During the 20th century, Mediterranean landscapes underwent extensive changes. In particular, decreasing grazing pressure combined with abandonment of agricultural uplands favoured the development of Mediterranean forests with a corresponding reduction of open habitats and landscape diversity. In some parts of the Mediterranean basin, including our study area, the reduction of open landscapes took place at the same time as a rapid decline in rabbits as a result of myxomatosis. This study assesses the impact of past changes in landscape and rabbit density on the density and the breeding performance of the eagle owl Bubo bubo, one of the largest predators of Mediterranean ecosystems. Eagle owl density, nest site structure and composition at the landscape level, diet and fecundity of 35 eagle owl nesting territories was analysed for 27 years. The study area is characterized by two distinct areas, the border of the massif and the interior, in which past changes acted differently. Eagle owls in the interior were simultaneously affected by the loss of rabbits, a shift to smaller prey, and by open areas reverting to forest. All of these factors reduced the foraging efficiency of the owls. When myxomatosis decimated the rabbit population, owls in the border area switched to other mammals, birds and fish, whereas in the interior, alternative prey were less abundant. The main impacts of reduced prey availability and landscape diversity on interior pairs were: (1) lower density of breeding pairs; (2) lower diversity in landscape structure and closer habitats; (3) lower richness and diversity in the diet; (4) later egg-laying dates; (5) lower productivity.