Sediment microbial fuel cell (sMFC) represents a variation of the typical configuration of a MFC in which energy can be harvested via naturally occurring electropotential differences. Moreover, constructed wetlands show marked redox gradients along the depth which could be exploited for energy production via sMFC. In spite of the potential application of sMFC to constructed wetlands, there is almost no published work on the topic. The main objective of the present work was to define the best operational and design conditions of sub-surface flow constructed wetlands (SSF CWs) under which energy production with microbial fuel cells (MFCs) would be maximized. To this aim, a pilot plant based on SSF CW treating domestic sewage was operated during six months. Redox gradients along the depth of SSF CWs were determined as function of hydraulic regime (continuous vs discontinuous) and the presence of macrophytes in two sampling campaigns (after three and six months of plant operation). Redox potential (EH) within the wetlands was analysed at 5, 15 and 25 cm. Results obtained indicated that the maximum redox gradient was between the surface and the bottom of the bed for continuous planted wetlands (407.7 ± 73.8 mV) and, to a lesser extent, between the surface and the middle part of the wetland (356.5 ± 76.7 mV). Finally, the maximum redox gradients obtained for planted wetlands operated under continuous flow regime would lead to a power production of about 16 mW/m2.