The toxicity of TiO2 engineered nanoparticles (NPs) to the marine microalga Nitzschia closterium was investigated by examining growth inhibition, oxidative stress and uptake. The results indicated that the toxicity of TiO2 particles to algal cells significantly increased with decreasing nominal particle size, which was evidenced by the 96 EC50 values of 88.78, 118.80 and 179.05 mg/L for 21 nm, 60 nm and 400 nm TiO2 particles, respectively. The growth rate was significantly inhibited when the alga was exposed to 5mg/L TiO2 NPs (21 nm). Measurements of antioxidant enzyme activities showed that superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) activities were first induced and subsequently inhibited following exposure to 5mg/L TiO2 NPs. The depletion of antioxidant enzymes with a concomitant increase in malondialdehyde (MDA) levels and reactive oxygen species (ROS) posed a hazard to membrane integrity. A combination of flow cytometry analysis, transmission electron microscopy and Ti content measurement indicated that TiO2 NPs were internalized in N. closterium cells. The level of extracellular ROS, which was induced by TiO2 NPs under visible light, was negligible when compared with the intracellular ROS level (accounting for less than 6.0% of the total ROS level). These findings suggest that elevated TiO2 nanotoxicity in marine environments is related to increased ROS levels caused by internalization of TiO2 NPs.
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