Abstract: Medium and large mammals are greatly affected by human activities, such as, habitat loss and hunting. In Colombia, these pressures have been most extensive in the dry and Andean forests. However, there is scare information available on the presence or abundance of these organisms for monitoring purposes. This study used photo-trapping cameras to determine the relative abundances of medium and large mammals in a mountain forest in the National Natural Park “Cueva de los Guacharos”, Huila (Colombia). Additionally, we examined whether trail use by humans could affect the distribution of these organisms. The cameras were located every 500 meters along the trails in strategic locations. We obtained the relative abundance of 11 species, being medium-sized mammals the most frequent (Cuniculus paca, Dasyprocta punctata and Mazama rufina). We recorded the presence of mountain tapir (Tapirus pinchaque), dwarf red brocket (Mazama rufina) and oncilla (Leopardus tigrinus), which are found in a high threat category according to the IUCN. Activity patterns were reported for 4 different species including daytime, nocturnal and cathemeral activities. Finally, we concluded that large mammals avoided trails of frequent use, and although the relative abundance of mammals is not very high, this National Park is still supporting relevant biodiversity.