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Vibrio anguillarum is an opportunistic pathogen of aquatic animals, biocontrol agents have been revealed effective in preventing this disease. Antarctic bacteria were used for the screening of antagonism against V. anguillarum and strain AN32 showed highest activity. AN32 was identified as Pseudoalteromonas according to its morphological, physiological and biochemical and 16S rDNA molecular properties. The antibacterial substance in AN32 culture supernatant was sensitive to pH, temperature and proteinases. The substance could not dissolve in organic solvents, but could be precipitated by ammonium sulfate. These characterizations indicated that the bioactive substance of strain AN32 would be some kind of protein or peptide. This study is a potential application in the control of aquiculture diseases.