Nitrogen removal performance of sulfur autotrophic denitrification under different S₂O₃^2- additions using isotopic fractionation of nitrogen and oxygen

The dual isotope fractionation of nitrogen (N) and oxygen (O) is an effective way to track the transformation of NO₃^--N in biological denitrification process. The Sulfur autotrophic denitrification combined with the different concentrations of S₂O₃^2- was investigated using the dual isotope fractionation of nitrogen (N) and oxygen (O) to reveal the nitrogen removal mechanism of the activated sludge. Based on successful autotrophic denitrification incubation, the modified Logistic model responded to the short-term effects of S₂O₃^2- addition on NO₃^--N removal and SO₄^2- generation. Under the S₂O₃^2- addition of 0.5, 1, 2 and 4 times of the incubation stage (49.29 mg/L-394.32 mg/L), the fractionation effect of N in NO₃^--N (¹⁵ε_NO3) decreased from 8.74 ± 1.81‰ to 2.08 ± 0.06‰, and the fractionation effect of O in NO₃^--N (¹⁸ε_NO3) declined from 11.34 ± 0.46‰ to 5.48 ± 0.46‰. The ¹⁵ε_NO3/¹⁸ε_NO3 was maintained at 0.46-0.94, indicating a negative correlation between addition amount and isotope effect, and the addition of high concentrations of S₂O₃^2- was not suitable for system stabilization. Moreover, the ¹⁸O-labeled H₂O (δ¹⁸O_H2O) tests significantly proved the presence of O exchange between NO₂^--N/NO₃^--N and H₂O (67%/97%) during the nitrogen removal process, while the reoxidation of NO₂^--N was explored in the autotrophic denitrification. The kinetic models coupled with isotope fractionation effectively revealed the nitrogen removal characteristics in the autotrophic denitrification systems, and verified the difference between the activated sludge-based wastewater treatment process and the natural ecosystem.