Insights into complete nitrate removal in one-stage nitritation-anammox by coupling heterotrophic denitrification

Nitritation-anammox has been considered to be the most promising process for nitrogen (N) removal from wastewater. However, the anammox reaction still produces an amount of nitrate, which cannot be removed further. This study hypothesizes that heterotrophic denitrification can be an appealing option to remove the residual nitrate in the one-stage nitritation-anammox process. Through monitoring N-removal performance and microbial community succession of a laboratory microaerobic reactor, the effect of four different levels of oxygen supply on nitrate removal was investigated. The reactor was continuously fed with real manure-free piggery wastewater containing ~240 mg NH₄⁺-N/L and chemical oxygen demand (COD)/total nitrogen (TN) ratio of less than 1 for 180 days. With a high influent loading rate of 0.7 kg N/(m³·d), efficient total nitrogen removal (>80 %) was achieved during stable operation of dissolved oxygen (DO) concentrations between 0.3 and 0.6 mg O₂/L, indicating N-removal via the nitritation-anammox pathway in the low-carbon wastewater treatment. At the same time, the effluent nitrate reduced with decreased oxygen supply and completely depleted at DO of 0.3 ± 0.1 mg O₂/L. In addition to oxygen, preventing ammonia nitrogen from falling to very low levels (<10 mg/L) could be also useful for the complete nitrate removal and stable nitritation-anammox. 16S rRNA gene-based analyses confirmed a complex microbial community including nitrifiers, denitrifiers and anammox bacteria in the biomass of the reactor. Collectively, this study provides new insights into high-level N-removal of a nitritation-anammox process by complete nitrate depletion.