The transmission of Yersinia pseudotuberculosis in the pork production chain was followed from farm to slaughterhouse by studying the same 364 pigs from different production systems at farm and slaughterhouse levels. In all, 1,785 samples were collected, and the isolated Y. pseudotuberculosis strains were analyzed by pulsed-field gel electrophoresis. The results of microbial sampling were combined with data from an on-farm observation and questionnaire study to elucidate the associations between farm factors and the prevalence of Y. pseudotuberculosis. Following the same pigs in the production chain from farm to slaughterhouse, we were able to show similar Y. pseudotuberculosis genotypes in live animals, pluck sets (containing tongue, tonsils, esophagus, trachea, heart, lungs, diaphragm, liver, and kidneys), and carcasses and to conclude that Y. pseudotuberculosis contamination originates from the farms, is transported to slaughterhouses with pigs, and transfers to pluck sets and carcasses in the slaughter process. The study also showed that the high prevalence of Y. pseudotuberculosis in live pigs predisposes carcasses and pluck sets to contamination. When production types and capacities were compared, the prevalence of Y. pseudotuberculosis was higher in organic production than in conventional production and on conventional farms with high rather than low production capacity. We were also able to associate specific farm factors with the prevalence of Y. pseudotuberculosis by using a questionnaire and on-farm observations. On farms, contact with pest animals and the outside environment and a rise in the number of pigs on the farm appear to increase the prevalence of Y. pseudotuberculosis.