With an area of 1,633 km2 and approximately 400 km of coastline, Lesvos is the third-largest Greek island and the eighth-largest in the Mediterranean. Situated in the northeastern part of the Aegean archipelago and bordering Turkey, Lesvos has been known to host 59 ant species and morphospecies. In this article, we present a list of 84 ant species and morphospecies collected in Lesvos. The checklist is a compilation of a literature data and material surveys of the materials collected in 2015, 2022, and 2024. Seven species are new to the East Aegean Islands, and 25 are recorded from Lesvos for the first time. Strongylognathus afer Emery, 1884, is a new species for the fauna of Greece.