Nitrate with enriched heavy oxygen isotope linked to changes in nitrogen source and transformation as groundwater table rises

Nitrate is a significant constituent of the total nitrogen pool in shallow aquifers and poses an escalating threat to groundwater resources, making it crucial to comprehend the source, conversion, and elimination of nitrogen using appropriate techniques. Although dual-isotope dynamics in nitrate have been widely used, uncertainties remain regarding the asynchronously temporal changes in delta 18O-NO3- and delta 15N-NO3 - observed in hypoxic aquifers. This study aimed to investigate changes in nitrogen sources and transformations using temporal changes in fieldbased NO3- isotopic composition, hydro-chemical variables, and environmental DNA profiling, as the groundwater table varied. The results showed that the larger enrichment in delta 18O-NO3- (+13%o) compared with delta 15N-NO3 (-2%o) on average during groundwater table rise was due to a combination of factors, including high 18O-based atmospheric N deposition, canopies nitrification, and soil nitrification transported vertically by rainfalls, and 18O-enriched O2 produced through microbial and root respiration within denitrification. The strong association between functional gene abundance and nitrogen-related indicators suggests that anammox was actively processed with nitrification but in small bacterial population during groundwater table rise. Furthermore, bacterial species associated with nitrogen-associated gradients provided insight into subsurface nitrogen transformation, with Burkholderiaceae species and Pseudorhodobacter potentially serving as bioindicators of denitrification, while Candidatus Nitrotogn represents soil nitrification. Fluctuating groundwater tables can cause shifts in hydro -chemical and isotopic composition, which in turn can indicate changes in nitrogen sources and trans-formations. These changes can be used to improve input sources for mixture models and aid in microbial remediation of nitrate.