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Background: The isoxazoline fluralaner is effective for prevention of Babesia canis transmission from infected Dermacentor reticulatus ticks to dogs for 84 days in a controlled environment. This study was designed to evaluate the effectiveness of fluralaner chewable tablets for sustained prevention of B. canis infection of dogs in endemic areas under natural conditions. Methods: In Europe, privately owned, clinically healthy pet dogs were enrolled and randomized either to receive fluralaner at 25–56 mg/kg (Bravecto® chewable tablets) on days 0 and 84, or to remain untreated during the D. reticulatus season. Blood samples were collected to evaluate B. canis exposure: on days 0 and 21 (exposure before day 0), during the study and at the end of the tick season (dogs suspected of having become infected after day 0). Efficacy was determined by the percentage reduction in B. canis transmission risk based on the difference in B. canis-positive tests in fluralaner-treated dogs compared with untreated dogs. In addition, ticks collected at monthly intervals throughout the study were identified to species level and females tested for B. canis DNA. Results: A total of 152 dogs were enrolled in the study, although nine dogs were excluded because they tested positive for B. canis DNA or antibodies within 21 days after enrollment. During the study period, no fluralaner-treated dog became positive for B. canis, resulting in calculated efficacy of 100%. However, babesiosis infection was diagnosed in five untreated control dogs (Fisher's exact test, left-sided, P = 0.0312). Tick analyses revealed that one sample collected in Hungary was infected with B. canis. Conclusion: Oral administration of Bravecto chewable tablets at the recommended dosage to dogs completely prevented B. canis transmission under field conditions in an endemic area for 12 weeks. [ABSTRACT FROM AUTHOR]