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Abstract The effects of acute sublethal doses of coumaphos, an acaricide used againstVarroa destructorinfestation in beekeeping, on the locomotor activities of four native honeybee subspecies of Türkiye including two ecotypes (Carniolan honeybee -A. m. carnica, Syrian honeybee -A. m. syriaca, Caucasian honeybee-A. m. caucasica, and Muğla and Yığılca ecotypes of Anatolian honeybeeA. m. anatoliaca) were investigated using an individual locomotor activity monitoring system. Analysis of locomotor activity data in the first 12-h, last 12-h, and 24-h time periods showed that bees fromcaucasicaandcarnicasubspecies were not affected by coumaphos at all three acute doses (1, 2, and 5 μg coumaphos in 10 μl sucrose syrup for each bee). In contrast, bees fromA. m. syriacasubspecies showed significantly elevated locomotor activity levels at 2 and 5 μg coumaphos doses within the first 12 h. Bees from both Muğla and Yığılca ecotypes ofanatoliacasubspecies also showed elevated locomotor activity levels at 5 μg coumaphos dose but the magnitude of increase was lower in these ecotypes compared to that seen insyriacasubspecies in the first 12-h period. In general, increasing doses of coumaphos resulted in increased locomotor activity (locomotor activity), with differences in sensitivity across honeybee populations. Possible mechanisms underlying this variance and suggestions for further studies are discussed.