The representatives of the genus Ametropus (Insecta, Ephemeroptera) were recorded 80 years ago in Georgia. Due to the taxonomic uncertainty, the recorded specimens were considered potentially new taxa. Since then, no other record of Ametropus has been documented from the whole South Caucasus and its nearby regions further south. In the present contribution, we provide a new record of Ametropus from the Alazani River (Kura River Basin, Georgia). Morphological and DNA barcoding of the COI gene fragment showed that the species belongs to the widespread Holarctic species A. fragilis. Morpho-anatomical characteristics also confirmed the high degree of similarity between the newly recorded specimens and those once recorded 80 years ago. In addition, the COI barcode showed a strikingly closer relationship between Caucasian A. fragilis and North American A. neavei (3.54% divergence) compared to Caucasian and European A. fragilis populations (4.3% divergence). The observed pattern clearly indicates the need for further and more thorough revision of the Ametropus species complex in the Holarctic region.