Current Microbiology

, Volume 62, Issue 1, pp 1–6 | Cite as

Diversity of the Formyltetrahydrofolate Synthetase (FTHFS) Gene in the Proximal and Mid Ostrich Colon

  • Hiroki MatsuiEmail author
  • Saori Yoneda
  • Tomomi Ban-Tokuda
  • Masaaki Wakita


We analysed fragments of the formyltetrahydrofolate synthetase (FTHFS) gene, which encodes a key enzyme in reductive acetogenesis, from the bacterial flora in the proximal (PC) and mid (MC) colon of three ostriches to assess and compare bacterial diversity in this organ. Two clone libraries of FTHFS fragments were constructed from DNA extracted from digesta of the PC and MC, and a total of 46 cloned sequences were analysed from each library. A wide variety of FTHFS sequences were recovered. The coverage of the PC and MC libraries was 90.0% and 83.3%, respectively. Shannon–Wiener index (H’) and Chao1 of the MC library were higher than those of PC library. The sequences from each library were classified into 15 operational taxonomic units (OTUs) and clusters. Only four OTUs in cluster I were distantly related to known acetogens from human feces and rumen, suggesting the presence of the novel acetogens. Phylogenetic analysis suggests that composition of FTHFS sequences differs for the PC and MC.


Methanogenesis Human Feces Acetogenesis Corn Gluten Meal Acetogenic Bacterium 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



The authors thank Professor Kazunari Ushida and his colleagues (Kyoto Prefectural University, Kyoto, Japan) for their instruction on DNA extraction. Funding for this study was provided by a Grant-in-Aid for Scientific Research, Japan Society for the Promotion of Science (17380157). Nucleotide sequencing was carried out at Life Science Research Center (Center for Molecular Biology and Genetics), Mie University (Tsu, Japan).


