Molecular detection of

Babesiosis is a tick-borne disease caused by protozoan parasites of the genus Babesia that can impair animal health, reduce livestock productivity, and result in economic losses. Sheep, an important livestock species in Indonesia, are particularly susceptible, especially under traditional farming systems. This study aimed to detect Babesia sp. infection in sheep from Sleman Regency using microscopic and molecular approaches and to characterize the detected isolate at the species level. A total of 35 blood samples were collected from smallholder sheep farms. Microscopic examination was performed using 10% Giemsa-stained blood smears, while molecular detection was conducted using Polymerase Chain Reaction (PCR) with panpiroF1 and panpiroR1 primers targeting the 18S rRNA gene. One sample tested positive for Babesia sp. by both microscopic and PCR examinations, with a low parasitemia level of 1.2%. The observed prevalence was 3.0% (95% Cl: 0.0–15%). Sequencing and phylogenetic analysis demonstrated that the detected isolate was closely related to Babesia motasi. Although based on a single detection, these findings provide preliminary molecular evidence of Babesia motasi infection in local sheep and underscore the importance of early detection for improving animal health and supporting effective disease surveillance in small ruminant farming systems.