Summary Background Fasciola hepatica (liver fluke) affects grazing animals including horses but the extent to which it affects UK horses is unknown. Objectives To define how liver fluke affects the UK horse population. Study design Descriptive, cross‐sectional, observational study. Methods An F. hepatica excretory‐secretory antibody detection ELISA with a diagnostic sensitivity of 71% and specificity of 97% was validated and used to analyse serum samples. An abattoir study was performed to determine prevalence. A case‐control study of 269 horses compared fluke exposure between horses with liver disease and controls. Data on clinical signs and blood test results were collected for sero‐positive horses. Genotyping of adult fluke was used to produce a multilocus genotype for each parasite. Results Four (2.2%) of 183 horses registered in the UK, sampled in the abattoir, had adult flukes in the liver, and the sero‐prevalence of F. hepatica was estimated as 8.7%. In the case‐control study, horses showing signs consistent with liver disease had significantly higher odds of testing positive for F. hepatica on ELISA than control horses. In 23 sero‐positive horses, a range of non‐specific clinical signs and blood test abnormalities was reported, with a third of the horses showing no signs. Genotypic analysis of liver flukes from horses provided evidence that these came from the same population as flukes from sheep and cattle. Main limitations Bias could have arisen in the prevalence and case‐control studies due to convenience sampling methods, in particular the geographic origin of the horses. Only a small number of horses tested positive so the data on clinical signs are limited. Conclusions Exposure to liver fluke occurs frequently in horses and may be an under‐recognised cause of liver disease. Flukes isolated from horses are from the same population as those found in ruminants. When designing and implementing parasite control plans, fluke should be considered, and horses should be tested if appropriate.