The aim of this study was to evaluate the occurrence of Salmonella spp. in fattening pigs in a slaughterhouse in the Czech Republic. Tonsils, mesenteric lymph nodes, and caecal contents were sampled from 120 pigs originating from eight farms. A total of 360 samples were examined. Cultivation methods were used to detect Salmonella. Suspect isolates were subjected to phenotypic identification. Serotyping was performed by slide agglutination method. The multiplex polymerase chain reaction (PCR) was used to detect genes encoding flagellar proteins. Salmonella spp. was isolated in 3 out of 360 samples (0.83%). The results proved the presence found a relatively low prevalence of Salmonella in fattening pigs at the Czech slaughterhouse, consistent implementation of Salmonella control program during pork production is critical in order to ensure the protection of consumer health.