The aim of this study was to estimate the seroprevalence of antibodies to Borrelia burgdorferi, Anaplasma phagocitophilum and tick-borne encephalitis (TBE) virus, and risk factors, in forestry rangers from the Friuli-Venezia-Giulia region in northeastern Italy. Sera from 181 forestry rangers were examined with two-tiered serological tests for TBE, Lyme borreliosis and ehrlichiosis. Information about risk factors such as job location, residence, number of tick bites and outdoor leisure activities was collected with a questionnaire. Seropositivity was 0.6% for TBE virus, 23.2% for Lyme borreliosis and 0.6% for ehrlichiosis. Lyme borreliosis positivity, as determined by Western blot, was associated with working in the foothills, with gardening in the northeastern part of the region, and with a history of yearly tick bites. Risk factors were similar when a case of Lyme borreliosis was defined either by Western blot positivity or by clinical history.