The honey bee (Apis mellifera) faces a significant threat from Varroa destructor, causing the losses of millions of colonies worldwide. While synthetic acaricides are widely used to control Varroa infestations, excessive application has led to resistant strains and poses side effects on the host. Consequently, there is an urgent need for a new acaricide that is both effective and affordable, yet safe to use on bees. One potential source of these acaricides is essential oils (EOs) and their constituents. This study evaluated the acaricidal properties of four essential oils (Eucalyptus globulus, Rosemary officinalis, Trachyspermum ammi (Ethiopian and Indian varieties), their constituents and mixture of constituents against V. destructor through the complete exposure method. Our finding showed that a 1:1 mixture of thymol and carvacrol (4 h-LC₅₀ = 42 μg/mL), thymol (4 h-LC₅₀ = 71 μg/mL), and T. ammi oil (4 h-LC₅₀ = 81–98 μg/mL) were the most toxic test samples against V. destructor. Honey bee behavior and selectivity were also assessed with one additional EO Thymus schimperi, indicating that T. schimperi, T. ammi, and their components were selective and did not affect the learning and memory of bees. In conclusion, the thymol and carvacrol (1:1) mixture was shown to be a promising replacement for synthetic acaricides, being three times more toxic than a commercial acaricide, fluvalinate (4 h-LC₅₀ = 143 μg/mL).

Honey bees (Apis mellifera) are major pollinators, playing a critical role in global food production, biodiversity, and ecosystem stability. However, their populations are increasingly threatened by multiple interacting stressors, including pesticide exposure. Among these, agricultural insecticides and anti-Varroa acaricides such as dinotefuran and amitraz can persist in hive matrices, resulting in chronic and combined exposure. This study investigates the low lethal (LC₁₀ and LC₃₀) effects of these compounds, individually and in combination, on honey bee survival, immune function, oxidative stress responses, detoxification pathways, and hypopharyngeal gland morphology. Both pesticides negatively affected honey bee health at low lethal concentrations, with dinotefuran showing higher toxicity. Exposure led to the reduced survival, suppression of vitellogenin expression, and dysregulation of genes related to antioxidant defense, immunity, and detoxification. Additionally, high concentrations of dinotefuran and its combination with amitraz impaired hypopharyngeal gland morphology. Notably, co-exposure resulted in synergistic toxic effects, exacerbating physiological damage beyond individual treatments. These findings emphasize the potential risks of combined exposure to agricultural and beekeeping pesticides. A more comprehensive risk assessment and stricter regulations are urgently needed.

The ectoparasitic mite Varroa destructor has become a major worldwide pest of the honey bees (Apis mellifera L). Intensive use of chemical acaricides has led to the development of acaricide resistance in mites, thereby reducing the efficacy of acaricides and causing contamination of hive products. Thus, natural products such as essential oils can become an important alternative to synthetic chemicals in controlling Varroa mites. In this study, essential oil from Thymus schimperi grown in the Ethiopian highland was investigated for its chemical composition and toxicity against Varroa mites through complete exposure tests. Hydrodistillation of T. schimperi leaves yielded a pale yellow essential oil (0.8% v/w) with a strong and pleasant odor. Analyzed by GC-MS, 29 compounds, comprising 96.67% of the oil, were identified. Carvacrol (34.84%), thymol (23.03%), and p-cymene (14.20%) were found to be the main constituents. In the toxicity assay, T. schimperi essential oil showed high acaricidal activity against Varroa mites, with a 4 h-LC₅₀ value of 109.16 µg/ml (5.46 mg oil/l air volume). Among the main constituents, thymol and carvacrol induced high toxicity to Varroa mites, with 4 h-LC₅₀ values of 70.71 µg/ml (3.54 mg thymol/l air volume) and 106.10 µg/ml (5.31 mg carvacrol/l air volume), respectively. However, p-cymene was less effective in toxicity testing (4 h-LC₅₀ > 5000 µg/ml). The results show for the first time that the essential oil of Thymus schimperi and its two main constituents, thymol and carvacrol, are highly acaricidal and can be used to control Varroa mite infestations.