Natural and uninterrupted water courses are important for biodiversity and fish population stability. Nowadays, many streams and rivers are obstructed by artificial migration barriers, often preventing the migration of fish. On the other hand, distribution of pathogens by migrating fishes is still a point of concern. Pathogen transport and transmission is a driving force in the dynamics of many infectious diseases. The aim of this study was to investigate the possible consequences of the removal of an artificial migration barrier for the upstream transport of Tetracapsuloides bryosalmonae, the causative agent of Proliferative Kidney Disease (PKD) in brown trout, by migrating fish. To test this question, a river system was selected with a migration barrier separating a PKD positive river from a PKD negative tributary. After removal of the barrier, PKD prevalence and pathology was examined during five years after elimination of the barrier. In the tributary, no PKD was recorded at any time of the survey. By means of unidirectional PIT (passive integrated transponder)-tagging, we confirmed upstream migration of adult brown trout into the tributary during the cold season, presumably for spawning. By eDNA, we confirmed presence of T. bryoalmonae and Fredericella sp., the definitive host, DNA in water from the PKD positive river stretch, but not in the PKD negative tributary. Our study illustrates the importance of the connectivity of streams for habitat maintenance. Although migration of brown trout from a PKD-positive river into a PKD-negative tributary, mainly for spawning, was confirmed, upstream spreading of PKD was not observed.