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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. Only screening a pregnant mother is not satisfactory to prevent mother-to-child transmission of HI