The occurrence of Thelazia gulosa and T. skrjabini in cattle in Italy, together with an update of the prevalence of Thelazia spp. in the southern Italy, is reported. On 764 slaughtered native cattle, fifty-five (7.2%) bovines were infected by Thelazia spp: T. rhodesi was found in 44 (80%) animals, T. gulosa was found in 19 (34.5%) cattle and only one (1.8%) cattle harboured T. skrjabini. This is the first report of T. gulosa and T. skrjabini in Southern Europe.