ABSTRACT: This study addresses the impacts of metabolites from different microbial taxa on the fabrication and multifunctional biological properties of spherical silver nanoparticles (Ag-NPs). Three microbial taxa, a bacterial (Bacillus cereus A1-5), actinomycetes (Streptomyces noursei H1-1), and fungal (Rhizopus stolonifer A6-2) strains were used for Ag-NPs biosynthesis, whereas Streptomyces noursei is demonstrated for the first time. These isolates were identified using either 16S rRNA or ITS gene sequencing. Characterization of Ag-NPs was done using color change analysis, Uv-Vis spectroscopy, FT-IR spectroscopy, XRD, TEM, SEM-EDX, DLS, and Zeta potential analysis. All biosynthesized NPs exhibited spherical shape with different sizes ranged from 6?50 nm, 6-30 nm and 6-40 nm for NPs obtained by A1-5, H1-1 and A6-2, respectively. The crystalline center cubic face of Ag-NPs was confirmed using XRD at 2? values 38.08o, 44.27o, 64.41o and 77.36o. FT-IR analysis revealed varied intense absorption peaks for biomolecules required for NPs synthesize by each microbial strain. The stability of spherical Ag-NPs was confirmed due to highly DLS negative surface charge of ?17.5mV, ?18.9mV, and ?15.6mV for NPs synthesized by strains A1-5, H1-1, and A6-2, respectively. Ag-NPs exhibited a broadspectrum of antibacterial activity against Gram-positive and Gram-negative bacteria with varied effectiveness. They also exhibited a cytotoxic effect against cancer cell line (caco-2) in a dose-dependent pattern with IC50 of 8.9 ± 0.5, 5.6 ± 3.0, 11.2 ± 0.5 ?g/ml for NPs synthesized by strains A1-5, H1-1, and A6-2, respectively. Moreover, these spherical Ag-NPs showed larvicidal activity against the 3rd instar larvae of the dengue vector Aedes aegypti.