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Applied and Environmental Microbiology, September 1999, p. 4189-4196, Vol. 65, No. 9
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

On the Occurrence of Anoxic Microniches, Denitrification, and Sulfate Reduction in Aerated Activated Sludge

Andreas Schramm,1,* Cecilia M. Santegoeds,1 Helle K. Nielsen,2 Helle Ploug,1 Michael Wagner,3 Milan Pribyl,4 Jiri Wanner,4 Rudolf Amann,1 and Dirk de Beer1

Max Planck Institute for Marine Microbiology, D-28359 Bremen,1 and Lehrstuhl für Mikrobiologie, Technische Universität München, D-80290 Munich,3 Germany; Department of Microbial Ecology, University of Aarhus, DK-8000 Aarhus C, Denmark2; and Department of Water Technology and Environmental Engineering, Prague Institute of Chemical Technology, CZ-166 28 Prague 6, Czech Republic4

Received 26 April 1999/Accepted 28 June 1999

A combination of different methods was applied to investigate the occurrence of anaerobic processes in aerated activated sludge. Microsensor measurements (O2, NO2-, NO3-, and H2S) were performed on single sludge flocs to detect anoxic niches, nitrate reduction, or sulfate reduction on a microscale. Incubations of activated sludge with 15NO3- and 35SO42- were used to determine denitrification and sulfate reduction rates on a batch scale. In four of six investigated sludges, no anoxic zones developed during aeration, and consequently denitrification rates were very low. However, in two sludges anoxia in flocs coincided with significant denitrification rates. Sulfate reduction could not be detected in any sludge in either the microsensor or the batch investigation, not even under short-term anoxic conditions. In contrast, the presence of sulfate-reducing bacteria was shown by fluorescence in situ hybridization with 16S rRNA-targeted oligonucleotide probes and by PCR-based detection of genes coding for the dissimilatory sulfite reductase. A possible explanation for the absence of anoxia even in most of the larger flocs might be that oxygen transport is not only diffusional but enhanced by advection, i.e., facilitated by flow through pores and channels. This possibility is suggested by the irregularity of some oxygen profiles and by confocal laser scanning microscopy of the three-dimensional floc structures, which showed that flocs from the two sludges in which anoxic zones were found were apparently denser than flocs from the other sludges.

* Corresponding author. Present address: Department of Ecological Microbiology, BITÖ, University of Bayreuth, D-95440 Bayreuth, Germany. Phone: 49 921 555-642. Fax: 49 921 555-799. E-mail: ancreas.schramm{at}bitoek.uni-bayreuth.de.

Applied and Environmental Microbiology, September 1999, p. 4189-4196, Vol. 65, No. 9
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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