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Applied and Environmental Microbiology, August 1999, p. 3690-3696, Vol. 65, No. 8
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Microscale Distribution of Populations and Activities of Nitrosospira and Nitrospira spp. along a Macroscale Gradient in a Nitrifying Bioreactor: Quantification by In Situ Hybridization and the Use of Microsensors

Andreas Schramm,1,* Dirk de Beer,1 Johan C. van den Heuvel,2 Simon Ottengraf,2 and Rudolf Amann1

Max Planck Institute for Marine Microbiology, D-28359 Bremen, Germany,1 and Department of Chemical Engineering, University of Amsterdam, NL-1018WV Amsterdam, The Netherlands2

Received 29 January 1999/Accepted 24 May 1999

The change of activity and abundance of Nitrosospira and Nitrospira spp. along a bulk water gradient in a nitrifying fluidized bed reactor was analyzed by a combination of microsensor measurements and fluorescence in situ hybridization. Nitrifying bacteria were immobilized in bacterial aggregates that remained in fixed positions within the reactor column due to the flow regimen. Nitrification occurred in a narrow zone of 100 to 150 µm on the surface of these aggregates, the same layer that contained an extremely dense community of nitrifying bacteria. The central part of the aggregates was inactive, and significantly fewer nitrifiers were found there. Under conditions prevailing in the reactor, i.e., when ammonium was limiting, ammonium was completely oxidized to nitrate within the active layer of the aggregates, the rates decreasing with increasing reactor height. To analyze the nitrification potential, profiles were also recorded in aggregates subjected to a short-term incubation under elevated substrate concentrations. This led to a shift in activity from ammonium to nitrite oxidation along the reactor and correlated well with the distribution of the nitrifying population. Along the whole reactor, the numbers of ammonia-oxidizing bacteria decreased, while the numbers of nitrite-oxidizing bacteria increased. Finally, volumetric reaction rates were calculated from microprofiles and related to cell numbers of nitrifying bacteria in the active shell. Therefore, it was possible for the first time to estimate the cell-specific activity of Nitrosospira spp. and hitherto-uncultured Nitrospira-like bacteria in situ.

* Corresponding author. Mailing address: Max Planck Institute for Marine Microbiology, Celsiusstraße 1, D-28359 Bremen, Germany. Phone: 49 421 2028 834. Fax: 49 421 2028 580. E-mail: aschramm{at}mpi-bremen.de.

Applied and Environmental Microbiology, August 1999, p. 3690-3696, Vol. 65, No. 8
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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