Antioxidant enzyme activities as biomarkers of fluvial biofilm to ZnO NPs ecotoxicity and the Integrated Biomarker Responses (IBR) assessment

• Toxicity of ZnO NPs on biofilm was studied.
• ZnO NPs reduced biofilm viability and increase ROS generation.
• Antioxidant enzymes are sensitive biomarkers of ZnO NPs toxicity.

The presence of ZnO nanoparticles (ZnO NPs) in natural waters has raised concerns about their environmental impacts, but the potential influences of ZnO NPs on fluvial biofilm have not been reported. In this study, the utility of antioxidant enzyme activities (AEA) as biomarkers of fluvial biofilm to ZnO NPs toxicity and a method that combines AEA into an index of “Integrated Biomarker Responses (IBR)” were studied. Compared with the absence of ZnO NPs, scanning electron microscopy (SEM) images revealed that a large amount of ZnO NPs were adsorbed onto biofilm and these NPs exerted adverse effects on the viability of bacteria in biofilm. The production of reactive oxygen species (ROS) with high concentrations (30 and 100 mg/L) of ZnO NPs exposure reached to 184% and 244% of the control, while no cell leakage and membrane damage were observed. After exposure to ZnO NPs for 0.25 and 3 days, the activities of catalase (CAT), superoxide dismutase (SOD) and glutathione reductase (GR), glutathione peroxidase (GSH-Px) were significantly increased, respectively. At the end of exposure period (21 days), the AEA with the presence of 1 mg/L ZnO NPs exposure were comparable to the control, while most of those in high concentrations of ZnO NPs were decreased. The results of IBR showed that the biofilm can adapt to 1 mg/L ZnO NPs exposure, while be seriously damaged by 30 and 100 mg/L ZnO NPs after 3 and 0.25 days. IBR can be used as an appropriate evaluation system of the toxicity effects of ZnO NPs on fluvial biofim.

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