The Microflora Structure Analysis In Digestive Tract Of One Local Chicken From Guizhou

The kirin chicken is a quality local chicken in china,a few of them exist in rural farm in mountain areas of Guizhou Province.As the feature of feather turn forward,the kirin chicken is lack the ability to keep body warm.So it is easy to infect the disease?grow slow and death by the change of environment.This is the main problem for large-scale feeding.To improve the benefit of kirin chicken feeding,this focus on the structure of the in the digestive tract of the kirin chicken,and to understand the changing process of bacterial flora structure in the digestive tract when the kirin chicken growing.The purpose of this research is to provide the reference for raising the feeding efficiency and enlarging the feeding scale of kirin chicken.1.Establishment of quantitative PCR method based on optical density of 16SrDNA Extract DH5 alpha Escherichia coli genome,after determining the concentration(1.29×107copies/?L),genome was gradient diluted with distilled water.PCR amplification based on the bacterial universal primers(16SrDNA),take the optical density value of electrophoretic band as a reliable indicator.The quantitative PCR method was the result established by the result,and the linear was made by the optical density value of five adjacent gradient dilution.The formula is:y=8.7562×-26.838(Y?29.78),the correlation coefficient R~2=0.996.The repeatability was good and the sensitivity can reach 166copies/L.2.Analysis of digestive tract bacteria contents in different days of kirin chicken5~10 chicken were used at 5d,10d,20d,40d,60d,90d and 120d respectively.Collect the amount of contents in crop,proventriculus,duodenum,jejunum,ileum,cecum and rectum,and using the PCR method to analysis bacterial of content.Result:With the increase of age,the number of contents bacteria were growth in different parts of the digestive tract of the kirin chicken.However,the contents of digestive tract bacteria were not changed much after20~40 days of age;As a whole,the crop,proventriculus and gizzard have the most amount of bacteria,and the number of bacteria in duodenum,jejunum and ileum was less.3.DGGE analysis of bacterial in digestive tract in different days of kirin chicken5~10 chicken were used at 5d,10d,20d,40d,60d,90d and 120d respectively.Collect the amount of contents in crop,proventriculus,duodenum,jejunum,ileum,cecum and rectum,and using the PCR-DGGE method to analysis structure of digestive tract flora.Result:A total of 132 bands were detected,the recycling bands were sequenced and compared on the GenBank.41 different sequence bands were found,and those belong to deformation bacteria,Firmicutes,actinomycetes door,intends to the phylum Bacteroidetes,mastigomycotina,joint deuteromycotina and Deinococcus-dwelling bacteria Phylum,it contains Lactobacillus,bacillus,and Bacillus,et al.Total 25 genus.The contents of the digestive tract belong to Lactobacillus,Lactobacillus and Staphylococcus in the 5~20 days kirin chicken.The contents of the digestive tract belong to lactobacillus,Lactobacillus,Mycobacterium and Staphylococcus in the 40~90 days kirin chicken,And belong to Mycobacterium and Staphylococcus in the 120 days.