Melioidosis is a severe disease caused by the environmental bacterium Burkholderia pseudomallei that affects both humans and animals throughout northern Australia, Southeast Asia and increasingly globally. While there is a considerable degree of genetic diversity amongst isolates, B. pseudomallei has a robust global biogeographic structure and genetic populations are spatially clustered in the environment. We examined the distribution and local spread of B. pseudomallei in Darwin, northern territory, Australia, which has the highest recorded urban incidence of melioidosis globally. We sampled soil and land runoff throughout the city centre and performed whole-genome sequencing (WGS) on B. pseudomallei isolates. By combining phylogenetic analyses, Bayesian clustering and spatial hot spot analysis our results demonstrate that some sequence types (STs) are widespread in the urban Darwin environment, while others are highly spatially clustered over a small geographic scale. this clustering matches the spatial distribution of clinical cases for one St. Results also demonstrate a greater overall isolate diversity recovered from drains compared to park soils, further supporting the role drains may play in dispersal of B. pseudomallei Sts in the environment. collectively, knowledge gained from this study will allow for better understanding of B. pseudomallei phylogeography and melioidosis source attribution, particularly on a local level. Burkholderia pseudomallei is an environmental Gram-negative bacillus and the causative agent of melioidosis, a potentially fatal infection of humans and animals 1. Regions of high B. pseudomallei endemicity predominantly include Southeast Asia and northern Australia, though the bacterium is also increasingly found in other tropical regions including the Indian subcontinent, Africa and the Americas 2. Melioidosis is typically considered noncommunicable with direct person-to-person transmission and zoonotic disease being remarkably rare 3,4. Nearly all B. pseudomallei infections are caused by a single direct exposure event to contaminated soil or surface water in the environment and individual cases of melioidosis are typically the result of infection by different strains of the bacterium 5. A limited geographic dispersal of B. pseudomallei strains has also been identified and the bacterium is now recognised as being ecologically established and spatially clustered in the environment 6-8. This is in spite of the frequent opportunities B. pseudomallei has to spread within the water table, via agricultural and migratory animals, or in transported soil 6,9-11. This restricted geographical distribution has resulted in distinct genetic populations of the bacterium, which are evident despite high levels of gene recombination and sequence type (ST)