In the present study, the larvicidal activity of ageing aqueous suspensions of spinosad against larvae of Culex pipiens biotype molestus, as well as their effect on the oviposition preferences of adult gravid females were evaluated in laboratory bioassays. Spinosad was applied at its label dose and the aqueous stock suspensions were stored for various ageing intervals up to 38 days. Untreated distilled water and diflubenzuron served as negative and positive control, respectively. Stock suspensions were taken after 0, 2, 6, 8, 16, 30 and 38 days of storage for diflubenzuron and after 0, 2, 6, 8, 20 and 27 days for spinosad, and were used for the bioassays. Furthermore, the effect of spinosad on the oviposition response of Cx. p. biotype molestus gravid females was investigated in two-choice oviposition preference bioassays. Spinosad was evaluated at half of its label dose and at its label dose, whereas diflubenzuron and distilled water served as positive and negative control, respectively. Results showed that both insecticides were found highly effective for the control of Cx. p. biotype molestus larvae, for ageing intervals up to 27 and 38 days for spinosad and diflubenzuron, respectively. Spinosad acted immediately after the preparation of the insecticidal solution (LT50 = 1.5 h), whereas for aged samples, LT50 values increased with the increase of the ageing interval (LT50 = 5 days for the 27 days old sample). For diflubenzuron, ageing time increased its insecticidal activity, as for aged diflubenzuron-treated solutions, lower LT50 values were achieved. In the oviposition preference bioassays, significantly fewer egg rafts were laid in water treated with spinosad at its label dose compared to control. However, this was not the case for water treated with spinosad at half of its label dose. Oviposition Activity Index (OAI) values were always comprised between −0.3 and 0.3, showing no relevant oviposition deterrence or attraction. The results of the present study contribute to our understanding of the effect of ageing on insecticidal solutions widely used in urban areas to control Cx. p. biotype molestus. Although an important vector of high public health importance, Cx. p. biotype molestus has been scarcely studied as target of environmentally and toxicologically reduced risk insecticides, such as spinosad.
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