Abstract

Burkholderia pseudomallei is a Gram-negative bacterium that causes the sapronotic disease melioidosis. An outbreak in 2003 in the state of Ceara, Brazil, resulted in subsequent surveillance and environmental sampling which led to the recognition of B. pseudomallei as an endemic pathogen in that area. From 2003 to 2015, 24 clinical and 12 environmental isolates were collected across Ceara along with one from the state of Alagoas. Using next-generation sequencing, multilocus sequence typing, and single nucleotide polymorphism analysis, we characterized the genomic diversity of this collection to better understand the population structure of B. pseudomallei associated with Ceara. We found that the isolates in this collection form a distinct subclade compared to other examples from the Western Hemisphere. Substantial genetic diversity among the clinical and environmental isolates was observed, with 14 sequence types (STs) identified among the 37 isolates. Of the 31,594 core single-nucleotide polymorphisms (SNPs) identified, a high proportion (59%) were due to recombination. Because recombination events do not follow a molecular clock, the observation of high occurrence underscores the importance of identifying and removing recombination SNPs prior to evolutionary reconstructions and inferences in public health responses to B. pseudomallei outbreaks. Our results suggest long-term B. pseudomallei prevalence in this recently recognized region of melioidosis endemicity.IMPORTANCEB. pseudomallei causes significant morbidity and mortality, but its geographic prevalence and genetic diversity are not well characterized, especially in the Western Hemisphere. A better understanding of the genetic relationships among clinical and environmental isolates will improve knowledge of the population structure of this bacterium as well as the ability to conduct epidemiological investigations of cases of melioidosis.

Highlights

  • Burkholderia pseudomallei is a Gram-negative bacterium that causes the sapronotic disease melioidosis

  • A phylogenetic tree based on single-nucleotide polymorphisms (SNPs) analysis of the draft genome sequence of each isolate indicates that all in our panel occur in the Western Hemisphere within a clade containing isolate BCC 215

  • ClonalFrameML estimated that 3 SNPs occurred for each recombination event, and the relative effect on recombination and mutation (r/m) was 6.53, which suggests that recombination introduced much more SNP changes than mutations did

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Summary

Introduction

Burkholderia pseudomallei is a Gram-negative bacterium that causes the sapronotic disease melioidosis. Using next-generation sequencing, multilocus sequence typing, and single nucleotide polymorphism analysis, we characterized the genomic diversity of this collection to better understand the population structure of B. pseudomallei associated with Ceara. Genetic diversity of the pathogen #Burkholderia #pseudomallei, which causes #melioidosis, is quite pronounced in the state of Ceara, Brazil, based on whole-genome sequence analysis described in this paper. Previous work to genetically characterize the isolates recovered in Ceara relied on pulsed-field gel electrophoresis (PFGE), ribotyping, and randomly amplified polymorphic DNA [4, 8] They provide some insight, these methods lack the high level of resolution available with whole-genome sequencing (WGS), which can provide near-complete genomic sequences. The appearance of relatedness by MLST in these instances can be overcome by analysis of the whole-genome data [11, 12]

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