Enhancing nitrogen removal in low C/N wastewater with recycled sludge-derived biochar: A sustainable solution

Denitrification is an important biological process in wastewater treatment plants (WWTPs). However, a low carbon-to-nitrogen (C/N) ratio limits the availability of organic carbon, potentially reducing denitrification efficiency. This study investigates the impact of sludge-derived biochar on the nitrogen removal of activated sludge for low C/N ratio municipal wastewater. Sludge-based biochar was characterized by its physicochemical properties, revealing that biochar prepared at 400 °C exhibited the highest specific surface area and the most favorable surface functional groups for electron transfer. The results from batch tests showed that adding 4 g/L of biochar dosage enhanced denitrification rates and total nitrogen (TN) removal efficiency the most. Sequencing batch reactors (SBRs) experiments further confirmed that biochar dosgae improved the removal efficiencies of COD, NH₄⁺-N, and TN, achieving stable values of 97.2 ± 1.2 %, 99.2 ± 0.6 %, and 83.8 ± 2.4 %, respectively. Metabolic and electrochemical analyses revealed that biochar addition enhanced the activity of denitrification enzymes, increasing the ammonia oxidation rate by 12.9 ± 0.7 %, nitrite oxidation rate by 14.7 ± 1.2 %, nitrate reduction rate by 36.9 ± 1.5 %, and nitrite reduction rate by 16.4 ± 0.8 %. The relative abundance of denitrification functional genes (amoA, nirS, nirK, narG, nosZ) increased, and the activities of the corresponding enzymes (AMO, NXR, NAP, NIR) rose by 23±6 %, 53±5 %, 260±15 %, and 55±7 %, respectively. This increase in enzyme activity suggested enhanced denitrification processes, which was further supported by the 60.1 ± 3.7 % increase in electron transfer system activity (ETSA), indicating that biochar acted as an electron shuttle. This study proposes a potential sustainable approach for sludge recycling and enhanced wastewater nitrogen removal under low C/N conditions.