Mosquitoes are rapidly advancing as vectors of several severe diseases. The increasing resistance of mosquitoes and the environmental harm caused by insecticides pose significant challenges for eradicating mosquito vectors. In this study, 18 plant extracts were tested for larvicidal properties against Aedes aegypti, Culex quinquefasciatus, and Anopheles stephensi larvae. Phyllanthus niruri (Pn) showed enhanced larvicidal activity in both laboratory and field trials. The biosynthesis of silver nanoparticles (AgNPs) using Pn leaf methanol extract (Pn-LME) was confirmed by UV-visible spectroscopy, X-ray diffraction, transmission electron microscopy, scanning electron microscopy, and Fourier-transform infrared spectroscopy. Among various concentrations, 3 mM AgNPs exhibited significant LC90 values of 0.83, 1.46, and 9.11 ppm compared to 9.25, 93.48, and 14.60 ppm of Pn-LME against A. stephensi, C. quinquefasciatus, and A. aegypti, respectively. This indicates the high mortality of mosquito vectors at low AgNP concentrations. Additionally, Pn-AgNPs showed enhanced antibacterial activity and no cytotoxicity in normal fibroblast cells (L929). Field trials demonstrated a 98.70% decrease in mosquito larval density at A. stephensi breeding sites, a 96.55% reduction at C. quinquefasciatus sites, and a 97.85% reduction at Ae. aegypti sites. This study presents an eco-friendly and cost-effective AgNP bio-pesticide synthesized from Pn leaves for controlling and preventing the transmission of filarial, dengue, and malaria vectors.