Ixodes ricinus ticks (20 males, 20 females and 20 nymphs) collected in Kosice, Slovakia were examined for the presence of Anaplasma phagocytophilum and Borrelia burgdorferi sensu 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-circulate in the enzootic sites of Eastern Slovakia and may cause co-infection in humans.