The relationship between humans and donkeys has ancient origins. In recent years, donkeys’ involvement in Animal-Assisted Intervention (AAI) has grown, raising major concerns about their welfare during these activities. We investigated the factors affecting donkeys’ behaviour during an AAI programme for migrants. We observed an effect of human distance from the animal, the interaction mode, and food presence on the animals’ behaviours and attitudes toward humans. Moreover, we provide preliminary evidence of the effect of AAI on migrants’ welfare and prosocial behaviours and offer insights into the issues to address in future studies.