Abstract Several life-history traits may increase vulnerability of species to extinction. Among snakes, ambush predation and dietary specialisation are factors that increase this vulnerability. European viper species, genus Vipera, display such traits and are categorised as endangered in several parts of its range. For their conservation management, a deeper knowledge of their ecology and habitat use is highly relevant. One of the species with less ecological data is the Lataste's viper Vipera latastei, a species which lives in the Iberian Peninsula and northwestern Africa. Here, we describe its diet based on the analysis of gut content of 435 museum specimens plus nine bibliographic data from the entire Iberian range. The species showed seasonal and ontogenetic shift in diet but no sexual variations. Feeding activity (percentage of vipers with prey) was low in accordance with its ambush predation tactics, being lower in spring than in summer and autumn. Prey spectrum included two main (reptiles and small mammals), and three sporadic, types of prey (arthropods, amphibians and birds). The consumption of reptiles and mammals was seasonal; the former decreased in occurrence from spring to autumn, whereas the latter showed an opposite pattern. There was an ontogenetic shift in the diet: juveniles fed mainly on reptiles and arthropods, whereas adult vipers progressively substitute this prey with insectivores, and the largest vipers primarily foraged on rodents and birds. Our results suggest that the seasonal variation in prey type was related to prey availability, whereas the ontogenetic shift was linked to gape limitation. The apparently wide prey spectrum of V. latastei must therefore be examined, taking into account that there are seasonal and ontogenetic dietary variations as well as geographic differences, the latter probably driven by climatic contrasts into the Iberian Peninsula. This new data of the endangered Iberian V. latastei can aid the effective conservation management of this species.