Diversity Of Gastrointestinal Microflora Of Tibetan Pig

Taken fresh feces of Landrace and Tibetan swine as materials, PCR/DGGE combinedwith16S rNDA sequence analysis were applied for biodiversity analysis of thegastrointestinal tract of Tibetan pig. DNA was directly extracted from feces of two swinespecies, and then Polymerase Chain Reaction (PCR) amplification was conducted to amplifyV6-V8region of16S rDNA gene. All the PCR products were separated by38%-60%Denaturing Gradient Gel Electrophoresis (DGGE) to compare the predominant bacterialcommunity structure. Then the advantaged marked bands of DGGE patterns were recoveredand subcloned into E. coli respectively. Colony PCR and double enzyme (EcoRâ… and Salâ… )digestion analysis were applied to select the positive clones. Then all the positive clones weresequenced and compared to the existed ones in NCBI and Quantity One software was appliedfor UPGMA analysis. The resultant27clone sequences in16S rDNA DGGE librariesrevealed that7clones show similarity over97%with that of database sequences, while therest present similarity in a range of90%-96%. The content of the new microbes in intestinalmicroflora of the Tibetan pigs is74.1%.14clones were similar to cellulose-decomposingmicroorganisms or hemicellulose-decomposing microorganisms. The main conclusions of thestudy were as follows:1. The dominant microbes of the Tibetan pigs are (more than30species)moremultiformity than The dominant microbes of the Landrace(about20species).2. The bacteria community structure of different species of pigs was similar to each otheronly0.44. The similarities within species were noticeably higher than that between species.3. Intestinal microflora of the Tibetan pigs include larger amount of unknown unculturedmicroorganisms(Succinivibrio dextrinosolvens,Lachnospiraceae bacterium,Clostridialesbact-erium,Clostridium sp.) than intestinal microflora of Landrace.4. The dominant microbes of the Tibetan pigs (Succinivibrio dextrinosolvens,Eubacterium rectale, Eubacterium eligens, Anaerobiospirillum succiniciproducens,Clostridiales bacterium,Lachnospiraceae bacterium and Lactobacillales bacterium) weresimilar to cellulose-decomposing microorganisms or hemicellulose-decomposingmicroorganisms.