Antibacterial activity of honey obtained from two different locations in Enugu State (Nsukka & Ugwuaji) Nigeria on Staphylococcus aureus, Escherichia coli and Streptococcus pyogens isolated from wound was studied. Agar well diffusion method was used to determine the antibacterial activity of the honey on the test microorganisms. The result revealed that the two honey samples have heavy antibacterial activities against the test organisms and zones of inhibition were obtained showing high antibacterial activity. The antibacterial activity increased with increase in the concentrations and honey from Nsukka produced a high antibacterial activity (clearer zone) on staphylococcus aureus and Escherichia coli at all concentration and moderately for streptococcus pyogens. The use of honey as a therapeutic substance has been rediscovered by the medical profession on more recent times, and it is gaining acceptance as an antibacterial agent for the treatment of ulcers and bed sores, and other infections resulting from burns and wounds.