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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Vydané v:Journal of medical microbiology Ročník 54; číslo Pt 11; s. 1093
Hlavní autori: Hayashi, Hidenori, Takahashi, Rei, Nishi, Takahiro, Sakamoto, Mitsuo, Benno, Yoshimi
Médium: Journal Article
Jazyk:English
Vydavateľské údaje: England 01.11.2005
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Abstract 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.
AbstractList 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.
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.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.
Author Takahashi, Rei
Benno, Yoshimi
Hayashi, Hidenori
Nishi, Takahiro
Sakamoto, Mitsuo
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  givenname: Hidenori
  surname: Hayashi
  fullname: Hayashi, Hidenori
  organization: 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
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  givenname: Rei
  surname: Takahashi
  fullname: Takahashi, Rei
  organization: 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
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  givenname: Takahiro
  surname: Nishi
  fullname: Nishi, Takahiro
  organization: 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
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  givenname: Mitsuo
  surname: Sakamoto
  fullname: Sakamoto, Mitsuo
  organization: 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
– sequence: 5
  givenname: Yoshimi
  surname: Benno
  fullname: Benno, Yoshimi
  organization: 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
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PublicationTitle Journal of medical microbiology
PublicationTitleAlternate J Med Microbiol
PublicationYear 2005
SSID ssj0017339
Score 2.2904668
Snippet Microbiota in gut contents of jejunum, ileum, caecum and recto-sigmoid colon obtained from three elderly individuals at autopsy were compared using 16S rRNA...
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pubmed
SourceType Aggregation Database
Index Database
StartPage 1093
SubjectTerms 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
Title Molecular analysis of jejunal, ileal, caecal and recto-sigmoidal human colonic microbiota using 16S rRNA gene libraries and terminal restriction fragment length polymorphism
URI https://www.ncbi.nlm.nih.gov/pubmed/16192442
https://www.proquest.com/docview/68644654
Volume 54
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