Mayfly emergence was studied in the Plitvice Lakes National Park (Croatia) monthly over a 2-year period in four habitats (springs, streams, mountainous rivers, tufa barriers) using monthly collections of emergence traps. A total of 12 mayfly taxa were recorded. Almost half of the collected specimens belonged to the genus Baetis Leach, 1815, which was recorded at every site, but we were unable to distinguish between two included species (B. rhodani and B. cf. nubecularis). Other abundant species were Centroptilum luteolum (Muller, 1776), Alainites muticus (Linnaeus, 1758), Habrophlebia lauta Eaton 1884, Paraleptophlebia submarginata (Stephens, 1835), Serratella ignita (Poda, 1761), Ephemera danica Muller, 1764 and Rhithrogena braaschi Jacob, 1974. The mayfly assemblages at all sites were dominated by species typical of the rhithral zone, but there was a shift in species composition along a longitudinal gradient (from 720 to 390 m a.s.l.) from dominance of eucrenal-epirhithral to metarhithral-hyporhithral elements and finally to appearance of metapotamal and littoral elements. Two environmental factors, maximum water temperature and mean pH, had the highest influence on the mayfly assemblages. Emergence mainly occurred between March and November and was related to the elevated water temperature. Emergence patterns of some species were in accordance with their typical Central European emergence patterns (e.g. S. ignita, H. lauta) while some others showed certain discrepancies (e.g. longer emergence period in Rh. braaschi and P. submarginata, one generation emergence in A. muticus and variable emergence patterns between the sites and between the two studied years in C. luteolum). The current study provides a significant contribution to the knowledge of mayfly ecology in karst freshwater habitats which forms a basis for further investigation and monitoring of mayflies in this area.