The study is focused on the removal of fecal coliforms from low-strength domestic wastewater using a vertical up-flow microbial fuel cell (MFC) reactor. Raw domestic wastewater was collected from the grit chamber of the sewage treatment plant, which is located inside the campus of the Indian Institute of Technology Guwahati. The MFC reactor was cylindrical with an effective volume of 4.5 L. Stainless steel mesh with graphite rod was used as the anode and graphite rod was used as the cathode in the MFC. The mean COD and fecal coliform of the raw wastewater during the study period were observed to be 177.8 mg/L and log 6.94 per 100 mL respectively. The lab scale MFC was operated with a retention time of 36 h and found to be efficient in COD and coliform removal. Mean COD removal in MFC was 91.34% and mean effluent fecal coliform was log 3.49 units. The MFC was operated in continuous vertical up-flow mode and stable operating voltage was obtained continuously under varying external resistances. Biomass activity study showed higher substrate utilization by reactor microbial population in summer compared to that of winter. Seasonal temperature variation was found to be a significant influencing factor for the removal of total and fecal coliforms inside the MFC reactor. Throughout the study, the first-order inactivation rate for fecal coliform in the MFC reactor was calculated and the seasonal effect on the variation of the rate coefficient was also monitored.