In ancient cultures humans and animals interacted in multiple ways, but tangible evidence illustrating this relationship is limited due to the nature of the archaeological record. Valued since Antiquity in military and civil life in the Mediterranean region, mules (female horse x male donkey) became an integral part of the animal world following the Roman conquest and occupation north of the Alps. Despite their importance, they remained largely invisible in Roman faunal assemblages. Here, for the first time, we investigated…