Effects of WS₂ Nanosheets on N₂-fixing Cyanobacteria: ROS overproduction, cell membrane damage, and cell metabolic...

The ecotoxicity of tungsten disulfide (WS₂) nanomaterials remains unclear so far. Here, the toxicity of WS₂ nanosheets on N₂-fixing cyanobacteria (Nostoc sphaeroides) was evaluated. Specifically, Nostoc were cultivated in media spiked with different concentrations of WS₂ nanosheets (0, 0.05, 0.1 and 0.5 mg/L) for 96 h. Relative to unexposed cells, WS₂ nanosheets at 0.5 mg/L significantly decreased cell density, content of total sugar and protein by 10.9 %, 0.43 %, and 6.1 %, respectively. Gas chromatography-mass spectrometry (GC-MS)-based metabolomics revealed that WS₂ nanosheets exposure altered the metabolite profile of Nostoc in a dose-dependent manner. Energy metabolism related pathways, including the Calvin-Benson-Bassham (CBB) cycle and tricarboxylic acid (TCA) cycle, were significantly inhibited. In addition, WS₂ nanosheets exposure resulted in downregulation (20-40 %) of S-containing amino acids (cystine, methionine, and cysteine) and sulfuric acid. Additionally, fatty acids and antioxidant-related compounds (formononetin, catechin, epigallocatechin, dehydroascorbic acid, and alpha-tocopherol) in Nostoc were drastically decreased by 4-50 % upon exposure to WS₂ nanosheets, which implies oxidative stress induced by the nanomaterials. Biochemical assays for reactive oxygen species (ROS) and malondialdehyde (MDA) confirmed that WS₂ nanosheets triggered ROS overproduction and induced lipid peroxidation. Taken together, WS₂ exposure perturbed carbon (C), nitrogen (N), and sulfate (S) metabolism of Nostoc, which may influence C, N, and S cycling, given the important roles of cyanobacteria in these processes. These results highlight the need for caution in the application and environmental release of WS₂ nanomaterials to prevent unintended environmental impacts due to their potential ecotoxicity.