Tick infestation and occurrence of Anaplasma phagocytophilum and piroplasms in cattle in the Republic of Serbia

Ticks transmit important pathogens affecting cattle such as intracellular bacteria of the genus Anaplasma or protozoa of the order Piroplasmida . This study aimed at assessing tick species present on pastures and cattle and determining occurrence of the tick-borne pathogens Anaplasma phagocytophilum and Babesia spp. in cattle. During May and June 2013, ticks and EDTA blood were collected from 185 dairy cows at six locations in Serbia. Ticks were also collected directly from the pastures at four of these locations. The occurrence of A . phagocytophilum was investigated by serological (immunofluorescence antibody test (IFAT)) and molecular methods (real-time PCR) and of piroplasms by traditional PCR followed by sequencing. The most prevalent tick species on animals was Ixodes ricinus , ( n = 167), followed by Haemaphysalis punctata ( n = 146) and Dermacentor marginatus ( n = 122). On the pasture, however, the most common species was H . punctata ( n = 41), I . ricinus ( n = 37), and D . marginatus ( n = 2). Altogether, 4 out of 163 (2.45%) serum samples of cows were positive for A . phagocytophilum -specific antibodies by IFAT. However, the 135 blood samples tested for A . phagocytophilum DNA were all negative. Altogether, 5 out of these 135 samples were positive for piroplasm DNA. PCR products were sequenced and identified as a benign Theileria spp. with 100% identity with GenBank entries from Italy ( Theileria sergenti ), China ( Theileria spp.), and Korea ( Theileria buffeli isolate HS252). The results provide evidence for the presence of several hard tick species infesting cattle in Serbia which can carry pathogens potentially influencing animal health, as well as evidence of contact with tick-borne pathogens Theileria spp. and A . phagocytophilum .