Tick-borne encephalitis (TBE) cases have been emerging in Europe. The Franche-Comte area, in northeastern France, borders Switzerland, but the two countries differ in their approach to TBE surveillance and prevention. Because family physicians (FPs) are in direct contact with the local population, at-risk of infected tick bites, they need to be well aware of TBE epidemiology and management. An observational survey was performed in 2019 in order to investigate Franche-Comte physicians' knowledge and vaccination practices with regard to TBE. Standardized online questionnaires were sent to a list of FPs practicing in Franche-Comte. The questionnaires included socio-demographic details, questions about TBE knowledge, symptomatology and vaccination. The response rate was 14.7%. FPs practicing in rural areas reported a significantly higher frequency of consultations for tick bites. While 81% of FPs indicated that they had some knowledge about TBE, only 20% were at ease with its clinical symptomatology. Thirty-one % of the FP participants performed TBE vaccinations. A general lack of knowledge about TBE and its clinical symptoms was observed in this survey. FPs play an essential role in screening and preventing TBE, especially those practicing in rural areas and in areas bordering Switzerland.