Neotropical cloud forests are one of the most biodiverse yet threatened ecosystems with deforestation and hunting creating major problems. We report a non-volant mammal species inventory for Cusuco National Park, Honduras, spanning 2006–2016. Data were collected using 4 methods: tracks and signs, camera trapping, direct sightings and live trapping. Recorded species of conservation importance included Central American red brocket (Mazama temama (Kerr, 1792)), Baird’s tapir (Tapirus bairdii (Gill, 1865)), margay (Leopardus wiedii (Schinz, 1821)), and jaguar (Panthera onca (Linnaeus, 1758)); the latter recorded for the first time. Cusuco National Park is a site of strategic geographical importance within the Central American jaguar corridor. In total, 43 species were recorded from 26 families. We demonstrate the impact of zonal protection measures with higher mammal abundance recorded in the park’s highly protected core compared to the less protected buffer zone. We advise continued monitoring to provide robust time-series of population trends.