Two flow patterns (plug flow (PF) and complete mixing (CM)) of microbial fuel cells (MFCs) with multiple anodes cathodes were compared in continuous flow mode for wastewater treatment and power generation. The results indicated that PF-MFCs had higher power generation and columbic efficiency (CE) than CM-MFCs, and the power generation varied along with the flow pathway in the PF-MFCs. The gradient of substrate concentrations along the PF-MFCs was the driving force for the power generation. In contrast, the CM-MFCs had higher wastewater removal efficiency than PF-MFCs, but had lower power conversion efficiency and power generation. This work demonstrated that MFC configuration is a key factor for enhancing power generation and wastewater treatment. Copyright (C) 2013, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved.