This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Zoetendal, E. G.
Right arrow Articles by de Vos, W. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Zoetendal, E. G.
Right arrow Articles by de Vos, W. M.
Agricola
Right arrow Articles by Zoetendal, E. G.
Right arrow Articles by de Vos, W. M.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, September 2002, p. 4225-4232, Vol. 68, No. 9
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.9.4225-4232.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Quantification of Uncultured Ruminococcus obeum-Like Bacteria in Human Fecal Samples by Fluorescent In Situ Hybridization and Flow Cytometry Using 16S rRNA-Targeted Probes

Erwin G. Zoetendal,1,2* Kaouther Ben-Amor,2,3 Hermie J. M. Harmsen,4 Frits Schut,5 Antoon D. L. Akkermans,2 and Willem M. de Vos1,2

Wageningen Centre for Food Sciences,1 Laboratory of Microbiology,2 Laboratory of Food Microbiology, Wageningen University, Wageningen,3 Department of Medical Microbiology, University of Groningen,4 Microscreen BV, Groningen, The Netherlands5

Received 12 February 2002/ Accepted 6 June 2002

A 16S rRNA-targeted probe was designed and validated in order to quantify the number of uncultured Ruminococcus obeum-like bacteria by fluorescent in situ hybridization (FISH). These bacteria have frequently been found in 16S ribosomal DNA clone libraries prepared from bacterial communities in the human intestine. Thirty-two reference strains from the human intestine, including a phylogenetically related strain and strains of some other Ruminococcus species, were used as negative controls and did not hybridize with the new probe. Microscopic and flow cytometric analyses revealed that a group of morphologically similar bacteria in feces did hybridize with this probe. Moreover, it was found that all hybridizing cells also hybridized with a probe specific for the Clostridium coccoides-Eubacterium rectale group, a group that includes the uncultured R. obeum-like bacteria. Quantification of the uncultured R. obeum-like bacteria and the C. coccoides-E. rectale group by flow cytometry and microscopy revealed that these groups comprised approximately 2.5 and 16% of the total community in fecal samples, respectively. The uncultured R. obeum-like bacteria comprise about 16% of the C. coccoides-E. rectale group. These results indicate that the uncultured R. obeum-like bacteria are numerically important in human feces. Statistical analysis revealed no significant difference between the microscopic and flow cytometric counts and the different feces sampling times, while a significant host-specific effect on the counts was observed. Our data demonstrate that the combination of FISH and flow cytometry is a useful approach for studying the ecology of uncultured bacteria in the human gastrointestinal tract.

* Corresponding author. Mailing address: Laboratory of Microbiology, Wageningen University, Hesselink van Suchtelenweg 4, 6703 CT Wageningen, The Netherlands. Phone: 31 317 483115. Fax: 31 317 483829. E-mail: egzoetendal{at}hotmail.com.

