Анотация: A survey to determine the occurrence of Mycobacterium avium subsp. paratuberculosis (MAP) in wild carnivores in Portugal was conducted by testing samples from road-killed animals between 2009 and 2012. Postmortem examinations were performed and tissues were collected from wild carnivores representing four families and six different species, with a total of 74 animals analyzed. Cultures were performed by using Lowenstein-Jensen and Middlebrook 7H11 solid media and acid-fast isolates were identified by polymerase chain reaction (PCR) and mycobactin dependency characteristics. Tissues were also screened for MAP by directly extracting DNA and testing for the MAP-specific sequences. The occurrence of infected animals (an animal had at least one tissue that was positive for culture or direct PCR) was 27.0 PROCENT (n = 20). MAP was isolated from culture of 25 tissue samples (3.8 PROCENT ) and was detected by direct PCR in 40 (6.0 PROCENT ) samples. Infection was recorded in 5/6 studied species: 7/49 (14.3 PROCENT ) red foxes (Vulpes vulpes), 3/3 (100 PROCENT ) beech martens (Martes foina), 2/4 (50.0 PROCENT ) Eurasian otters (Lutra lutra), 7/15 (46.7 PROCENT ) Egyptian mongooses (Herpestes ichneumon), and 1/1 (100 PROCENT ) European badger (Meles meles). These species represent three different taxonomic families: Canidae (14.3 PROCENT were positive), Mustelidae (75.0 PROCENT were positive), and Herpestidae (46.7 PROCENT were positive). The results of this study confirm the presence of MAP infection in wild carnivores in Portugal