Silk fibroin hydrogels are promising materials for controlled drug delivery device due to their aqueous process ability, biocompatibility, and biodegradability. The research work is aimed to prepare silk fibroin (SF) hydrogels loaded with Ag nanoparticles and to evaluate its antifungal activities. The Silk fibroin hydrogels were formulated at 37°C using 2% (w/v) silk fibroin aqueous solution either by treating 50% (v/v) of ethanol, or 50% (v/v) of propanol, or 50% (v/v) of glycerol, respectively. Above these, the rate of gelation was sufficiently accelerated by addition of glycerol. The silk fibroin hydrogels and prepared silver nanoparticles (AgNPs) were characterized by using Scanning Electron Microscopy (SEM), Thermo Gravimetrical Analysis (TGA). The encapsulation efficiency and release profile of AgNPs were studied by UV-vis spectrometry. The particle size of AgNPs was measured by Malvern Zetasizer Nano and found 93±5 nm. The encapsulation efficiency and morphology of the hydrogels was affected by the formulation conditions. The in vitro release profile showed an initial burst release of AgNPs followed by controlled release for next 20 hours. The antifungal activity of AgNPs loaded SF hydrogels showed a positive response to Aspergillus Niger pathogen. Therefore, silk fibroin hydrogels might be an effective biopolymeric matrix for antifungal applications.