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Applied and Environmental Microbiology, September 2003, p. 5472-5482, Vol. 69, No. 9
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.9.5472-5482.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Growth and Methane Oxidation Rates of Anaerobic Methanotrophic Archaea in a Continuous-Flow Bioreactor

Peter R. Girguis, Victoria J. Orphan,{dagger} Steven J. Hallam, and Edward F. DeLong*

Monterey Bay Aquarium Research Institute, Moss Landing, California 95039

Received 28 March 2003/ Accepted 25 June 2003

Anaerobic methanotrophic archaea have recently been identified in anoxic marine sediments, but have not yet been recovered in pure culture. Physiological studies on freshly collected samples containing archaea and their sulfate-reducing syntrophic partners have been conducted, but sample availability and viability can limit the scope of these experiments. To better study microbial anaerobic methane oxidation, we developed a novel continuous-flow anaerobic methane incubation system (AMIS) that simulates the majority of in situ conditions and supports the metabolism and growth of anaerobic methanotrophic archaea. We incubated sediments collected from within and outside a methane cold seep in Monterey Canyon, Calif., for 24 weeks on the AMIS system. Anaerobic methane oxidation was measured in all sediments after incubation on AMIS, and quantitative molecular techniques verified the increases in methane-oxidizing archaeal populations in both seep and nonseep sediments. Our results demonstrate that the AMIS system stimulated the maintenance and growth of anaerobic methanotrophic archaea, and possibly their syntrophic, sulfate-reducing partners. Our data demonstrate the utility of combining physiological and molecular techniques to quantify the growth and metabolic activity of anaerobic microbial consortia. Further experiments with the AMIS system should provide a better understanding of the biological mechanisms of methane oxidation in anoxic marine environments. The AMIS may also enable the enrichment, purification, and isolation of methanotrophic archaea as pure cultures or defined syntrophic consortia.

* Corresponding author. Mailing address: Monterey Bay Aquarium Research Institute, 7700 Sandholdt Rd., Moss Landing, CA 95039. Phone: (831) 775-1843. Fax: (831) 775-1620. E-mail: delong{at}mbari.org.

{dagger} Present address: NASA Ames Research Center, Moffett Field, CA 94035-1000.

Applied and Environmental Microbiology, September 2003, p. 5472-5482, Vol. 69, No. 9
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.9.5472-5482.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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