Microbial fuel cells (MFCs) are bioelectrochemical transducers that can be used to produce electrical power under the activity of microbes during the wastewater treatment processes. In the present study, the electrode spacing was considered as a parameter to investigate the influence on the performance of MFCs. The electrode spacing was defined as the distance of the anode electrode plate to the polymer exchange membrane in the MFCs. Three values were set at 0.0, 3.0, and 6.0 cm, respectively. In addition, a flow device, like a honeycomb type flow straightener, was introduced and implemented in the anode chamber for creating a uniform flow. The inner diameter of the honeycomb was 0.7 cm. Results showed that a higher limiting current density with 4108.7 mA/m(2)and a lower resistance with 2.51 omega can be found in the case of the 0.0 cm electrode spacing. These results also indicated that the shorter electrode spacing with flow straightener devices would improve the performance of MFCs, leading to lower internal resistance and higher power density. In addition, the scanning electron microscopy was employed to analyze the biofilm thickness for MFCs with different electrode spacing. It was also found that the biofilm thickness with 0 cm electrode spacing was larger than the other two cases, leading to a lower internal resistance in the MFCs.