Applied and Environmental Microbiology, September 2002, p. 4225-4232, Vol. 68, No. 9
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.9.4225-4232.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Turroni, F., Foroni, E., Pizzetti, P., Giubellini, V., Ribbera, A., Merusi, P., Cagnasso, P., Bizzarri, B., de'Angelis, G. L., Shanahan, F., van Sinderen, D., Ventura, M. (2009). Exploring the Diversity of the Bifidobacterial Population in the Human Intestinal Tract. Appl. Environ. Microbiol. 75: 1534-1545 [Abstract] [Full Text]  
  • Liu, C., Finegold, S. M., Song, Y., Lawson, P. A. (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: 1896-1902 [Abstract] [Full Text]  
  • Derrien, M., Collado, M. C., Ben-Amor, K., Salminen, S., de Vos, W. M. (2008). The Mucin Degrader Akkermansia muciniphila Is an Abundant Resident of the Human Intestinal Tract. Appl. Environ. Microbiol. 74: 1646-1648 [Abstract] [Full Text]  
  • Swidsinski, A, Goktas, O, Bessler, C, Loening-Baucke, V, Hale, L P, Andree, H, Weizenegger, M, Holzl, M, Scherer, H, Lochs, H (2007). Spatial organisation of microbiota in quiescent adenoiditis and tonsillitis. J. Clin. Pathol. 60: 253-260 [Abstract] [Full Text]  
  • Klaassens, E. S., de Vos, W. M., Vaughan, E. E. (2007). Metaproteomics Approach To Study the Functionality of the Microbiota in the Human Infant Gastrointestinal Tract. Appl. Environ. Microbiol. 73: 1388-1392 [Abstract] [Full Text]  
  • Dinoto, A., Marques, T. M., Sakamoto, K., Fukiya, S., Watanabe, J., Ito, S., Yokota, A. (2006). Population Dynamics of Bifidobacterium Species in Human Feces during Raffinose Administration Monitored by Fluorescence In Situ Hybridization-Flow Cytometry. Appl. Environ. Microbiol. 72: 7739-7747 [Abstract] [Full Text]  
  • Kuhbacher, T, Ott, S J, Helwig, U, Mimura, T, Rizzello, F, Kleessen, B, Gionchetti, P, Blaut, M, Campieri, M, Folsch, U R, Kamm, M A, Schreiber, S (2006). Bacterial and fungal microbiota in relation to probiotic therapy (VSL#3) in pouchitis. Gut 55: 833-841 [Abstract] [Full Text]  
  • Maukonen, J., Satokari, R., Matto, J., Soderlund, H., Mattila-Sandholm, T., Saarela, M. (2006). Prevalence and temporal stability of selected clostridial groups in irritable bowel syndrome in relation to predominant faecal bacteria.. J Med Microbiol 55: 625-633 [Abstract] [Full Text]  
  • Mueller, S., Saunier, K., Hanisch, C., Norin, E., Alm, L., Midtvedt, T., Cresci, A., Silvi, S., Orpianesi, C., Verdenelli, M. C., Clavel, T., Koebnick, C., Zunft, H.-J. F., Dore, J., Blaut, M. (2006). Differences in Fecal Microbiota in Different European Study Populations in Relation to Age, Gender, and Country: a Cross-Sectional Study. Appl. Environ. Microbiol. 72: 1027-1033 [Abstract] [Full Text]  
  • van Tongeren, S. P., Slaets, J. P. J., Harmsen, H. J. M., Welling, G. W. (2005). Fecal Microbiota Composition and Frailty. Appl. Environ. Microbiol. 71: 6438-6442 [Abstract] [Full Text]  
  • de Vos, W. M. (2005). Lipotechoic acid in lactobacilli: D-Alanine makes the difference. Proc. Natl. Acad. Sci. USA 102: 10763-10764 [Full Text]  
  • Ben-Amor, K., Heilig, H., Smidt, H., Vaughan, E. E., Abee, T., de Vos, W. M. (2005). Genetic Diversity of Viable, Injured, and Dead Fecal Bacteria Assessed by Fluorescence-Activated Cell Sorting and 16S rRNA Gene Analysis. Appl. Environ. Microbiol. 71: 4679-4689 [Abstract] [Full Text]  
  • Lay, C., Rigottier-Gois, L., Holmstrom, K., Rajilic, M., Vaughan, E. E., de Vos, W. M., Collins, M. D., Thiel, R., Namsolleck, P., Blaut, M., Dore, J. (2005). Colonic Microbiota Signatures across Five Northern European Countries. Appl. Environ. Microbiol. 71: 4153-4155 [Abstract] [Full Text]  
  • Swidsinski, A., Weber, J., Loening-Baucke, V., Hale, L. P., Lochs, H. (2005). Spatial Organization and Composition of the Mucosal Flora in Patients with Inflammatory Bowel Disease. J. Clin. Microbiol. 43: 3380-3389 [Abstract] [Full Text]  
  • Ott, S J, Musfeldt, M, Wenderoth, D F, Hampe, J, Brant, O, Folsch, U R, Timmis, K N, Schreiber, S (2004). Reduction in diversity of the colonic mucosa associated bacterial microflora in patients with active inflammatory bowel disease. Gut 53: 685-693 [Abstract] [Full Text]  
  • Zoetendal, E. G., Collier, C. T., Koike, S., Mackie, R. I., Gaskins, H. R. (2004). Molecular Ecological Analysis of the Gastrointestinal Microbiota: A Review. J. Nutr. 134: 465-472 [Abstract] [Full Text]  
  • Yanagita, K., Manome, A., Meng, X.-Y., Hanada, S., Kanagawa, T., Tsuchida, T., Mackie, R. I., Kamagata, Y. (2003). Flow cytometric sorting, phylogenetic analysis and in situ detection of Oscillospira guillermondii, a large, morphologically conspicuous but uncultured ruminal bacterium. Int. J. Syst. Evol. Microbiol. 53: 1609-1614 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»