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The adverse effects of synthetic acaricides on humans, animals, non-target organisms and the ecosystem are serious problems. Thus, there is a new trend to use nanotechnology for developing new, natural, bio and safe acaricides for mite control in green-pest management. This is the first work for preparing a nanoformulation of rosemary essential oil (EO) and evaluating its effect against the two-spotted spider mite Tetranychus urticae Koch. GC/MS analysis of rosemary EO showed that 1,8 cineole (31.45%), borneol (11.07%), α-pinene (10.91%), D-limonene (9.19%), L-linalool (8.86%), D-camphor (7.32%), γ-terpinene (3.92%), linalyl acetate (3.37%), α-terpineol (3.32%), and p-cymene (1.82%) were the major components. After 6 min of sonication, a nanoemulsion of rosemary EO was formulated with a droplet size of 139.9 nm. The balance between oil (lyophilic) and surfactant (hydrophilic) was correlated with the droplet size and the stability of the nanoemulsion. Spray application of rosemary nanoemulsion showed high acaricidal activity against immature and adult two-spotted spider mites T. urticae with LC50 723.71 and 865.68 μg · ml−1 and the toxicity increased by 54.15 and 52.69% for immature and adult mites, respectively. There were no toxic effects or mortality of rats treated with rosemary nanoemulsion. High acaricidal activity, stability, and safety of rosemary nanoemulsion make this nanoformulation a possible green and nano-acaricidal product. Further studies under field conditions are necessary to study the acaricidal efficiency of rosemary nanoemulsion against two-spotted spider mites and the toxic effect on predacious mites.
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