Freshwater mussels are one of the most threatened taxonomic groups in the world, and many species are on the brink of local or global extinction. Human activities have altered mussel living conditions in a plethora of ways. One of the most destructive human-induced impacts on running waters is the catastrophic spill of harmful substances, which results in massive die-offs. Even though Finland is regarded as the world's top country in terms of environmental regulation quality, riverine systems are not safe. In 2014, River Kokemäenjoki in western Finland experienced the worst NiSO4 spill in the country's history, visibly affecting the mussel community -including protected Unio crassus -along the river. Because freshwater mussel toxicology is grossly understudied (particularly in Europe), any pollution -linked die-offs offer valuable opportunities to study the issue in natural environment. Here, we report the mussel investigations from 2014 and a follow-up study conducted in 2017 in order to assess the variation in species sensitivity on nickel pollution. In total, 104 sites were sampled, and over 20 000 mussels were identified and counted. Our results 2 indicate that the most impacted species (i.e. that which experienced the highest spill-induced mortality) was Anodonta anatina (62%), followed by Unio pictorum (32%), U. crassus (24%) and Unio tumidus (9%). The underlying reason for the sensitivity of A. anatina is not resolved, hence more research is urgently needed. The low mortality among most of the species in 2017 highlights the temporal nature of the pollution impact and the recovery potential of the mussel community. However, the case is more complex with U. crassus population, which may be experiencing delayed impacts of the spill. Because nickel is one of the most commonly produced industrial metals in the world (hence the pollution incident risk is high) and River Kokemäenjoki hosts mussel community typical for European rivers, our results may benefit many researchers and stakeholders dealing with riverine environments.Main finding: The freshwater mussels exhibit differences in NiSO4 pollution tolerance. Anodonta anatina is the most sensitive species, followed by Unio pictorum, Unio crassus and Unio tumidus.