Honey produced by honeybee (Apis mellifera) which is used in herbal medicine was examined for its chemical constituents and antimicrobial activity. The phytochemical analysis of honey showed the presence of alkaloids, flavonoids, saponins, steroids, reducing sugar and glycosides. Antimicrobial activity of honey on fresh hospital isolates: Staphylococcus aureus, Escherichia coli, and Candida albicans obtained from Glanson Medical Laboratory Awka were determined using well diffusion method. The result shows that the honey produced by mellifera has strong antimicrobial activity against E. coli and S. aureus but not against C. albicans. The result obtained shows that the honey produced a zone of clearance of 45 and 34 mm on S. aureus and E. coli, respectively. The result of minimum inhibitory concentration (MIC) determined on liquid culture was 20%v/v for both S. aureus and E. coli while the minimum bactericidal concentration (MBC) determination of the sample showed 20% and 30%v/v for S. aureus and E. coli, respectively. Our result shows that honey, apart from their role as food additives and supplements, may also be utilized as effective and cheap sources of antibacterial agents for the treatments of bacterial infections.