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The aim of our work was to find the positivity rate of antibodrrelian antibodies (IgG) in wild-living rodents in a locality situated in north Moravia, Czech Republic. Results of a survey for heart rinses (172) and sera (2) antibodies to Borrelia burgdorferi sensu lato (s.l.) from 6 species of 174 wild rodents from the northern part of the Czech Republic are presented. Samples were obtained in 2001-2002 at one locality (Studenka, 49 degrees 44′, 18 degrees 05′). Host samples included yellow-necked mouse (Apodemus flavicollis, n = 106), wood mouse (A. sylvaticus, n = 170) and striped field mouse (A. agrarius, n = 13) from Muridae and bank vole (Clethrionomys glareolus, n = 3) and common/field vole (Microtus sp., n = 5) from Microtidae families. An indirect enzyme linked immunosorbent assay (ELISA) antibody test was used for testing heart rinses. Goat immunoglobulins against mouse were used as a conjugate. Antibodies to B. burgdorferi s.l. were found in all species. The highest positivity rate (58.8 % ) was recorded for wood mouse (58.8 % ), bank vole (45.5 % ), and yellow-necked mouse (44.3 % ). Mean positivity rate for both years of collection was 43.7 %, mean annual positivity rate was 60.6 % in 2001, and the value of 21.3 % in 2002 appeared significantly different. Three times as many of the Muridae as of the family Microtidae were caught and the actual number of seropositive Muridae was not significantly higher than Microtidae. Positivity of compared males and females was not significantly different. Results indicate that spirochaetes were widely represented in the northern part of the Czech Republic.