Camels are the most adapted species to the harsh ecosystems of the African continent where they represent 80% of world's population. However, they can be susceptible to many pathogens such as Trypanosoma evansi. This parasite is widely distributed in Africa and represents a real threat to animal health and production. In Tunisia, despite its importance, this disease is underestimated and only a few data are available. The present study aims to investigate the prevalence of the disease by combining microscopic and serological tests. A cross-sectional study was conducted on 307 dromedary camels randomly selected from Southern Tunisia. Collected blood samples were microscopically examined and tested with Card Agglutination Trypanosomiasis Test to detect anti-T. evansi antibodies. The microscopic and serological prevalence were estimated at 2.93 and 62.54%, respectively. Microscopic examination showed morphological features of T. evansi. All positives samples were collected from aged males (>5years old). Seroprevalence was similar for both sexes but it was higher in dry season and in the southwest of Tunisia. The present study revealed high prevalence of T. evansi infection among dromedary camels. This highlights the need for adequate control measures based on the detection and treatment of infected animals and vector control. We recommend investigating the prevalence of infection in other domestic animals living in the same environment.