Honeybee (Apis mellifera) reproduction is polyandrous: the queen obtains millions of spermatozoid by mating with several drones outside the colony. Fertility problems of honeybee queens are reported where failure of the production and quality of sperm drones are suspected. Several factors can affect sperm quality drones include pesticides. The aim of this study is to determinate the in vitro effect of fipronil, ethiprole, imidacloprid, thiamethoxam, cypermethrin, and coumaphos at different concentrations from 0.1 to 100 μM on the viability and the energetic state, through ATP content, of spermatozoids of honeybee drones. Exposure during 24 h showed that all the active ingredients used in this test increase the ATP levels. Four of them i.e. fipronil, ethiprole, imidacloprid, and thiamethoxam reduced significantly the viability of spermatozoids. Hence, pesticides could affect the drone’s spermatozoids which may have negative impact on semen quality and further queen fecundity.