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Bacterial flora in gut contents of sea cucumber Apostichopus japonicus at different culture periods

The bacterial flora in gut contents was studied in cultured sea cucumber Apostichopus japonicus collected on June,September and October of 2009,and March,April and May of 2010 by PCR-DGGE in which variable regional fragments of 16 S r DNA-V3 were amplified by specific primers of bacteria. There were 49,49,54,53,52 and 51 bands in the samples collected on March,April,May,June,September and October,respectively,in the DGGE gels which were cut,recovered,cloned,and sequenced. The maximal similarity( 67.9%) in bacterial flora was observed in the samples collected on March and September,followed by the samples( 63.8%) collected on between April and October. The sequencing analysis revealed that 11 nucleic acid sequences were similar to uncultured bacteria,and 32 sequences were similar to the sequences of 7 groups of bacterial including α-proteobacteria,γ-proteobacteria,δ-proteobacteria,Flavobacteria,Cyanobacteria,Chloroflexi and Verrucomicrobia in the 43 major bands. The dominant groups in the bacterial flora in different culture periods were found to be α-proteobacteria,γ-proteobacteria and Flavobacteria. It was found that γ- proteobacteria had the maximal proportion,varying from 25.00% to 35.71%. There was higher relative count in Flavobacterium than α-proteobacteria in most samples except that on March and September. The findings indicated that similar flora succession was observed in spring and autumn,the proportion of α-proteobacteria being declined while that of γ-proteobacteria and Flavobacterium elevated.