rGO outperforms GO in generating oxidative stress and DNA strand breaks in zebrafish liver cells.

Graphene oxide (GO) and reduced graphene oxide (rGO) are both widely applicable and there is a massive production throughout the world which imply in inevitable contamination in the aquatic environment by their wastes. Nevertheless, information about their interaction at the cellular level in fish is still scarce. We investigated the metabolic activity, reactive oxygen species (ROS) production, responses of antioxidant defenses, and total antioxidant capacity (TAC) as well as oxidative stress and DNA integrity in zebrafish liver cells (ZFL) exposed to (0.001, 0.01, 0.1 and 1 µg mL-1) of GO and rGO after two exposure period (24 and 72 h). Higher ROS production and no significant changes in the antioxidant defenses resulted in lipid peroxidation in cells exposed to rGO. Cells exposed to GO increased the activity of antioxidant defenses sustaining the TAC and avoiding lipid peroxidation. Comet assay showed that both, GO and rGO, caused DNA strand breaks after 24 h of exposure; however, only rGO caused DNA damage after 72 h of exposure. The exposure to rGO was significantly more harmful to ZFL cells than GO, even at very low concentrations. The cells showed a high capacity to neutralize ROS induced by GO preventing genotoxic effects and metabolic activity, thus sustaining cell viability. The time of exposure had different impacts for both nanomaterials, GO caused more changes in 24 h showing recovery after 72 h, while cells exposed to rGO were jeopardized at both exposure times. These results indicate that the reduction of GO by removal of the oxygen functional groups (rGO) increased toxicity leading to adverse effects in the cells, even at very low concentrations.

Concentration- and time-dependence toxicity of graphene oxide (GO) and reduced graphene oxide (rGO) nanosheets upon zebrafish liver cell line.

Graphene oxide (GO) and reduced graphene oxide (rGO) are carbon-based nanomaterials that have a wide range of applicability. Therefore, it is expected that their residual traces reach the aquatic environment, accumulate, and interact with its different compartments and the biota living in them. The concentration- and time-dependency response to GO and rGO in aquatic organisms are still poorly known. In the present study, the effects of GO and rGO on zebrafish hepatocytes were investigated using in vitro assays performed with established liver cell lines from zebrafish (ZFL). GO and rGO nanosheets were applied on ZFL cells at a concentration range of 1-100 µg mL-1 for 24 and 72 h. The internalization of GO and rGO nanosheets, reactive oxygen species (ROS) production, cell viability, and cell death were evaluated. The internalization of GO increased as the concentrations of GO increased. The rGO nanosheets were smaller than GO nanosheets, and their hydrophobic characteristic favors their interaction with the cell membrane. However, the rGO nanosheets were not observed in the uptake assay. Exposure for 72 h was found to cause harmful effects in ZFL cells, causing higher ROS production in cells exposed to rGO and stopping cell replication. Nevertheless, GO did not stop cell replication, but exposed cells had higher levels of apoptosis and necrosis. After 72 h, both GO and rGO were toxic, but with different mechanisms of toxicity.

Reactive oxygen species and other biochemical and morphological biomarkers in the gills and kidneys of the Neotropical freshwater fish, Prochilodus lineatus, exposed to titanium dioxide (TiO2) nanoparticles.

This study investigated the action of titanium dioxide nanoparticles (TiO2-NPs), on the gills and kidneys of Neotropical freshwater fish, Prochilodus lineatus, with emphasis on reactive oxygen species (ROS) production, antioxidant responses, and morphological changes. Fish were exposed to 1, 5, 10, and 50 mg L-1 nominal TiO2-NPs suspended into water for 2 or 14 days. In gills, ROS decreased and glutathione (GSH) increased after 2 days, while ROS and GSH increased and superoxide dismutase activity decreased after 14 days. In kidneys, GSH and lipoperoxidation increased after 2 days and catalase activity decreased after 14 days. Common histopathologies in gills were epithelium hyperplasia, cellular hypertrophy, proliferation of mitochondria-rich cells (MRC), and lamellar stasis; in kidneys, there were cellular and nuclear hypertrophy, focal tubule degeneration, necrosis, and melanomacrophage (MM) proliferation. Although environmentally unlikely, high-dose exposures clarified biological effects of TiO2-NPs, such as ROS formation and MRC responses in the gills, which may impair ionic balance. It was also found that MM are likely responsible for eliminating NPs in the kidney. These findings will help to regulate TiO2-NP disposal, but longer-term studies are still needed.

Changes in the blood parameters of the Amazonian manatee (Trichechus inunguis) after long-distance transportation / Alterações nos padrões sanguíneos do peixe-boi da Amazônia (Trichechus inunguis) após transporte de longa distância

In this study we report the hematological, biochemical and hormonal parameters in a juvenile male Amazonian manatee measured before transport, immediately after transport, and during adaptation to a new facility. The animal was transported from Manaus, Amazonas State, Brazil, to São Paulo, São Paulo State, Brazil, (2,733 km) within 6 hours. Among all blood parameters analyzed, we observed obvious neutrophilia, lymphopenia, and increases in the neutrophil/lymphocyte ratio and serum glucose and aspartate aminotransferase (AST) levels, but these parameters subsequently returned to normal. These results suggest that transport and changes in the environment are temporary stressful events for Amazonian manatees. We, therefore, recommend monitoring the hematological and biochemical parameters before and after translocation to minimize the effects of handling stressors in this species.

Neste estudo, são relatados os parâmetros hematológicos, bioquímicos e hormonais em um peixe-boi da Amazônia macho jovem, medidos antes do transporte, imediatamente depois, e durante a sua adaptação a um novo recinto. O animal foi transportado de Manaus, Estado do Amazonas, Brasil, para São Paulo, Estado de São Paulo, Brasil, (2.733 km) durante 6h de viagem. Entre os parâmetros sanguíneos analisados, foi observada clara neutrofilia, linfopenia e aumento na relação neutrófilo/linfócito e nos níveis de glicose e aspartato aminotransferase (AST), mas estes parâmetros posteriormente voltaram aos valores normais. Estes resultados sugerem que o transporte e as mudanças no ambiente são eventos estressantes para o peixe-boi da Amazônia. Desta forma, é recomendável o monitoramento dos parâmetros hematológicos e bioquímicos antes e após a translocação para minimizar os efeitos de agentes estressores nesta espécie.