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Contribution of ammonia-oxidizing archaea and ammonia-oxidizing bacteria to ammonia oxidation in two nitrifying reactors


In this study, two laboratory nitrifying reactors (NRI and NRII), which were seeded by sludge from different sources and operated under different operating conditions, were found to possess distinct dominant ammonia-oxidizing microorganisms. Ammonia-oxidizing archaeal (AOA) amoA genes outnumbered ammonia-oxidizing bacterial (AOB) amoA genes in reactor NRI, while only AOB amoA genes were detectable in reactor NRII. The AOA amoA gene sequences retrieved from NRI were characterized within the Nitrososphaera sister cluster of the group 1.1b Thaumarchaeota. Two inhibitors for ammonia oxidation, allylthiourea (ATU) and 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (PTIO), were applied individually and as a mixture to observe the ammonia-oxidizing activity of both microorganisms in the reactors’ sludge. The results indicated that AOA and AOB jointly oxidized ammonia in NRI, while AOB played the main role in ammonia oxidation in NRII. DNA-stable isotope probing with labeled 13C–HCO3 was performed on NRI sludge. Incorporation of 13C into AOA and AOB implied that both microorganisms may perform autotrophy during ammonia oxidation. Taken together, the results from this study provide direct evidence demonstrating the contribution of AOA and AOB to ammonia oxidation in the nitrifying reactors.

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The authors are deeply grateful of the Thailand Research Fund (grant no. RSA5780036) and the 90th Anniversary of the Chulalongkorn University Fund (Ratchadaphiseksomphot Endowment Fund) for their financial support of the work.

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Correspondence to Tawan Limpiyakorn.

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Responsible editor: Gerald Thouand

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Srithep, P., Pornkulwat, P. & Limpiyakorn, T. Contribution of ammonia-oxidizing archaea and ammonia-oxidizing bacteria to ammonia oxidation in two nitrifying reactors. Environ Sci Pollut Res 25, 8676–8687 (2018).

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  • Ammonia-oxidizing archaea
  • Ammonia-oxidizing bacteria
  • DNA-stable isotope probing
  • Nitrifying reactor
  • Wastewater treatment
  • Selective inhibitors for ammonia oxidation