As protected areas increasingly compete with other land uses, their ability to generate economic value through tourism has become central to both public acceptance and long-term sustainability. However, the effects of tourism on wildlife remain understudied, particularly in African protected areas. Mole National Park, Ghana’s largest protected area, is critical for both biodiversity conservation and ecotourism. With plans to expand tourism into its less-visited areas, we assessed the impacts of tourism on medium- to large-sized mammals using multiple metrics. Camera traps were deployed across zones with low, medium and high tourism use, recording 24 mammal species over 6,050 trap-days. Although overall species richness and abundance were similar across zones, certain species showed higher relative abundance in high-use areas (e.g. kob Kobus kob, African buffalo Syncerus caffer) and others were more abundant in low-use areas (e.g. western hartebeest Alcelaphus buselaphus major, African savannah elephant Loxodonta africana). The most consistent effect of higher tourism levels was a shift in daily activity patterns towards more nocturnal behaviour, probably in response to human presence. Olive baboons Papio anubis and common warthogs Phacochoerus africanus, however, maintained peak activity around midday. Juvenile presence was highest in low-tourism zones, suggesting possible reproductive sensitivity to disturbance. These behavioural and demographic responses may have longer-term consequences for population dynamics. Our findings raise concerns about the expansion of tourism into currently undisturbed areas, as wildlife is already responding to existing pressure. We recommend designating no-access zones as wildlife breeding areas, implementing buffer zones based on species activity and educating tourists on responsible wildlife viewing. Finally, we highlight the need for broader research on tourism impacts across conservation landscapes in Africa.