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Applied and Environmental Microbiology, June 2009, p. 3656-3662, Vol. 75, No. 11
0099-2240/09/$08.00+0     doi:10.1128/AEM.00067-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Enrichment and Molecular Detection of Denitrifying Methanotrophic Bacteria of the NC10 Phylum{triangledown}

Katharina F. Ettwig,* Theo van Alen, Katinka T. van de Pas-Schoonen, Mike S. M. Jetten, and Marc Strous{dagger}

Department of Microbiology, Institute for Water and Wetland Research, Radboud University Nijmegen, NL-6525 ED Nijmegen, The Netherlands

Received 12 January 2009/ Accepted 23 March 2009

Anaerobic methane oxidation coupled to denitrification was recently assigned to bacteria belonging to the uncultured phylum NC10. In this study, we incubated sediment from a eutrophic ditch harboring a diverse community of NC10 bacteria in a bioreactor with a constant supply of methane and nitrite. After 6 months, fluorescence in situ hybridization showed that NC10 bacteria dominated the resulting population. The enrichment culture oxidized methane and reduced nitrite to dinitrogen gas. We assessed NC10 phylum diversity in the inoculum and the enrichment culture, compiled the sequences currently available for this bacterial phylum, and showed that of the initial diversity, only members of one subgroup had been enriched. The growth of this subgroup was monitored by quantitative PCR and correlated to nitrite-reducing activity and the total biomass of the culture. Together, the results indicate that the enriched subgroup of NC10 bacteria is responsible for anaerobic methane oxidation coupled to nitrite reduction. Due to methodological limitations (a strong bias against NC10 bacteria in 16S rRNA gene clone libraries and inhibition by commonly used stopper material) the environmental distribution and importance of these bacteria could be largely underestimated at present.

* Corresponding author. Mailing address: Department of Microbiology, Institute for Water and Wetland Research, Radboud University Nijmegen, Heyendaalseweg 135, NL-6525 AJ Nijmegen, The Netherlands. Phone: 31-24-3652658. Fax: 31-24-3652830. E-mail: k.ettwig{at}science.ru.nl

{triangledown} Published ahead of print on 27 March 2009.

{dagger} Present address: Max Planck Institute for Marine Microbiology, Celsiusstrasse 1, D-28359 Bremen, Germany.

Applied and Environmental Microbiology, June 2009, p. 3656-3662, Vol. 75, No. 11
0099-2240/09/$08.00+0     doi:10.1128/AEM.00067-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

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