In Vitro Control of Anisakis Larvae Using Nanoemulsified Thyme, Garlic, and Grape Seed Essential Oils Combined with Brines

Introduction: Anisakiasis is a zoonotic parasitic disease transmitted through consumption of fish, with increasing global cases. While thorough cooking kills Anisakis larvae, thermo-stable allergens can still cause allergic reactions. Traditional Mediterranean seafood methods such as marinating, salting, and brining, which lack heat treatment, pose a high risk of anisakiasis. Materials and methods: This study investigated the antilarval effects of nanoemulsions made from thyme, garlic, and grape seed essential oils combined with brines on Anisakis L3 larvae in vitro. Results: Larvae were collected from mackerel, chub mackerel, anchovy, horse mackerel, and sardine, with 1746 larvae examined, mostly Anisakis type I and molecularly identified as A. pegreffii. Thyme oil’s main components, carvacrol and thymol, showed strong antimicrobial and antioxidant effects and superior larvicidal activity at lower doses compared to garlic and grape seed oils. Garlic oil, rich in organosulfur compounds, was more effective in Lakerda samples. High salt brine alone inactivated larvae within 24 h. The thyme oil nanoemulsion had droplet sizes below 90 nm, enhancing stability, whereas garlic and grape seed oils had larger droplets. Conclusion: These results indicate that nanoemulsified essential oils, especially thyme oil, are promising natural agents to control Anisakis larvae in traditional sea-food products.