Photochemical degradation increases polycyclic aromatic hydrocarbon (PAH) toxicity to the grouper Epinephelus marginatus as assessed by multiple biomarkers.

The effects of halogen-light-irradiated and non-irradiated PAHs on the grouper Epinephelus marginatus were assessed through biomarkers including morphometric parameters, liver histopathology, biliary PAH concentration, genetic alterations, and enzyme activity modulation. E. marginatus juveniles were divided into three groups: control (C), non-irradiated PAHs (PAHs1), and irradiated PAHs (PAHs2). Test groups were exposed for 14 days to a 0.5 ppm PAH solution in the semi-static system. After this period, fish were anesthetized with benzocaine (2%) and peripheric blood was collected by caudal puncture. Blood smears were prepared and stained with propidium iodide. Fish livers were collected, fixed in McDowell's solution, embedded in paraplast, thin-sectioned, and stained with hematoxylin-eosin (H&E). For biochemical analyses including superoxide dismutase, catalase, and glutathione S-transferase activities, fish livers were collected and preserved in liquid nitrogen. Water samples were analyzed using gas chromatography-mass spectrometry (GC-MS) and bile synchronous fluorescence spectroscopy. Fish in the PAHs2 group had micronuclei (MN) in blood cells, as well as significant differences in nuclear morphology (NMA). Significant morphological alterations were observed in the livers from fish exposed to PAHs as well as inhibition of the catalase activity. Our results show that irradiation altered the bioavailability of PAHs, especially benzanthracene, which has great impact in aquatic ecosystems. Among the consequences of physical and chemical changes to PAHs, we observed a significant increase in NMA and MN incidence in E. marginatus erythrocytes, indicating the potential initiation of mutagenic and carcinogenic processes.

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.

Antioxidant defense responses in Mytella guyanensis (Lamarck, 1819) exposed to an experimental diesel oil spill in Paranaguá Bay (Paraná, Brazil).

We evaluated the effects of diesel oil on the bivalve Mytella guyanensis using biomarkers of oxidative stress (glutathione S-transferase, glutathione peroxidase, and reduced glutathione) after an experimental in situ spill in a mangrove area in southern Brazil. A linear model was developed for the Multiple Before-After Control-Impact (MBACI) experimental design to assess the significance of biological responses. Control and impacted sites were sampled seven and two days before as well as two and seven days after the spill. With the exception of a late response of reduced glutathione (GSH) levels on day seven, none of the biomarkers were significantly altered by the impact. This result was attributed to the high environmental variability of the experimental sites combined with a low sensitivity of Mytella guyanensis to diesel oil at short time-scales. The high resistance of M. guyanensis suggests that its antioxidant response is triggered only after a medium- to long-term exposure to contaminants.

Saxitoxins induce cytotoxicity, genotoxicity and oxidative stress in teleost neurons in vitro.

The aim of this study was establish a protocol for isolation and primary culture of neurons from tropical freshwater fish species Hoplias malabaricus for assessment of the effects of neurotoxic substances as saxitoxins (STXs). Cells from brain of H. malabaricus were treated with different concentrations of trypsin, dispase and papain for tissue dissociation. Cells type was separated by cellular gradient and basic fibroblast growth factor (bFGF) supplement nutrition media were added. The dissociated cells were plated with medium and different STXs concentrations and the toxic cellular effects such as oxidative stress, neurotoxicity, and genotoxicity and apoptosis process were evaluated. Cultures treated with bFGF showed the greatest adherence, survival and cellular development. STXs increased specific activity of glutathione peroxidase and lipoperoxidation levels, were cytotoxic and genotoxic indicated by the comet assay. Although the STXs effects due the blockage of sodium channels is reported to be reversible, the time exposure and concentration of STXs suggested cellular injuries which can lead to neuropathology. The establishment of primary neuronal culture protocol enables new applications for neurotoxicological assessments.