Long term performance and dynamics of microbial biofilm communities performing sulfur-oxidizing autotrophic denitrification in a moving-bed biofilm reactor

Sulfide-oxidizing autotrophic denitrification (SOAD) implemented in a moving-bed biofilm reactor (MBBR) is a promising alternative to conventional heterotrophic denitrification in mainstream biological nitrogen removal. The sulfide-oxidation intermediate – elemental sulfur – is crucial for the kinetic and microbial properties of the sulfur-oxidizing bacterial communities, but its role is yet to be studied in depth. Hence, to investigate the performance and microbial communities of the aforementioned new biosystem, we operated for a long term a laboratory-scale (700 d) SOAD MBBR to treat synthetic saline domestic sewage, with an increase of the surface loading rate from 8 to 50 mg N/(m²·h) achieved by shortening the hydraulic retention time from 12 h to 2 h. The specific reaction rates of the reactor were eventually increased up to 0.37 kg N/(m³·d) and 0.73 kg S/(m³·d) for nitrate reduction and sulfide oxidation with no significant sulfur elemental accumulation. Two sulfur-oxidizing bacterial (SOB) clades, Sox-independent SOB (SOB_I) and Sox-dependent SOB (SOB_II), were responsible for indirect two-step sulfur oxidation (S²⁻→S⁰→SO₄ ²⁻) and direct one-step sulfur oxidation (S²⁻→SO₄ ²⁻), respectively. The SOB_II biomass-specific electron transfer capacity could be around 2.5 times greater than that of SOB_I (38 mmol e⁻/(gSOB_II·d) versus 15 mmol e⁻/(gSOB_I·d)), possibly resulting in the selection of SOB_II over SOB_I under stress conditions (such as a shorter HRT). Further studies on the methods and mechanism of selecting of SOB_II over SOB_I in biofilm reactors are recommended. Overall, the findings shed light on the design and operation of MBBR-based SOAD processes for mainstream biological denitrification.