A biochemical oxygen demand (BOD) monitoring system, based on electrochemically-active bacteria in combination with a microbial fuel cell, has been developed for the purpose of on-site, on-line and real-time monitoring of practical wastewater. A microbial fuel cell that had been enriched with electrochemically-active bacteria was used as the basis of the measurement system. When synthetic wastewater was fed to the system, the current generation pattern and its Coulombic yield were found to be dependent on the BOD5 of the synthetic wastewater. A linear correlation between the Coulombic yields and the BOD5 of the synthetic wastewater were established. Real wastewater obtained from a sewage treatment plant also produced a highly linear correlation between the Coulombic yield and BOD5 in the system. To examine on-site, on-line and real-time monitoring capability, the BOD monitoring system was installed at a sewage treatment plant. Over 60 days, the measurement system was successfully operated with high accuracy and good stability with the measuring period for a sample being 45 min. This application showed that the application of the measurement system was a rapid and practical way for the determination of BOD5 in water industries.