larvicidal activity of Agave sisalana against L4 larvae of Aedes aegypti is mediated by internal necrosis and inhibition of nitric oxide production

Dengue is a viral disease that affects about 50 million people per year around the world. The aim of this study was to investigate the larvicidal activity of Agave sisalana crude extract in order to develop a new insecticide against Aedes aegypti. In larvicidal activity assays, fourth-stage Ae. aegypti larvae were exposed to different concentrations of A. sisalana crude extract for 3, 6, 12, and 24 h for determining the LC₅₀. Next, we explored its cytotoxic activity by flow cytometry. Furthermore, histological alterations were confirmed by histopathological analysis, and the nitric oxide (NO) production by hemocytes was checked after different periods of exposure to A. sisalana crude extract. The LC₅₀was 4.5 ± 0.07 mg/mL. In addition, flow cytometry revealed an increase of cellular necrosis (21 and 16.5 % after 12 and 24 h, respectively) in larvae that were exposed to A. sisalana crude extract. The histological analysis revealed cell lysis and destruction of the peritrophic membrane. Furthermore, there was a reduction in the concentration of NO in the hemolymph from larvae exposed to A. sisalana crude extract after 3, 6, and 24 h (5.3 ± 4.3 vs. 22.7 ± 5.2 μM, 4.3 ± 5.5 vs. 25.4 ± 6.6 μM, and 6 ± 1.7 vs. 37.1 ± 7.8 μM, respectively). Our findings show that A. sisalana crude extract constitutes an effective larvicidal agent against Ae. aegypti larvae due to its necrotizing activity in hemocytes and inhibition of the NO production.