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Applied and Environmental Microbiology, July 2005, p. 3725-3733, Vol. 71, No. 7
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.7.3725-3733.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Growth and Population Dynamics of Anaerobic Methane-Oxidizing Archaea and Sulfate-Reducing Bacteria in a Continuous-Flow Bioreactor

Peter R. Girguis,1 Aaron E. Cozen,2 and Edward F. DeLong3*

Monterey Bay Aquarium Research Institute, 7700 Sandholdt Road, Moss Landing, California 95039,1 University of California, Santa Cruz, 225 Sinsheimer, Santa Cruz, California 95064,2 Massachusetts Institute of Technology, Department of Civil & Environmental Engineering, 15 Vassar St., 48-427, Cambridge, Massachusetts 02139-43073

Received 9 November 2004/ Accepted 8 February 2005

The consumption of methane in anoxic marine sediments is a biogeochemical phenomenon mediated by two archaeal groups (ANME-1 and ANME-2) that exist syntrophically with sulfate-reducing bacteria. These anaerobic methanotrophs have yet to be recovered in pure culture, and key aspects of their ecology and physiology remain poorly understood. To characterize the growth and physiology of these anaerobic methanotrophs and the syntrophic sulfate-reducing bacteria, we incubated marine sediments using an anoxic, continuous-flow bioreactor during two experiments at different advective porewater flow rates. We examined the growth kinetics of anaerobic methanotrophs and Desulfosarcina-like sulfate-reducing bacteria using quantitative PCR as a proxy for cell counts, and measured methane oxidation rates using membrane-inlet mass spectrometry. Our data show that the specific growth rates of ANME-1 and ANME-2 archaea differed in response to porewater flow rates. ANME-2 methanotrophs had the highest rates in lower-flow regimes (µANME-2 = 0.167 · week–1), whereas ANME-1 methanotrophs had the highest rates in higher-flow regimes (µANME-1 = 0.218 · week–1). In both incubations, Desulfosarcina-like sulfate-reducing bacterial growth rates were approximately 0.3 · week–1, and their growth dynamics suggested that sulfate-reducing bacterial growth might be facilitated by, but not dependent upon, an established anaerobic methanotrophic population. ANME-1 growth rates corroborate field observations that ANME-1 archaea flourish in higher-flow regimes. Our growth and methane oxidation rates jointly demonstrate that anaerobic methanotrophs are capable of attaining substantial growth over a range of environmental conditions used in these experiments, including relatively low methane partial pressures.

* Corresponding author. Mailing address: Massachusetts Institute of Technology, Department of Civil & Environmental Engineering, 15 Vassar St., 48-427, Cambridge, MA 02139-4307. Phone: (617) 253-5271. Fax: (617) 258-8850. E-mail: delong{at}mit.edu.

Applied and Environmental Microbiology, July 2005, p. 3725-3733, Vol. 71, No. 7
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.7.3725-3733.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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