Ixodes ricinus ticks (20 males, 20 females and 20 nymphs) collected inKosice, Slovakia were examined for the presence of Anaplasma phagocytophilum and Borrelia burgdorferisensu lato (s.l.) by PCR. 38.3 % of the tested ticks carried single infection of B. burgdorferi s.l.and 8.3 % were infected with A. phagocytophilum. Double infection of both pathogens was detected in 5% of tested ticks. These results indicate that both B. burgdorferi s.l. and A. phagocytophilum co-circulatein the enzootic sites of Eastern Slovakia and may cause co-infection in humans.