Development in microbial fuel cell is increasing day by day due to utilizing waste material into useful energy and fulfilling future requirement of energy. Due to nature of microbial fuel cell for converting organic material into bioelectricity using microorganism get part economy of the world, due to waste material disposing and handling problem. The main purpose of this work is to investigate the cow dung manure as substrate for degrading and generation of electricity, different substrate percentage, oxygen, and pH on electricity generation. The maximum voltage were measured using 50%w/v of cow dung manure 801±03mv/l and minimum at 25% about 488±02mv/l, and for as other parameter were concerned 455mv/l at pH 6, and 290mv/l at 35ml/min for air flow rate to cathode chamber for oxidation of proton. It was concluded from the whole work that microbial fuel cell has ability to utilize microbes from cow dung for power generation and presents cleaner technology.