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Ships and boats may transport whole communities of non-indigenous species (NIS) through hull biofouling, some members of which may become invasive. Several studies have evaluated the diversity of these communities, but very few have analyzed the survival of organisms after their voyages into different and potentially inhospitable conditions. This factor is important to consider because the last port of call approach for risk assessments assumes that if the conditions observed in the last port of call are different from those observed in a receiving port, risks are diminished or null. Using an innovative experimental system, we tested the survival and recovery of the marine blue mussel (Mytilus edulis) and the freshwater zebra mussel (Dreissena polymorpha) by exposing them to adverse salinity conditions at varying temperatures to simulate ships and boats transiting to ports or marinas with contrasting environmental conditions. Both mussel species, which are well-known for their adaptability to new environments as aquatic NIS, survived better at colder temperatures, with blue mussels surviving up to 14 days in freshwater, and zebra mussels up to 8 days in marine water. This highlights the importance of considering the resistance of fouling organisms to adverse conditions in vector and species risk assessments.