Mainstream double-anammox driven by nitritation and denitratation using a one-stage step-feed bioreactor with real municipal wastewater

A novel double-anammox process for advanced mainstream nitrogen removal was established using step-feed sequencing batch reactor (SBR) system with integration of suspend sludge and biofilms. Following optimization of influent distribution ratio, the effluent total inorganic nitrogen (TIN) was < 10.2 mg N/L, with influent TIN of 43.4 mg N/L, and anammox contributed 71.4% to TIN removal. Biological processes and batch tests revealed that gradient C/N reduction promoted denitratation/anammox in anoxic stage, and simultaneous nitritation and anammox were achieved in oxic stage. Specially, anammox maintained on biofilms with abundance over 109 copies/ (g dry sludge). High-throughput sequencing revealed that Thauera and Nitrosomonas were enriched in flocs. Furthermore, metagenomic sequencing confirmed that Thauera owns narG and napA (NO3--> NO2-) and Nitrosomonas owns amoA (NH4+-> NO2-), support stable NO2- supply for double-anammox. This mainstream anammox-dominant process could potentially be used for stable nitrogen removal in municipal wastewater treatment plants.