Abstract: Predation on livestock is a cause of serious and long-lasting conflict between farmers and wildlife, promoting negative public attitudes and endangering conservation of large carnivores. However, while large carnivores, especially the grey wolf (Canis lupus), are often blamed for killing sheep and other farm animals, free-ranging dogs may also act as predators. To develop appropriate measures for livestock protection, reliable methods for identifying predator species are critical. Identification of predators from visual examination of livestock wounds can be ambiguous and genetic analysis is strongly preferable for accurate species determination. To estimate the proportion of wolves and dogs implicated in sheep predation, we developed a sensitive genetic assay to distinguish between wolves and domestic dogs. A total of 183 predator saliva samples collected from killed sheep in Estonia were analysed. The assay identified the predator species in 143 cases (78%). Sheep were most often killed by wolves (81%); however, predation by dogs was substantial (15%). We compared the molecular results with field observations conducted by local environmental officials and recorded some disagreement, with the latter underestimating the role of dogs. As predator saliva samples collected from prey are often of poor quality, we suggest using mitochondrial DNA as a primary tool to maximise the number of successfully analysed samples. We also suggest adopting forensic DNA analysis more widely in livestock predation assessments as a legislative measure since misidentification that is biased against wolves can be counterproductive for conservation by enhancing conflict with society and leading to increased culling and poaching.