Calotropis gigantea leaf extract and Bacillus thuringiensis were tested first to fourth-instar larvae and pupae of Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus. The medicinal plants were collected from the area around Bharathiar University, Coimbatore, India. Calotropis gigantea leaf was washed with tap water and shade-dried at room temperature. An electrical blender powdered the dried plant materials (leaves). The powder 500 g of the leaf was extracted with 1.5 L of organic solvents of methanol for 8 h using a Soxhlet apparatus and filtered. The crude leaf extracts were evaporated to dryness in a rotary vacuum evaporator. The plant extract showed larvicidal and pupicidal effects after 24 h of exposure; no mortality was observed in the control group. For Calotropis gigantea, the median lethal concentration values (LC(50)) observed for the larvicidal and pupicidal activities against mosquito vector species Anopheles stephensi I to IV larval instars and pupae were 73.77, 89.64, 121.69, 155.49, and 213.79 ppm; Aedes aegypti values were 92.27, 106.60, 136.48, 164.01, and 202.56 ppm; and Culex quinquefasciatus values were 104.66, 127.71, 173.75, 251.65, and 314.70 ppm, respectively. For B. thuringiensis, the LC(50) values of I to IV larval instars and pupae of Anopheles stephensi were 37.24, 45.41, 57.82, 80.09, and 98.34 ppm; Aedes aegypti values were 42.38, 51.90, 71.02, 96.17, and 121.59 ppm; and Culex quinquefasciatus values were 55.85, 68.07, 94.11, 113.35, and 133.87 ppm, respectively. The study proved that the methanol leaf extract of Calotropis gigantea and bacterial insecticide B. thuringiensis has mosquitocidal property and was evaluated as target species of mosquito vectors. This is an ideal ecofriendly approach for the control of vector control programs.