A novel polyvalent attenuated live bacterial vaccine for preventing and curing vibriosis of cultivated fish is provided. The vaccine mainly comprises attenuated deletion strain of Vibrio anguillarum without marker gene, which has significant low toxicity, but remains immunogenicity against wild type strain of V. anguillarum, as compared with wild type strain MVM425. Moreover, the vaccine strain has excellent cross immunoprotection against Vibrio alginolyticus. The attenuated live vaccine made from the vaccine strain is effective to prevent and cure vibriosis of fish resulted from wild type strain of V. anguillarum and V. alginolyticus.