Aspergillus flavus F5 was used to reduce AgNO3 to form silver nanoparticles (Ag-NPs) that were monitored by a color change from colorless to yellowish-brown. The characterizations were achieved by UV-Vis spectroscopy, FT-IR, TEM, SEM-EDX, and XRD. Data showed that there was a successful formation of crystalline, spherical shape Ag-NPs with a particle average size of 12.5 ± 5.1 nm. The FT-IR clarified the role of various functional groups in the reducing/capping process. EDX-SEM revealed that the main component of the as-formed sample was set to be mainly Ag with a weight percentage of 46.1%. The synthesized Ag-NPs exhibit antibacterial and anti-Candida activity against Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, Escherichia coli, Candida albicans, C. glabrata, C. tropicalis, and C. parapsilosis, with inhibition zones ranging between 9.3 ± 0.5 to 20.8 ± 0.3 nm based on concentrations used and MIC values between 6.25 to 25 ppm. The mortality percentages of Tyrophagus putrescentiae mite species due to the mixing of their diet with different Ag-NPs concentrations of 0.5, 1.0, and 1.5 mg were 55.7 ± 2.1, 73.3 ± 1.5, and 87.4 ± 1.6% respectively after 20 days post-treatment. The catalytic activity of Ag-NPs to degrade methylene blue (MB) was investigated in the presence and absence of light irradiation. Data showed that a high photocatalytic degradation of MB compared with dark conditions at various times and concentrations. At a concentration of 70 mg/30 mL after 200 min., the dye removal percentages were 86.4 ± 0.4% in the presence of light irradiation versus 66.5 ± 1.1% in dark conditions.