The economic losses caused by gastrointestinal nematodes of small ruminants are increasing today due to the development of anthelmintic resistance. Therefore, researchers from different parts of the world are searching for new, alternative strategies to control these parasites. As a valuable natural resource, medicinal plants and their products have emerged as a viable option. The aim of this study was to evaluate in vitro ovicidal activity of two chemotypes of the yarrow (Achillea millefolium L.) essential oil against sheep gastrointestinal nematodes. For this purpose, nematode eggs were collected from naturally infected sheep of two farms located in Southern Italy to perform the egg hatch test. In both farms, the coproculture examination identified the presence of four genera of sheep GINs: Haemonchus, Trichostrongylus, Teladorsagia and Chabertia. Main components of the yarrow essential oil identified by GC-MS analysis were 1,8-cineole (41.69%), camphor (8.37%) and trans-chrysanthenyl acetate (4.90) for the type 1 and β-pinene (28.53%), β-caryophyllene (18.71%) and 1,8-cineole (11.69%) for the type 2. In vitro ovicidal activity was evaluated at six different concentrations (50, 12.5, 3.125, 0.781, 0.195 and 0.049 mg/mL), whereby the inhibitory effect of the essential oil (Achillea millefolium L.) on egg hatchability varied from 46.5-99.5% (type 1) and 69.6-97.25% (type 2) and all concentrations tested showed a significantly higher efficacy compared to the negative control (p<0.0001). Furthermore, the inhibitory effect on egg hatching was similar (p>0.05) to the positive control (98.0%) at concentrations of 50 mg/mL (99.5%), 12.5 mg/mL (98.0%) and 3.125 (95.25%) of the type 1, and at concentrations of 50 mg/mL (97.25%) and 12.5 mg/mL (90.0%) of the type 2. The obtained results suggested that the A. millefolium essential oil has high anthelmintic potential, especially azulene-free chemotype rich in 1,8-cineole and camphor, which requires confirmation in further in vivo studies.
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