A strategy for fast anammox biofilm formation under mainstream conditions

One of the bottlenecks to applying anaerobic ammonium oxidation (Anammox) is the long start-up time, especially under mainstream conditions. This study proposed a strategy for fast anammox biofilm formation under mainstream conditions. By first cultivating an aerobic heterotrophic biofilm, and then transferring to anoxic conditions, a pre-cultivated heterotrophic biofilm can be formed in 12 days. The pre-cultivated heterotrophic biofilm then functions as a “glue” to accelerate anammox bacteria adhesion and biofilm formation. Secondary settled effluent with externally added 15–30 mg-N·L⁻¹ ammonium and nitrite was applied as reactor influent. With a single inoculation of suspended growth anammox-laden biomass and no bioaugmentation, an anammox-enriched biofilm formed after 5 months of operation under uncontrolled temperature of 15–20 °C. Both the nitrogen removal rate and specific anammox activity exponentially increased over the course of study, corresponding to an estimated anammox doubling time of 10.8 days. The biofilm thickness on primed carriers was 2–3 times higher than on the non-primed carriers over the first 5 months of operation, and the hszA gene copy number in primed biofilms revealed was consistently 1 to 2 times higher than the non-primed carrier biofilm, indicating that biofil m carrier priming via selection for a pre-cultivated heterotrophic biofilm base can effectively improve the anammox enrichment rate at early stages of reactor operation. Time, rather than the type of biofilm (primed versus non-primed), had a stronger influence on microbial community structure over the full 230 days of reactor operation. Candidatus Brocadia was the only detected anammox bacteria genus. Overall, pre-cultivation of heterotrophs on biofilm carriers provides a simple route to accelerate anammox-enriched biofilm formation under mainstream conditions.