Evaluating the role of recalcitrant dissolved organic matter in bacterial community dynamics in urbanized freshwater ecosystems

Dissolved organic matter (DOM) and recalcitrant dissolved organic matter (RDOM) play distinct roles in shaping microbial communities. However, characterizing these roles is difficult, especially in ecosystems subjected to varying degrees of anthropogenic influence. This study investigated the molecular compositions and ecological impacts of DOM and RDOM in the Fen River, Shanxi Taiyuan, comparing pristine upstream regions with highly urbanized downstream areas. Using 16S rRNA gene sequencing and LC-MS-based metabolomics, we observed significant shifts in microbial community composition, diversity, and metabolic functions. Upstream communities, characterized by higher diversity, were dominated by Bacteroidota, Proteobacteria, and Cyanobacteria, while downstream communities, influenced by pollution, exhibited increased expression of genes related to amino acid metabolism. Fluorescence spectroscopy and Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) revealed that upstream DOM contained higher proportions of complex, high molecular weight compounds, including significant proportions of carboxyl-rich alicyclic molecules (CRAM) and island of stability (IOS) compounds, which play key roles in long-term carbon storage and microbial carbon sequestration. In contrast, downstream DOM was characterized as having lower aromaticity and more saturated compounds, with reduced proportions of CRAM and IOS, reflecting the impact of anthropogenic activities. These findings underscored the critical roles of CRAM and IOS in regulating DOM stability and microbial communities, further highlighting the need for targeted pollution control strategies to preserve ecosystem function in urbanized water bodies.