Spieck, Eva, Hartwig, Christine, McCormack, Iris, Maixner, Frank, Wagner, Michael, Lipski, Andre and Daims, Holger (2006) Selective enrichment and molecular characterization of a previously uncultured Nitrospira-like bacterium from activated sludge. Environmental Microbiology, 8 (3). pp. 405-415. DOI 10.1111/j.1462-2920.2005.00905.x.

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Abstract

Previously uncultured nitrite-oxidizing bacteria affiliated to the genus Nitrospira have for the first time been successfully enriched from activated sludge from a municipal wastewater treatment plant. During the enrichment procedure, the abundance of the Nitrospira-like bacteria increased to approximately 86% of the total bacterial population. This high degree of purification was achieved by a novel enrichment protocol, which exploits physiological features of Nitrospira-like bacteria and includes the selective repression of coexisting Nitrobacter cells and heterotrophic contaminants by application of ampicillin in a final concentration of 50 µg ml−1. The enrichment process was monitored by electron microscopy, fluorescence in situ hybridization (FISH) with rRNA-targeted probes and fatty acid profiling. Phylogenetic analysis of 16S rRNA gene sequences revealed that the enriched bacteria represent a novel Nitrospira species closely related to uncultured Nitrospira-like bacteria previously found in wastewater treatment plants and nitrifying bioreactors. The enriched strain is provisionally classified as ‘Candidatus Nitrospira defluvii’.

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