Microbial Fuel Cells (MFCs) use bacteria as biocatalyst to convert biodegradable substrates into electricity. The natural cow dung was found more suitable as it generated 150.9 mV Open Circuit Voltage (OCV) against 3.2 mV OCV generated by the sterile cow dung in the H-shaped MFC. On configuration, a MFC with 50 % cow dung and a salt bridge (5 cm × 2 cm) containing a mixture of 10 % sodium chloride and 5 % agar, electrodes of 32.20 cm2 surface area and phenol red (0.1 %) as exogenous mediator was found ideal. Furthermore, five electrogenic bacteria were isolated from the cow dung and individually studied for their electrogenic properties. The bacterial isolate CD64 was found best among the isolates, as it generated 710.7 mV OCV at 37 ºC, pH 7.0 with LB medium as anolyte against vinegar as catholyte. Based on the phenotypic characteristics and 16S rDNA sequencing, isolate CD64 was identified as Bacillus sp.