Varroa destructor is one of the major parasites threatening the eusocial species Apis mellifera. Current treatments remain constrained by the difficulty of targeting the mite without compromising the host. Although lactic acid is already in-use by small apiaries in the beekeeping industry as an organic treatment compatible with integrated pest management, its mode of action on the parasite remains unclear. We investigated how lactic acid affects attachment-dependent behaviours in V. destructor. Particular attention was given to the arolia, the adhesive pads located at the distal end of the legs that enable the mite to grip and maintain contact with the bee cuticle through a fluid-mediated adhesion. Our innovative approach combines behavioural assays with imaging, including Raman microscopy, to characterise the potential role of the arolia fluid during the dispersal phase. Under controlled laboratory conditions, mites were exposed to lactic acid via contact with treated substrates to mimic hive application procedures used by beekeepers. Grip performance was quantified using a rotating machine (Rotavar) and complementary locomotor assays to test dynamic attachment capacity. High-resolution optical microscopy was employed to analyse arolia morphology and detect structural alterations. Raman microscopy enabled, for the first time, the characterisation of the chemical composition of the arolia fluid. In parallel, nutritional status and reproductive output were measured to evaluate potential fitness costs resulting from impaired attachment. We demonstrated that lactic acid does not induce acute mortality in V. destructor but instead disrupts attachment- dependent behaviours. Acute topical exposure caused morphological alterations of the arolia and modification of the associated fluid, leading to reduced grip performance and locomotor activity. Nutritional status remained unaffected, while reproductive output showed a potential decline. Altogether, these results reveal that lactic acid primarily acts locally by destabilising the attachment system of V. destructor rather than through direct toxicity. By identifying attachment as a behavioural and functional target, this work provides a mechanistic framework for understanding and optimising mite control strategies while improving the specificity of organic treatments.