This study investigated the effects of different doses of 17-beta-estradiol (E(2)) in Rhamdia quelen. Groups of males exposed to different doses of E(2) (0.1 mg kg(-)(1), 1 mg kg(-)(1) and 10 mg kg(-)(1)) were compared with non-exposed male and female fish groups. Among the considered biomarkers, no significant differences were observed for micronuclei test, reduced glutathione concentration and lipid peroxidation. All E(2)-treated individuals had decreased glutathione S-transferase activity. Increased catalase and superoxide dismutase activities, increased vitellogenin expression and decreased metallothionein concentration were observed in males treated with the highest dose. Liver of all test groups showed necrotic areas, but cytoplasm vacuolization was again found only in the individuals exposed to highest dose. E(2) causes deleterious hepatic effects to R. quelen, and vitellogenin expression, catalase and superoxide dismutase activity and metallothionein concentration represent appropriate biomarkers for studying E(2) effects. Additionally, the response of some biomarkers was similar in males exposed to E(2) and unexposed females, and therefore exposure to endocrine disruptors may cause consequences for fish populations.
Many tropical freshwater ecosystems are impacted by cyanobacteria blooms increasing the risk of cyanotoxins exposure to aquatic organisms while human populations may be exposed by eating fish, drinking water, or dermal swimming. However, few toxicological data are available on the influence of cyanobacteria blooms in particular, cylindrospermopsin (CYN) on Brazilian neotropical fish. A number of studies demonstrated the ability of CYN to bioaccumulate in freshwater organisms and consequently enter the human food chain. The aim of the current study was to examine the effects of CYN following single intraperitoneal injection (50 µg/kg) of purified CYN (CYNp) or aqueous extract of CYN-producing cyanobacteria extract (CYNex) after 7 or 14 days. Biomarkers such as histopathology (liver), oxidative stress (liver and brain), and acetylcholinesterase (AChE) activity (muscle and brain) were utilized in order to assess the influence of CYN on Hoplias malabaricus. In terms of AChE activity, administration of CYNex and CYNp both muscle and brains were used as target tissues. In brain an increase of glutathione S-transferase (GST) activity and lipid peroxidation (LPO) levels was noted suggesting an imbalance in redox cycling. The majority of biomarkers did not present significant alterations in liver, but an elevation in superoxide dismutase (SOD) and glucose 6 phosphate dehydrogenase (G6PDH) activities was found. Different profiles of GST activity were observed in both studied groups (CYNex and CYNp) while LPO (CYNex and CYNp) and protein carbonylation (PCO) (CYNp) levels increased after exposure to CYN. The incidence of necrosis, melanomacrophages centers, and free melanomacrophages were detected as evidence of cell death and immune responses. Nonprotein thiols (NPT) levels were not markedly affected in both exposed groups. Data demonstrated that in vivo exposure to CYN produced biochemical and morphological disturbances in liver and brain of H. malabaricus.
Human hepatoma cells (HepG2) were exposed to purified cylindrospermopsin (CYN), a potent toxicant for eukaryotic cells produced by several cyanobacteria. Exposure to 10 μg l of CYN for 24 h resulted in alteration of expression of 48 proteins, from which 26 were identified through mass spectrometry. Exposure to 100 μg l of CYN for 24 h affected nuclear area and actin filaments intensity, which can be associated with cell proliferation and toxicity. The proteins are implicated in different biological processes: protein folding, xenobiotic efflux, antioxidant defense, energy metabolism and cell anabolism, cell signaling, tumorigenic potential, and cytoskeleton structure. Protein profile indicates that CYN exposure may lead to alteration of glucose metabolism that can be associated with the supply of useful energy to cells respond to chemical stress and proliferate. Increase of G protein-coupled receptors (GPCRs), heterogeneous nuclear ribonucleoproteins (hnRNP), and reactive oxygen species (ROS) levels observed in HepG2 cells can associate with cell proliferation and resistance. Increase of MRP3 and glutathione peroxidase can protect cells against some chemicals and ROS. CYN exposure also led to alteration of the expression of cytoskeleton proteins, which may be associated with cell proliferation and toxicity.
The knowledge concerning associations between chronic chemical exposure and many disorders with complex etiology involving gene-environment interactions is increasing, and new methods must be developed to improve water quality monitoring. The complexity of chemical mixtures in polluted aquatic environments makes the evaluation of toxic potential in those sites difficult, but the use of biomarkers and bioindicators has been recognized as a reliable tool to assess risk of exposure to biota and also the human population. In order to evaluate the use of fish and biomarkers to assess toxic potential and bioavailability of chemicals in human-related hydric resources, an in situ experiment was accomplished in two water reservoirs designated for human supply, which were previously evaluated by the local environmental regulatory agency through a set of physical, chemical, and classical biological parameters. Molecular, biochemical, and morphological biomarkers were performed in caged Oreochromis niloticus kept for 6 months in the studied reservoirs to assess potentially useful biomarkers to evaluate the quality of water for human supply. Chemical analysis of toxic metals in liver and muscle and polycyclic aromatic hydrocarbons (PAHs) in bile was considered to assess the bioavailability of pollutants and highlight human activity impact. The reservoir previously classified by a governmental agency as less impacted presented more risk of exposure to biota. These results were supported by chemical analysis, vitellogenin expression, histopathological findings (gonads, liver, and gills), as well as indicators of neurotoxic effects and oxidative stress in liver. The inclusion of some biomarkers as parameters in regulatory monitoring programs in reservoirs designated for human supply is strongly suggested to evaluate the risks of exposure to the human population. Thus, a revision of the traditional biological and physicochemical analysis utilized to establish the conditions of water quality is necessary.
Endocrine-disrupting chemicals (EDCs) are widespread used and can interfere on hormone regulation with adverse consequences for both biota and human. Vitellogenin (vtg) is a yolk precursor protein synthesized by the liver in response to estrogen. In order to characterize the vtg of tropical fish Rhamdia quelen and establish a molecular biomarker, adult male individuals were exposed to 17-β-estradiol (E) for vtg induction and anti-R. quelen vtg polyclonal antibodies production. Vitellogenic female fish were used as positive control group. E-induced vtg was characterized as a glycolipophosphoprotein of high molecular mass with peptide mass fingerprint very similar in E-exposed male and vitellogenic female fish. A polyclonal serum containing anti-R. quelen vtg antibodies was produced and showed high specificity and sensibility to detect the vtg of three fish species: R. quelen, Piaractus mesopotamicus and Prochilodus lineatus. Wildlife and laboratory studies reported that EDCs released into the environment may alter the levels of plasma vtg in male fish, making this protein a valuable biomarker of xenoestrogens exposure. Then, we propose the use of anti-R. quelen vtg as a tool for biomonitoring studies and water quality assessment in Brazil and South American countries where the three fish species occur.
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