A number of pathogenic microorganisms, including, several NIAID Priority Pathogens, secrete virulence factors utilizing a novel pathway referred to as the type VI secretion system (T6SS). The main objective of this project is to develop inhibitors of T6SS that could lead to a new class of antimicrobial agents. Burkholderia thailandensis will be used to identify T6SS inhibitors compounds in a high throughput screen and these will be tested for interference with secretion in several category A-C select agents, including Burkholderia pseudomallei and Burkholderia mallei, causative agents of melioidosis and glanders, respectively. The candidate inhibitors of T6SS and their structural derivatives will be used in subsequent studies, which include demonstration of their efficacy in an animal model of melioidosis. The T6SS components, targeted by the active compounds will be identified. The work proposed in this project represents a logical pre-clinical development strategy for therapeutic agents targeting several agents of biological warfare and bioterrorism.