In this study, four dual-chambered microbial fuel cells (MFC1-4) were constructed and filled with different ratios of mature fine tailings and oil sands process-affected water to test the feasibility of MFCs to simultaneously generate electricity and treat oil sands tailings. After 800 h of operation, the maximum voltage was observed in MFC4 at 0.726 V with 1.2 kΩ external resistance loaded. The maximum power density reached 392 ± 15 mW/m2 during the 1700 h of MFC4 operation. With continuous electricity generation, MFC4 removed 27.8% of the total COD, 81.8% of the soluble COD and 32.9% of the total acid extractable organics. Moreover, effective removal of eight heavy metals, includes 97.8% of 78Se, 96.8% of Ba, 94.7% of 88Sr, 81.3% for 66Zn, 77.1% of 95Mo, 66.9% of 63Cu, 44.9% of 53Cr and 32.5% of Pb, was achieved.