Myotis alcathoe has been recorded in the territory of Slovakia for the first time. On 2 August 2001, two individuals of the species were netted at the entrance to the Stĺpová jaskyňa cave in the basaltic plateau of the Cerová vrchovina Mts (48° 12’ 23” N, 19° 54’ 37” E; 525 m a. s. l; ca. 2 km south of the village of Šurice). The species identification was confirmed by sequencing part of the mitochondrial gene ND1 in two specimens. Measurements and description of external and cranial characters, as well as description of the baculum of these two reference specimens are given for further comparisons of this poorly known species.