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Applied and Environmental Microbiology, March 2005, p. 1276-1282, Vol. 71, No. 3
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.3.1276-1282.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Influence of Starvation on Potential Ammonia-Oxidizing Activity and amoA mRNA Levels of Nitrosospira briensis

Annette Bollmann,1,2* Ingo Schmidt,3,{dagger} Aaron M. Saunders,1,{ddagger} and Mette H. Nicolaisen1,§

Department of Microbial Ecology, Institute of Biological Sciences, University of Aarhus, Aarhus, Denmark,1 Department for Microbial Ecology, NIOO-KNAW Centre for Limnology, Nieuwersluis,2 Department of Microbiology, University of Nijmegen, Nijmegen, The Netherlands3

Received 10 June 2004/ Accepted 7 October 2004

The effect of short-term ammonia starvation on Nitrosospira briensis was investigated. The ammonia-oxidizing activity was determined in a concentrated cell suspension with a NOx biosensor. The apparent half-saturation constant [Km(app)] value of the NH3 oxidation of N. briensis was 3 µM NH3 for cultures grown both in continuous and batch cultures as determined by a NOx biosensor. Cells grown on the wall of the vessel had a lower Km(app) value of 1.8 µM NH3. Nonstarving cultures of N. briensis showed potential ammonia-oxidizing activities of between 200 to 250 µM N h–1, and this activity decreased only slowly during starvation up to 10 days. Within 10 min after the addition of fresh NH4+, 100% activity was regained. Parallel with activity measurements, amoA mRNA and 16S rRNA were investigated. No changes were observed in the 16S rRNA, but a relative decrease of amoA mRNA was observed during the starvation period. During resuscitation, an increase in amoA mRNA expression was detected simultaneously. The patterns of the soluble protein fraction of a 2-week-starved culture of N. briensis showed only small differences in comparison to a nonstarved control. From these results we conclude that N. briensis cells remain in a state allowing fast recovery of ammonia-oxidizing activity after addition of NH4+ to a starved culture. Maintaining cells in this kind of active state could be the survival strategy of ammonia-oxidizing bacteria in nature under fluctuating NH4+ availability.

* Corresponding author. Present address: Department of Biology, Northeastern University, 134 Mugar Life Science Building, 360 Huntington Ave., Boston, MA 02115. Phone: (617) 373 3229. Fax: (617) 373 3724. E-mail: a.bollmann{at}neu.edu.

{dagger} Present address: University of Bayreuth, Department of Microbiology, 95447 Bayreuth, Germany.

{ddagger} Present address: Advanced Wastewater Management Centre, The University of Queensland, 4072 Brisbane, Australia.

§ Present address: Royal Veterinary and Agricultural University, Institute of Ecology, Section of Genetics and Microbiology, 1871 Frederiksberg C, Denmark.

Applied and Environmental Microbiology, March 2005, p. 1276-1282, Vol. 71, No. 3
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.3.1276-1282.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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