Context. Habitat degradation caused by roads and wildlife–vehicle collisions are two of the main drivers of terrestrial vertebrate mortality. Spatial collision patterns are influenced by habitat and road characteristics and the species’ ecology and biology, including its life history. Costa Rica has a network of 10,000 km of paved roads; however, there is a knowledge gap on road ecology and its impact to wildlife. Understanding the interconnectedness of roadkill with spatial and temporal patterns is crucial to mitigate this threat in the country.

Aims. Here, we describe spatial and temporal patterns in bird and mammal–vehicle collisions on a 45 km section of the Interamerican Highway and identify the critical zones that require attention.

Methods. Surveys were conducted by car between Río Macho Forest Reserve and Los Santos Forest Reserve for a period of 8 months. We recorded bird and mammal individuals and road and landscape features in 100 m sections to identify collision hotspots. We used the road and landscape information to try to explain the hotspot segments using generalized linear models.

Key results. We found 148 carcasses that belonged to 16 species of bird and five species of mammal. There was no temporal pattern in frequency of collisions for both birds and mammals. We identified 34 collision hotspots, five for birds and 29 for mammals, and seven hotspots for both groups combined. Bird collision frequency was explained by the 150 m scale forest area, distance to rivers, and road sinuosity. Mammal collisions were not explained by the evaluated variables.

Conclusions. Roadkill of birds and mammals was influenced by biological traits and environmental factors. The identification of specific environmental factors and collision hotspots are an initial step on collision monitoring in the Talamanca mountain range in Costa Rica.

Implications. This research has increased our knowledge on some of the factors that influence roadkill frequencies and vulnerability of species. Our results identified segments on the Interamerican Highway where roadkill mitigation measures should be implemented. Our findings can also serve as a reference point to analyze other segments of the highway or other roads with similar features in the country.