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Applied and Environmental Microbiology, March 2001, p. 1351-1362, Vol. 67, No. 3
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.3.1351-1362.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Community Structure and Activity Dynamics of Nitrifying Bacteria in a Phosphate-Removing Biofilm

Armin Gieseke,1,* Ulrike Purkhold,2 Michael Wagner,2 Rudolf Amann,1 and Andreas Schramm1,3

Molecular Ecology Group, Max Planck Institute for Marine Microbiology, D-28359 Bremen,1 Department of Microbiology, Technical University Munich, D-85350 Freising,2 and Department of Ecological Microbiology, BITOEK, University of Bayreuth, D-95440 Bayreuth,3 Germany

Received 11 September 2000/Accepted 19 December 2000

The microbial community structure and activity dynamics of a phosphate-removing biofilm from a sequencing batch biofilm reactor were investigated with special focus on the nitrifying community. O2, NO2-, and NO3- profiles in the biofilm were measured with microsensors at various times during the nonaerated-aerated reactor cycle. In the aeration period, nitrification was oxygen limited and restricted to the first 200 µm at the biofilm surface. Additionally, a delayed onset of nitrification after the start of the aeration was observed. Nitrate accumulating in the biofilm in this period was denitrified during the nonaeration period of the next reactor cycle. Fluorescence in situ hybridization (FISH) revealed three distinct ammonia-oxidizing populations, related to the Nitrosomonas europaea, Nitrosomonas oligotropha, and Nitrosomonas communis lineages. This was confirmed by analysis of the genes coding for 16S rRNA and for ammonia monooxygenase (amoA). Based upon these results, a new 16S rRNA-targeted oligonucleotide probe specific for the Nitrosomonas oligotropha lineage was designed. FISH analysis revealed that the first 100 µm at the biofilm surface was dominated by members of the N. europaea and the N. oligotropha lineages, with a minor fraction related to N. communis. In deeper biofilm layers, exclusively members of the N. oligotropha lineage were found. This separation in space and a potential separation of activities in time are suggested as mechanisms that allow coexistence of the different ammonia-oxidizing populations. Nitrite-oxidizing bacteria belonged exclusively to the genus Nitrospira and could be assigned to a 16S rRNA sequence cluster also found in other sequencing batch systems.

* Corresponding author. Mailing address: Molecular Ecology Group, Max Planck Institute for Marine Microbiology, Celsiusstraße 1, D-28359 Bremen, Germany. Phone: 49 421 2028 836. Fax: 49 421 2028 690. E-mail: agieseke{at}mpi-bremen.de.

Applied and Environmental Microbiology, March 2001, p. 1351-1362, Vol. 67, No. 3
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.3.1351-1362.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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