Anammox enrichment and constructed wetland inoculation for improvement of wastewater treatment performance


1 Department of Biologically Active Substances, Pharmacy and Biotechnology, Lviv Polytechnic National University, Lviv, Ukraine AND Department of Environmental Biotechnology, Helmholtz Centre for Environmental Research, Leipzig, Germany

2 Kurdistan Environmental Health Research Center, Kurdistan University of Medical Sciences, Sanandaj, Iran AND Department of Environmental Biotechnology, Helmholtz Centre for Environmental Research, Leipzig, Germany

3 Department of Biologically Active Substances, Pharmacy and Biotechnology, Lviv Polytechnic National University, Lviv, Ukraine


This study contributes to the improvement of low-cost biotechnology for wastewater treatment in constructed wetlands (CWs). Constructed wetlands are energy efficient engineered systems that mimic the treatment processes of natural wetlands, removing polluting organic matter, nutrients, and pathogens from water. The aim of this study was to investigate the advisability of the inoculation of horizontal subsurface flow constructed wetlands with the enriched biomass of anaerobic ammonium oxidation (anammox) bacteria to enhance nitrogen removal. Contaminants removal in constructed wetlands occurs mainly due to the biological transformations caused by indigenous water-borne microorganisms. However, the role of different microbial mechanisms is still unknown. To estimate the role of the anammox process in wetlands the laboratory-scale fixed bed reactor planted with Juncus effusus was inoculated with enriched biomass of anammox bacteria and fed with synthetic wastewater containing ammonium-nitrogen as the main contaminant. In order to obtain the active enriched culture of anammox bacteria, an upflow anaerobic fixed bed reactor inoculated with activated sludge from a municipal wastewater treatment plant was run. The reactor was fed with enrichment medium containing ammonium and nitrite in high concentrations. After 270 days of operation, nitrite was not found in measurable levels, the concentration of ammonium had slightly increased, and the concentration of nitrate in the reactor had significantly dropped compared to its level at the initial phase. The microbial association, which had developed in the enrichment reactor, allowed continuous removal of ammonium and nitrite. The anammox bacteria abundance in the reactor accounted for approximately 95% of total biomass.


  1. Garc?a J, Rousseau DP, Lesage E, Matamoros V, Bayona JM. Contaminant Removal Processes in Subsurface-Flow Constructed Wetlands: A Review. Critical Reviews in Environmental Science and Technology 2010; 40(7): 561-661.
  2. Vymazal J. Constructed Wetlands for Wastewater Treatment. Water 2010; 2(3): 530-49.
  3. Rousseau L, Vanrolleghem PA, Pauw ND. Constructed wetlands in Flanders: a performance analysis. Ecological Engineering 2004; 23(3): 151-63.
  4. Wu MY, Franz EH, Chen S. Oxygen fluxes and ammonia removal efficiencies in constructed treatment wetlands. Water Environ Res 2001; 73(6): 661-6.
  5. Vymazal J. Removal of nutrients in various types of constructed wetlands. Sci Total Environ 2007; 380(1-3): 48-65.
  6. Kadlec RH, Wallace S. Treatment Wetlands, Second Edition. 2nd ed. New York, NY: Taylor & Francis; 2008.
  7. Tanner CC. Nitrogen Removal Processes in Constructed Wetlands. In: Wong MH, Editor. Wetlands Ecosystems in Asia: Function and Management: Function and Management. Philadelphia, PA: Elsevier; 2004. p. 331-46.
  8. Kadlec RH. Vegetation Effects on Ammonia Reduction in Treatment Wetlands. In: Vymazal J, Editor. Natural and constructed wetlands: nutrients, metals and management. Winschoten, Netherlands: Backhuys Publishers; 2005. p. 233-60.
  9. Vymazal J, Kr?pfelov? L. Removal of nitrogen in constructed wetlands with horizontal sub-sureface flow: a review. Wetlands 2009; 29(4): 1114-24.
  10. Stottmeister U, Wiessner A, Kuschk P, Kappelmeyer U, Kastner M, Bederski O, et al. Effects of plants and microorganisms in constructed wetlands for wastewater treatment. Biotechnol Adv 2003; 22(1-2): 93-117.
  11. Wallace S, Austin D. Emerging models for nitrogen removal in treatment wetlands. J Environ Health 2008; 71(4): 10-6.
  12. Zhu G, Wang S, Feng X, Fan G, Jetten MS, Yin C. Anammox bacterial abundance, biodiversity and activity in a constructed wetland. Environ Sci Technol 2011; 45(23): 9951-8.
  13. Erler DV, Eyre BD, Davison L. The contribution of anammox and denitrification to sediment N2 production in a surface flow constructed wetland. Environ Sci Technol 2008; 42(24): 9144-50.
  14. Paredes D, Kuschk P, Stange F, Muller RA, Koser H. Model experiments on improving nitrogen removal in laboratory scale subsurface constructed wetlands by enhancing the anaerobic ammonia oxidation. Water Sci Technol 2007; 56(3): 145-50.
  15. Dong Z, Sun T. A potential new process for improving nitrogen removal in constructed wetlands Promoting coexistence of partial-nitrification and ANAMMOX. Ecological Engineering 2007; 31(2): 69-78.
  16. Rusten B, Eikebrokk B, Ulgenes Y, Lygren E. Design and operations of the Kaldnes moving bed biofilm reactors. Aquacultural Engineering 2006; 34(3): 322-31.
  17. Kappelmeyer U, Wie?ner A, Kuschk P, K?stner M. Operation of a Universal Test Unit for Planted Soil Filters - Planted Fixed Bed Reactor. Engineering in Life Sciences 2002; 2(10): 311-5.
  18. Wiessner A, Rahman KZ, Kuschk P, Kastner M, Jechorek M. Dynamics of sulphur compounds in horizontal sub-surface flow laboratory-scale constructed wetlands treating artificial sewage. Water Res 2010; 44(20): 6175-85.
  19. Spiegelman D, Whissell G, Greer CW. A survey of the methods for the characterization of microbial consortia and communities. Can J Microbiol 2005; 51(5): 355-86.
  20. Strous M, Heijnen JJ, Kuenen G, Jetten MS. The sequencing batch reactor as a powerful tool for the study of slowly growing anaerobic ammonium-oxidizing microorganisms. Applied Microbiology and Biotechnology 1998; 50(5): 589-96.