Trypanosomosis is the most important single cause of economic losses in camels. The present study was conducted to determine the prevalence of camel trypanosomosis and its vectors in Al-Kharj, Central Province, Saudi Arabia. One hundred and seven serum samples were examined to detect the presence of T. evansi infection by CATT test. Eight Nzi traps for sampling of biting flies were deployed in camel farms, including water station, people residence, cactus farm (Aloe vera), irrigated areas, open Range, cowsheds, sheep pens, and control. Camel trypanosomosis prevalence was 21.5% during the study period. The highest catches of flies were around water station and people residence. The blood biting flies caught were Stomoxys calcitrans and some mosquito's species. Stomoxys calcitrans may be an important mechanical vector of camel trypanosomosis in the study area. The peak of muscidae abundance was observed in March, April and October.