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Tropospheric ozone (O 3) is a naturally occurring gas in the atmosphere. However, the concentration of O 3 increased in the 20 th century. Although the effects of O 3 on vegetation have been extensively studied since the 1950s, limited information exists regarding the effects of O 3 on insect herbivores. In particular, evidence is lacking regarding the effects of O 3 on the biology of insect herbivores. Agelastica coerulea (Baly, 1874) is a coleopteran species that grazes on Betulaceae plants. In this study, to investigate the effects of O 3 on A. coerulea biology for the first time, female adult insects were collected from Japanese white birch trees grown in a Free Air Controlled Exposure System (FACE) in Sapporo, Japan. These beetles inhabited trees exposed either to ambient or to elevated O 3 for 23 days. After 2 collection, the enzyme activities in the beetles were measured. Elevated O 3 led to a greater total antioxidant activity and lower α-and β-esterase activities, a phenomenon that may suggest an increased resistance of the beetles to stress. Our results are further discussed with regard to biological and toxicological aspects. Collectively, our findings indicate that total antioxidants and α-and β-esterase activities can serve as effective O 3 biomarker systems in this beetle species. This adaptive response of the beetle, which was induced by moderate O 3 exposure, should be further tested across generations and for its protection against greater exposure.

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Solid lipid nanoparticles second generation, nanostructure lipid carrier (NLC), is one of the most important biodegradable nanoparticles. Nanostructure Lipid carrier (NLC) was used to encapsulate methylene blue (MB) dye, carvacrol and citronellal and their efficacy as insecticidal against Culex pipiens (Cx. pipiens) were distinguished. The prepared nanoformulation revealed very good physicochemical properties, especially the homogeneity of the particle size. Transmission electron microscope showed spherical shaped nanoparticles within range less than 200 nm. The prepared NLC-MB-MT system showed a very competitive insecticidal activity and high virulence against the mosquito larvae with higher mortality rate of LC50 of 0.141 µl/mL, in addition to high level of Oxidative stress parameters obtained through all the tested enzymes including hydrogen peroxide (4.8 ppm), protein carbonyl amount (0.12 OD/mg protein), ascorbic acid (0.15 mg) and Superoxide dismutase (SOD) showed strong increasing (0.09 OD/mg protein/min) at 6 µg/mL, respectively. Whereas paradoxical results of the oxidative stress enzymes were obtained from different concentration of nanoformulation that introduce a convenient reason for their potential insecticidal effect. The cytotoxic effect of NLC-MB-MT was evaluated using WI38 human lung cell lines, the LC50 was 6.4 mg/mL. The low cytotoxic reactivity towards the tested cell line makes the NLC-MB-MT nanoformulation has its promising insecticidal efficacy. Molecular docking study for each component were done against acetylcholine esterase protein and accepted binding modes achieved by the three compounds.

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