Honey is an ancient remedy for the treatment of infected wounds, which has recently been rediscovered by the medical profession, particularly where conventional modern therapeutic agents are failing . Honey samples were collected from two different geographical regions , i.e Hill (kodaikkanal) and plain (Mannargudi) . The antimicrobial activity of honey was performed by well diffusion assay against pathogens i.e, bacteria [Bacillus subtilis, Bacillus cereus, Pseudomonas aeruginosa Enterobacter aerogenes, klebsiella], Fungi [Aspergillus niger, Aspergillus flavus, Trichoderma viride], yeast [Saccharomyces cerevisiae, candida], Actinomycetes [Streptomyces] and antibiotic sensitivity test was also performed for these pathogens using some antibiotics [Gentamycin, Auxamine, Tetracycline, Ampicillin].