Expression of luminescence in the Penaeus monodon pathogen Vibrio harveyi is regulated by an intercellular quorum sensing mechanism involving the synthesis and detection of two signaling molecules, one of which is N-hydroxy butanoyl-L-homoserine lactone and the other of which is uncharacterized. Indirect evidence has suggested that virulence, associated with a toxic extracellular protein, and luminescence in V. harveyi are coregulated. In this study the effects of an acylated homoserine lactone antagonist produced by the marine alga Delisea pulchra on luminescence and toxin production in a virulent strain of V. harveyi were analyzed. Luminescence and toxin production were both inhibited by the signal antagonist at concentrations that had no impact on growth. Toxin production was found to be prematurely induced in V. harveyi cultures incubated in a 10% conditioned medium. Additionally, a significant reduction in the toxicity of concentrated supernatant extracts from V. harveyi cultures incubated in the presence of the signal antagonist, as measured by in vivo toxicity assays in mice and prawns, was observed. These results suggest that intercellular signaling antagonists have potential utility in the control of V. harveyi prawn infections.Vibrio harveyi is a gram-negative, luminescent, marine bacterium isolated both in a free-living state (16,17,20) and as a commensal organism in the enteric contents of marine animals (11,22). Recognized as a primary pathogen of many commercially cultured invertebrate species, such as the black tiger prawn (Penaeus monodon), V. harveyi can cause up to 100% mortality of larvae in the hatchery stage of penaeid culture (12,16).Virulence in V. harveyi (strain 47666-1) has been attributed to the production of an extracellular protein referred to as toxin T1 with a molecular mass of approximately 100 kDa (10,19). The extracellular protein is produced during the midexponential phase of growth and has sequence similarity to virulence-associated proteins in Salmonella, Shigella, and Bacillus species (10).As suggested by the name of the disease phenomenon commonly referred to as luminous vibriosis, the expression of luminescence in V. harveyi has long been associated with virulence in pathogenic strains of this organism (14). Expression of the luminescence phenotype in V. harveyi is controlled at the transcriptional level by an atypical quorum sensing system (1). In the model quorum sensing system of Vibrio fischeri, a diffusible N-acylated-L-homoserine lactone (AHL) molecule is employed to link expression of the luminescence phenotype to bacterial population density. AHL, synthesized by the LuxI protein, accumulates throughout the population and at high concentrations interacts with a regulatory protein (LuxR), which then transcriptionally activates expression of the structural genes (luxCDABEG) encoding the luminescence phenotype (23). Other AHL quorum sensing systems, such as those regulating expression of virulence factors in Pseudomonas aeruginosa and Erwinia carotovora, also rely on L...