Enhanced nitrite accumulation under mainstream conditions by a combination of free ammonia-based sludge treatment and low dissolved oxygen: reactor performance and microbiome analysis

Partial nitritation under mainstream conditions is one of the major bottlenecks for the application of deammonification processes to municipal wastewater treatment plants. This study aimed at evaluating the combination effect of a side-stream free ammonia (FA) treatment and low dissolved oxygen (0.2 ± 0.1 mg L−1) on inhibiting nitrite oxidizing bacteria (NOB) from enhancing nitrite accumulation in long-term lab-scale experiments. Two continuous floccular sludge reactors treating low-strength synthetic wastewater (60 mg N–NH4+ L−1 without COD) with a fixed nitrogen loading rate of 0.22 ± 0.03 g N per L per day were operated in a varied temperature range of 7–31 °C, with one acting as the experimental reactor and the other as the control. Side-stream sludge treatment with a stepwise elevation of FA concentration (65.2–261.1 mg NH3 L−1) was carried out every day in the experimental reactor; the nitrite accumulation ratio (NAR, (NO2–N/(NO2−–N + NO3−–N) × 100%)) in the experimental reactor was always about twice that in the control one. Quantitative PCR (q-PCR) and high-throughput sequencing analyses showed the dominant NOB was mostly Nitrobacter, while there was an alternating trend between Nitrobacter and Nitrospira. Even though the whole microbial communities of each experimental stage between the two reactors were relatively clustered due to an incomplete NOB washout, three abundant metabolisms (amino acid metabolism, pyruvate metabolism and nitrogen metabolism) and key functional genes of nitrification predicted by PICRUSt in the experimental reactor were enriched, providing a better understanding of nitrite accumulation. These results have demonstrated that the positive hybrid effects of FA side-stream sludge treatment and a low DO could enhance nitrite accumulation. It is expected that a complete washout of NOB would be achieved after further process optimization.