Small plastic particles, microplastics (MPs) and nanoplastics (NPs) represent a major threat in aquatic environments. Freshwater organisms are exposed to MPs and NPs, particularly in industrial and urban areas. The present study aimed to compare the toxicity between polystyrene NPs (PS NPs) and environmental microplastics (ENV MPs) and nanoplastics (ENV NPs) generated from macro-sized debris collected in the Garonne River on the freshwater bivalve C. fluminea. The organisms were exposed to the different plastic particles at three environmentally relevant concentrations: 0.008, 10, and 100 μg L−1 for 21 days. The biological responses of organisms were assessed using a multi-biomarker approach from the sub-individual to the individual level. The results demonstrated that: i) ENV NPs triggered more effects on detoxification processes and immune response, confirming that using manufactured NPs for laboratory exposure can lead to misleading conclusions on the risks posed by plastic particles; ii) effects of ENV MPs were less marked than ENV NPs, emphasizing the importance of testing a size continuum of plastic particles from NPs to MPs; iii) some effects were only observed for the low and/or intermediate concentrations tested, underlining the importance of using environmentally relevant concentrations. In light of these results, laboratory studies should be continued by exposing aquatic species to environmental MPs and NPs. The properties of these particles have to be characterized for a better risk assessment of environmental plastic particles.< Réduire