Most studies concerning parasitic infections in European bison have been performed on free-ranging animals: comparatively little is known about the abomasal nematodes of captive wisents, which are widely used in reintroduction programmes. The aim of the study was to determine the infection level and species composition of abomasal nematodes in captive European bison in enclosures (including zoos) and breeding centres compared to free-ranging individuals. It also includes a morphological analysis of the parasites based on figures and measurement data. Altogether, 11 species of nematodes were detected, with both captive and free-ranging animals demonstrating similar species compositions. Among those, 2 species of blood-sucking nematodes were detected, including Ashworthius sidemi and Haemonchus contortus. Interestingly, A. sidemi was found in almost all free-roaming animals, but only in 1 captive European bison. In addition, H. contortus was predominant in captive animals. The morphological identification was confirmed molecularly for 5 nematode species: A. sidemi, H. contortus, Ostertagia kolchida, O. ostertagi and Spiculopteragia boehmi. The identification was performed using small subunit ribosomal rDNA. The study provides the first available set of specular lengths of the gastric nematodes of European bison, and the first molecular data of O. kolchida and S. boehmi derived from the same host species. Our findings may simplify the morphometrical and molecular identification of Trichostrongylidae species infecting European bison, and can be useful in developing new management strategies for populations of this near-threatened species in Europe.