A lot of cultured flounders died by ascitic disease blooming during the summer season in 2002, southern parts of Korea. This study was conducted to investigate the microbial flora for the ascitic cultivated olive flounder (Paralichthys olivaceus) which were collected in the Koe-je island in the southern of Korea from July 2002 to October 2003. Three Genera (Vibrio, Photobacterium and Edwardsiella), seven species of bacteria (V. harveyi, V. alginolyticus, V. parahaemolyticus, V. carcariae, V. metschenikovii, P. damsela and E. tarda) and a fish pathogenic birnavirus (marine birnavirus, MABV) were isolated from the liver, dropsy, spleen and identified by biochemical and molecular biological characterization.