Understanding how obligate freshwater organisms colonize seemingly isolated ecosystems has long fascinated ecologists. While recent investigations reveal that fish eggs can survive the digestive tract of birds and successfully hatch once deposited, evidence for avian zoochory in natura are still lacking. Here, we used a multiple lines and levels of evidence approach to demonstrate plausible bird-mediated colonization of lakes by the European perch (Perca fluviatilis). We studied a set of newly-formed artificial lakes that the public is either prohibited to access because of gravel extraction or allow to access (mainly for angling). The motivating observation is that a large proportion of prohibited-access lakes (>80%) were colonized by European perch while stocking by anglers and managers never occurred. Three supplementary lines of evidence supported avian zoochory. First, European perch spawning occurs when waterfowl abundance is very high. Second, European perch lays sticky eggs at shallow depths where they can be eaten by waterfowls or attached to their bodies. Third, genetic analyses suggested that European perch actually migrate among lakes, and that distance moved matches with daily flight range of foraging waterfowls. Together, multiple lines of evidence point to avian zoochory as a pathway for fish colonizing remote or newly-formed freshwater ecosystems.