Human intervention is the main cause of the decline of the Critically Endangered European mink Mustela lutreola. In this study we analysed the main causes of direct human-caused mortality of the species in Spain. A total of 47 mortality records were obtained for the period 1950–1989, and 145 for 1990–2008. There was temporal variation in the cause of death, with trapping and shooting being the most common causes of mortality in 1950–1989 and road-kills in 1990–2008. In the case of road-kills there was variation related to road type. Males were more affected by road-kills than females, especially during the mating season when they range more widely. Our results indicate that there has been a change in human social behaviour and in people's awareness of the species, with a reduction in European mink captured and shot but an increase in mortality on roads.