BackgroundAnaplasma marginale, Babesia bovis and Babesia bigemina cause huge economic losses to the world's cattle industry every year. At present, there are limited investigations on A. marginale, B. bovis and B. bigemina in Hainan. The purpose of this study was to investigate the infection of A.marginale, B.bovis and B.bigemina in different breeds of cattle (Leiqiong and Wagyu), buffaloes and Rhipicephalus microplus in the northern part of Hainan.MethodsIn this study, 63 R. microplus and 151 samples of bovine blood were collected from cattle in the northern part of Hainan. A. marginale, B. bovis and B. bigemina were detected by PCR and nPCR. ResultsThe results showed that the infection rates of A. marginale, B. bovis and B. bigemina in 214 samples were 10.75% (23/214), 4.21% (9/214) and 9.35% (20/214), respectively. In Leiqiong cattle, the total infection rate of several pathogens was the highest, which was 73.33% (22/30), and there was cross-infection of several pathogens, the infection rate was 46.67% (14/30). Among them, A. marginale and B. bigemina had the highest cross-infection rate, which was 26.67% (8/30). No mixed infection was detected in R. microplus, buffalo and Wagyu. Among buffalo, the infection rates of A. marginale, B. bovis and B. bigemina were 2.17% (1/46), 6.52% (3/46) and 4.35% (2/46), respectively. A. marginale infection was only found in Wagyu, with an infection rate of 2.67% (2/75). A. marginale and B. bigemina infections were found in R. microplus, but no B. bovis infection was found. ConclusionsThe results of this study provide a reference for the formulation of prevention and control strategies for A. marginale, B. bigemina and B. bovis in Hainan.