Advancing fluorescence tracing with 3D-2D spectral conversion: A mixed culture on microbial degradation mechanisms of DOM from a large-scale watershed

Abstract

Fluorescence tracing, known for its precision, rapid application, and cost-effectiveness, faces challenges due to the microbial degradation of dissolved organic matter (DOM) in aquatic environments, altering its original spectral fingerprint. This study conducted a 15-day microcosm experiment to examine the effects of biodegradation on the spectral properties of DOM from various sources: livestock excrement (EXC), urban sewage (URB), industrial wastewater (IND), and riparian topsoil (tDOM). Our findings show that while the spectral structures of DOM from different sources change during 15 days of microbial degradation, these changes do not overlap or interfere with each other. However, distinguishing between tDOM and URB in the presence of both IND and EXC is only possible at high resolution. Spectral index calculations revealed significant fluctuations and interference in FI and BIX indices among samples from different sources due to microbial degradation. In contrast, the HIX index exhibited independent fluctuations and remained a reliable spectral index for tracing. LEfSe (Linear discriminant analysis Effect Size) identified characteristic bio-indicators (CBI) for each DOM source. The CBI for tDOM and URB differed significantly; tDOM showed a marked CBI only within the first four days of microbial degradation, with a sharp decline in abundance thereafter, while URB's CBI remained abundant for 12 days. Similarly, IND's CBI maintained high relative abundance for the first 12 days. EXC's CBI was unique, showing a distinct and stable community only after six days of degradation, likely due to its high bioavailability and initial rapid microbial utilization. This study addresses the temporal variability in spectral tracing techniques caused by pollutant biodegradation. We developed a combined spectral-biological tracing technique using the “three-dimensional to two-dimensional” method along with bio-indicators, enhancing the accuracy and timeliness of spectral tracing.