Nitrogen removal performance of anammox immobilized fillers in response to seasonal temperature variations and different operating modes: Substrate utilization and microbial community analysis.

Four anaerobic ammonium oxidation (anammox) immobilized filler reactors (R1: 33 °C-normal, R2: seasonal temperature-normal, R3: seasonal temperature-feast, R4: seasonal temperature-starvation) were established to study the response of anammox immobilized fillers to seasonal temperature changes and different operating modes. The results showed that the anammox immobilized filler could better adapt to the seasonal temperature drop and maintain the activity potential by adjusting the hydraulic retention time (HRT). During the temperature rise phase, R2 activity increased rapidly with the highest nitrogen removal rate reaching 1.26 kgN·(m·d), which was equivalent to control sample R1 (1.33 kgN·(m·d)). However, feasting and famine conditions severely impaired anammox performance and changed stoichiometric ratios; feasting, in particular, significantly lowered the nitrogen removal potential of R3. The specific anammox activity of R2, R3 and R4 was 92.2%, 52.6% and 67.9%, respectively, that of R1, respectively, where the accumulation of functional bacteria was the reason for the higher activity of R2. Degradation kinetics and NO-N inhibition curves showed that R3 was less sensitive to high concentrations of NH-N, while R4 responded earlier to low concentrations of NH-N, and the reduction of IC50 at low temperature was the reason for the inhibition of R3 activity. Furthermore, seasonal temperature fluctuations had little effect on the microbial community structure but had a considerable impact on bacteria abundance. The anammox functional bacteria Candidatus Kuenenia was found to be the dominant genus in R1-R4; however, the relative abundance of most bacteria, including anammox bacteria, decreased in R3, while the proportion of fermentation bacteria and denitrifying bacteria increased in R4. These findings highlight the necessity of rational regulation of HRT for the adaptation of anammox immobilized fillers to seasonal temperature changes, which could enhance our understanding of the synergistic effect of seasonal temperature changes and different operating modes on nitrogen removal.