The aim of the present work was to determine whether Dermacentor reticulatus (Fabricius), tick species common in eastern Poland could be infected with Toxoplasma gondii (Nicolle et Manceaux, 1908). A total of 664 unfed D. reticulatus ticks were collected from six localities of Lublin province (eastern Poland) within the framework of study for the presence of bacterial, viral and parasitological infections, with use of PCR and confirmed by sequencing analysis. The prevalence of T. gondii DNA of B1 gene in the total examined D. reticulatus ticks was 3.2%. The infection varies greatly depending on the locality of tick collection (0-16.7%). Preliminary identification of clonal type (I or II/III) by Restriction Fragments Length Polymorphism PCR (RFLP-PCR) with use B1 gene showed that all isolates of T. gondii belonged to type I. RFLP analysis using genetic markers SAG1, 5'-SAG2, 3'-SAG2, SAG3, and GRA6 on B1-positive samples showed that only a single isolate proved to be type I with all five markers, another type was classified to type I according to four markers, while another five isolates had only type I alleles at GRA6, which cannot be regarded as type I confirmation. It must be pointed out that the used DNA isolation method by boiling with ammonium hydroxide enables to receive the total DNA from ticks, but may be not quite suitable for genotyping. In conclusion, this study indicates that besides Ixodes ricinus (Linnaeus), also D. reticulatus should be considered as a potential vector of T. gondii. The presumption of tick-borne transmission as an alternative pathway of disease spreading could well explain the high prevalence of toxoplasmosis among the herbivorous mammals and birds. However, this hypothesis needs verification by further experimental and ecological studies.