Epizootic haemorrhagic disease virus (EHDV) was detected for the first time in Tunisia and in other Northern African countries in 2006. The objective of the present study was to investigate whether EHDV circulated in Tunisian livestock before and after the officially-reported outbreak of 2006. Thus, serum samples from cattle and dromedaries collected in different time periods (before and after 2006) and from different regions of Tunisia were screened for the presence of EHDV antibodies. Serological investigations conducted on cattle and dromedary sera collected in 2000 and 2001 demonstrated no virus circulation on these dates. However, viral circulation was evidenced in 2012 and 2013, although no EHDV cases were officially reported in these years. Serum-neutralization assessed on few ELISA positive samples, confirmed the presence of antibodies against EHDV serotype 6, which was the serotype involved in the EHDV outbreak in the Maghreb region in 2006.