LAUSR

Each case of melioidosis results from a single event when a human is infected by the environmental bacterium Burkholderia pseudomallei. Darwin, in tropical northern Australia, has the highest incidences of melioidosis globally, and the Darwin Prospective Melioidosis Study (DPMS) commenced in 1989, documenting all culture-confirmed melioidosis cases. ABSTRACT Each case of melioidosis results from a single event when a human is infected by the environmental bacterium Burkholderia pseudomallei. Darwin, in tropical northern Australia, has the highest incidences of melioidosis globally, and the Darwin Prospective Melioidosis Study (DPMS) commenced in 1989, documenting all culture-confirmed melioidosis cases. From 2000 to 2019, we sampled DPMS patients’ environments for B. pseudomallei when a specific location was considered to be where infection occurred, with the aim of using genomic epidemiology to understand B. pseudomallei transmission and infecting scenarios. Environmental sampling was performed at 98 DPMS patient sites, where we collected 975 environmental samples (742 soil and 233 water). Genotyping matched the clinical and epidemiologically linked environmental B. pseudomallei for 19 patients (19%), with the environmental isolates cultured from soil (n = 11) and water (n = 8) sources. B. pseudomallei isolates from patients and their local environments that matched on genotyping were subjected to whole-genome sequencing (WGS). Of the 19 patients with a clinical-environmental genotype match, 17 pairs clustered on a Darwin core genome single-nucleotide polymorphism (SNP) phylogeny, later confirmed by single sequence typing (ST) phylogenies and pairwise comparative genomics. When related back to patient clinical scenarios, the matched clinical and environmental B. pseudomallei pairs informed likely modes of infection: percutaneous inoculation, inhalation, and ingestion. Targeted environmental sampling for B. pseudomallei can inform infecting scenarios for melioidosis and dangerous occupational and recreational activities and identify hot spots of B. pseudomallei presence. However, WGS and careful genomics are required to avoid overcalling the relatedness between clinical and environmental isolates of B. pseudomallei.