The Gruta da Aroeira (Torres Novas, Portugal), with evidence of human occupancy dating back ∼ 400,000 years, is one of very few Middle Pleistocene cave sites to provide a fossil hominin cranium in association with Acheulean bifaces and the by-products of fire usage. Zooarchaeological, taphonomic and tooth-wear analyses suggest that the accumulation of the faunal remains and their modification are anthropogenic. Large game constituted the basis of subsistence, with equids and cervids being preferentially targeted. Woodland and open landscapes formed the ecosystems supporting the populations of the mammals that were preyed upon by the inhabitants of the site. Most of the animal carcasses were carried to, and fully butchered at the site, which was used as a residential base camp. The features of the Aroeira faunal assemblage foreshadow the subsistence strategies developed by the hunter-gatherers of the Middle and the Upper Palaeolithic and testify to their very ancient roots.