Background: Silver nanoparticles play a profound role in the field of biology and medicine due to its attractive physiochemical properties. Objective: The present work was aimed to develop green approach for the synthesis of silver nanoparticles (DRAgNPs) using D. zibethinus rind aqueous extract and determination of its antimicrobial and cytotoxic effect against brine shrimp. Materials and Methods: Aqueous extract of D. zibethinus rind was used to reduce silver nitrate to silver nanoparticles. The various reaction parameters were optimized, and DRAgNPs were characterized for size, shape, and nature. Results: Surface plasmon resonance confirmed the formation of DRAgNP's with maximum absorbance at λmaxof 418 nm. Scanning transmission electron microscopy and transmission electron microscope images revealed the morphology of the DRAgNPs were spherical with size range of 20 and 60 nm. Atomic force microscopy images confirmed the average particles size of DRAgNPs was to be 55 nm. The stability of the nanoparticles was also confirmed by the zeta potential which was found to be −15.82 mV. X-ray powder diffraction and energy-dispersive X-ray spectroscopy analysis confirmed the nature and the presence of Ag. DRAgNPs showed considerable antimicrobial activity against Salmonella typhimurium, Escherichia coli, Salmonella typhi, Staphylococcus aureus, Staphylococcus haemolyticus, and Bacillus subtilis, and exhibited better cytotoxicity against brine shrimp (LC50 =2.55 mg/mL). Conclusion: Based on the present study, it can be concluded that the green synthesis of silver nanoparticles using D. zibethinus rind is an eco-friendly and inexpensive method, and DRAgNPs could be used in the field of water treatment, pharmaceuticals, biomedicine, biosensor, and nanotechnology in the near future.