The cholinesterases are enzymes involved in severe biological functions in mammals; however, it was not yet study in dairy cows during the transition period. In this period, the animals have high energetic demands for milk production, and the supplementation with mineral can be considered an important approach to fulfill this demand. Thus, the aim of this study was to evaluate the activities of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) in dairy cows supplemented with minerals based on selenium, copper, phosphorus, potassium, and magnesium intramuscularly in the transition period. The mineral supplementation in cows stimulates the increase of AChE postpartum (on days 2 and 7 postpartum, p < 0.05), as well as stimulates the BChE before and after the partum (on day 5 prepartum, and day 2 and 7 postpartum, p < 0.05). Calves of dairy cows supplemented showed low AChE activity when compared to calves of non-supplemented dairy cows. Thus, the supplementation with mineral did influence participates indirectly or directly especially in the modulation of cholinesterases that may have anti-inflammatory effect and contribute to improve on hematopoiesis and neurotransmission.