was capable of conjugative transfer. These results support the hypothesis that EHEC O121 acquired an ESBL-producing plasmid from E. coli O86 during the outbreak. This report demonstrates the importance of implementing preventive measures during EHEC outbreaks to control both secondary infection and the spread of antimicrobial resistance factors.

Highlights

  • Enterohaemorrhagic Escherichia coli (EHEC) is an important human pathogen worldwide

  • EHEC isolates are collected in the National Institute of Infectious Diseases, Japan, and nation-wide surveillance of the bacterium has been performed using multilocus variablenumber tandem-repeat analysis for serogroups O157, O26, O111, O91, O103, O121, O145 and O165, and pulsed-field gel electrophoresis (PFGE) for the other serogroups [4, 5]

  • Our results demonstrate the importance of monitoring antimicrobial resistance during EHEC outbreaks

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Summary

Introduction

Enterohaemorrhagic Escherichia coli (EHEC) is an important human pathogen worldwide. EHEC produces Shiga toxin (Stx), and infections with EHEC strains present clinical symptoms such as diarrhoea, haemorrhagic colitis and lifethreatening haemolytic uraemic syndrome (HUS) [1]. EHEC isolates are collected in the National Institute of Infectious Diseases, Japan, and nation-wide surveillance of the bacterium has been performed using multilocus variablenumber tandem-repeat analysis for serogroups O157, O26, O111, O91, O103, O121, O145 and O165, and pulsed-field gel electrophoresis (PFGE) for the other serogroups [4, 5]. We investigated the isolates in more detail by whole-genome sequence (WGS) analysis, which has been widely implemented in EHEC surveillance [6, 8, 14]. High-resolution phylogenetic analysis and the complete sequence of an extended-spectrum β-lactamase (ESBL) plasmid provided direct evidence of plasmid transfer from a non-toxigenic E. coli strain to an EHEC strain

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