The suitability of carbon nanofibers (CNF) based cathodes as alternative to the platinum (Pt)-based cathode in single chamber microbial fuel cells (SCMFCs) were extensively studied over 3-month operational period. MFCs were fed with two solutions: synthetic wastewater (phosphate buffer (PBS) plus sodium acetate) and real wastewater (mixed liquor suspendedsolid (MISS) solution). CNFs were chemically activated using HNO3 and then hot pressed on a carbon cloth support to increase surface area. The cathode polarization showed a better behavior of the clean Pt-based cathode in abiotic conditions. The activation of the nanofibers (ACNFs) gave an advantage to the cathode performances compared to the raw CNFs. The SCMFCs fed with PBS showed four times higher power generation compared to MLSS solution. All the cathodes showed a decrease in performances over time, and the advantage of the Pt over CNF/ACNF disappeared. CNF/ACNF cathodes showed more stability in performances in long time operations. Biofilm formation, salt precipitations on the cathode, and the presence of hydrogen sulfide decreased the activity of Pt cathodes. A degradation and Pt detachment were noticed on Pt cathodes over time. In contrast, CNF/ACNF cathodes exhibited less deterioration throughout the operational period, which demonstrated a great potential as cost-effective cathodes for long-term operation. (C) 2013 Elsevier B.V. All rights reserved.