Co-exposure effects of lead and TiO2 nanoparticles in primary kidney cell culture from the freshwater fish Hoplias malabaricus.

This study evaluated the effects of Lead (Pb) and titanium dioxide nanoparticles (TiO2 NPs) alone or in combination in anterior kidney macrophages of the freshwater fish Hoplias malabaricus, naïve or stimulated with 1 ng.mL-1 lipopolysaccharide (LPS). Pb (1 ×10-5 to 1 ×10-1 mg.mL-1) or TiO2 NPs (1.5 ×10-6 to 1.5 ×10-2 mg.mL-1) reduced cell viability despite LPS stimulation, especially Pb 10-1 mg.mL-1. In combination, lower concentrations of NPs intensified Pb-induced cell viability reduction while higher concentrations restored the cell viability independently of LPS stimulation. Basal and LPS- induced NO production was reduced by both TiO2 NPs and Pb isolated. The combination of both xenobiotics avoided this reduction of NO production by the isolated compounds at lower concentrations but the protective effect was lost as the concentrations increased. None xenobiotic increase DNA fragmentation. Therefore, at specific conditions, TiO2 NPs may have a protective effect over Pb toxicity, may also provide additional toxicity at higher concentrations.

Co-exposure to titanium dioxide nanoparticles (NpTiO2) and lead at environmentally relevant concentrations in the Neotropical fish species Hoplias intermedius.

Growing production and utilization of titanium dioxide nanoparticles (NpTiO2) invariably lead to their accumulation in oceans, rivers and other water bodies, thus increasing the risk to the welfare of this ecosystem. The progressive launch of these nanoparticles in the environment has been accompanied by concern in understanding the dynamics and the toxic effect of these xenobiotic in different ecosystems, either on their own or in tandem with different contaminants (such as organic compounds and heavy metals), possibly altering their toxicity. Nevertheless, it remains unknown if these combined effects may induce damage in freshwater organisms. Therefore, this study aimed to analyze the consequences caused by NpTiO2, after a waterborne exposure of 96 h to a Neotropical fish species Hoplias intermedius, as well as after a co-exposure with lead, whose effects for fish have already been well described in the literature. The characterization of NpTiO2 stock suspension was carried out in order to provide additional information and revealed a stable colloidal suspension. As a result, NpTiO2 showed some genotoxic effects which were observed by comet assay in gill, kidney and brain cells. Also, the activity of brain acetylcholinesterase (AChE) has not changed, but the activity of muscle AChE decreased in the group exposed only to PbII. Regarding the hepatic antioxidant system, catalase (CAT) did not show any change in its activity, whereas that of superoxide dismutase (SOD) intensified in the groups submitted only to PbII and NpTiO2 alone. As for lipid peroxidation, there was a decrease in the group exposed to the NpTiO2 alone and to the co-exposed group (NpTiO2+PbII). As far as metallothionein is concerned, its concentration rose for the co-exposed group (NpTiO2+PbII) and for the group exposed to PbII alone. Overall, we may conclude that NpTiO2 alone caused DNA damage to vital tissues. Also, some impairment related to the antioxidant mechanism was described but it is probably not related to the DNA damage observed, suggesting that the genotoxic effect observed may be due to a different mechanism instead of ROS production.

Biochemical responses in freshwater fish after exposure to water-soluble fraction of gasoline.

The water-soluble fraction of gasoline (WSFG) is a complex mixture of mono-polycyclic aromatic hydrocarbons. The study aimed to evaluate the effects of WSFG diluted 1.5% on freshwater fish. Astyanax altiparanae were exposed to the WSFG for 96 h, under a semi-static system, with renewal of 25% of the gasoline test solution every 24 h. In addition, a decay of the contamination (DC) was carried out. During DC, the fish was exposed to the WSFG for 8 d, followed by another 7 d with renewal of 25% of volume aquaria with clean water every 24 h. For depuration, fish were transferred to aquaria with clean water, and in addition, 25% of the water was replaced every 24 h. The liver and kidney biotransformation, antioxidant defenses and lipid peroxidation (LPO) levels were evaluated. In the liver, the WSFG 1.5% caused reduction of glutathione S-transferase (GST) after 96 h and DC. In the kidney, only in depuration an increased GST activity was observed, and after DC a higher LPO levels. An increase of the superoxide dismutase (SOD) activity occurred at 96 h in both tissues; however, in the liver was also observed during the depuration. In WSFG 96 h, the glutathione peroxidase (GPx) activity in the kidney increased. As biomarkers of neurotoxicity, the brain and muscle acetylcholinesterase activities were measured, but the WSFG 1.5% did not change them. Therefore, this study brought forth more data about WSFG effects on freshwater fish after lower concentrations exposure and a DC, simulating an environmental contamination.

Effects of anti-inflammatory drugs in primary kidney cell culture of a freshwater fish.

The non-steroidal anti-inflammatory drugs are emerging contaminants in aquatic ecosystems. This study aimed to evaluate toxic effects of some representative drugs of this pharmaceutical group on primary culture of monocytic lineage of Hoplias malabaricus anterior kidney. The effects of diclofenac, acetaminophen and ibuprofen in cell viability, lipopolysaccharide (LPS)-induced NO production and genotoxicity were evaluated. Cytometry analysis CD11b(+) cells showed 71.5% of stem cells, 19.5% of macrophages and 9% of monocytes. Cell viability was lower in the ficoll compared to percoll separation. LPS-induced NO production by these cells was blocked after treatment with dexamethasone and NG-Methyl-L-Arginine (L-NMMA). Exposure of the cells to diclofenac (0.2-200 ng/mL), acetaminophen (0.025-250 ng/mL) ibuprofen (10-1000 ng/mL) reduced basal NO production and inhibited LPS-induced NO production at all concentrations after 24 h of exposure. Genotoxicity occurred at the highest concentration of diclofenac and at the intermediary concentration of acetaminophen. Genotoxicity was also observed by ibuprofen. In summary, the pharmaceuticals influenced NO production and caused DNA damage in monocytic cells suggesting that these drugs can induce immunosuppression and genotoxicity in fish.