Infection with the gram-negative bacterium Burkholderia pseudomallei can result in a life-threatening disease known as melioidosis. Historically, melioidosis was a common infection in military forces serving in Southeast Asia, and it has the potential to have a serious impact on force health readiness. With the U.S. Department of Defense's increasing strategic and operational focus across the Pacific Theater, melioidosis is an increasingly important issue from a force health protection perspective. U.S. Marines deploy annually to Darwin, Australia, a "hyperendemic" region for B. pseudomallei, to engage in training exercises. In an effort to assess the risk of B. pseudomallei infection to service personnel in Australia, 341 paired samples, representing pre- and post-deployment samples of Marines who trained in Australia, were analyzed for antibodies against B. pseudomallei antigens. Serological evidence of possible deployment-related infection with B. pseudomallei was found in 13 Marines. Future prospective studies are required to further characterize the risk to service members deployed to melioidosis endemic areas.