Abstract: A total of 100 seawater samples were collected by sterile sampler at Lingang sea area of shanghai, and 33 Vibrio alginolyticus isolates were obtained after cultured,separated and identified by physiological biochemical test and PCR rapid detection method.All isolates were tested for susceptibility to 20 kinds of antibiotics by improved Kirby -Bauer method. Susceptibility test showed that all strains were resistant to tetracycline(100.0%),followed by ampicillin(97.0%) and carbenicillin(93.9%). However,they were sensitive to cefepime,aztreonam,erythromycin,ciproffloxacin,norfloxacin and chloramphenicol. All isolates showed multiple resistance to no less than 3 antibiotics ,15 (45.5% ) isolates showed multiple resistance to no less than 6 antibiotics,and 6(18.2%) isolates showed multiple resistance to no less than 8 antibiotics,indicating that all the V. alginolyticus which we isolated have strong multiple resistance to many antibiotics. This study can provide reference for controlling the occurrence of foodborne diseases,and establish the theoretical base to illustrate the antibiotic resistance mechanism of Vibrio alginolyticus.