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Applied and Environmental Microbiology, August 2003, p. 4463-4473, Vol. 69, No. 8
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.8.4463-4473.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Diversity and Seasonal Variability of ß-Proteobacteria in Biofilms of Polluted Rivers: Analysis by Temperature Gradient Gel Electrophoresis and Cloning

I. H. M. Brümmer, A. Felske, and I. Wagner-Döbler*

Division of Microbiology, German Research Center for Biotechnology, 38124 Braunschweig, Germany

Received 12 February 2003/ Accepted 29 April 2003

The ß-subgroup of the Proteobacteria has been shown to be important in aquatic habitats and was investigated in depth here by molecular 16S rRNA techniques in biofilms of the Elbe River and its polluted tributary, the Spittelwasser River. The bacterial 16S rRNA genes were cloned from each site, screened for ß-proteobacterial clones and sequenced. River biofilm clones from both rivers grouped into 9 clusters (RBFs). RBFs 1, 2, and 3 fell into the recently described ßI cluster of cosmopolitan freshwater bacteria, where they represented new species related to Rhodoferax, Aquaspirillum, and Hydrogenophaga. RBFs 4 to 7 affiliated with Aquabacterium commune, Ideonella dechloratans, and Sphaerotilus natans, respectively. The two remaining RBFs were uncultivated clusters, one of them being distantly related to Gallionella ferruginea. Seasonal changes in the relative intensity of the ß-proteobacterial 16S rRNA genes of biofilms harvested monthly for 1 year were determined by specific amplification and separation by temperature gradient gel electrophoresis (TGGE). Bands were identified by comparison of clones to community fingerprints by TGGE. Eight of 13 identified bands were shared by both habitats but showed different relative abundance and seasonal variability in the two rivers, probably caused by differences in temperature and pollutants. The data indicate new not-yet-cultivated clusters of river biofilm organisms, some of them probably distributed globally. They confirm the importance of certain known freshwater genera in river biofilms. The high phylogenetic resolution obtained by clone library analysis combined with the high temporal resolution obtained by TGGE suggest that the observed microdiversity in the river biofilm clone libraries might be caused by phylogenetically closely related microbial populations which are adapted to ecological parameters.

* Corresponding author. Mailing address: German Research Center for Biotechnology, Division of Microbiology, Mascheroder Weg 1, D-38124 Braunschweig, Germany. Phone: 49 531 6181 408. Fax: 49 531 6181 411. E-mail: iwd{at}gbf.de.

Applied and Environmental Microbiology, August 2003, p. 4463-4473, Vol. 69, No. 8
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.8.4463-4473.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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