Influence of Photosynthetic Cathodes on Anodic Microbial Communities in Acetate-Fed Microbial Fuel Cells Pre-Enriched Under Applied Voltage

Electrical stimulation is increasingly explored as a strategy to accelerate the development of electroactive biofilms in microbial fuel cells (MFCs), yet its integration with photosynthetic MFCs (pMFCs) remains insufficiently understood. This study evaluated how short-term anodic stimulation (0.5–5 V, 4 days) affects biofilm formation and COD removal, and how subsequent operation with photosynthetic cathodes—Chlorella sp., Arthrospira platensis and Tetraselmis subcordiformis—modulates anodic microbial communities and functional potential. Stimulation at 1 V yielded the best activation effect, resulting in the highest voltage output, power density and fastest COD removal kinetics, whereas 5 V inhibited biofilm development. During pMFC operation, Chlorella produced the highest voltage (0.393 ± 0.064 V), current density (0.14 ± 0.02 mA·cm−2) and Coulombic efficiency (~19%). Arthrospira showed moderate performance, while Tetraselmis generated no current despite efficient COD removal. 16S rRNA sequencing revealed distinct cathode-driven community shifts: Chlorella enriched facultative electroactive taxa, Arthrospira promoted sulfur-cycling bacteria and Actinobacteria, and Tetraselmis induced strong methanogenic dominance. Functional prediction and qPCR confirmed these trends, with Chlorella showing increased pilA abundance and Tetraselmis displaying enriched methanogenic pathways. Overall, the combined use of optimal anodic stimulation and photosynthetic cathodes demonstrates that cathodic microalgae strongly influence anodic redox ecology and energy recovery, with Chlorella-based pMFCs offering the highest electrochemical performance.