There is an urgent need to develop alternate antimicrobial agents and huge amount of research is underway for the search for a novel antimicrobial compounds. With increase in microbes acquiring resistance to multiple antibiotics, an alternate stratergy has to be developed. The new development in the field of nanotechnology is expected to influence the field of therapeutics. The objective of the present investigation was to synthesize silver nanoparticles by fungal cultures under stress condition, evaluate, and compare its antimicrobial property on different bacterial pathogens. Production of nanoparticles using biological systems has gained more importance in the present scientific world. Four different fungal colonies were isolated from coalmine soil sample and cultured in Potato Dextrose Agar medium, Sabouraud's Dextrose Agar and Rose Bengal Agar. Among which Fusarium equiseti and Fusarium solani were chosen for synthesis of silver nanoparticles. Morphological identification and 18S rRNA gene sequencing was conducted. The brown colouration of the culture broth after 3-4 d indicates the production of silver nanoparticles. The silver nanoparticles produced were analyzed by UV-Vis Spectrophotometer, X-ray Diffraction (XRD), Atomic Force Microscopy (AFM) and screened for the antimicrobial property against pathogens. A comparative study of their antimicrobial property was performed. Copyright © 2012 American Scientific Publishers All rights reserved.