Skip to main content

Advertisement

Log in

Enrichment of Anammox from Activated Sludge and Its Application in the CANON Process

  • Published:
Microbial Ecology Aims and scope Submit manuscript

Abstract

A microbial culture capable of actively oxidizing ammonium to dinitrogen gas in the absence of oxygen, using nitrite as the electron acceptor, was enriched from local activated sludge (Western Australia) in <14 weeks. The maximum anaerobic ammonium oxidation (i.e., anammox) activity achieved by the anaerobic culture was 0.26 mmol NH +4 (g biomass)−1 h−1 (0.58 kg total-N m−3 day−1). Qualitative FISH analysis (fluorescence in situ hybridization) confirmed the phylogenetic position of the enriched microorganism as belonging to the order Planctomycetales, in which all currently identified anammox strains fall. Preliminary FISH analysis suggests the anammox strain belongs to the same phylogenetic group as the Candidatus ‘Brocadia anammoxidans’ strain discovered in the Netherlands. However, there are quite a few differences in the target sites for the more specific probes of these organisms and it is therefore likely to represent a new species of anammox bacteria. A small amount of aerobic ammonium-oxidizing biomass was inoculated into the anammox reactor (10% v/v) to initiate completely autotrophic nitrogen removal over nitrite (the CANON process) in chemostat culture. The culture was always under oxygen limitation and no organic carbon was added. The CANON reactor was operated as an intermittently aerated system with 20 min aerobiosis and 30 min anaerobiosis, during which aerobic and anaerobic ammonium oxidation were performed in sequential fashion, respectively. Anammox was not inhibited by repeated intermittent exposure to oxygen, allowing sustained, completely autotrophic ammonium removal (0.08 kg N m−3 day−1) for an extended period of time.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Figure 1
Figure 2
Figure 3
Figure 4

Similar content being viewed by others

References

  1. K Egli U Fanger PJJ Alvarez H Siegrist JR Meer ParticleVan der AJB Zehnder (2001) ArticleTitleEnrichment and characterisation of an anammox bacterium from a rotating biological contactor treating ammonium-rich leachate Arch Microbiol 175 198–207 Occurrence Handle10.1007/s002030100255 Occurrence Handle11357512

    Article  PubMed  Google Scholar 

  2. AE Greenberg LS Clesceri AD Eaton (1992) Standard Methods for the Examination of Water and Wastewater, EditionNumber18 American Public Health Association Washington, DC

    Google Scholar 

  3. MSM Jetten M Strous KT Pas-Schoonen ParticleVan de J Schalk UGJM Dongen ParticleVan AA Graaf ParticleVan de S Logemann G Muyzer MCM Loosdrecht ParticleVan GJ Kuenen (1999) ArticleTitleThe anaerobic oxidation of ammonium FEMS Microbiol Rev 22 421–437 Occurrence Handle10.1016/S0168-6445(98)00023-0

    Article  Google Scholar 

  4. AD Sliekers S Haaijer M Schmid H Harhangi K Verwegen JG Kuenen MSM Jetten (2004) ArticleTitleNitrification and anammox with urea as the energy source Syst Appl Microbiology 27 IssueID3 271–278 Occurrence Handle10.1078/0723-2020-00259

    Article  Google Scholar 

  5. S Juretschko G Timmermann M Schmid KH Schleifer A Pommereningroser HP Koops (1998) ArticleTitleCombined molecular and conventional analyses of nitrifying bacterium diversity in activated sludge — Nitrosococcus mobilis and Nitrospira-like bacteria as dominant populations Appl Env Microbiol 64 3042–3051

    Google Scholar 

  6. L Kuai W Verstraete (1998) ArticleTitleAmmonium removal by the oxygen-limited autotrophic nitrification–denitrification system Appl Env Microbiol 64 4500–4506

    Google Scholar 

  7. A Mulder AA Graaf ParticleVan de LA Robertson JG Kuenen (1995) ArticleTitleAnaerobic ammonium oxidation discovered in a denitrifying fluidised bed reactor FEMS Microbiol Ecol 16 177–183 Occurrence Handle10.1016/0168-6496(94)00081-7

    Article  Google Scholar 

  8. A Neef R Amann H Schlesner KH Schleifer (1998) ArticleTitleMonitoring a widespread bacterial group: in situ detection of planctomycetes with 16S rRNA-targeted probes Microbioloy 144 3257–3266

    Google Scholar 

  9. N Puznava M Payraudeau D Thornberg (2001) ArticleTitleSimultaneous nitrification and denitrification in biofilters with real time aeration control Water Sci Tech 43 IssueID1 269–276

    Google Scholar 

  10. CW Randall JL Barnard DH Stensel (1992) Design and Retrofit of Wastewater Treatment Plants for Biological Nutrient Removal Technomic Lancaster, PA

    Google Scholar 

  11. M Schmid U Twachtmann M Klein M Strous S Juretschko M Jetten JW Metzger KH Schleifer M Wagner (2000) ArticleTitleMolecular evidence for genus level diversity of bacteria capable of catalyzing anaerobic ammonium oxidation Syst Appl Microbiol 23 93–106 Occurrence Handle10879983

