Study On Diversity And Function Of Interstinal Bacterial Flora From African Lions Of Different Seasons

The African Lions are the greatest Catamount;it belonging to Panthera,Carnivoraand,Felidae,There is the extinction of small populations because of habitat destruction.While,The diseases of African Lions occur in mouth and digestive tract.Thus,study on diversity of intestinal bacterial flora from African Lion of different seasons is very necessary for changing the constitution of intestinal bacterial flora,adjusting the quantity of dominant bacterial community and preventing and treating of intestinal diseases.Firstly,in this study we used PCR-DGGE combining with RT-PCR technology to research diversity of intestinal bacterial flora from African Lions of different seasons.Through the hapmap of PCR-DGGE we found that the winter and spring intestinal flora have higher similarity,and the summer and autumn intestinal flora have higher diversity.We obtained 13 different bands,further sequencing result showed that Firmicutes and Proteobacteria is the dominant intestinal microflora.Devising 5specific primers among the 13 bands to precede RT-PCR,the result showed that the 5bacteria are all existed in intestinal flora of different seasons.But the numbers of bacteria had significant difference.We also used metagenome sequencing technology to explore diversity of intestinal bacterial flora from African Lions.The dominant bacterium are Firmicutes,Proteobacteria and Fusobacteria.But there were abundant Bacteroidetes in summer intestinal bacterial,while there were a little Chloroflexi in autumn and spring intestinal bacterial.At the genus level,the dominant bacterium was Collinsella and Lachnospiraceae in spring,Lachnospiraceae and Campylobacter in summer;while Lachnospiraceae was replaced with Campylobacter becaming the dominant bacteria.And in the winter had much Escherichia-Shigella.Finally in order to study the function of different strains,we used traditional cultured method to screen out function of active strain and processing further verification with them.The result showed that both Subtilis-partial and Bacillus-safensis in Bacillus genus had cellulose,amylase and protease functions.While only detected in metagenome sequencing,the Sporosarcina-globispora had cellulose and amylase functions.Sporosarcina-saromensis in Sporosarcina genus onlyhad protease function.Enterococcus-faecium and Enterococcus-mundtii in Enterococcus genus both had lipase and ligninase functions.Detected in PCR-DGGE and as dominant bacteria in metagenome sequencing,Bacillus-safensis had cellulose and protease.Enzyme-producing conditions optimization of Bacillus-safensis showed that the highest production cellulose activity at 35 ℃ for 84 h with 1%inoculation quantity and pH8.0.The highest production protease activity at 40℃ for 48 h with 5%noculation quantity and pH4.0.From this study,the results showed structural and functional characteristics of strains in different seasons intestinal bacterial flora from African Lions.This provides a theoretical basis for future research of intestinal flora diversity and screening functional dominant strains.