Distribution Analysis of Six Predominant Bacteroides Species in Normal Human Feces Using 16S rDNA-Targeted Species-Specific Primers

Article English OPEN
Miyamoto, Yukiko ; Watanabe, Koichi ; Tanaka, Ryuichiro ; Itoh, Kikuji (2011)
  • Publisher: Microbial Ecology in Health and Disease
  • Journal: Microbial Ecology in Health and Disease (issn: 1651-2235)
  • Related identifiers: doi: 10.3402/mehd.v14i3.8239
  • Subject:
    mesheuropmc: food and beverages | fluids and secretions

In order to analyze the predominant Bacteroides species in normal human feces, species-specific primer sets for Bacteroides eggerthii, B. fragilis, B. ovatus, B. thetaiotaomicron, B. uniformis and B. vulgatus were designed. Normal human feces were cultured on Bacteroides selective agar (neomycin brilliant green taurocolic acid (NBGT) agar plates). We identified the colonies with the aforementioned species-specific primers. B. vulgatus and B. ovatus were found to be predominant in all volunteers, and B. thetaiotaomicron and B. uniformis varied between volunteers. However, B. fragilis and B. eggerthii were not detected in any volunteers using this culture method. When we used the direct PCR method with extracted bacterial DNA from feces, B. vulgatus and B. ovatus were detected from all samples, and B. thetaiotaomicron, B. uniformis and B. fragilis from four or five out of six samples, but no B. eggerthii was detected. This is the first report that analyzes the predominant Bacteroides species in normal human intestine using a systematic molecular identification method.Keywords: Bacteroides, 16S rRNA, species-specific primer, human, feces.
  • References (17)
    17 references, page 1 of 2

    1. Mitsuoka T, Sega T, Yamamoto S. Improved methodology of qualitative and quantitative analysis of the intestinal ora of man and animals. Zentralbl Bakteriol 1965; 195: 455 - 69.

    2. Moore WE, Holdeman LV. Human fecal ora: the normal ora of 20 Japanese - Hawaiians. Appl Microbiol 1974; 27: 961 - 79.

    3. Paster BJ, Dewhirst FE, Olsen I, Fraser GJ. Phylogeny of Bacteroides, Prevotella, and Porphyromonas spp. and related bacteria. J Bacteriol 1994; 176: 725 - 32.

    4. Miyamoto Y, Itoh K. Design of cluster-speci c 16S rDNA oligonucleotide probes to identify bacteria of the Bacteroides subgroup harbored in human feces. FEMS Microbiol Lett 1999; 177: 143 - 9.

    5. Finegold SM, Attebery HR, Sutter VL. Effect of diet on human fecal ora: comparison of Japanese and American diets. Am J Clin Nutr 1974; 27: 1456 - 69.

    6. Kreader CA. Design and evaluation of Bacteroides DNA probes for the speci c detection of human fecal pollution. Appl Environ Microbiol 1995; 61: 1171 - 9.

    7. Tran T, Flynn MJ, Chen C, Slots J. Absence of Porphy - romonas asaccharolytica, Bacteroides fragilis and Chlamydia pneumoniae in human subgingival plaque. Oral Microbiol Immunol 1997; 12: 377 - 8.

    8. Wang RF, Cao WW, Campbell WL, Hairston L, Franklin W, Cerniglia CE. The use of PCR to monitor the population abundance of six human intestinal bacterial species in an in vitro semicontinuous culture system. FEMS Microbiol Lett 1994; 124: 229 - 37.

    9. Wang RF, Cao WW, Cerniglia CE. PCR detection and quantitation of predominant anaerobic bacteria in human and animal fecal samples. Appl Environ Microbiol 1996; 62: 1242 - 7.

    10. Tateno Y, Gojobori T. DNA Data Bank of Japan in the age of information biology. Nucleic Acids Res 1997; 25: 14 - 7.

  • Metrics
    No metrics available
Share - Bookmark