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Abstract Mites of the species Lynxacarus radovskyi, which are commonly found on domestic cats in Brazil, can cause discomfort, itching, and alopecia. The development of new, safer and more effective treatments with a broad spectrum of activity, including the use of isoxazolines, is needed. The purpose of this study was to assess the efficacy of transdermal fluralaner in domestic cats naturally infested with L. radovskyi. Twenty cats were evaluated by trichograms and divided into two groups of 10 animals. The control group was not treated, while the treated group was given a single topical dose of fluralaner, as per the manufacturer’s instructions. The cats were reassessed for the presence of L. radovskyi eggs and mites on days D+7, D+14, D+28, D+42, D+56, D+70, D+84, and D+98. As of D+42, all the animals (100%) tested negative for mites, and remained parasite-free until the end of the study, while the control group tested positive throughout the experiment. It can be concluded that a single dose of fluralaner applied topically was effective in treating cats naturally infested with L. radovskyi.