Source identification of nitrous oxide on autotrophic partial nitrification in a granular sludge reactor

Emission of nitrous oxide (N₂O) during biological wastewater treatment is of growing concern since N₂O is a major stratospheric ozone-depleting substance and an important greenhouse gas. The emission of N₂O from a lab-scale granular sequencing batch reactor (SBR) for partial nitrification (PN) treating synthetic wastewater without organic carbon was therefore determined in this study, because PN process is known to produce more N₂O than conventional nitrification processes. The average N₂O emission rate from the SBR was 0.32 ± 0.17 mg-N L⁻¹ h⁻¹, corresponding to the average emission of N₂O of 0.8 ± 0.4% of the incoming nitrogen load (1.5 ± 0.8% of the converted NH₄⁺). Analysis of dynamic concentration profiles during one cycle of the SBR operation demonstrated that N₂O concentration in off-gas was the highest just after starting aeration whereas N₂O concentration in effluent was gradually increased in the initial 40 min of the aeration period and was decreased thereafter. Isotopomer analysis was conducted to identify the main N₂O production pathway in the reactor during one cycle. The hydroxylamine (NH₂OH) oxidation pathway accounted for 65% of the total N₂O production in the initial phase during one cycle, whereas contribution of the NO₂⁻ reduction pathway to N₂O production was comparable with that of the NH₂OH oxidation pathway in the latter phase. In addition, spatial distributions of bacteria and their activities in single microbial granules taken from the reactor were determined with microsensors and by in situ hybridization. Partial nitrification occurred mainly in the oxic surface layer of the granules and ammonia-oxidizing bacteria were abundant in this layer. N₂O production was also found mainly in the oxic surface layer. Based on these results, although N₂O was produced mainly via NH₂OH oxidation pathway in the autotrophic partial nitrification reactor, N₂O production mechanisms were complex and could involve multiple N₂O production pathways.