Final report of ENGAGE ‐ Establishing Next Generation sequencing Ability for Genomic analysis in Europe

The ENGAGE project (http://www.engage-europe.eu/) was a collaboration between eight institutions across Europe. The aim was to boost the scientific cooperation to use whole genome sequencing (WGS) analysis in food safety and public health protection. ENGAGE focused on Escherichia coli (commensal E. coli) and different Salmonella spp. serotypes. A total of 3,360 genomes, 778 and 2,582 of E. coli and Salmonella, respectively, were produced. These genomes were stored and shared among partners in a temporary repository to be submitted to the European Nucleotide Archive by the end of the project. Generated genomes were used for benchmarking exercises to assess the possibility of replacing conventional typing with WGS for outbreak investigation. For the analysed strains, the benchmarking exercises showed that SPAdes assembly performed better than Velvet and that, by using different bioinformatics tools, WGS Salmonella serotyping and antimicrobial resistance genes detection, were largely in concordance with phenotypic data. Discrepancies were related to sequence quality and phenotype misclassification rather than to limitations of the bioinformatics tools. All partners were able to infer the expected phylogeny for the Salmonella and Campylobacter isolates in benchmarking exercises. Two WGS proficiency tests (assessing different genomic quality markers) were conducted among partners with satisfactory results. Guidelines including available bioinformatics tools and standard operating procedures (wet and dry lab) were prepared and posted online. Workshops, training courses and twinning programmes were conducted. The training focused on

[1]  Rene S. Hendriksen,et al.  Emergence of a Clonal Lineage of Multidrug-Resistant ESBL-Producing Salmonella Infantis Transmitted from Broilers and Broiler Meat to Humans in Italy between 2011 and 2014 , 2015, PloS one.

[2]  F. Weill,et al.  Human infections due to Salmonella Napoli: a multicountry, emerging enigma recognized by the Enter-net international surveillance network. , 2009, Foodborne pathogens and disease.

[3]  Leigh A Knodler,et al.  Salmonella enterica. , 2019, Trends in microbiology.

[4]  Vitali Sintchenko,et al.  Proficiency testing for bacterial whole genome sequencing: an end-user survey of current capabilities, requirements and priorities , 2015, BMC Infectious Diseases.

[5]  Andrew J. Page,et al.  Roary: rapid large-scale prokaryote pan genome analysis , 2015, bioRxiv.

[6]  Sergey I. Nikolenko,et al.  SPAdes: A New Genome Assembly Algorithm and Its Applications to Single-Cell Sequencing , 2012, J. Comput. Biol..

[7]  Daniel H. Huson,et al.  Dendroscope: An interactive viewer for large phylogenetic trees , 2007, BMC Bioinformatics.

[8]  Paramvir S. Dehal,et al.  FastTree 2 – Approximately Maximum-Likelihood Trees for Large Alignments , 2010, PloS one.

[9]  Herman Goossens,et al.  Identification of a novel plasmid-mediated colistin-resistance gene, mcr-2, in Escherichia coli, Belgium, June 2016. , 2016, Euro surveillance : bulletin Europeen sur les maladies transmissibles = European communicable disease bulletin.

[10]  P. Ashton,et al.  Distributed under Creative Commons Cc-by 4.0 Most: a Modified Mlst Typing Tool Based on Short Read Sequencing , 2022 .

[11]  E. Myers,et al.  Basic local alignment search tool. , 1990, Journal of molecular biology.

[12]  A. Hoszowski,et al.  Identification of common, non-typable and autoagglutinating Salmonella strains with Premi®Test Salmonella assay. , 2013, Acta veterinaria Hungarica.

[13]  Jessica C. Chen,et al.  Comparative Analysis of Extended-Spectrum-β-Lactamase CTX-M-65-Producing Salmonella enterica Serovar Infantis Isolates from Humans, Food Animals, and Retail Chickens in the United States , 2017, Antimicrobial Agents and Chemotherapy.

[14]  I. Szabó,et al.  VIM-1-producing Salmonella Infantis isolated from swine and minced pork meat in Germany , 2017, The Journal of antimicrobial chemotherapy.

[15]  Richard Myers,et al.  SnapperDB: A database solution for routine sequencing analysis of bacterial isolates , 2017, bioRxiv.