Abstract

BackgroundSurveillance of commensal Escherichia coli, a possible reservoir of antimicrobial resistance (AMR) genes, is important as they pose a risk to human and animal health. Most surveillance activities rely on phenotypic characterisation, but whole genome sequencing (WGS) presents an alternative.AimIn this retrospective study, we tested 515 E. coli isolated from pigs to evaluate the use of WGS to predict resistance phenotype.MethodsMinimum inhibitory concentration (MIC) was determined for nine antimicrobials of clinical and veterinary importance. Deviation from wild-type, fully-susceptible MIC was assessed using European Committee on Antimicrobial Susceptibility Testing (EUCAST) epidemiological cut-off (ECOFF) values. Presence of AMR genes and mutations were determined using APHA SeqFinder. Statistical two-by-two table analysis and Cohen’s kappa (k) test were applied to assess genotype and phenotype concordance.ResultsOverall, correlation of WGS with susceptibility to the nine antimicrobials was 98.9% for test specificity, and 97.5% for the positive predictive value of a test. The overall kappa score (k = 0.914) indicated AMR gene presence was highly predictive of reduced susceptibility and showed excellent correlation with MIC. However, there was variation for each antimicrobial; five showed excellent correlation; four very good and one moderate. Suggested ECOFF adjustments increased concordance between genotypic data and kappa values for four antimicrobials.ConclusionWGS is a powerful tool for accurately predicting AMR that can be used for national surveillance purposes. Additionally, it can detect resistance genes from a wider panel of antimicrobials whose phenotypes are currently not monitored but may be of importance in the future.

Highlights

  • Escherichia coli in animals comprise a diverse range of strains

  • Values, interpreted using epidemiological cut-off (ECOFF), with antimicrobial resistance (AMR) genotypes in whole genome sequencing (WGS) data from 515 E. coli isolated from pooled caecal contents of healthy pigs collected at abattoir from farms across the United Kingdom (UK) from 2014 to Isolates were selected on Brilliance UTI Agar (Oxoid, Basingstoke, UK) plates containing either 1 mg/L

  • AMR genes or cSNP-AMR present in our database, with the remaining isolates having between one and nine acquired AMR genes and/or cSNP-AMR associated with the antimicrobials tested

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Summary

Introduction

Escherichia coli in animals comprise a diverse range of strains They are a reservoir and indicator of antimicrobial resistance (AMR) genes that can be mobilised to other bacteria including zoonotic pathogens, and pose a risk to human and animal health. Antimicrobial resistance is traditionally measured using phenotypic methods, for example, the minimal inhibitory concentration (MIC), and interpreted using breakpoints to determine resistance or susceptibility. The. Surveillance of commensal Escherichia coli, a possible reservoir of antimicrobial resistance (AMR) genes, is important as they pose a risk to human and animal health. Results: Overall, correlation of WGS with susceptibility to the nine antimicrobials was 98.9% for test specificity, and 97.5% for the positive predictive value of a test.

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