The Succession Change Research Of Bifidobacterium Probiotics In Bama Minipig Fecal Flora

In this experiment,we selected four Chinese characteristics resources Bama minipigs,all are male,littermates born.Keeping the environment in the same condition within 90 days followed up. Expect to obtain the minipigs succession changes of Bifidobacterium probiotics in the fecal through sampling completely.These studies is valuable to Bama minipigs and human intestinal flora intestinal flora research in the similar,and the promotion of special resources in Bama minipigs fecal provide data in microecology research.1.As the specificity of microbial flora of Bama minipig in fecal.It is necessary to optimizate the components for the 16SrDNA.First the nucleic acid extraction methods were selected for Bama minpigs microflora.Then through the application of PCR-DGGE technology to four Bama minpigs Bifidobacterium flora.Cluster analysis of the diversity of flora research found that fauna flora changes were relatively unstable during weaning.Finaly established 16S rDNA clone library on the basis.2.The study of fecal Bifidobacterium flora will contribute to the understanding of intestinal microecology,and PCR-DGGE method is the important way to examine the livestock and manure Bifidobacterium flora.As a result of bifidobacterium flora is particularity in Bama minipigs,it is necessary to optimize each component in PCR reactions.The experimental conditions of the system is optimized first,PCR amplification products tested by gel imaging system and Quantity One.The results showed that the curve drawn by the signal to noise ratio of each component concentration. Then the best reaction obtained:25ul system,the DNA template is 20-40ng(15nM),primer concentration 7.SpM,Mg²⁺(25mM) the addition is 1.5uL;dNTP(10mM) added to the 2.0uL,and compared to conventional conditions,the results is better.The PCR product increased.Amplification stage selected 35 cycles,annealing temperature to 58℃is the best results.At the same time,the examination by denaturing gradient gel,and finally the trial shows that the scope of DGGE gel variability 50-80%can be achieved the best results.The establishment of the 16S rDNA based for fecal Bifidobacterium flora diversity provides an effective method in molecular biology research technology.Compared to conventional PCR conditions the results is better,increasing the DGGE bands,obtaining more stable results for Bama minipig Bifidobacterium fecal flora.The results also show that animals fecal Bifidobacterium flora is relative stability befor weaning,followed by a lot of changes for weaning behavior in fecal Bifidobacterium flora.Dynamic tracking and cluster analysis of similarity found in the in fecal samples bacterial flora showed that significant instability in the different stages to different trends,however the whole time divided into three stages.But dramaticly obvious differences between individuals.This study clearly show that the dynamics in the succession in the piglet,weaning is a greater stress.The succession of the dynamic changes of Bifidobacterium flora was not obvious to all the piglets.Weaning stress is not a significant impact on some individuals for the rapid establishment of a stable post-weaning,and a significant factor may be the late weaning time relatively,and animals can adapt quickly to changes in diet..3.To intensively study fecal Bifidobacterium flora,we established Bama minipig fecal 16S rDNA library of bacteria.BLAST analysis revealed that,90 minipigs clones for the Bama,the 56 sequence results are similar to Bifidobacterium adolescentis(T) and Bifidobacterium breve are 99%, accounting for 62.2%percent of the library;14 sequence close to Bifidobacterium infantis; Bifidobacterium longum(T) are 99%,accounting for 15.6%;4 sequence similar to Bifidobacterium animals(T);Bifidobacterium angulatum(T) to 99%,accounting for 4.4%.Bama minpigs,as an experimental animal models,further research is still needed to chose the best approach.for the more probiotic flora data access,...