Understanding how wildlife responds to road and traffic is essential for effective conservation. Yet, not many studies have evaluated how roads influence wildlife in protected areas, particularly within the large iconic African National Parks where tourism is mainly based on sightings from motorized vehicles with the consequent development and intense use of roads. To reduce this knowledge gap, we studied the behavioral response and local spatial distribution of impala Aepyceros melampus along the heterogeneous (with variation in road surface type and traffic intensity) road-network of Kruger National Park (KNP, South Africa). We surveyed different types of roads (paved and unpaved) recording the occurrence of flight responses among sighted impala and describing their local spatial distribution (in relation to the roads). We observed relatively few flight responses (19.5% of 118 observations), suggesting impalas could be partly habituated to vehicles in KNP. In addition, impala local distribution is apparently unaffected by unpaved roads, yet animals seem to avoid the close proximity of paved roads. Overall, our results suggest a negative, albeit small, effect of traffic intensity, and of presence of pavement on roads on the behavior of impala at KNP. Future studies would be necessary to understand how roads influence other species, but our results show that even within a protected area that has been well-visited for a long time, wildlife can still be affected by roads and traffic. This result has ecological (e.g., changes in spatial distribution of fauna) and management implications (e.g., challenges of facilitating wildlife sightings while minimizing disturbance) for protected areas where touristic activities are largely based on driving.