Microbial activity and biomass are important factors that determine nutrient and carbon fluxes in freshwater ecosystems and, therefore are also related to both water quality and climate change induced stressors. This study aimed at assessing the feasibility of a power assisted Microbial Fuel Cell (MFC)-based biosensors for the continuous monitoring of microbial activity and biomass concentrations in saturated freshwater ecosystems. For this purpose, four lab-scale reactors were constructed and operated for 30 weeks. Reactors were fed with four different organic matter concentrations to promote a suite of microbial activity and biomass conditions. The reactors consisted of 3.8 L PVC vessels filled with 23 extractable gravel- sockets, used for microbial activity and biomass assessment, and 1 MFC granular-graphite socket, for biosensing assessment. Microbial activity was determined by the ATP content and the hydrolytic enzymatic activity, and the biomass content was assessed as the volatile solids attached to the gravel. Very significant linear relationships could be established between the parameters studied and the current density produced by the MFC with a very short detection time: 10 min for the ATP content (R2 = 0.88) and 1 h for the enzymatic activity (R2 = 0.78) and biomass (R2 = 0.74). Moreover, the power assisted MFC-based biosensing tool demonstrated to be functional after a long operation time and under a wide range of organic loading conditions. Overall, the results highlight the feasibility to develop a power assisted MFC-based biosensor for on-line monitoring of the microbial activity and biomass of a given ecosystem (either natural or artificial) even in remote locations.