Enhancement of nitrogen removal in constructed wetlands through boron mud and elemental sulfur addition: Regulation of sulfur and oxygen cycling

Constructed wetlands (CWs) utilizing elemental sulfur (S0) as an electron donor are effective for nitrate (NO3–-N) removal from low C/N wastewater. However, the limited bioavailability of chemical S0 (chem-S0) due to low water solubility and inadequate mass transfer impedes complete denitrification. In this study, boron mud waste and S0 (S0&BM) were processed and combined to convert chem-S0 to biological S0 (bio-S0) in CWs. The employment of S0&BM improved NO3–-N and total nitrogen (TN) removal rates by 15.36 % and 10.55 % compared to S0 alone. The 15N isotope tracing experiment unveiled the pathways of N2O production and demonstrated that S0&BM facilitated full denitrification, resulting in a higher percentage of nitrogen being degraded in a non-polluting form (54.70 %). Releasing trace amounts of boron and magnesium in S0&BM mitigated the excessive acidification caused by the sulfur autotrophic process and promoted biofilm formation, thereby pronouncing vertical oxygen fluctuations. Meanwhile, oxygen vacancies generated in the S0&BM promoted the closed-loop sulfur cycle, starting from the oxidation of chem-S0 to sulfate (SO42-), the important process of SO42--reducing bacteria using partial organic compounds as electron donors to reduce SO42- to sulfide (S2-) was realized, and finally the S2- was oxidized to bio-S0 with better properties. S0&BM enriched functional microbial communities in the biofilm related to nitrogen and sulfur transformation. Functional gene analysis showed an increase in the narG, nirS, norB, and nosZ genes supporting denitrification. The dsrA and sqr genes responsible for the sulfur cycle exhibited the highest abundance. This realization allowed for the utilization of waste materials for treatment, enhancing the energy efficiency of carbon reduction and pollution reduction in CWs.