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Continuous flow processes for the in situ determination of N2O emissions during low C/N municipal wastewater treatment have rarely been reported. The anaerobic/aerobic/anoxic (AOA) process has recently shown promising potential in energy savings and advanced nitrogen removal, but it still needs to be comprehensively explored in relation to N2O emissions for its carbon reduction advantages. In this study, a novel gas-collecting continuous flow reactor was designed to comprehensively evaluate the emissions of N2O from the gas and liquid phases of the AOA process. Additionally, the measures of enhancing endogenous denitrification (ED) and self-enriching anaerobic ammonium oxidation (Anammox) were employed to optimize nitrogen removal and achieve N2O reduction in the anoxic zone. The results showed that enhanced ED coupled with Anammox led to an increase in the nitrogen removal efficiency (NRE) from 67.65 to 81.96%, an enhancement of the NO3–removal rate from 1.76 mgN/(L h) to 3.99 mgN/(L h), and the N2O emission factor in the anoxic zone decreased from 0.28 to 0.06%. Impressively, ED eliminated 91.46 ± 2.47% of the dissolved N2O from the upstream aerobic zone, and the dissolved N2O in the effluent was reduced to less than 0.01 mg/L. This study provides valuable strategies for fully evaluating N2O emissions and N2O reduction from the AOA process.

An innovative partial nitrification, in situ fermentation, and Anammox (PNFA) system was developed to achieve mature landfill leachate and waste activated sludge simultaneous treatment. Three separate sequencing batch reactors (SBRs) were used for partial nitrification (PN-SBR), integrated fermentation–denitrification (IFD-SBR), and partial nitrification–Anammox (PNA-SBR). After 200 days of continuous operation, a satisfactory nitrogen removal efficiency (NRE) of 99.2 ± 0.1% was obtained, with an effluent total nitrogen (TN) of 15.2 ± 3.2 mg/L. In IFD-SBR, the volatile fatty acids generated from fermentation drove efficient denitrification, obtaining sludge and nitrogen reduction rates of 4.2 ± 0.7 and 0.61 ± 0.04 kg/m3·day, respectively. Furthermore, unwanted fermentation metabolites (134.1 mg/L NH4+-N) were further treated by PNA-SBR using a combination of step-feed and intermittent aeration strategies. In PNA-SBR, Anammox significantly contributed to 82.1% nitrogen removal, and Anammox bacteria (Candidatus Brocadia, 2.3%) mutually benefited with partially denitrifying microorganisms (Thauera, 4.2%), with 66.3% of generated nitrate reduced to nitrite and then reutilized in situ by Anammox. Compared with the conventional nitrification–denitrification process, PNFA reduced oxygen energy consumption, external carbon source dosage, and CO2emission by 21.3, 100, and 38.9%, respectively, and obtained 50.1% external WAS reduction efficiency.

Anammox granulation is an efficient solution proffered to enrich slow-growing anammox bacteria (AnAOB), but the lack of effective granulation strategies for low-strength domestic wastewater impedes its application. In this study, a novel granulation model regulated by Epistylisspp. for highly enriched AnAOB was revealed for the first time. Notably, anammox granulation was achieved within 65 d of domestic wastewater treatment. The stalks of Epistylisspp. were found to act as the skeleton of granules and provide attachment points for bacterial colonization, and the expanded biomass layer in turn provided more area for the unstalked free-swimming zooids. Additionally, Epistylisspp. exerted much less predation stress on AnAOB than on nitrifying bacteria, and AnAOB tended to grow in aggregates in the interior of granules, thus favoring the growth and retention of AnAOB. Ultimately, the relative abundance of AnAOB reached up to a maximum of 8.2% in granules (doubling time of 9.9 d) compared to 1.1% in flocs (doubling time of 23.1 d), representing the most substantial disparity between granules and flocs. Overall, our findings advance the current understanding of interactions involved in granulation between protozoa and microbial communities and offer new insight into the specific enrichment of AnAOB under the novel granulation model.

The novel partial denitrification-driven anammox (PD/A) is an energy-efficient method for nitrogen removal from wastewater. However, its stability and efficiency are impeded by the competition between heterotrophic denitrifying bacteria and relatively slow-growing anammox bacteria. In this study, a PD/A granular sludge system was developed, which achieved a nitrogen removal efficiency of 94% with 98% anammox contribution, even as the temperature dropped to 9.6 °C. Analysis of bacterial activity in aggregates of different sizes revealed that the largest granules (>2.0 mm) exhibited the highest anammox activity, 2.8 times that of flocs (<0.2 mm), while the flocs showed significantly higher nitrite production rates of PD, more than six times that of the largest granules. Interestingly, fluorescent in situ hybridization (FISH) combined with confocal laser scanning microscopy (CLSM) revealed a nest-shaped structure of PD/A granules. The Thaueragenus, a key contributor to PD, was highly enriched at the outer edge, providing substrate nitrite for anammox bacteria inside the granules. As temperature decreased, the flocs transformed into small granules to efficiently retain anammox bacteria. This study provides multidimensional insights into the spatiotemporal assembly and immigration of heterotrophic and autotrophic bacteria for stable and high-rate nitrogen removal.