Porcine lymphotropic herpesviruses 1, 2 and 3 (PLHV-1, PLHV-2 and PLHV-3) are DNA viruses belonging to the genus Macavirus and the subfamily Gammaherpesvirinae within the family Herpesviridae. PLHV was detected in domestic pigs in Germany in 1999, with subsequent outbreaks in Spain, Brazil, Italy and Ireland, which was the trigger for our preliminary study to investigate its occurrence in Croatian pig herds. According to previous studies, natural infections with PLHV in domestic pigs do not cause clinical signs of disease. However, PLHV-1 has been found to cause lymphoproliferative disorders in domestic pigs after bone marrow transplantation that are similar to those described in humans infected with human herpesvirus 4 (HHV-4), which originates from individuals after organ transplantation. HHV-4 is the causative agent of mononucleosis and is the first virus described to have oncogenic potential. HHV-8 causes Kaposi’s sarcoma and contributes to the development of lymphoproliferative disorders in humans, such asprimary effusion lymphoma and multicentric Castleman’s disease. In this study, blood and spleen samples from domestic pigs were analysed using real-time polymerase chain reaction, which has been shown to be an excellent method for the detection of PLH viruses as it is rapid, highly specific and sensitive. The presence of all three PLHV strains in domestic pigs in Croatia was confirmed for the first time with a prevalence of 55.8% regardless of breeding conditions. The most dominant strain was PLHV-1 and the most frequent co-infection was PLHV-1 with PLHV-3. The virus was detected in 10 Croatian counties, with the highest prevalence found in Vukovar-Srijem County. Although herpesviruses are generally species-specific, the close genetic relationship of PLHV with HHV-4 and HHV-8 may indicate a possible zoonotic potential, particularly in immunocompromised human recipients following xenotransplantation. Further investigation of PLHV will contribute to a better understanding of its importance in maintaining the health of pigs and will include genotyping to identify origin of the viruses and potential public health risks.