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Applied and Environmental Microbiology, October 2005, p. 6134-6141, Vol. 71, No. 10
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.10.6134-6141.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Identification of Bacteria Potentially Responsible for Oxic and Anoxic Sulfide Oxidation in Biofilters of a Recirculating Mariculture System

Eddie Cytryn,1,2 Jaap van Rijn,1 Andreas Schramm,3,5 Armin Gieseke,4 Dirk de Beer,4 and Dror Minz2*

Department of Animal Sciences, Faculty of Agricultural, Food, and Environmental Quality Sciences, The Hebrew University of Jerusalem, Rehovot,1 Institute of Soil, Water, and Environmental Sciences, Agricultural Research Organization, The Volcani Center, Bet-Dagan, Israel,2 Department of Microbiology, University of Aarhus, Aarhus, Denmark,3 Max Planck Institute for Marine Microbiology, Bremen,4 Department of Ecological Microbiology, University of Bayreuth, Bayreuth, Germany5

Received 22 February 2005/ Accepted 6 May 2005

Bacteria presumably involved in oxygen- or nitrate-dependent sulfide oxidation in the biofilters of a recirculating marine aquaculture system were identified using a new application of reverse transcription-PCR denaturing gradient gel electrophoresis (DGGE) analysis termed differential-transcription (DT)-DGGE. Biofilter samples were incubated in various concentrations of sulfide or thiosulfate (0 to 5 mM) with either oxygen or nitrate as the sole electron acceptor. Before and after short-term incubations (10 to 20 h), total DNA and RNA were extracted, and a 550-bp fragment of the 16S rRNA genes was PCR amplified either directly or after reverse transcription. DGGE analysis of DNA showed no significant change of the original microbial consortia upon incubation. In contrast, DGGE of cDNA revealed several phylotypes whose relative band intensities markedly increased or decreased in response to certain incubation conditions, indicating enhanced or suppressed rRNA transcription and thus implying metabolic activity under these conditions. Specifically, species of the gammaproteobacterial genus Thiomicrospira and phylotypes related to symbiotic sulfide oxidizers could be linked to oxygen-dependent sulfide oxidation, while members of the Rhodobacteraceae (genera Roseobacter, Rhodobacter, and Rhodobium) were putatively active in anoxic, nitrate-dependent sulfide oxidation. For all these organisms, the physiology of their closest cultured relatives matches their DT-DGGE-inferred function. In addition, higher band intensities following exposure to 5 mM sulfide and nitrate were observed for Thauera-, Hydrogenophaga-, and Dethiosulfovibrio-like phylotypes. For these genera, nitrate-dependent sulfide oxidation has not been documented previously and therefore DT-DGGE might indicate a higher relative tolerance to high sulfide concentrations than that of other community members. We anticipate that DT-DGGE will be of general use in tracing functionally equivalent yet phylogenetically diverse microbial populations in nature.

* Corresponding author. Mailing address: Institute of Soil, Water, and Environmental Sciences, Agricultural Research Organization, The Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel. Phone: 972-3-9683316. Fax: 972-3-9604017. E-mail: minz{at}volcani.agri.gov.il.

Applied and Environmental Microbiology, October 2005, p. 6134-6141, Vol. 71, No. 10
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.10.6134-6141.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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