The pollution is a result of drastically rising fossil fuel usage due to increased global population and industrialization. Today’s world needs to replace fossil fuels with new and renewable energy sources. Microbial Fuel Cells (MFCs) are devices that convert the chemical energy to direct electricity by microbial metabolic activity. Microbial growth was actualized at the anodic chamber of MFC and must be controlled carefully by microbial methods. The main aim of this study is optimizing the growth parameters of microorganisms in two-chambered MFC with optical density, dried and wet weight of microorganisms that were grown at the anode chamber of MFC. Moreover, the Chemical Oxygen Demand (COD) values of medium were determined at fed cycles of MFC. The total volume of MFC was 0.7 L and the connection was made with the salt bridge between anode and cathode compartments. The anode chamber was used for bacterial growth that was taken from Akkaya Dam slime, was fed with molasses medium at 5 days intervals. The OD values were determined with a spectrophotometer at 600 nm, COD values were determined with the standard method and wet-dry weight also determined as a function of the incubation period.