Microbial carbon metabolic functions of biofilms on plastic debris influenced by the substrate types and environmental factors

[Display omitted] •Microbial functioning of biofilms on plastics and natural substrates were studied.•Biofilms on plastics showed higher biomasses than that of natural substrates.•Distinct carbon metabolic activities of biofilms inhabited on plastics were observed.•Environmental factors dominated the metabolism differentiation over substrate types.•Plastic debris are new microbial niche affecting carbon cycling in aquatic ecosystems. 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 substrate types on the carbon metabolic functions of biofilms were studied by in situ cultivation of biofilms on plastics (polyvinyl chloride, PVC and polyethylene, PE) and natural substrate (cobblestone) for 44 days in two rivers (the Niushoushan River and the Qinhuai River) and two lakes (Donghu Lake and Xuanwu Lake). Biofilms on plastics showed higher biomasses than those on natural substrates in all ecosystems. Variations in the micro-structure and compactness of biofilms developed under different substrates were observed from scanning electron microscope and confocal laser scanning microscope image analyses. The carbon metabolic activities of the biofilms evaluated by BIOLOG EcoPlate were different between plastics (PVC and PE) and natural substrate (cobblestone) in the four freshwater ecosystems. In the Niushoushan River, PE-associated biofilms had different capacity in using carbon sources from cobblestone-associated biofilms as illustrated by the Shannon-Wiener diversity index and Shannon evenness index. Additionally, the metabolic functional diversity profiles of biofilms on PVC were significantly different from those on cobblestone in the other three aquatic ecosystems. Moreover, results from variation partitioning analysis suggested that the impact of environmental factors (contribution: 21%) on microbial carbon metabolic functions was much greater than that of substrate types (contribution: 6%). These findings illustrated distinct microbial functions of biofilms inhabited on plastics, and