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...

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Veröffentlicht in:Journal of medical microbiology Jg. 54; H. Pt 11; S. 1093
Hauptverfasser: Hayashi, Hidenori, Takahashi, Rei, Nishi, Takahiro, Sakamoto, Mitsuo, Benno, Yoshimi
Format: Journal Article
Sprache:Englisch
Veröffentlicht: England 01.11.2005
Schlagworte:
Aged
Bacteria - classification
Bacteria - genetics
Cecum - microbiology
Colon, Sigmoid - microbiology
Deoxyribonuclease HpaII - metabolism
Deoxyribonucleases, Type II Site-Specific - metabolism
DNA Fingerprinting
DNA, Bacterial - analysis
DNA, Bacterial - chemistry
DNA, Bacterial - isolation & purification
DNA, Ribosomal - genetics
Female
Humans
Ileum - microbiology
Intestines - microbiology
Jejunum - microbiology
Male
Molecular Sequence Data
Polymorphism, Restriction Fragment Length
RNA, Ribosomal, 16S - genetics
Sequence Analysis, DNA
ISSN:0022-2615
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Zusammenfassung: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.
Bibliographie:ObjectType-Article-2
SourceType-Scholarly Journals-1
ObjectType-Feature-1
content type line 23
ISSN:0022-2615
DOI:10.1099/jmm.0.45935-0