Larger scale microbial fuel cells (MFCs) require compact architectures to efficiently treat wastewater. We examined how anode-brush diameter, number of anodes, and electrode spacing affected the performance of the MFCs operated in fed-batch and continuous flow mode. All anodes were initially tested with the brush core set at the same distance from the cathode. In fed-batch mode, the configuration with three larger brushes (25 mm diameter) produced 80% more power (1240 mW m-2) than reactors with eight smaller brushes (8 mm) (690 mW m-2). The higher power production by the larger brushes was due to more negative and stable anode potentials than the smaller brushes. The same general result was obtained in continuous flow operation, although power densities were reduced. However, by moving the center of the smaller brushes closer to the cathode (from 16.5 to 8 mm), power substantially increased from 690 to 1030 mW m-2 in fed batch mode. In continuous flow mode, power increased from 280 to 1020 mW m-2, resulting in more power production from the smaller brushes than the larger brushes (540 mW m-2). These results show that multi-electrode MFCs can be optimized by selecting smaller anodes, placed as close as possible to the cathode. © 2013 Elsevier B.V. All rights reserved.