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This study investigated the bacterial diversity of gut content of sea cucumber (Apostichopus japonicus) and its habitat surface sediment in a bottom enhancement area using PCR-based denaturing gradient gel electrophoresis (DGGE) technique. Bacte-rial diversity evaluation showed that the value of the Shannon-Wiener index of gut content in different intestinal segments of A. ja-ponicus varied between 2.88 and 3.00, lower than that of the surrounding sediment (3.23). Phylogenetic analysis showed that bacte-rial phylotypes in gut content and the surrounding sediment of A. japonicus were closely related to Proteobacteria includingγ-,α-,δ-andε-proteobacteria, Bacteroidetes, Firmicute, and Actinobacteria, of whichγ-proteobacteria were predominant. These results sug-gested that the sea cucumber A. japonicus was capable of feeding selectively, and PCR-DGGE was applicable for characterizing the bacterial community composition in gut content and the surrounding sediment of sea cucumber. Further investigation targeting longer 16S rDNA gene fragments and/or functional genes was recommended for obtaining more information of the diversity and function of bacterial community in the gut content of sea cucumber.