The role of the combined nitrogen-sulfur-carbon cycles for efficient performance of anammox-based systems

The combined anammox/mixotrophic denitrification process was conducted in two granular sequencing batch reactors (SBRs) during a 200-day operation. Both reactors were fed with synthetic medium, but SBR2 was enriched with additional sulfate (SO₄^2-) which influenced sulfate reduction ammonium oxidation (SRAO) and heterotrophic reduction of SO₄^2- by sulfate reducing bacteria. It was hypothesized that the addition of SO₄^2- could positively impact the removal rates of N-S-C compounds. A low C/N ratio (0.4-1.6) was maintained to prevent inhibition of anaerobic ammonium oxidizing bacteria (AnAOB), and alternating chemical oxygen demand (COD) on/off conditions were used to regenerate AnAOB during COD-off phases and heterotrophic denitrifiers during COD-on phases. Stoichiometric analysis showed that introducing SO₄^2- in SBR2 enhanced the ammonium utilization rate, which was approximately 10 % higher compared to SBR1 in the final stage of the experiment (25.8 vs. 22.8 mg N/(g VSS·h)). The total nitrogen removal efficiencies ranged from 62 % to 99 % in both reactors, with SBR2 consistently exhibiting approximately 4 % higher efficiency than SBR1. In SBR2, the maximum overall SO₄^2- utilization efficiency reached 27 % under COD-off conditions, while overall COD utilization was almost complete under COD-on conditions. A strong correlation (R² = 0.98) was observed between SO₄^2- production and COD utilization. The key players responsible for N and S transformations in response to SO₄^2- addition were Candidatus Brocadia and Chloroflexi - Anaerolineae. This study highlights the potential to enhance the overall efficiency of N-S-C removal by implementing an integrated anammox/mixotrophic denitrification process. The combination of cycles emerges as a sustainable approach for treating wastewater rich in N-S-C compounds.