A study of ticks on wildlife was carried out in the area of Levice, Bratislava, Stupava and Vrbovce (south-western Slovakia) during the years 2021-2022. Overall, 512 ticks were collected from 51 wild mammalian individuals of six species. In total, eight tick species were identified, namely Dermacentor reticulatus, D. marginatus, Haemaphysalis inermis, H. concinna, Ixodes ricinus, I. hexagonus and two Ixodes spp. Ixodes hexagonus were collected from Northern white-breasted hedgehogs (Erinaceus roumanicus), females belonging to Ixodes spp. were collected from a red fox (Vulpes vulpes) and nymphs from a Eurasian badger (Meles meles). Ixodes hexagonus and the Ixodes spp. were identified morphologically and molecularly based on sequences of fragments of two mitochondrial genes, COI and 16S rRNA. Molecular analysis of Ixodes spp. confirmed the identity of Ixodes kaiseri Arthur, 1957 and I. canisuga (Johnston, 1849). Sequence analyses show that the I. kaiseri isolate from Slovakia is identical to I. kaiseri isolates from Romania, Poland, Germany, Turkey, and Croatia. We demonstrate for the first time the presence of I. kaiseri in Slovakia using both morphological and molecular methods.