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As an artificial type of microbial carrier, plastic debris has been widely detected in freshwater habitats, and the potential impacts of the plastisphere (biofilms colonized on plastics) in aquatic ecosystems have drawn increasing attention. Distinct community compositions and structures of biofilms in plastic and natural substrates have been recorded in freshwater environments. However, the microbial metabolic functioning of the plastisphere was underestimated, especially in freshwater environments. In this study, the effects of subst