Ammonia half-saturation constants of sludge with different community compositions of ammonia-oxidizing bacteria

Kayee, Pantip;Rongsayamanont, Chaiwat;Kunapongkiti, Pattaraporn;Limpiyakorn, Tawan

  • Received : 2015.10.26
  • Accepted : 2016.02.13
  • Published : 2016.06.30


Owing to the kinetic differences in ammonia oxidation among ammonia-oxidizing microorganisms (AOM), there is no standard set of kinetic values that can be used as a representative set for nitrifying wastewater treatment plant (WWTP) design. As a result, this study clarified a link between the half-saturation constants for ammonia oxidation (Ks) and the dominant ammonia-oxidizing bacterial (AOB) groups in sludge from full-scale WWTPs and laboratory-scale nitrifying reactors. Quantitative polymerase chain reaction analyses revealed that AOB affiliated with the Nitrosomonas oligotropha cluster were the dominant AOM groups in the sludge taken from the low-ammonia-level WWTPs, while AOB associate with the Nitrosomonas europaea cluster comprised the majority of AOM groups in the sludge taken from the high-ammonia-level WWTPs and nitrifying reactors. A respirometric assay demonstrated that the ammonia Ks values for the high-ammonia-level WWTPs and nitrifying reactors were higher than those of the low-ammonia-level plants. Using the Ks values of available AOM cultures as a reference, the Ks values of the analyzed sludge were mainly influenced by the dominant AOB species. These findings implied that.different sets of kinetic values may be required for WWTPs with different dominant AOM species for more accurate WWTP design and operations.


Ammonia-oxidizing bacteria;Ammonia half-saturation constant;Wastewater treatment plant


  1. Dionisi HM, Layton AC, Harms G, Gregory IR, Robinson KG, Sayler GS. Quantification of Nitrosomonas oligotropha-like ammonia- oxidizing bacteria and Nitrospira spp. from full-scale wastewater treatment plants by competitive PCR. Appl. Environ. Microbiol. 2002;68:245-253.
  2. Harms G, Layton AC, Dionisi HM, et al. Real-time PCR quantification of nitrifying bacteria in a municipal wastewater treatment plant. Environ. Sci. Technol. 2003;37:343-351.
  3. Limpiyakorn T, Shinihara Y, Kurisu F, Yagi O. Communities of ammonia-oxidizing bacteria in activated sludge of various sewage treatment plants in Tokyo. FEMS Microbiol. Ecol. 2005;54:205-117.
  4. Bai Y, Sun Q, Wen D, Tang X. Abundance of ammonia-oxidizing bacteria and archaea in industrial and domestic wastewater treatment systems. FEMS Microbiol. Ecol. 2012;80:323-330.
  5. Limpiyakorn T, Furhacker M, Haberl R, Chodanon T, Srithep P, Sonthiphand P. amoA-encoding archaea in wastewater treatment plants: A review. Appl. Microbiol. Biotechnol. 2013;97:1425-1439.
  6. Widdel F, Bak F. The Prokaryotes. In: Ballows A, Truper HG, Dworkin M, Harder W, Schleifer K-H, eds. A Handbook on the Biology of Bacteria: Ecophysiology, Isolation, Identification, Application. 2nd ed. New York: Springer; 1992. p. 3352-3378.
  7. Francis CA, Roberts KJ, Beman, JM, Santoro AE, Oakley, BB. Ubiquity and diversity of ammonia-oxidizing archaea in water columns and sediments of the ocean. Proc. Natl. Acad. Sci. U.S.A. 2005;102:14683-14688.
  8. Rotthauwe JH, Witzel KP, Liesack W. The ammonia monooxygenase structural gene amoA as a functional marker: molecular fine-scale analysis of natural ammonia-oxidizing populations. Appl. Environ. Microbiol. 1997;63:4704-4712.
  9. Norton JM, Alzerreca JJ, Suwa Y, Klotz MG. Diversity of ammonia monooxygenase operon in autotrophic ammonia oxidizing bacteria. Arch. Microbiol. 2002;177:139-149.
  10. Lim J, Do H, Seung GS, Hwang S. Primer and probe sets for group-specific quantification of the genera Nitrosomonas and Nitrosospira using real-time PCR. Biotechnol. Bioeng. 2008;99: 1374-1383.
  11. Aakra A, Utaker JB, Nes IF. RFLP of rRNA genes and sequencing of the 16S-23S rDNA intergenic spacer region of ammonia-oxidizing bacteria: a phylogenetic approach. Int. J. Syst Bacteriol. 1999;49:123-130.
  12. Spanjers H, Vanrolleghem P. Respirometry as a tool for rapid characterization of wastewater and activated sludge. Water Sci. Technol. 1995;31:105-114.
  13. Chandran K, Smets BF. Single-step nitrification models erroneously describe batch ammonia oxidation profiles when nitrite oxidation becomes rate limiting. Biotechnol. Bioeng. 2000;68:396-406.<396::AID-BIT5>3.0.CO;2-S
  14. You SJ, Hsu CL, Chuang SH, Ouyang CF. Nitrification efficiency and nitrifying bacteria abundance in combined AS-RBC and A2O systems. Water Res. 2003;37:2281-2290.
  15. Matsumoto S, Terada A, Aoi Y, Tsuneda S, Alpkvist E, Picioreanu C, Loosdrecht, MCM. Experimental and simulation analysis of community structure of nitrifying bacteria in a membrane-aerated biofilm. Water Sci. Technol. 2007;55:283-290.
  16. Vejmelkova D, Sorokin DY, Abbas B, et al. Analysis of ammonia-oxidizing bacteria dominating in lab-scale bioreactors with high ammonium bicarbonate loading. Appl. Microbiol. Biotechnol. 2011;93:401-410.

Cited by

  1. Photosynthetic oxygenation for urine nitrification pp.1996-9732, 2018,
  2. Kinetics of nitrification and acrylamide biodegradation by Enterobacter aerogenes and mixed culture bacteria in sequencing batch reactor wastewater treatment systems vol.24, pp.2, 2018,


Supported by : Chulalongkorn University