Analysis Of Bacterial Diversity In The Intestine Of Takifugu Rubripes By 16S RDNA

In this study,we use 2216E media to isolate the cultured bacterial in intestine of Dalian farmed Takifugu rubripes.The isolated and purified microorganisms were identified based on bacterial morphs,physiological-biochemical characteristic and 16S rDNA molecular biology to find out the bacterial composiotion of cultured microflora in intestine.and we also investigated the intestinal bacterial diversity of Takifugu rubripes using 16S ribosomal DNA gene(rDNA)clone library.In this study,we use 2216E media to isolate and identificate the cultured bacterial in inte Stineof Dalian farmed Takifugu rubripes.The isolated and purified microorganisms were identified based on bacterial morphs,physiological-biochemical characteristic and 16S rDNA molecular biology to find out the bacterial composiotion of cultured microflora in intestine of Dalian fanned Takifugu rubripes.The result showed that 12 bacteria were identified and purified from intestine of Dalian farmed Takifugu rubripes.Among them 12 different kinds of genera were identified as:Enterobacter cloacae,Pseudomonas azotoformans,Shewanella putrefaciens,Raoultella terriqena,Sphinqobacterium,Bizionia,Shewanella fidelis,Bacillus cereus,Vibrio scophthalmi,Vibrio harveyi,Staphylococcus sciuri,Aliivibrio.The environmental total DNAs was extracted from intestinal contents in Takifugu rubripes,and the intestinal bacterial diversity was investigated by 16S rDNA gene libraries were generated by using PCR and TA cloning.Constructing the 16S rDNA clone library,as many as 100 positive clones of 16S rDNA clone library were selected and digested with the restriction endonucleases Hae III and Hinf I.The patterns of the bands were then used to classify clones into operational taxonomic units(OTUs),and 60 of individual purified PCR products were selected for sequencing and classified into 29 operational taxonomy units(OTUs).The coverage of the library was 71%.The results show that the majority sequences were compared by using the basic local alignmentsearch tool(BLAST)in GenBank databases),the homology from 88%to 99%.There are eight OTUs sequence homology less than 98%.Four major phylogenetic lineages were identified:?-Proteobacteria(44.8%),CFB group(6.9%),Firmicutes(13.8%),Unclassified group(34.5%),y-Proteobacteria is the dominating group.The sequences from the libraries indicating a high richness in the intestinal contents microbial communities.These results suggested that Vibrio,Psychrobacter,Pseudomonas,Bacillus,Staphylococcus,Aliivibrio,uncultured bacterium,Raoultella,Exiguobacterium,Weissella,Sphingobacterium,Bizionia and Mycoplasma,the Vibrio,Shewanella and Photobacterium are the predominant bacteria in the intestine of Takifugu rubripes.