Objective: To assess the in vitro antimicrobial activity of eight types of local honeys from Green Mountain province, Libya on different microorganisms. Methods: 100% of different types (Undiluted) of honey were studied in vitro using, Escherichia coli, Streptococcus pyogenes Pseudomonas aeruginosa and Candida albicans, to determine the zone of inhibition against each type of microorganism. The antimicrobial activity of honey samples was compared with standard antibiotics like Ciprofloxacin, Amoxil/clavulunic acid, Tetracycline, Gentamycin, Ampicillin/cloxacillin, Erythromycin and Nitrofurantoin was determined by the disc diffusion method. Results: The diameter of zone inhibition of honey has various values tested ranged from 11-18.5 mm for Pseudomonas aeruginosa, 0-12 mm for both Streptococcus pyogenes and Escherichia coli, while with no effect for Candida albicans. Conclusions: The eight types of Libyan honey have a various antibacterial activity in-vitro.