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Abstract

The aim of this study was to investigate interactions between conventional antifungal drug and essential oils against isolates of Malassezia pachydermatis. Antifungal activity of Cinnamomum cassia, Melaleuca alternifolia, Mentha piperita, Origanum vulgare and Syzygium aromaticum essential oils were tested against 19 strains of M. pachydermatis isolated from healthy dogs and reference strain M. pachydermatis CBS 1879. The checkerboard assay was used to search for in- teractions. Synergism was observed for the combination of clotrimazole with Melaleuca alternifolia essential oil, Mentha piperita and Origanum vulgare. The combinations of Cinnamomum cassia and Syzygium aromaticum essential oils with clotrimazole showed indifferent effect. Additive antimicrobial activity was observed for the combination of clotrimazole with Syzygium aromaticum and Melaleuca alternifolia essential oils against reference strain. The obtained results showed synergistic interactions between essential oils and clotrimazole which could improve effectiveness of this antifungal drug.
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Abstract

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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