Four hundred and eighty Ixodes ricinus (278 female and 202 male) samples collected in the natural biotopes of 13 administrative districts in southwest Byelarus were studied via DSK-H medium (Sigma) inoculation. Twenty-four spirochetes isolates (3, 2, 1, and 18 in the Brest, Gomel, Mogilev, and Grodno regions, respectively) were obtained on the territory of Byelarus. After adapted to the medium, most isolates (as many as 7 x 10(6)-5 x 10(7) microbial cells per ml) in the stationary phase. All the obtained isolates were cryoconserved at the level of 2-6 passages and after -70 degrees C storage during 4-6 months (a followup period) they were able to recover their initial reproductive activity in the fresh BSK-H medium. Proceeding from preidentification using a comparative electrophoretic analysis of the molecular mass of polypeptides, the pattern of their specific reactivity with polyclonal serum antibodies from the rabbit immunized with cultured Borrelia afzelii (Ip21 strain), in immunoblotting and indirect immunofluorescence assay, I. ricinus spirochetes were referred to Borrelia burgdorferi sensu lato isolates.
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