New human cases of tick-borne encephalitis (TBE) have recently been recorded outside the recognised foci of this disease, i.e. in the province of Trento in northern Italy. In order to predict the highest risk areas for increased TBE virus activity, we have combined cross-sectional serological data, obtained from 459 domestic goats, with analysis of the autumnal cooling rate based on Moderate Resolution Imaging Spectroradiometer (MODIS) land surface temperature (LST) data. A significant relationship between finding antibodies against the virus in serum (seroprevalence) in goats and the autumnal cooling rate was detected, indicating that the transmission intensity of the virus does not only vary spatially, but also in relation to climatic factors. Virus seroprevalence in goats was correlated with the occurrence of TBE in humans and also with the average number of forestry workers' tick bites, demonstrating that serological screening of domestic animals, combined with an analysis of the autumnal cooling rate, can be used as early-warning predictors of TBE risk in humans.