Production potential of cow dung for the generation of electricity was investigated using microbial fuel cell (MFC). Cow dung was collected from FUTA farm and Ilesha Garage farm in Akure. Voltage and current was measured per day for 21 days and the electrode used for the set up are carbon-carbon and carbon-aluminium electrode. Proximate, physico-chemical and mineral composition were determined using standard methods. Isolation and identification of microorganisms present in the cow dung were determined before and after generation of voltage and current using microbiological techniques. The microorganisms isolated were Providencia alcalifaciens, P. rettgeri, P. stuartti, Escherichia coli, E. fergusonii, Morganella morganii, Staphylococcus haemolyticus, S. aureus, Micrococcus luteus, Fusarium solani, Saccharomyces cerevisiae and Mucor mucedo. The highest voltage 0.737±0 mV and electric current 1.265±0 mA were generated from FUTA cow dung. The pH ranged between 7.3 to 9.9 and the temperature ranged between 25 ˚C to 33 ˚C during generation of voltage and current. This study has shown that cow dung is a potential substrate for generation of electric current using fabricated double chamber MFC.