Abstract
seasons. Roe deer were sampled during the summer and the fall in 10 districts (Czarnkow, Krzykosy, Oborniki, Margonin, Miedzychod, Murowana-Goslina, Poniec, Sierakow, Trzcian- ka, Wielen ´ ), whereas samples from red and fallow deer were obtained only during the fall in nine (listed above with the exception of Krykosy) and two districts (Margonin, Murowa- na-Goslina), respectively. In total, 238 partially or fully engorged female and 63 non-engorged male I. ricinus ticks were obtained from 51 animals (20 fallow deer, 18 roe deer and 13 red deer). Total DNA was extracted from blood and I. ricinus ticks using the genomic Mini AX Blood (A&A Biotechnology, Gdynia, Poland) and Sherlock AX kit (A&A Biotechnology), respec- tively. Blood and tick samples were analysed for the presence of A. phagocytophilum DNA by PCR using three different primer sets: EHR521-EHR747, LA6-LA1 and HGE396-HGE921, based on the sequences of the 16S rDNA, ankA and hge-44 genes, respectively (4,5). PCR products were considered positive when A. phagocytophilum DNA was detected by at least two primer sets. Additionally, selected PCR-positive samples tested with the primers LA6-LA1 specific for a 444-bp region of the ankA gene were sequenced with the ABI PRISM 3100 Genetic Analyser (Applied Biosystem 850, Foster City, CA, USA) according to the manufacturer's protocol. Sequences were edited and compared with gene sequences deposited in the GenBank database using the NCBI BLAST program (U.S. National Institute of Health, Bethesda, Maryland).
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