Abstract

Wastewater treatment plants are known to be important point sources for nitrous oxide (N2O) in the anthropogenic N cycle. Biofilm based treatment systems have gained increasing popularity in the treatment of wastewater, but the mechanisms and controls of N2O formation are not fully understood. Here, we review functional groups of microorganism involved in nitrogen (N) transformations during wastewater treatment, with emphasis on potential mechanism of N2O production in biofilms. Biofilms used in wastewater treatment typically harbour aerobic and anaerobic zones, mediating close interactions between different groups of N transforming organisms. Current models of mass transfer and biomass interactions in biofilms are discussed to illustrate the complex regulation of N2O production. Ammonia oxidizing bacteria (AOB) are the prime source for N2O in aerobic zones, while heterotrophic denitrifiers dominate N2O production in anoxic zones. Nitrosative stress ensuing from accumulation of NO2 − during partial nitrification or denitrification seems to be one of the most critical factors for enhanced N2O formation. In AOB, N2O production is coupled to nitrifier denitrification triggered by nitrosative stress, low O2 tension or low pH. Chemical N2O production from AOB intermediates (NH2OH, HNO, NO) released during high NH3 turnover seems to be limited to surface-near AOB clusters, since diffusive mass transport resistance for O2 slows down NH3 oxidation rates in deeper biofilm layers. The proportion of N2O among gaseous intermediates (NO, N2O, N2) in heterotrophic denitrification increases when NO or nitrous acid (HNO2) accumulates because of increasing NO2 −, or when transient oxygen intrusion impairs complete denitrification. Limited electron donor availability due to mass transport limitation of organic substrates into anoxic biofilm zones is another important factor supporting high N2O/N2 ratios in heterotrophic denitrifiers. Biofilms accommodating Anammox bacteria release less N2O, because Anammox bacteria have no known N2O producing metabolism and reduce NO2 − to N2, thereby lowering nitrosative stress to AOB and heterotrophs.

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