Road-kills are the greatest source of direct human-induced wildlife mortality, especially in amphibians. Country roads could act as the most important source of mortality when main roads act as strong barriers hampering the migration movements of some species. Mortality patterns of amphibians on country roads (1380 km) were studied in Salamanca (Spain) in order to quantify the mortality levels, to test the effects of sex and age factors on road-kills, to determine the spatial distribution patterns of road-kills, and to identify routes of migration through a friction map and hotspots of road-kills. From a total of 819 records of amphibians, 38.1% were road-killed and 61.9% were live. Fourteen amphibian species were recorded during the surveys (10 anurans and four urodeles). The species more affected by road-kills were the anurans Bufo calamita, Pelobates cultripes and B. bufo (38.5, 23.4 and 11.9%, respectively). Females had higher incidence of road-kills than males, due to the differential activity patterns of both sexes during the reproductive period. Adults were the most common age period and also the most road-killed. The spatial distribution patterns of live and road-killed records were clustered. On the sampled roads, there were 0.23 road-kills per kilometre and 52 hotspots of road-kills. The friction map showed that most of the road-killed and live specimens were located on migration routes crossing suitable habitats. Conservation measures should be implemented in these areas, as these mortality patterns may be causing significant negative impacts at the population level.