Previous studies have found that the relationship between wildlife road mortality and traffic volume follows a threshold effect on low traffic volume roads. We aimed at evaluating the response of several species to increasing traffic intensity on highways over a large geographic area and temporal period. We used data of four terrestrial vertebrate species with different biological and ecological features known by their high road-kill rates: the barn owl (Tyto alba), hedgehog (Erinaceus europaeus), red fox (Vulpes vulpes) and European rabbit (Oryctolagus cuniculus). Additionally, we checked whether road-kill likelihood varies when traffic patterns depart from the average. We used annual average daily traffic (AADT) and road-kill records observed along 1000 km of highways in Portugal over seven consecutive years (2003–2009). We fitted candidate models using Generalized Linear Models with a binomial distribution … Palabras clave: Mortality