Salmonella infection in horses can be subclinical and they can also manifest various clinical signs, especially fever, colic, diarrhea and leukopenia. The main problem in recognizing and preventing the spread of salmonellosis in horses are subclinical infected horses which occasionally shed bacteria in feces. Developing clinical salmonellosis in subclinical infected horses depends on a variety of predisposing factors such as transportation, overcrowding, sudden changes in diet, surgery, gastrointestinal disorders and antibiotic treatment. There were no researches conducted in non-specific infections salmonella in horses so far in Croatia and there are no data on the frequency of subclinical infection nor on diseases. The aim of this study was to determine the prevalence of Salmonella shedding in horses treated in clinics in the Veterinary Faculty and horses with clinical signs of colic treated in the field. There were 162 faecal samples collected from 62 horses. Samples were taken from each horse in the frequency of 24 hours. They were cultured using enrichment techniques and selective media for the isolation of Salmonella from feces. Suspicious colonies were further identified by conventional biochemical methods. Salmonella sp. was isolated from 1/62 (1.61%) examined horses. The relatively low prevalence of shedding determined in this study is possibly due to the fact that it was possible to sample the recommended five samples of feces only from a small number of examined horses. A minimum of five samples is required to achieve a satisfactory sensitivity of the examination, because Salmonella sheds intermittently. This research includes the first isolated Salmonella sp. from the faeces of horses in Croatia, and as it is isolated from the horse with typical clinical signs of salmonellosis (fever, colic, leukopenia), is also the first time that an objective diagnosis of the disease was determined. The results should serve to raise awareness about the presence of the disease to veterinarians and to lay the foundation for the implementation of biosecurity measures in the veterinary institutions with receiving horses in which they are present risk factors for shedding of Salmonella, especially horses in which there is a clinical suspicion of salmonellosis.