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Applied and Environmental Microbiology, September 2003, p. 5503-5511, Vol. 69, No. 9
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.9.5503-5511.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Filamentous "Epsilonproteobacteria" Dominate Microbial Mats from Sulfidic Cave Springs

Annette Summers Engel,1* Natuschka Lee,2 Megan L. Porter,3 Libby A. Stern,1 Philip C. Bennett,1 and Michael Wagner4

Research Group for Microbial Geochemistry, Department of Geological Sciences, University of Texas at Austin, Austin, Texas 78712,1 Lehrstuhl für Mikrobiologie, Technische Universität München, D-85350 Freising, Germany,2 Department of Integrative Biology, Brigham Young University, Provo, Utah 84602,3 Department of Microbial Ecology, Institute of Ecology and Conservation Biology, Vienna University, A-1090 Vienna, Austria4

Received 6 February 2003/ Accepted 2 May 2003

Hydrogen sulfide-rich groundwater discharges from springs into Lower Kane Cave, Wyoming, where microbial mats dominated by filamentous morphotypes are found. The full-cycle rRNA approach, including 16S rRNA gene retrieval and fluorescence in situ hybridization (FISH), was used to identify these filaments. The majority of the obtained 16S rRNA gene clones from the mats were affiliated with the "Epsilonproteobacteria" and formed two distinct clusters, designated LKC group I and LKC group II, within this class. Group I was closely related to uncultured environmental clones from petroleum-contaminated groundwater, sulfidic springs, and sulfidic caves (97 to 99% sequence similarity), while group II formed a novel clade moderately related to deep-sea hydrothermal vent symbionts (90 to 94% sequence similarity). FISH with newly designed probes for both groups specifically stained filamentous bacteria within the mats. FISH-based quantification of the two filament groups in six different microbial mat samples from Lower Kane Cave showed that LKC group II dominated five of the six mat communities. This study further expands our perceptions of the diversity and geographic distribution of "Epsilonproteobacteria" in extreme environments and demonstrates their biogeochemical importance in subterranean ecosystems.

* Corresponding author. Mailing address: Department of Geological Sciences, University of Texas at Austin, 1 University Station C1100, Austin, TX 78712-0254. Phone: (512) 471-5413. Fax: (512) 471-5766. E-mail: aengel{at}mail.utexas.edu.

Applied and Environmental Microbiology, September 2003, p. 5503-5511, Vol. 69, No. 9
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.9.5503-5511.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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