We reported a complete check-list of Vertebrates for the “Tenuta dei Massimi” nature reserve, a protected area well representing a remnant agro-forest heterogeneous mosaic landscape of the “Campagna Romana”. Data were obtained from a large original and bibliographic data-set on a large time-span (1991-2012). This reserve represents an area with a high value of species richness when compared to the surrounding metropolitan areas, also including a large number of species of conservation concern at different scales. We obtained evidences for the occurrence of 141 species (one fish, six amphibians, 12 reptiles, 101 birds, 21 mammals). Among them, some species are of conservation concern at European, regional or local scale. Further research should be focused on a set of fragmentation-sensitive species to verify the long-term viability of their local small populations [<em>e.g. Hyla intermedia</em> Boulenger, 1882, <em>Bufo lineatus</em> Ninni, 1879, <em>Muscardinus avellanarius</em> (Linnaeus, 1758), <em>Sorex samniticus</em> Altobello, 1926)].