The present research was undertaken to investigate the antimicrobial activity of ethanol extract of Pouzolzia zeylanica (L.) Benn. (Family: Urticaceae) as well as to determine the phytochemical,cytotoxicity,antifungal activity of the extracts. In phytochemical screening, it was found that the ethanol extract contains alkaloids, glycosides, tannins and flavonoids. From the present study, it was found that the ethanol extract posseses significant cytotoxic activity with the LC50 value of 6.1 μg/ml and the LC90 value of 12.2μg/ml. The antibacterial activity of the ethanol extract were evaluated by disc diffusion method against 3 gram positive and 4 gram negative bacteria using amoxicillin as standard. The ethanol extract shows varying degrees of antibacterial activities with zone of inhibition ranging from 11.5-35.75 mm .The antifungal activity of the ethanol extract were evaluated by disc diffusion method against 6 fungi using griseofulvin as standard.The ethanol extract showed strong antifungal activity with zone of inhibition of 7-15 mm for 250μg/disc and 15-26 mm for 500 μg/disc respectively.