Groundwater contamination by per- and polyfluoroalkyl substances (PFAS) poses a persistent environmental and public health concern. This study evaluates a two-chambered bioelectrochemical system (BES) inoculated with the West Branch Consortium (WBC-2) for PFAS remediation. Under an applied cathodic potential of −450 mV (versus Ag/AgCl), the BES with active WBC-2 achieved >99.0% perfluorooctanesulfonic acid (PFOS) removal within 21 days in deionized water with culture medium and > 98.9% removal of PFOS, perfluorooctanoic acid (PFOA), perfluorohexanoic acid (PFHxA), and perfluorohexanesulfonic acid (PFHxS) in contaminated groundwater after 102 days. Intermediate formation (e.g., PFOA, 6:2 fluorotelomer sulfonate (6:2 FTS), perfluoropropionic acid (PFPrA), perfluorobutanoic acid (PFBA)) and background-corrected fluoride release were consistent with PFOS transformation under anaerobic reducing conditions potentially involving defluorination. Following repeated PFOS spikes (100 μg/L on Days 0, 50, and 399), PFOA, PFPrA, and PFBA accumulated over 664 days. Despite being the dominant accumulated compound, PFOA accounted for <1.8% of the total spiked PFOS mass. Minimal PFOS transformation occurred in controls without active WBC-2, highlighting the importance of microbial metabolism. Biofilm analysis revealed dense colonization of rod-shaped bacteria on carbon fiber brushes. Enrichment of Bacillus, Agrobacterium, and other low-abundance taxa suggests selective adaptation to BES and PFAS conditions. These findings highlight BES driven by electrochemically stimulated microbial activity as a promising strategy for PFAS remediation.