The epizootic of Bluetongue virus (BTV) throughout Europe in 2006 revealed insufficient knowledge regarding seasonal activities, endo/exophilic and endo/exophagic behaviour of the species of the Culicoides genus (Diptera: Ceratopogonidae). Although several studies have been carried out in Western Europe, bringing new knowledge of the above mentioned topics, there was still a gap to fill in Central Europe. Therefore, this study investigated seasonal variations in the indoor/outdoor Culicoides activity observed in the south-eastern Slovakia from 2012 to 2014, using a light-trapping method. In total, 52,741 Culicoides were collected and identified; majority of them were caught outdoors (82.6%), with the highest activity period being mid-June. The C. obsoletus/C. scoticus and C.punctatus species predominated, altogether representing 88.8 and 94.1% of the total indoor and outdoor collections, respectively. Positive correlation was observed between the temperature and the abundance, whereas no correlation with other studied factors was detected. In autumn, Culicoides activity was observed outdoors at temperatures ranging between-1.5°C and 9.3°C, whereas the indoor activity was detected at temperatures ranging between-3.9°C and 0°C. This demonstrated that the most significant BTV vectors in the studied area are C. obsoletus/C. scoticus. The C. dewulfi and C. chiopterus vectors appeared to be of lower significance. The period with the highest risk of the BTV transmission in Eastern Slovakia appeared to be mid-June. The autumnal Culicoides activity inside the cowsheds may be important for the assessment of virus overwintering. This study is the first to provide the data on indoor/outdoor behaviour of biting midges in Slovakia.