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An efficient and stable combined denitrification-partial nitrification-Anammox process with a dual recycling system was used to remove nitrogen from mature landfill leachate. After 155 d of operation, the NO3− as the PN-Anammox byproduct was almost treated with biodegradable organic carbon in raw wastewater in a pre-denitrification reactor by external recycling system. When raw landfill leachate with NH4+-N concentration of 1900 mg/L was treated, an integrated reactor with airlift recycling was combined with the PN and Anammox reactions to efficiently remove NH4+ from the inflow. The total nitrogen concentration of effluent stabilized at 20 mg/L and total nitrogen removal efficiency was 99%. The maximum NO2− production rate in the aerobic zone was 2.2 kg/(m3·d) and the maximum nitrogen removal rate in the anaerobic zone was 21.4 kg/(m3·d). The most common phyla among the nitrification and the Anammox functional bacteria were Nitrosomonas, Candidatus Kuenenia, and Candidatus Brocadia after landfill leachate treatment.

The effects of salinity changes on nitrogen transformation efficiency and recoverability were studied by using a partial nitration (PN)- anaerobic ammonium oxidation (Anammox) integrated reactor. The changes of microbial community structure and population abundance during the increase and decrease of salinity were also analyzed by 16S rRNA gene high-throughput sequencing. The results showed that when the salinity was increased to 1.35%, the combined PN-Anammox process achieved the maximum stimulated and total nitrogen removal rate (TNRR) arrived at 1.1kg/(m3·d). When the salinity was higher than 1.35%, the activities of AOB and Anammox bacteria began to be inhibited. When the salinity reached 2.4%, the TNRR decreased to 60%. TNRR was fast restored, when salinity was reduced to 0.11%. The genes of AOB and Anammox bacteria indicated that the TNRR of the reactor was restored after salinity inhibition, but the functional microbial community structure and abundance had relatively large, irreversible changes.

For municipal wastewater with low temperature and ammonium, conventional oxygen-limited have difficulty achieving long-term stable inhibition of nitrite oxidizing bacteria (NOB) and stable nitritation. So a partial nitrification-anaerobic ammonium oxidation integrated reactor with independent partitions was used to investigate the feasibility of adding an auto-recycling system to promote low exposure of nitrite in the aerobic zone and to inhibit the NOB activity. The results showed that nitrite produced in the aerobic zone could be timely transported to the anaerobic zone for Anammox utilization, and the nitrite nitrogen concentration was diluted to keep within 1 mg/L in the aerobic zone by the effluent recycling. NOB growth was inhibited by nitrite deficiency. The maximum nitrogen removal rate of the reactor was 0.29 kg/(m3·d), and the nitrate nitrogen production rate of NOB was controlled within 0.04 kg/(m3·d). Nitrosomonas and Candidatus Kuenenia were found as functional species of ammonia-oxidizing bacteria and Anammox bacteria, respectively.

The differences in the physiological characteristics between AOB and ANAMMOX bacteria lead to suboptimal performance when used in a single reactor. In this study, aerobic and anaerobic zones with different survival environments were constructed in a single reactor to realize partitioned culture of AOB and ANAMMOX bacteria. An external air-lift reflux system was formed which used the exhaust from the aeration zone as power to return the effluent to the aeration zone. The reflux system effectively alleviated the large pH fluctuations and promoted NO2--N to rapidly use by ANAMMOX bacteria, effectively inhibiting the activity of NOB. After 95d of running, the nitrogen removal rate increased from the initial 0.21kg/(m3·d) to 3.1kg/(m3·d). FISH analyses further demonstrated that AOB and ANAMMOX bacteria acquired efficient enrichment in the corresponding zone. Thus, this type of integrative reactor may create the environments needed for the partial nitritation-ANAMMOX processing.

In this study, three upflow column reactors were compared for anaerobic ammonium oxidation (Anammox) process start-up time with different ferrous iron concentration in feeding. Continuous experiments indicated that the start-up time of Anammox process could be shortened from 70 to 58d in R2 (0.06mM Fe(2+)) and 50d in R3 (0.09mM Fe(2+)). The Anammox activity appeared after 16days operation in R3. Quantitative PCR (q-PCR) analysis demonstrated a significant increase in quantity of Anammox bacteria in R3 compared with the other two reactors during entire operation. At the Fe(II) concentration of 0.09mM, the heme c levels inside Anammox cell and hydrazine dehydrogenase (HDH) activity increased dramatically, which could be the trigger of fast Anammox start-up.

In this study, we explored the effects of Fe(II) on the activity of anammox biomass both in short and long terms. With the influent Fe(II) at 0.09 and 0.12 mM, continuous experiments demonstrated that the nitrogen removal rates increased 32.2% and 29.9% compared to that with normal Fe(II) level (0.03 mM). Influent Fe(II) variation could affect the total Fe, heme c levels and hydrazine dehydrogenase activity. At the Fe(II) concentration of 0.09 mM, the total Fe, heme c levels inside anammox cell and hydrazine dehydrogenase activity could increase about 2.0, 2.1 and 2.35 folds as much as that with normal Fe(II) level. However, when the Fe(II) elevated to 0.18 mM, it would cause a mal-effect on anammox bacteria and further deteriorate nitrogen removal performance. It was indicated that the appropriate increase of Fe(II) was beneficial for more heme c synthesis, enhancement of hydrazine dehydrogenase activity, the growth of anammox bacteria.

This study examined long-term effect of MnO(2) powder (average diameter of 4-7 μm) on nitrogen removal in anammox process. Two lab-scale up-flow anammox reactors were operated for 380 days, one with and one without MnO(2) powder addition. During the period when only substrate concentrations varied, the maximum nitrogen removal rate in the reactor with MnO(2) addition reached 920.9 g-N/m(3)/d. This value was 2-folds higher than that (464.6 g-N/m(3)/d) of the reactor without MnO(2) addition. The crude enzyme activities of the anammox biomass from the two reactors was measured as 0.531±0.019 and 0.298±0.007 μmol cytochrome c reduced/mg protein/min, respectively. Transmission electron microscopy observation demonstrated more undefined particles existing inside anammox bacterial cell in the reactor with MnO(2) powder addition. Furthermore, filament-like structures inside anammoxosome were observed, which formed a net-like structure with particles as the connecting nodes. The experiment results demonstrated that MnO(2) improved nitrogen removal performance of anammox process.