Bioactivity and Safety Evaluations of Cupressus Sempervirens Essential Oil, Its Nanoemulsion and Main Terpenes Against Culex Quinquefasciatus Say

The essential oil (EO) of Cupressus sempervirens was obtained by hydrodistillation and analysed using gas chromatography-flame ionization detection (GC-FID) and gas chromatography-mass spectrometry (GC-MS). Two monoterpenes, α-pinene (49.1%) and δ-3-carene (21.4%), and one sesquiterpene hydrocarbon, α-cedrol (5.1%), were isolated as the EO major terpenes. An oil-in-water nanoemulsion (particle size 71.2 nm) was produced from the EO through a low-energy method. The EO, its nanoemulsion and its main constituents showed mosquitocidal and biochemical effects against Culex quinquefasciatus Say, the common vector of lymphatic filariasis parasites. All treatments showed dose-dependent bioactivity, and adults were more susceptible to the EO products than the larvae. The nanoemulsion showed superior activity, followed by the crude EO and α-cedrol. At 40 μg/ml, the nanoemulsion caused 100% larval mortality, while the EO and α-cedrol required twice this concentration to achieve the same larval mortality. The LC values were 8.4, 16.1, 15.1, 30.7 and 53.4 μg/ml at 24 h after exposure for the nanoemulsion, crude oil, α-cedrol, δ-3-carene and α-pinene, respectively. For adults, 20.0 μl/l nanoemulsion caused 100% mortality, while twice this concentration of the EO was required to achieve the same effect. The LC's against adults ranged between 6.2 and 40.4 μl/l. EO products prominently repelled mosquitoes at concentrations between 0.75 and 6.0 μl/cm. The EO products caused remarkable inhibition of Cx. quinquefasciatus acetylcholinesterase activity but were safer towards the non-target aquatic species Gambusia affinis. These results recommend the use of C. sempervirens EO, its nanoemulsion and main terpenes as natural tools to control Cx. quinquefasciatus.