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Microbial community changes in biological phosphate-removal systems on altering sludge phosphorus content

Institute of Environmental Engineering, National Central University, Chungli 32054, Taiwan¹
Center for Microbial Ecology, Michigan State University, East Lansing, MI 48824, USA²
Department of Urban Engineering, University of Tokyo, Tokyo 113, Japan³
National Institute of Bioscience and Human Technology, Agency of Industrial Science and Technology, 1-1 Tsukuba, Ibaraki, 305-8566 Japan⁴
Center for Ecological and Evolutionary Studies, University of Groningen, The Netherlands⁵

Author for correspondence: Wen-Tso Liu. Tel: +886 3422 7151 ext 4683. Fax: +886 3426 9401. e-mail: liuwt{at}cc.ncu.edu.tw

Biomarkers (respiratory quinones and cellular fatty acids) and denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rRNA genes were used to characterize the microbial community structure of lab-scale enhanced biological phosphate-removal (EBPR) systems in response to altering sludge phosphorus (P) content. All the data suggest that the microbial community structures of sludge samples with a P content between 8 and 12·3% (sludge dry weight) (i.e. good EBPR activity) were very similar, but differed from those with 2% P content (i.e. no EBPR activity). For all samples analysed, ubiquinones Q-8 and Q-10, menaquinone MK-8(H₄), and fatty acids C_16:0, C_{16:1 9c} and C_{18:1 11c} were the major components. The dominance of Q-8, Q-10 and MK-8(H₄) suggested that large numbers of organisms belonging to the ß and subclasses of the Proteobacteria and the Actinobacteria from the high G+C Gram-positive bacteria, respectively, were present. DGGE analysis revealed at least 7–9 predominant DNA bands and numerous other fragments in each sample. Five major DGGE fragments from each of the 2% and 12% P-containing sludge samples, respectively, were successfully isolated and sequenced. Phylogenetic analysis of the sequences indicated that both 2% and 12% P-containing sludge samples shared three common phylotypes that were separately affiliated with a novel bacterial group from the subclass of the Proteobacteria, two MK-8(H₄)-containing actinobacteria previously isolated from the 2% P-containing sludge, and a Caulobacter spp. in the subclass of the Proteobacteria. The phylogenetic analysis also revealed phylotypes unique to both sludge samples. Changes in sludge P content therefore had an effect on the composition and abundance of the predominant microbial populations, though specific phylotypes could not be unequivocally associated with EBPR.

Keywords: Activated sludge, biological phosphate removal, biomarker, DGGE, 16S rDNA

Abbreviations: DGGE, denaturing gradient gel electrophoresis; EBPR, enhanced biological phosphate removal; PHA, polyhydroxyalkanoate

The GenBank/EMBL/DDBJ accession numbers for the sequences obtained in this report are AF109792 (strain Lpha5), AF109793 (strain Lpha7) and AF124650 to AF124659.

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