Mixotrophic denitrification using thiocyanate as an electron donor: Role of thiocyanate under different C/N conditions

Thiocyanate (SCN − ) is a highly toxic reducing inorganic compound commonly found in various nitrogen-rich wastewater and is also a promising electron donor for mixotrophic denitrification. However, its extent of involvement in mixotrophic denitrification under conditions of carbon limitation or excess remains unclear. In this study, five reactors were constructed to investigate the participation and microbial mechanisms of SCN − in mixotrophic denitrification under high C/N and low C/N conditions. The results showed that under low C/N conditions, SCN⁻ synergizes with organic electron donors to enhance denitrification, with mixotrophic systems exhibiting higher electron utilization efficiency and achieving 1.12 times the TN removal rate of autotrophic and heterotrophic systems combined. Under high C/N conditions, an ample supply of electron donors achieve 100% nitrate removal; however, SCN⁻ also participates in denitrification, competing with organic electron donors for NO 3 − -N, which leads to the waste of organic matter. Additionally, high-throughput sequencing revealed that under low C/N mixotrophic conditions, SCN⁻ effectively promoted the growth of SCN⁻-degrading microorganisms such as Thiobacillus , with its abundance increasing from 0% to 8.86%, approximately 1.85 times higher than under autotrophic conditions. This enhancement strengthened the sulfur and nitrogen metabolic capabilities of the microbial community, enabling the system to utilize SCN − more fully for denitrification. This study provides novel insights for reducing the addition of external organic matter to nitrogen-rich wastewater containing SCN − , offering theoretical and technical support for energy-saving and emission reduction in denitrification processes of actual industrial wastewater treatment.