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Seaports are introduction hotspots for invasive alien species (IAS). This is especially true for rodents, which have accompanied humans around the globe since the earliest days of ocean-going voyages. The rapid spread of IAS soon after arrival in a new environment is facilitated by further human-mediated transport or landscape features, like roads. By measuring genetic diversity and structure to investigate dispersal pathways, we gained insight into the transport, spread and establishment stages of a biological invasion, leveraging the most common rodent species (R. norvegicus) in this setting. We characterized the genetic structure of three Norway rat populations along a busy industrial road used by trucks to access the Port area in Paranaguá city (Brazil). A total of 71 rats were genotyped using 11 microsatellite markers. The results revealed a pattern of gene flow contrary to the expected stepping-stone model along the linear transect, with the two furthest apart populations being clustered together. We hypothesize that the observed outcome is explained by natural dispersal along the corridor being lower than human-mediated transport. The sampled area furthest from the port is a gas station frequented by trucks which are considered the most likely mode of transportation. In terms of management strategies, we suggest more emphasis should be put on cargo surveillance to lower the risk of Norway rat dispersal, not only for biosecurity, but also for sanitary reasons, as this port is a major grain trading point.

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