The health of 34 different Fraxinus taxa in association with the pathogenic fungus Hymenoscyphus fraxineus was assessed in four Slovak arboreta. Averaged across all arboreta, nearly one-quarter (24.9%) of all evaluated trees showed ash dieback symptoms. The damage was most serious on the common ash F. excelsior, a native species. The percentage of dead trees did not exceed 2% for all evaluated trees. Generally, ash trees of all ages were affected, though the intensity of the damage varied among the sites. The identity of H. fraxineus was confirmed by conventional PCR targeting the internal transcribed spacer (ITS) sequences of the nuclear ribosomal DNA, as well as the 18S gene/ITS-2 region of the rDNA operon. In Slovakia, the pathogen has expanded its host range from native species not only to their ornamental cultivars, but also to introduced North American (F. cinerea, F. latifolia, F. pennsylvanica, F. quadrangulata) and Asian (F. bungeana, F. chinensis ssp. rhynchophylla, F. mandshurica) ash species. H. fraxineus was also observed on the previous year’s leaf petioles of the native European species F. ornus, considered a weakly susceptible host. In Slovak arboreta, H. fraxineus was found on 23 Fraxinus taxa; 21 of them represent first records for the country. F. bungeana is recorded as a new host species of H. fraxineus.