Melioidosis is caused by the environmental pathogen Burkholderia pseudomallei. Among 1,331 patients with melioidosis during 1989-2023 in the Darwin Prospective Melioidosis Study in Australia, we identified 6 locally acquired cases caused by B. pseudomallei sequence type 46. Because of global transmission and expansion of endemicity, clinicians should increase awareness of melioidosis.