bioRxiv Science⌕ Search

bioRxiv · 10.1101/535914

Global phylogenomics of multidrug resistant Salmonella enterica serotype Kentucky ST198

Abstract

Salmonella enterica serotype Kentucky (S. Kentucky) can be a common causative agent of salmonellosis, usually associated with consumption of contaminated poultry. Antimicrobial resistance (AMR) to multiple drugs, including ciprofloxacin, is an emerging problem within this serotype. We used whole-genome sequencing (WGS) to investigate the phylogenetic structure and AMR content of 121 S. Kentucky ST198 isolates from five continents. Population structure was inferred using phylogenomic analysis and whole genomes were compared to investigate changes in gene content, with a focus on acquired AMR genes. Our analysis showed that multidrug resistant (MDR) S. Kentucky isolates belonged to a single lineage, which we estimate emerged circa 1989 following the acquisition of the AMR-associated Salmonella genomic island 1 (variant SGI1-K) conferring resistance ampicillin, streptomycin, gentamicin, sulfamethoxazole, and tetracycline. Phylogeographic analysis indicates this clone emerged in Egypt before disseminating into Northern, Southern and Western Africa, then to the Middle East, Asia and the European Union. The MDR clone has since accumulated various substitution mutations in the quinolone resistance determining regions (QRDR) of DNA gyrase (gyrA) and DNA topoisomerase (parC), such that most strains carry three QRDR mutations which together confer resistance to ciprofloxacin. The majority of AMR genes in the S. Kentucky genomes were carried either on plasmids or SGI structures. Remarkably, each genome carried a different SGI1-K derivative structure; this variation could be attributed to IS26-mediated insertions and deletions, which appear to have hampered previous attempts to trace the clone's evolution using sub-WGS resolution approaches. Several different AMR plasmids were also identified, encoding resistance to chloramphenicol, third-generation cephalosporins, carbapenems, and/or azithromycin. These results indicate that most MDR S. Kentucky circulating globally result from the clonal expansion of a single lineage that acquired chromosomal AMR genes 30 years ago, and has continued to diversify and accumulate additional resistances to last-line oral antimicrobials.

Source connections

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Hawkey, J., Le Hello, S., Doublet, B., Granier, S., Hendriksen, R., Fricke, W. F., Ceyssens, P.-J., Gomart, C., Billman-Jacobe, H., Holt, K. E., Weill, F.-X.. 2019-02-03. Global phylogenomics of multidrug resistant Salmonella enterica serotype Kentucky ST198. https://doi.org/10.1101/535914

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

A Staphylococcus phage recombination cassette that enables recombineering across distant bacteria

Modifying bacterial genomes with phage recombinases, a process termed recombineering, has enabled precise genome editing, but its reach is limited by host specificity: each new organism generally requires its own recombinase to be identified and optimized. Using known phage recombination genes as synteny anchors, we mined Staphylococcus genomes and recovered a diverse collection of phage recombination cassettes, spanning the RAD52-like annealase superfamily (Sak, Erf) and the RAD51/RecA-like Sak4 family. Functional testing of representatives in S. aureus identified several highly efficient systems that support recombineering. We engineered the most efficient of these into a single-plasmid tool that yields markerless editing in S. aureus in four days. Notably, where other recombineering systems rely on host-specific factors for nuclease evasion and ssDNA loading, this cassette encodes both activities itself: a predicted broad-spectrum nuclease inhibitor and a dedicated single-stranded DNA-binding protein. We therefore tested the cassette across phylogenetically distant hosts, and observed recombination in Streptococcus pyogenes, Enterococcus faecium, Francisella novicida, and Escherichia coli. These results identify a self-sufficient class of phage recombination systems and provide a portable tool for

microbiology↗

Phenotypic and Molecular Characterization of Potentially New Shigella boydii Serotype in Bangladesh

Objective: Shigella is a major cause of bacillary dysentery among children under 5 years of age, especially in low- and middle-income countries. At Dhaka hospital of icddr,b, every year around 3% of Shigella isolates from patients with diarrhea are identified as Shigella-like organisms (SLOs). These isolates do not react with any of the existing and provisional Shigella antisera. The major aims of the present study were to characterize a unique cluster of SLOs using phenotypic and genotypic methods. Methods: A cluster of 6 SLOs was characterized using biochemical tests, serotyping, antimicrobial susceptibility, and virulence assay by using the Sereny test, PCR and compared their pulsed-field gel electrophoresis (PFGE) patterns with known serotypes of Shigella spp. Antisera against the isolate were raised in a rabbit model to determine cross-reactivity with known serotypes. Results: All six SLOs were mannitol fermenting, have invasive properties, positive for ipaH and Shigella enterotoxin 2 gene along with possession of virulence plasmid of ~ 140 MDa. All isolates were resistant to ampicillin, tetracycline, sulfamethoxazole-trimethoprim, but were sensitive to mecillinam, nalidixic acid, and ciprofloxacin. Plasmid and PFGE profiles showed that the genomic patterns of these isolates were identical but were different from all other S. boydii serotypes. Antiserum prepared against one of the isolates agglutinated with all isolates but did not cross-react with other Shigella and E. coli reference strains. Conclusion: This unique cluster of isolates might belong to a new provisional serovar of S. boydii and further analysis is required to incorporate it into the stereotypic scheme of Shigella.

microbiology↗

Long-read shotgun metagenomics identifies distinct urogenital microbial community states associated with functional potential and urinary tract infection during pregnancy

Urinary tract infections (UTIs) are among the most common bacterial infections during pregnancy and are associated with adverse maternal and neonatal outcomes. Although previous studies used enhanced culture, 16S rRNA gene sequencing, and/or short-read shotgun metagenomic sequencing to characterize the urogenital microbiome of adult females, few have assessed the urogenital microbiome during pregnancy and thus this microbiome remains poorly understood. Here, we applied Oxford Nanopore long-read shotgun metagenomic sequencing to characterize urogenital microbial composition, functional potential, metagenome-assembled genomes (MAGs), and the urogenital virome of pregnant females. Taxonomic profiling identified three distinct microbial community states predominated by Lactobacillus crispatus, Gardnerella species, and L. iners. The Gardnerella-predominated community was associated with a higher frequency of UTI during pregnancy. Functional profiles strongly reflected these community states, demonstrating that taxonomic differences were accompanied by distinct functional potential. Genome-resolved analysis recovered 415 MAGs, including 29 high-quality genomes, revealing substantial genomic diversity among urogenital microbes. Viral sequences, predominantly bacteriophages, also were detected in most samples, highlighting an additional component of the pregnancy-associated urogenital microbiome. These findings demonstrate that pregnant females possess distinct urogenital microbial community states and show that they differ not only in taxonomic composition but also in their functional potential.

microbiology↗