This article reports the first record of the Eptesicus serotinus (Schreber, 1774) in the Early and Mid Holocene in Southern Europe. Fragments of mandibles, maxilla, rostrum, humerii and radii from this species were collected in Mesolithic and Neolithic layers of the cave of Sarakenos (Boeotia, Greece). This provides evidence of the presence of the serotine bat in Greece during the Holocene. The relevance of this new record increases the areal distribution of serotine bat in the postglacial Europe.