Molecular screening of Babesia, Hepatozoon, and Theileria (Apicomplexa: Piroplasmida) in ticks (Acari: Ixodidae) infesting farm ruminants in Peninsular Malaysia

Ticks are obligate hematophagous arachnids that feed on both humans and animals. Despite the extensive research on detection of bacteria in ixodid ticks in Malaysia, there remains limited knowledge about the detection of protozoa in these ectoparasites, especially in those that feed on farm ruminants. In this study, 1,241 ticks belonging to four species (Rhipicephalus microplus, R. haemaphysaloides, Haemaphysalis bispinosa, and H. wellingtoni) were collected from 674 farm ruminants across Peninsular Malaysia. The ticks were pooled and subjected to DNA extraction, followed by protozoal screening using 18S rRNA gene polymerase chain reaction (PCR). Of the 130 tick pools tested, 15 were positive for Babesia (11.54%) and ten for Theileria (7.69%). No Hepatozoon protozoa were detected. All positive pools consisted solely of R. microplus ticks, with no protozoa found in the other three tick species. BLAST analyses revealed that the Babesia sequences were identical to Babesia bigemina, while the Theileria sequences closely resembled Theileria orientalis and Theileria sinensis. This paper presents the first nationwide screening of Babesia, Hepatozoon, and Theileria in ticks infesting farm ruminants from Peninsular Malaysia.