In this study, an endophytic actinomycetes strain of Streptomyces viridodiastaticus strain MGR23 was isolated from the roots of the medicinal plant Centella asiatica. The biosynthesized Ag-NPs were characterized using UV-Vis spectroscopy, Fourier transform infrared (FT-IR), Scanning electron microscopy (SEM), X-ray diffraction (XRD) and Zeta potential, which confirmed the successful formation of crystalline, spherical silver nanoparticles. Biosynthesized AgNPs significantly inhibited and reduced the growth of medically important survival microorganisms (Streptomyces viridodiastaticus MGR 23 against Streptomyces toxytricini D2, Acidothermus cellulolyticus CA16, Streptomyces sp. MGR 21, Aciditerrimonas ferrireducens SM7 and Streptomyces tritici D5). The biosynthesized Ag-NPs exhibited broad-spectrum antihelminthic activity. Our data elucidated that the biosynthesized Ag-NPs had a highly cytotoxic effect against the cancerous L6 cell line under in vitro conditions, revealing a maximum IC50 value of 1.0 μg/mL and no known anticancer drug was used as a positive regulation. AgNPs have a remarkable synergistic impact that points to their considerable potential in nanomedicine for future clinical use as a combination therapy.