In this study, Vallisneria natans plants were exposed to 5 and 20 nm of titanium dioxide nanoparticles (TiO2 NPs) anatase and 600-1000 nm of bulk at 5 and 20 mg/L for 30 days. SEM images and EDX spectra revealed that epiphytic biofilms were more prone to TiO2 NPs adhesion than bare plant leaves. TiO2 NPs injured plant leaf cells, ruptured epiphytic diatoms membranes and increased the ratio of free-living microbes. The TN, NH4+-N and NO3--N concentrations significantly decreased, respectively, by 44.9%, 33.6%, and 23.6% compared to bulk treatments after 30 days due to macrophyte damage and a decline in diversity of epiphytic bacterial community and abundance of nitrogen cycle bacteria. TiO2 NPs size-dependent decrease in bacterial relative abundance was detected, including phylum Cyanobacteria, Planctomycetes, and Verrucomicrobia. Although TiO2 NPs increased eukaryotic diversity and abundance, abundances of Bacillariophyceae and Vampyrellidae classes and Gastrotricha and Phragmoplastophyta phylum decreased significantly under TiO2 NPs exposure compared to bulk and control. TiO2 NPs reduced intensities of interaction relationships among epiphytic microbial genera. This study shed new light on the potential effects of TiO2 NPs toxicity toward aquatic plants and epiphytic microbial communities and its impacts on nitrogen species removal in wetlands.
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