Background and Aim: Several studies have been done that showed honey has been therapeutic effects on infection disease like Pseudomonas infections. Our aim of this study is evaluation of the antibacterial activity of mono floral and multi floral honeys from different origin of Iran on growth of Pseudomonas aeruginosa and compare their activities with artificial honey and antibiotics. Materials and Methods: Antimicrobial effect of honey was determined by disc diffusion and broth dilution method on 5 different concentration of honey. Results: The highest inhibition zone (16 ± 1/52 mm) was recorded from persimmon honey in disc diffusion method. In this study، the minimum inhibitory concentration (MIC) for manna honey and other natural honeys obtained 25% and 12.5% respectively. While P. aeruginosa was inhibited at concentration of 50% (ml/ml) of artificial honey. Conclusions: This study showed that honey has a significant antibacterial effect on P. aeruginosa. There is a direct link between the concentration of honey and inhibition zone.