Molecular analysis of jejunal, ileal, caecal and recto-sigmoidal human colonic microbiota using 16S rRNA gene libraries and terminal restriction fragment length polymorphism.
Microbiota in gut contents of jejunum, ileum, caecum and recto-sigmoid colon obtained from three elderly individuals at autopsy were compared using 16S rRNA gene libraries and terminal restriction fragment length polymorphism (T-RFLP). Random clones of 16S rRNA gene sequences were isolated after PCR amplification with universal primer sets of total genomic DNA extracted from each sample of gut contents. An average of 90 randomly selected clones were partially sequenced (about 500 bp). T-RFLP analysis was performed using the 16S rRNA gene amplified from each sample. The lengths of the terminal restriction fragments were analysed after digestion with HhaI and MspI. The jejunal and ileal microbiota consisted of simple microbial communities of streptococci, lactobacilli, 'Gammaproteobacteria', the Enterococcus group and the Bacteroides group. Most of the species were facultative anaerobes or aerobes. The Clostridium coccoides group and the Clostridium leptum subgroup, which are the most predominant groups in human faeces, were not detected in samples from the upper gastrointestinal tract. The caecal microbiota was more complex than the jejunal and ileal microbiota. The C. coccoides group, the C. leptum subgroup and the Bacteroides group were detected in the caecum. The recto-sigmoidal colonic microbiota consisted of complex microbial communities, with numerous species that belonged to the C. coccoides group, the C. leptum subgroup, the Bacteroides group, 'Gammaproteobacteria', the Bifidobacterium group, streptococci and lactobacilli, and included more than 26 operational taxonomic units. The results showed marked individual differences in the composition of microbiota in each region.
Hayashi
Hidenori
H
Microbe Division/Japan Collection of Microorganisms, RIKEN BioResource Center, Saitama 351-0198, Japan 2Departments of Pathology and Tumor Biology, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan 3Department of Medicine, Shiga Health Insurance Hospital, Otsu, Shiga 520-0846, Japan.
Takahashi
Rei
R
Microbe Division/Japan Collection of Microorganisms, RIKEN BioResource Center, Saitama 351-0198, Japan 2Departments of Pathology and Tumor Biology, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan 3Department of Medicine, Shiga Health Insurance Hospital, Otsu, Shiga 520-0846, Japan.
Nishi
Takahiro
T
Microbe Division/Japan Collection of Microorganisms, RIKEN BioResource Center, Saitama 351-0198, Japan 2Departments of Pathology and Tumor Biology, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan 3Department of Medicine, Shiga Health Insurance Hospital, Otsu, Shiga 520-0846, Japan.
Sakamoto
Mitsuo
M
Microbe Division/Japan Collection of Microorganisms, RIKEN BioResource Center, Saitama 351-0198, Japan 2Departments of Pathology and Tumor Biology, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan 3Department of Medicine, Shiga Health Insurance Hospital, Otsu, Shiga 520-0846, Japan.
Benno
Yoshimi
Y
Microbe Division/Japan Collection of Microorganisms, RIKEN BioResource Center, Saitama 351-0198, Japan 2Departments of Pathology and Tumor Biology, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan 3Department of Medicine, Shiga Health Insurance Hospital, Otsu, Shiga 520-0846, Japan.
D000368:Aged; D001419:Bacteria / Q000145:classification* / Q000235:genetics; D002432:Cecum / Q000382:microbiology; D012809:Colon, Sigmoid / Q000382:microbiology; D016172:DNA Fingerprinting; D004269:DNA, Bacterial / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000235:genetics; D019003:Deoxyribonuclease HpaII / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D005260:Female; D006801:Humans; D007082:Ileum / Q000382:microbiology; D007422:Intestines / Q000382:microbiology*; D007583:Jejunum / Q000382:microbiology; D008297:Male; D008969:Molecular Sequence Data; D012150:Polymorphism, Restriction Fragment Length; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D017422:Sequence Analysis, DNA
D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D019003:Deoxyribonuclease HpaII; D015252:Deoxyribonucleases, Type II Site-Specific; C037130:GCGC-specific type II deoxyribonucleases