Synergistic Regulation by Long- and Short-Chain Quorum Sensing Signaling Molecules Enhances Sulfamethoxazole Metabolism in Electroactive Biofilms within a Microbial Electrolysis Cell Coupled Anaerobic Digestion System
High-strength sulfamethoxazole (SMX) wastewater severely inhibits anaerobic microorganisms, reducing organic degradation and methane yield. This study investigated the effects of short-chain (C6-HSL) and long-chain (C12-HSL) N-acyl-homoserine lactone (AHL) signaling molecules, individually and in combination, on the construction, performance, and antibiotic resistance gene (ARG) profiles of anaerobic electroactive biofilms within a microbial electrolysis cell coupled anaerobic digestion (MEC-AD) system. Compared to the control (no AHLs), SMX removal efficiency increased by 9.26%, 7.44%, and 10.67% for C6-HSL (T1), C12-HSL (T2), and combined (T3) treatments, respectively. Methane production rates rose by 20.4%, 16.9%, and 23.1% for T1, T2, and T3, respectively. AHLs promoted extracellular polymeric substance secretion, enhancing electroactive microbe attachment to the anode. Microbial community analysis revealed increased diversity and modulated key functional genera. Notably, Georgenia abundance increased by 16.77% (T1) and 36.47% (T3) but decreased by 15.99% (T2). ARG analysis showed that single AHLs elevated intI1, sul1, and sul2 abundances, whereas combined AHLs (T3) exhibited a milder response, with sul2 abundance reduced by 4.92% relative to control. This suggests synergistic AHLs suppress ARG host proliferation. This study first demonstrates that combined short- and long-chain AHLs enhance electroactive biofilm formation, maintain microbial community stability, and modulate ARG dissemination risk, offering a quorum sensing-based strategy for antibiotic wastewater treatment and risk management.