Cerium oxide nanoparticles (CeO 2 NPs) were synthesized using Aspergillus niger culture filtrate. The mycosynthesized CeO 2 NPs were characterized by UVVisible (UV-Vis), Fourier Transform Infrared (FT-IR), X-ray diffraction (XRD), Micro Raman, Thermogravimetric/Differential Thermal Analysis (TG/DTA), Photoluminescence, and Transmission Electron Microscopy (TEM) analyses. UV-Vis spectrum exhibited a corresponding absorption peak for CeO 2 NPs at 296 nm, and the functional groups present in the fungal filtrate responsible for the synthesis of NPs were analyzed by FT-IR. The further characterization of the mycosynthesized CeO 2 NPs revealed particles of the cubic structure and spherical shape, with the particle sizes ranging from 5 to 20 nm. The antibacterial activity of CeO 2 NPs was examined in respect of two Gram-positive (G?) bacteria (Streptococcus pneumoniae, Bacillus subtilis) and two Gram-negative (G-) bacteria (Proteus vulgaris, Escherichia coli) by disk diffusion method. The test results for CeO 2 NPs at a concentration of 10 mg/mL showed higher activities on the zone of inhibition of up to 10.67 ± 0.33 and 10.33 ± 0.33 mm against Streptococcus pneumonia and Bacillus subtilis, respectively, The CeO 2 NPs caused 100 % mortality on first instar of Aedes aegypti at 0.250 mg/L concentration after 24-h exposure. The mycosynthesis of CeO 2 NPs is a simple, cost-effective and eco-friendly approach and it will also potentially helpful to control pathogenic bacteria and dengue vector.