An Study Of Intestinal Microflora In Healthy People In Kunming And The Difference Between Fecesand And Colonic Mucosa Microflora

objective:Studies have shown that there are correLMtions and differences between luminal microbiota(LM)and mucosal-associated microbiota(MAM),which are important components of intestinal microbiotain healthy people.However,intestinal microbiotais provenly affected by different genetic background,social environment and living and eating habits.In this study,High through put sequencing of the 16SrDNA gene was used to characterize LM and MAM's bacterial communities,which both of healthy popuLMtion in Yunnan Province,and their difference were analyzed.To further underst and the general composition of intestinal mucosa and fecal microecology of healthy people in Yunnan,and the normal difference between LM and MAM.It providesa theoretical basis for improving the health status of the popuLMtion by adjusting the abnormal differences between the two groups in the future.Methods:A total of 28 healthy people were enrolled from the First Affiliated Hospital of Kunming Medical University since September to December 2016.All subject swere excluded from other systemic diseases.Andthesex,age,nationality,BMI,smoking history and appendix surgery of all participants were recorded.The feces and mucosa samples were collected an ddividedinto two groups:the intestinal mucosa sample group and the fecal sample group.The samples were constructed into a Paired-End(PE)sequencing library,and the V3-V4region was detected by PE300 strategy.Basedon Illumina MiSeq sequencing pLMtform,bacterial flora structure of feces and mucosa samples from 28 healthy controls was detected.The sequences plicing software FLMSH splie in greads was Tags.The sample quality statisties showed that 16SrDNA sequencing was of good quality.Through OTU species clusteringand species an notation,the abundanee of individual bacteria in the samples was obtained.The differences betweenL A and MAM microeco system composition were obtained by Alphad iversity analysis(including Chao index,Ace index,Shannon index and Simpson index of calcuLMting OTU number in sequencing results).Results:1.By comparing the OTU species annotations after grouping the samples,632 OTUs were found in the fecal sample group(FH)of healthy people.There were 842 OUTs in the mucosal sample group(MH).A total of 569 strains of 905 OTUs were common in feces and intestinal mucosa of FH and MH groups.63 strains of OUT were unique to FH and 273 were unique to intestinal mucosa of MH.2.On the phylum level,the bacteria isolated from intestinal mucosa(MAM)and feces(LM)in this study can be classified into 18 phylum categories.Bacteroides and sclerenchyma aeeount for more than half of the total flora,and the ratio of sclerenchyma to Bacteroides is less than 1.Proteus in healthy population of Kunming.Proteobacteria,Actinobacteria,Fusobacteria and are relatively less abundant bacteria,accounting for a relatively large proportion of mucosa-related bacteria than fecal bacteria.The relative abundance of Clostridium in mucosa and feces of volunteers No.20 and 34 was higher than that of other volunteers.3.At the level of Class,the strains analyzed by MAM and LM in this study can be classified into 32 classes,and the relative abundance of Bacteroides in these two classes is more than that of the other 31 classes.In addition,Clostridium,,Bacili,Chlamydia and Proteus alpha,Proteus beta,Proteus Delta and Proteus epsilon account for more than half of the intestinal flora of healthy people.Among the 32 bacterial classes,the relative abundance of flexible membranes,actinomycetes,ferricobacterium,verrucous microorganisms,Termoleophilia,acidobacterium,bacillus and anomalous cocci was relatively low.The proportion of MAM is relatively larger than that of LM.4.At the order level,63 orders were identified in MAM and LM.LMctobacilLMles and Bacteroidales are the most abundant of the 63 identified orders.Secondly,887 species of Clostridium with abundance more than 1%consisted of BacilLMles,Selenomonadales,Pseudomonas,Enterobacteriales,Burkholderiales,Erysipelotrichales and Spirochaetales.15 orders of bacteria in intestinal mucosa were significantly higher than those in feces(23.8%),and the relative abundance of 2 families was higher than that of Enterobacteriales and BacilLMles.The relative abundance of bacteria in intestinal mucosa was more than 1%.5.At the Family level,of the 117 families identified by MAM and LM,Streptococcaceae and PrevotelLMceae account for the highest abundance of 117 families identified at the Family level.Secondly,Bacteroidaceae,Leuconostocaceae,PLMnococcaceae,Ruminococcaceae,LMctobacilLMceae,VeillonelLMceae,MoraxelLMe,Comamonaceae,Enterococceae,Enterobacteriaceae were more than 1%abundant.Cteriaceae,Erysipelotrichaceae and Peptostreptococcaceae.The relative abundance of 32 bacteria(27.4%)in mucosa was significantly higher than that in feces,including Bacillaceae,Streptococcaceae,Enterobacteriaceae and Porphyromonaceae.The relative abundance of Furtsobacteriaceae and EggehelLMceae in feces was more than 1%.P values were 0.041 and 0.047 respectively.6.At genus level,241 strains identified in MAM and LM.Bacteroides,Clostridium,Lactobacillus and Untyped Bacteria constitute the majority of intestinal flora in 241 identified genera.At the generic level,the relative abundance of 49 bacteria(20.3%)in mucosa was significantly higher than that in feces.The relative abundance of Lactococcus,Escherichia,Parabacteroides and Lachnoclostridium was more than 1%.Conclusions:1.The total number of gene sequences detected in intestinal tract of healthy people in Kunming was 905 OTUs,and the diversity of intestinal flora may be superior to that of healthy population in some developed western areas reported in relevant literature,2.Although the diversity of intestinal mucosa-related flora and intestinal cavity flora needs to be confirmed by enlarging the sample size.However,through this study,we can conclude that there are significant differences in the composition and structure of intestinal mucosa-related microbial flora between healthy people and intraluminal microbial flora,which should be studied separately in this study.3.The intestinal bacteria can be classified into 18 categories,of which Bacterioides and Thickwalled bacteria are dominant,and there is no significant difference between East and West.4.At the order level,the relative abundance advantages of Lactobacilus(LactobacilLMles)and Bacteroidales may be protective factors for metabolic-related diseases,inflammatory bowel diseases and irritable bowel syndrome;at the Family level,the relative abundance of Enterobacteriaceae and Fusobacteriaceae may increase as well.The high incidence of IBD in Kunming may be a risk factor for IBD.