To prepare and characterize silver nanoparticles from Guava leaves (Psidium guajava), extracts (chloroform and ethanol). The fabricated silver nanoparticles were tested on resistant fungal strains, Aspergillus niger and Candida glabrata. Psidium guajava leaf extracts were obtained by cold maceration method, by using Chloroform and Ethanol. To one mL of extract 2mM silver nitrate solution was added. The mixture was kept overnight in a dark chamber at room temperature. The solution was centrifuged at 10,000 rpm for 15 minutes and filtered by wattman filter paper after centrifugation. Psidium guajava leaf extract based silver nanoparticles were prepared and evaluated for anti-fungal activity by microtitre well plate method on resistant strains Aspergillus niger and Candida glabrata. The silver nanoparticles showed significant activity at 120 µL, Nystatin was used as standard.