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Prevalence of Leishmania spp. infection was studied in stray cats in two military bases in Southern Israel during a cutaneous leishmaniosis (CL) human outbreak caused by Leishmania major. Human CL cases increased from 0/100 in 2008 to 1.28/100 in 2022 in camp #1, and from 0.17/100 in 2008 to 6.4/100 in 2022, in camp #2. Eight out of 29 cats sampled were Leishmania-seropositive (28 %) and 7/29 (24 %) were internal transcribed spacer 1 (ITS1) PCR-positive, out of which four (14 %) were positive for L. major and three (10 %) for L. infantum. Five positive-cats had skin lesions including ulcers, alopecia and scabs, and five had eye lesions. This is the first report of L. major infection in cats in Israel and one of the first descriptions in felines worldwide. A larger cohort of cats and vector studies are necessary to determine if felids may act as reservoirs or sentinels of human L. major infection.