Enhanced heterotrophic denitrification in groundwater using pretreated Ginkgo biloba leaves: Optimized carbon...

Ginkgo biloba leaves (Gbl), as abundant agricultural and forestry residues which contains the quercetin that plays an important role in mediating electron transfer, represent a promising heterotrophic denitrification carbon source. Nonetheless, challenges persist due to concerns over nitrate leaching. This study pioneers the application of pretreated Gbl as external carbon sources for heterotrophic denitrification, with a focus on enhancing carbon bioavailability and mitigating nitrate leaching risks. Among the pretreatment strategies employed, the extraction process effectively eliminated NO₃^--N leaching, while the fermentation process reduced it by 52.8%. The saturated total organic carbon (TOC) concentration per unit mass of fermented Gbl was marginally lower compared to untreated leaves, yet the secondary kinetic reaction constant increased from 10.94 to 12.91 mg/(g·h·L), indicating an accelerated organic carbon release rate. Fermentation with Eurotium cristatum disrupted the rigid lignocellulose structure, thereby enhancing carbon source bioavailability. This resulted in a significant increase in alcohols in the leaching solution, from 27.0% to 68.6%, and a substantial reduction in aromatic compounds, from 20.2% to 0.2%, which alleviated microbial toxicity. In terms of denitrification performance, fermented Ginkgo biloba leaves (Fl) outperformed Ginkgo biloba extract residue leaves (Erl), which in turn surpassed untreated Gbl. Both Fl and Erl demonstrated robust adaptability across a broad pH range of 5.0-11.0. Under neutral conditions, the Fl system exhibited the highest primary kinetic constant for nitrate removal, reaching 0.0494 h⁻¹. Microbial community revealed that all three carbon sources harbored denitrification and lignocellulose degradation capabilities. Notably, the Fl and Erl systems exhibited enhanced carbohydrate transport (G), amino acid transport (E), and inorganic ion transport (P), underscoring the potential pretreatments to optimize carbon source utilization. Collectively, these findings affirm the viability of Gbl as a carbon source for heterotrophic denitrification, providing valuable insights for its application in addressing nitrate pollution in aquatic environments.