The ALSA(CE)TIQUE study aimed to estimate the incidence of three tick-borne diseases in Alsace: Lyme disease (LD), human granulocytic anaplasmosis, and tick-borne encephalitis (TBE). Between 2014 and 2015, the annual number of LD cases was estimated at approximately 2,200, bringing the average annual incidence rate to 117 cases per 100,000 inhabitants [95% CI: 109–126]. This rate remained stable over the two-year study period. Case characteristics were consistent with data from the literature, notably with nearly 80% of cases presenting as erythema migrans. Forests and public or private gardens were the most frequently reported tick bite sites among cases presenting with erythema migrans. The garden as the sole site of tick bite was cited in 25% of cases. The presence of a tick-borne encephalitis outbreak in Alsace was confirmed, even though the number of reported cases remained low (an average of 7 cases per year). Human granulocytic anaplasmosis was a rare condition, with 3 cases reported during the study. These results show that it remains important for public health messages regarding tick-borne diseases—including both information for healthcare professionals and prevention guidance for the general public—to be disseminated as closely as possible to local communities.