Mosquitoes act as a vector for most of the life-threatening diseases like malaria, yellow fever, dengue fever, chikungunya fever, filariasis, encephalitis, West Nile Virus infection, etc. Under the Integrated Mosquito Management, emphasis was given on the application of alternative strategies in mosquito control. The continuous application of synthetic insecticides causes development of resistance in vector species, biological magnification of toxic substances through the food chain, and adverse effects on environmental quality and nontarget organisms including human health. Application of active toxic agents from plant extracts as an alternative mosquito control strategy was available from ancient times. These are nontoxic, easily available at affordable prices, biodegradable, and show broad-spectrum target-specific activities against different species of vector mosquitoes. In the present study, the larvicidal activity of silver nanoparticles (AgNPs) synthesized using Sida acuta plant leaf extract against late third instar larvae of Culex quinquefasciatus, Anopheles stephensi, and Aedes aegypti was determined. Range of concentrations of synthesized AgNPs (10, 20, 30, 40, and 50 μg/mL) and aqueous leaf extract (50, 100, 150, 200, and 250 μg/mL) were tested against the larvae of C. quinquefasciatus, A. stephensi and A. aegypti. The synthesized AgNPs from S. acuta leaf were highly toxic than crude leaf aqueous extract in three important vector mosquito species. The results were recorded from UV-Vis spectrum, Fourier transform infrared spectroscopy, scanning electron microscopy, transmission electron microscopy, and energy-dispersive X-ray spectroscopy analysis. Larvae were exposed to varying concentrations of aqueous crude extract and synthesized AgNPs for 24 h. Considerable mortality was evident after the treatment of S. acuta for all three important vector mosquitoes. The LC50 and LC90 values of S. acuta aqueous leaf extract appeared to be most effective against A. stephensi (LC50, 109.94 μg/mL and LC90, 202.42 μg/mL) followed by A. aegypti LC50 (119.32 μg/mL and LC90, 213.84 μg/mL) and C. quinquefasciatus (LC50, 130.30 μg/mL and LC90, 228.20 μg/mL). Synthesized AgNPs against the vector mosquitoes of A. stephensi, A. aegypti, and C. quinquefasciatus had the following LC50 and LC90 values: A. stephensi had LC50 and LC90 values of 21.92, and 41.07 μg/mL; A. aegypti had LC50 and LC90 values of 23.96, and 44.05 μg/mL; C. quinquefasciatus had LC50 and LC90 values of 26.13 and 47.52 μg/mL. These results suggest that the use of S. acuta synthesized silver nanoparticles can be a rapid, environmentally safer biopesticide which can form a novel approach to develop effective biocides for controlling the target vector mosquitoes. This is the first report on the mosquito larvicidal activity of the plant aqueous extract and synthesized nanoparticles.