In Europe, at least 6 agents of zoonotic vector-borne diseases have been identified in which ticks play an important role in transmission (Rickettsia sp., Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato, Francisella tularensis, Crimean-Congo hemorrhagic fever virus, tick-borne encephalitis virus). Our preliminary investigations indicate the presence of A. phagocytophilum, F. tularensis, Rickettsia sp. and five genospecies of the B. burgdorferi complex in ixodid ticks in Serbia. According to epidemiological data, tick-borne encephalitis and Crimean-Congo hemorrhagic fever viruses are present in natural foci on the territory of Serbia, but no systematic studies on their presence in ticks have been performed so far. Research in this project included regular collection of material (ticks from vegetation and hosts, serum samples and host tissues), molecular detection of pathogens in samples, and cultivation, characterization and genotyping of these microorganisms. The research focused on potential natural hosts of these zoonotic diseases. The research results provided a clearer picture of the distribution, genetic diversity, and pathogenic potential of tick-borne microorganisms.