Dataset Information

Development of food safety risk assessment tools based on molecular typing and WGS of Campylobacter jejuni genome.

ABSTRACT: The 'learning-by-doing' EU-FORA fellowship programme in the development of risk assessment tools based on molecular typing and WGS of Campylobacter jejuni genome was structured into two main activities: the primary one focused on training on risk assessment methodology and the secondary one in starting and enhancing the cooperation between the hosting and home organisations, or other joint activities. The primary activities had three subsequent work packages (WPs): WP1 data organisation, WP2 cluster and association analyses, and WP3 development of risk assessment models. The secondary activities have branched into one workshop and the initiation of a cooperation programme between the hosting and home organisations. In the last quarter, the fellow had contributed to the characterisation of some pathogens in possible response to a changing climate, part of the CLEFSA project. The fellow attended various forms of training: online and on-site courses, and also participated at several conferences and meetings for improving his knowledge and skills, contributing to performing the Campylobacter risk assessment and source attribution.

SUBMITTER: Istituto Zooprofilattico Sperimentale dell’ Abruzzo e del Molise “G. Caporale”, Teramo, Italy

PROVIDER: S-EPMC7015486 | biostudies-literature | 2019 Sep

REPOSITORIES: biostudies-literature

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Development of food safety risk assessment tools based on molecular typing and WGS of Campylobacter jejuni genome.

Ardelean A I AI Calistri P P Giovannini A A Garofolo G G Di Pasquale A A Conte A A MorelliD D D

EFSA journal. European Food Safety Authority 20190917 Suppl 2

The 'learning-by-doing' EU-FORA fellowship programme in the development of risk assessment tools based on molecular typing and WGS of Campylobacter jejuni genome was structured into two main activities: the primary one focused on training on risk assessment methodology and the secondary one in starting and enhancing the cooperation between the hosting and home organisations, or other joint activities. The primary activities had three subsequent work packages (WPs): WP1 data organis ...[more]

PMID: 32626461

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Project description:Rapid developments in the field of whole genome sequencing (WGS) make in silico antimicrobial resistance (AMR) a target within reach. Campylobacter jejuni is a leading cause of foodborne infections in Israel with increasing rates of resistance. We applied WGS analysis to study the prevalence and genetic basis of AMR in 263 C. jejuni human and veterinary representative isolates retrieved from a national collection during 2003-2012. We evaluated the prediction of phenotypic AMR from genomic data. Genomes were screened by the NCBI AMRFinderPlus and the BioNumerics tools for acquired AMR genes and point mutations. The results were compared to phenotypic resistance determined by broth microdilution. The most prevalent resistant determinants were the multi-drug efflux transporter gene cmeABC (100%), the tetracycline resistance tet(O) gene (82.1%), the quinolone resistance gyrA T861 point mutation (75.7%), and the aadE streptomycin resistance gene. A variety of 12 known β lactam resistance genes (bla OXA variants) were detected in 241 (92%) isolates, the most prevalent being bla OXA-193, bla OXA-461, and bla OXA-580 (56, 16, and 7%, respectively). Other aminoglycoside resistance genes and the macrolide resistance point mutation were rare (<1%). The overall correlation rate between WGS-based genotypic prediction and phenotypic resistance was 98.8%, sensitivity, specificity, positive, and negative predictive values being 98.0, 99.3, 99.1, and 98.5%, respectively. wgMLST-based phylogeny indicated a high level of clonality and clustering among the studied isolates. Closely related isolates that were part of a genetic cluster (single linkage distance ≤ 15 alleles) based on wgMLST phylogeny mostly shared a homogenous AMR determinant profile. This was observed in 18 of 20 (90.0%) clusters within clonal complex-21, suggesting clonal expansion of resistant isolates. Strong association to lineage was noted for the aadE gene and the various bla OXA genes. High resistance rates to tetracycline and quinolones and a low resistance rate to macrolides were detected among the Israeli C. jejuni isolates. While a high genotypic-phenotypic correlation was found, some resistance phenotypes could not be predicted by the presence of AMR determinants, and particularly not the level of resistance. WGS-based prediction of antimicrobial resistance in C. jejuni requires further optimization in order to integrate this approach in the routine workflow of public health laboratories for foodborne surveillance.

Project description:We present an optimized multilocus sequence typing (MLST) scheme with universal primer sets for amplifying and sequencing the seven target genes of Campylobacter jejuni and Campylobacter coli. Typing was expanded by sequence determination of the genes flaA and flaB using optimized primer sets. This approach is compatible with the MLST and flaA schemes used in the PubMLST database and results in an additional typing method using the flaB gene sequence. An identification module based on the 16S rRNA and rpoB genes was included, as well as the genetic determination of macrolide and quinolone resistances based on mutations in the 23S rRNA and gyrA genes. Experimental procedures were simplified by multiplex PCR of the 13 target genes. This comprehensive approach was evaluated with C. jejuni and C. coli isolates collected in Switzerland. MLST of 329 strains resulted in 72 sequence types (STs) among the 186 C. jejuni strains and 39 STs for the 143 C. coli isolates. Fourteen (19%) of the C. jejuni and 20 (51%) of the C. coli STs had not been found previously. In total, 35% of the C. coli strains collected in Switzerland contained mutations conferring antibiotic resistance only to quinolone, 15% contained mutations conferring resistance only to macrolides, and 6% contained mutations conferring resistance to both classes of antibiotics. In C. jejuni, these values were 31% and 0% for quinolone and macrolide resistance, respectively. The rpoB sequence allowed phylogenetic differentiation between C. coli and C. jejuni, which was not possible by 16S rRNA gene analysis. An online Integrated Database Network System (SmartGene, Zug, Switzerland)-based platform for MLST data analysis specific to Campylobacter was implemented. This Web-based platform allowed automated allele and ST designation, as well as epidemiological analysis of data, thus streamlining and facilitating the analysis workflow. Data networking facilitates the exchange of information between collaborating centers. The described approach simplifies and improves the genotyping of Campylobacter, allowing cost- and time-efficient routine monitoring.

Dataset Information

Development of food safety risk assessment tools based on molecular typing and WGS of Campylobacter jejuni genome.

Publications

Development of food safety risk assessment tools based on molecular typing and WGS of <i>Campylobacter</i> <i>jejuni</i> genome.

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