The microorganisms in anodic biofilms of a microbial fuel cell (MFC) oxidize substrates to generate electrons, protons, and metabolic products. This study started up two single-chamber MFCs at different temperatures (25 degrees C for MFC A and 15 degrees C for MFC B); after successful startup, the cell temperatures were swapped. The MFC A had peak voltage at 540 mV at 25 degrees C, which was decreased rapidly as fed substrate was consumed. At 15 degrees C, the MFC A yielded a nearly constant voltage of 500 mV over complete feed cycle. Conversely, the MFC B produced higher maximum power than MFC A, and can deliver nearly constant voltage over the entire feed cycle at either 15 or 25 degrees C. Electrochemical analysis revealed that the MFC B had lower internal resistance than MFC A, with the former having much lower anodic resistance than the latter. Microbial analysis showed that the MFC started up at low temperatures had anodic biofilm enriched with psychrophilic bacteria Simplicispira psychrophila LMG 5408(T)[AF078755] and Geobacter psychrophilus P35(T)[AY653549]. This study suggests the strategy to promote the development of anodic biofilms at low temperatures that are capable of yielding electricity at constant voltage. Copyright (c) 2012, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved.