Due to their potential application in various fields of science and technology, the eco-friendly bio-synthesis of silver (Ag) nanoparticles (NPs) is a growing area for researchers. In this study, we report the green synthesis of Ag nanoparticles and their characterization by using various techniques. For the preparation of Ag particles, aqueous plant extract of ailanthus altissima was used as a reducing medium for Ag+ ions of silver nitrate to Ag0. UV-Vis spectrophotometry was used to trace the formation of Ag particles by noting their surface plasmon resonance peaks (400 nm to 440 nm). Fourier transform infrared spectroscopy (FT-IR) was employed to reveal the chemical composition of Ag nanoparticles which were capped by plant extract. Scanning electron microscopy (SEM) was used to get the lattice image, morphology and average size of Ag particles. The average size distribution of Ag NPs dispersed in aqueous media was also measured using dynamic light scattering (DLS). It was found that DLS results are in good agreement with those obtained from SEM. The synthesized particles were then subjected to the antibacterial and antifungal activities by studying them against various species, such as bacillus cereus, staphylococcus aureus, pseudomonas aeruginosa, E. coli and A. parasiticus, A. niger and A. flavus fungi. It was noted from the growth curves of both bacteria and fungi that in the presence of silver nanoparticles they show more in-zone growth as compared to the plant extract.