  1. 1.
    Altschul SF, Madden TL, Schäffer AA et al (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402CrossRefPubMedGoogle Scholar
  2. 2.
    De Graeve KG, Grivet JP, Durand M et al (1994) Competition between reductive acetogenesis and methanogenesis in the pig large-intestinal flora. J Appl Bacteriol 76:55–61PubMedGoogle Scholar
  3. 3.
    Fievez V, Mbanzamihigo L, Piatton F et al (2001) Evidence for reductive acetogenesis and its nutritional significance in ostrich hindgut as estimated from in vitro incubactions. J Anim Physiol A Anim Nutr 85:271–280CrossRefGoogle Scholar
  4. 4.
    Fukatsu T (1999) Acetone preservation: a practical technique for molecular analysis. Mol Ecol 8:1935–1945CrossRefPubMedGoogle Scholar
  5. 5.
    Hattori K, Matsui H (2008) Diversity of fumarate reducing bacteria in the bovine rumen revealed by culture dependent and independent approaches. Anaerobe 14:87–93CrossRefPubMedGoogle Scholar
  6. 6.
    Hayashi H, Shibata K, Bakir MA et al (2007) Bacteroides coprophilus sp. nov., isolated from human faeces. Int J Syst Evol Microbiol 57:1323–1326CrossRefPubMedGoogle Scholar
  7. 7.
    Juottonen H, Galand PE, Yrjälä K (2006) Detection of methanogenic Archaea in peat: comparison of PCR primers targeting the mcrA gene. Res Microbiol 157:914–921CrossRefPubMedGoogle Scholar
  8. 8.
    Kitahara M, Sakamoto M, Ike M et al (2005) Bacteroides plebeius sp. nov. and Bacteroides coprocola sp. nov., isolated from human faeces. Int J Syst Evol Microbiol 55:2143–2147CrossRefPubMedGoogle Scholar
  9. 9.
    Larkin MA, Blackshields G, Brown NP et al (2007) Clustal W and Clustal X version 2.0. Bioinformatics 23:2947–2948CrossRefPubMedGoogle Scholar
  10. 10.
    Leaphart AB, Lovell CR (2001) Recovery and analysis of formyltetrahydro-folate synthetase gene sequences from natural populations of acetogenic bacteria. Appl Environ Microbiol 67:1392–1395CrossRefPubMedGoogle Scholar
  11. 11.
    Liu C, Finegold SM, Song Y et al (2008) Reclassification of Clostridium coccoides, Ruminococcus hansenii, Ruminococcus hydrogenotrophicus, Ruminococcus luti, Ruminococcus productus and Ruminococcus schinkii as Blautia coccoides gen. nov., comb. nov., Blautia hansenii comb. nov., Blautia hydrogenotrophica comb. nov., Blautia luti comb. nov., Blautia producta comb. nov., Blautia schinkii comb. nov. and description of Blautia wexlerae sp. nov., isolated from human faeces. Int J Syst Evol Microbiol 58:1896–1902CrossRefPubMedGoogle Scholar
  12. 12.
    Ljungdahl LG (1986) The autotrophic pathway of acetate synthesis in acetogenic bacteria. Annu Rev Microbiol 40:415–450CrossRefPubMedGoogle Scholar
  13. 13.
    Matsui H, Kojima N, Tajima K (2008) Diversity of formyltetrahydrofolate synthetase gene (fhs), a key enzyme for reductive acetogenesis, in the bovine rumen. Biosci Biotechnol Biochem 72:3273–3276CrossRefPubMedGoogle Scholar
  14. 14.
    Matsui H, Kato Y, Chikaraishi T, Moritani M, Ban-Tokuda T, Wakita M (2010) Microbial diversity in ostrich ceca as revealed by 16S ribosomal RNA gene clone library and detection of novel Fibrobacter species. Anaerobe 16:83–93CrossRefPubMedGoogle Scholar
  15. 15.
    Ohashi Y, Igarashi T, Kumazawa F et al (2007) Analysis of acetogenic bacteria in human feces with formyltetrahydrofolate synthetase sequences. Biosci Microflora 26:37–40Google Scholar
  16. 16.
    Ohashi Y, Andou A, Kanaya M et al (2009) Acetogenic bacteria mainly contribute to the disposal of hydrogen in the colon of healthy Japanese. Biosci Microflora 28:17–19Google Scholar
  17. 17.
    Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425PubMedGoogle Scholar
  18. 18.
    Schloss PD, Handelsman J (2005) Introducing DOTUR, a computer program for defining operational taxonomic units and estimating species richness. Appl Environ Microbiol 71:1501–1506CrossRefPubMedGoogle Scholar
  19. 19.
    Song YL, Liu CX, McTeague M et al (2004) Clostridium bartlettii sp. nov., isolated from human faeces. Anaerobe 10:179–184CrossRefPubMedGoogle Scholar
  20. 20.
    Swart D, Mackie RI, Hayes JP (1993) Fermentative digestion in the ostrich (Struthio camelus var. domesticus), a large avian species that utilizes cellulose. S Afr J Anim Sci 23:127–135Google Scholar
  21. 21.
    Wolin MJ, Miller TL (1983) Interactions of microbial populations in cellulose fermentation. Fed Proc 42:109–113PubMedGoogle Scholar
  22. 22.
    Wolin MJ, Miller TL, Collins MD et al (2003) Formate-dependent growth and homoacetogenic fermentation by a bacterium from human feces: description of Bryantella formatexigens gen. no., sp.nov. Appl Environ Microbiol 69:6321–6326CrossRefPubMedGoogle Scholar
  23. 23.
    Wang RF, Cao WW, Cerniglia CE (1996) PCR detection and quantification of predominant anaerobic bacteria in human and animal fecal samples. Appl Environ Microbiol 62:1242–1247PubMedGoogle Scholar
  24. 24.
    Xu J, Bjursell MK, Himrod J et al (2003) A genomic view of the human Bacteroides thetaiotaomicron symbiosis. Science 299:2074–2076CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Hiroki Matsui
    • 1
    Email author
  • Saori Yoneda
    • 1
  • Tomomi Ban-Tokuda
    • 1
  • Masaaki Wakita
    • 1
  1. 1.Graduate School of BioresourcesMie UniversityTsuJapan

Personalised recommendations