Human-animal interactions occur in many sectors of livestock production. This study aims to determine whether human manipulation during the critical period could disrupt their long-term spatial memory and learning functions. Forty-four pups rats (22 males and 22 females), were divided into two groups (handling, non-handling: corresponding to the control group). Subsequently, each group was subdivided into two subgroups: male subgroup (Handling Males, H ♂) and a female subgroup (Handling Female, H ♀), which were handled daily for 5 min from birth to weaning. On postnatal day 81, they were subjected to Morris Water Maze test (MWM). The results showed that in both sexes, the manipulated group had spent more time in the quadrants where was the platform. Therefore, human handling during the neonatal period of rats could induce in long term better learning, and greater spatial memory without any negative effects on behavior.