The Diversity Of Lactobacillus Community By PCR-DGGE Method And Its Application In Intestinal Flora Of Cured Dogs And Tapping Posion Dogs

This article established a kind of rapid,accurate analysis method of Lactobacil lus diversity by specific primer.Then we use this method to monitoring the diversi ty of Lactobacillus of cured dogs and tapping poison dogs.We want to know the diversity of Lactobacillus in intestinal flora of healthy dogs and diarrhea dogs,esta blishing the system of environmental indicators and providing a theoretical basis of probiotics of Lactobacillus.The main content of this article and the results are as f ollows:1.105 species or subspecies Lactobacillus sequence and other intestinal bacteria sequence comparison by using the Check Probe,ARB,and BLAST programs confirmed that the targeted region was conserved only among all of the 16S rDNA sequences of Lactobacillus.This result is proved by primer specific experiments.2.16 weaned dogs were randomly divided into 4 groups which were two control group,and two test groups.The control group A fed diets added with Bacillus (0.1%, 1.1×106CFU/g),and the control group B fed black diets.The treatment group fed diets added with Bacillus (0.1%,1.1×106CFU/g) and Salmonella(105CFU/mL).The tapping group fed diets added with Salmonella(105 CFU/mL).Each group had 4 replicates, The parallel feeding tests lasted for 9 days.To studying the differences dietary due to the variation of the diversity of Lactobacillus,and applied PCR-DGGE to analysis puppies at different stages of experime-ntal intestinal flora structure and diversity.The experimental results showed that groups of not fed Bacillus had emerged diarrhea,compared in only one of the fed Bacillus groups,which indicated that the Pathological model built successful (P <0.05).3.The DGGE results indicated that:①DGGE band pattern similarity indices of all groups in the first experimental day were above 57%.There had individual diff erence in the structure on intestinal microflora and its diversity of puppies.②The Dogs Intestinal Microflora diversity of the Control group A was improves gradually from 0 day to 9 days,4 replicates results showed that the bands of dominant mic roflora were increased,and the different bands were decreased,these results indicat e that diets of dogs with Bacillus had the proliferation action to intestinal available bacteria.The result of dendrogram shows that similarity of the Control group A ever y body's DGGE profiles similarity index of 0 day is 66-88%,3 day is 71-90%,6 day is 77-89%,9day is 62-74%.③Infected Salmonella after 9day of the tapping p oison group Lactobacillus diversity appeared to decline significantly indicate that sal monella can reduce the diversity of Lactobacillus. From average band number of ch annel group,the number is 5.3 after poison attack,compared with Oday declined 2 7.4%,compared with 3day and 6day declined 39%,35%.④Infected Salmonella after 9day of the treatment group Lactobacillus diversity appeared to decline significantly From average band number of channel group,the number is 7.3 after poison atta ck,compared with Oday declined 8%,compared with 3day declined 35.3%,compa red with 6day declined 18.9%.But the protection of bacillus,the average number of band is more than tapping poison group indicate that bacillus can increase diarrhe a dog Lactobacillus diversity.