Effects of Nanoplastics on Freshwater Biofilm Microbial Metabolic Functions as Determined by BIOLOG ECO Microplates

Nanoplastic (NP) contamination is becoming a pervasive issue as NPs, originating from microplastic particles, pose potentially harmful environmental impacts on aquatic ecosystems. The environmental hazards of NPs on microorganisms have been well documented in recent studies; however, little is known about their ecotoxicity effects on freshwater biofilms, which serve as important primary producers and decomposers and are highly connected with other ecosystem components. We investigated the effects of NPs on the microbial metabolic functions of freshwater biofilms in terms of carbon source utilization ability. Biofilm samples were collected, cultivated in a hydrodynamic flume for six weeks, and then exposed in polystyrene (PS) beads (100 nm in size) with different NP concentrations (1, 5, and 10 mg/L). BIOLOG ECO microplates were used to quantify carbon source utilization characteristics. The data were analyzed using average well-color development (AWCD), functional diversity indices, and principle component analysis (PCA). Results showed that the total carbon metabolic functions (represented by AWCD) remained constant (<i>p</i> > 0.05) with elevated NP concentrations, but some specific carbon sources (e.g., esters) changed in their utilization ability (<i>p</i> < 0.05). The microbial functional diversity (Shannon-Wiener diversity index, Simpson diversity index, and Shannon evenness index) was significantly reduced under 10 mg/L NPs (<i>p</i> < 0.05), indicating an inhibiting effect of NPs on biofilm metabolic diversity. This study examined NP ecotoxicity effects on microbial metabolic activities at the community level, but further studies are required to fully understand the mechanisms driving this change.