Abstract Varroa destructor is an ectoparasitic mite that affects colonies of honey bee Apis mellifera worldwide. In the last years, substances of botanical origin have emerged as natural alternative acaricides to diminish the population levels of the mite. In the present work, the bioactivity of propolis from different geographical locations of Pampean region from Argentina on V. destructor was evaluated. Fourteen propolis samples were organoleptic and physico- chemically characterized and, by means topical applica- tions, their activity was tested on mites. All propolis had a homogeneous composition and the bioactivity levels against mites were comparable among the different propolis samples. The percentage of mites killed by the treatments ranged between 60.5% and 90% after 30 s of exposure. Thus, V. destructor was highly susceptible to propolis. Moreover, the mites remained anesthetized during the first hours after topical treatment. The results suggest that