Using biological treatment to deal with harmful algal blooms is highly potential over the physical and chemical methods due to its species specificity and eco-friendly characteristics. In this study, algicidal broth were produced from a Vibrio sp. co-culture composed mainly of V. brasilliensis and V. tubiashii. The productivity of the algicidal compounds was optimized under a dilution rate of 0.1 h with a minimum algicidal broth dosage of 0.3% for 100% algal lysis. The algicidal threshold and EC of the spray-dried algicidal broth were 0.17 and 0.68 g/L, respectively. Treatment with the algicidal agents led to an increase in cellular reactive oxygen species (ROS) level that causes membrane damage as supported by the increase in Malondialdehyde (MDA) levels. and a further inhibition to the antioxidant system as indicated by a sharp decrease in the catalase (CAT) activity. The algicidal compound was identified as hexahydro pyrrolo[1,2-a] pyr azine-1,4-dione.