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This study was aimed to evaluate the antimicrobial resistance and molecular resistance mechanisms of 87 Vibrio parahaemolyticus isolates from cultured sea cucumbers (Apostichopus japonicus). The results showed that all isolates were resistant to ampicillin and cephazolin, fewer of them were resistant to streptomycin (43·7%), cefuroxime sodium (18·4%), tetracycline (4·6%), sulphamethoxazole/trimethoprim (2·3%) and four quinolones (2·3%). More than half (56·2%) of the isolates displayed multiple resistance to at least three antimicrobials. The resistance genes were detected in all antimicrobial-resistant isolates except two tetracycline-resistant isolates. Among all these tested resistance genes, blaTEM, sul2, strA and strB were predominant, and none of blaSHV, blaCTX-M, blaOXA, sul1, sul3, tetA, tetM and tetQ genes was detected. Point mutations were found in quinolone resistance-determining regions of gyrA and parC genes in quinolone-resistant isolates. All isolates harboured class 1 integrons but only one carried gene cassette without any resistance genes, and none of them was positive to class 2, 3 integrons and SXT constins. These results indicate that the antimicrobial-resistant V. parahaemolyticus isolates from sea cucumbers and resistance genes could be potential risks to public health or other environments.Significance and Impact of the Study: This study is the first report on characterization of antimicrobial resistance of Vibrio parahaemolyticus from sea cucumbers (Apostichopus japonicus). Our findings reveal a high level of resistance to some antimicrobials and prevalence of the resistance genes in V. parahaemolyticus isolates from sea cucumbers and underline the need for prudent use of antimicrobials in aquaculture to minimize the spread of antimicrobial-resistant V. parahaemolyticus.