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A cross-sectional sero-epidemiological study was conducted on forestry workers, a high risk population for Lyme borreliosis. The prevalence of seropositive forestry workers (indirect immuno fluorescence assay) is 15.2% (n = 211). Almost 70% of participants reported a history of tick bite. Among districts, high seroprevalences were observed in Fontainebleau and Rambouillet; two sites of high risk for Lyme borreliosis. However, clinical prevalence of the disease is very low. During the investigation no active Lyme borreliosis was observed. Thus, asymptomatic infection predominates.