    PubMed  Google Scholar 

  12. A Schramm LH Larsen NP Revsbech RI Amann (1997) ArticleTitleStructure and function of a nitrifying biofilm as determined by microelectrodes and flourescent oligonucleotide probes Wat Sci Technol 36 263–270 Occurrence Handle10.1016/S0273-1223(97)00333-8

    Article  Google Scholar 

  13. A Schramm D Beer ParticleDe M Wagner RI Amann (1998) ArticleTitleIdentification and activities in situ of Nitrosospira and Nitrospira spp. as dominant populations in a nitrifying fluidised bed reactor Appl Env Microbiol 64 3480–3485

    Google Scholar 

  14. OA Sliekers N Derwort JL Campos Gomez M Strous GJ Kuenen MSM Jetten (2002) ArticleTitleCompletely autotrophic nitrogen removal over nitrite in one single reactor Wat Res 36 2475–2482 Occurrence Handle10.1016/S0043-1354(01)00476-6

    Article  Google Scholar 

  15. OA Sliekers KA Third W Abma GJ Kuenen MSM Jetten (2003) ArticleTitleCANON and anammox in a gas-lift reactor FEMS Microbiol Lett 218 339–344 Occurrence Handle10.1016/S0378-1097(02)01177-1 Occurrence Handle12586414

    Article  PubMed  Google Scholar 

  16. M Strous E Gerven GIJ Kuenen M Jetten (1997) ArticleTitleEffects of aerobic and microaerobic conditions on anaerobic ammonium-oxidizing (anammox) sludge Appl Env Microbiol 63 2446–2448

    Google Scholar 

  17. M Strous JJ Heijnen JG Kuenen MSM Jetten (1998) ArticleTitleThe sequencing batch reactor as a powerful tool for the study of slowly growing anaerobic ammonium-oxidizing microorganisms Appl Microbiol Biotechnol 50 589–596 Occurrence Handle10.1007/s002530051340

    Article  Google Scholar 

  18. M Strous JG Kuenen MSM Jetten (1999) ArticleTitleKey physiology of anaerobic ammonium oxidation Appl Env Microbiol 65 3248–3250

    Google Scholar 

  19. Strous M (2000) Microbiology of anaerobic ammonium oxidation. PhD Thesis, Department of Microbiology, TU Technical University, Delft, The Netherlands

  20. M Strous JG Kuenen JA Fuerst M Wagner MSM Jetten (2002) ArticleTitleThe anammox case—a new experimental manifesto for microbiological eco-physiology Ant Leeuwen 81 693–702 Occurrence Handle10.1023/A:1020590413079

    Article  Google Scholar 

  21. KA Third OA Sliekers GJ Kuenen MSM Jetten (2001) ArticleTitleThe CANON system (Completely Autotrophic Nitrogen-Removal Over Nitrite) under ammonium limitation: interaction and competition between three groups of bacteria Syst Appl Microbiol 24 588–596 Occurrence Handle11876366

    PubMed  Google Scholar 

  22. KA Third N Burnett R Cord-Ruwisch (2003) ArticleTitleSimultaneous nitrification and denitrification using stored substrate (PHB) as the electron donor in a sequencing batch reactor Biotechnol Bioeng 83 706–720 Occurrence Handle10.1002/bit.10708 Occurrence Handle12889035

    Article  PubMed  Google Scholar 

  23. SK Toh NJ Ashbolt (2002) ArticleTitleAdaptation of anaerobic ammonium-oxidising consortium to synthetic coke-ovens wastewater Appl Microbiol Biotechnol 59 344–352 Occurrence Handle10.1007/s00253-002-1007-7 Occurrence Handle12111169

    Article  PubMed  Google Scholar 

  24. SK Toh RI Webb NJ Ashbolt (2002) ArticleTitleEnrichment of autotrophic anaerobic ammonium-oxidizing consortia from various wastewaters Microb Ecol 43 154–167 Occurrence Handle10.1007/s00248-001-0033-9 Occurrence Handle11984637

    Article  PubMed  Google Scholar 

  25. AA Van de Graaf P De Bruijn LA Robertson MSM Jetten JG Kuenen (1996) ArticleTitleAutotrophic growth of anaerobic ammonium-oxidizing microorganisms in a fluidized bed reactor Microbiology 142 2187–2196

    Google Scholar 

  26. U Dongen ParticleVan M Loodsdrecht ParticleVan M Jetten (2001) ArticleTitleThe SHARON-Anammox process for treatment of ammonia rich wastewater Wat Sci Technol 44 153–160

    Google Scholar 

Download references

Acknowledgments

Murdoch University thanks the Microbiology Department at the University of Nijmegen for their technical guidance during the FISH analyses and for their critical review of this paper.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to R. Cord-Ruwisch.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Third, K.A., Paxman, J., Schmid, M. et al. Enrichment of Anammox from Activated Sludge and Its Application in the CANON Process. Microb Ecol 49, 236–244 (2005). https://doi.org/10.1007/s00248-004-0186-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00248-004-0186-4

Keywords

Navigation