Spatial difference in nitrogen removal pathways and microbial functional diversity in an EGSB reactor during the start-up of PD/Anammox.

The start-up of a relatively high nitrogen load PD/Anammox in an EGSB reactor was achieved through strategies of bioaugmentation, mass transfer enhancement, and COD/NO-N control, with NRR of 5.2 g N/L/d. Longitudinal heterogeneity in EGSB reactor induced divergent nitrogen conversion pathways and enriched different functional microorganisms between stratified sludge. Along the elevation of the reactor, the proportion of removed nitrogen through anammox increased continuously from bottom, middle and up, which were 65.0 %, 79.8 %, and 84.1 %, respectively, consistent with the trend of ex-situ activities calculated with Gompertz model. The bottom zone played a role in mixed nitrogen conversion to provide NO-N accumulation and nitrogen removal, with higher abundance of Thauera, Denitratisoma and Ignavibacterium. The middle part was enriched Candidatus_Kuenenia (12.51 %), and up inhibited completed denitrification, together forming the anammox dominant zone. The proposed functional zones in the EGSB reactor provided approaches for the optimisation of high-load PD/Anammox systems.