10 resultados para EROD activity
em Repositório Institucional UNESP - Universidade Estadual Paulista "Julio de Mesquita Filho"
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Pós-graduação em Química - IBILCE
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Pós-graduação em Genética - IBILCE
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Among the sexual hormones the estrogens are receiving major attention due to their biological activity. Such biological response is atributed to the best conformation recognized by their receptors, resulting in maximum responses. The estrogens are also considered responsible for most of disruptor´s effects caused by their presence in aquatic ecosystems. The 17β-estradiol hormone is produced by vertebrates during the reproductive phase of their lives and its presence has been detected in superficial waters. The objective of this study was to evaluate the cause-effect of tilapia exposition to the hormone 17β-estradiol through the analysis of biotransformation enzymes in liver and gills, complemented with the quantification of 17β-estradiol and estrone in water samples collected from fish ponds integrated to swine production. The present study was conducted under laboratory conditions, in a randomized experimental design with three levels of 17β-estradiol (E2) (0, 5, 15 µg L-1), with three replicates. After 7 days of exposure time, liver and gills were extracted to analyze three isoforms of cytochrome P450: EROD, BROD, PROD and the activity of Glutathione S-Transferase (GST). The results showed that the EROD activity (CYP1A), normally induced by the metabolism of aromatic compounds, did not present statistical differences among the treatments exposed to E2, what means that the hormone did not induce isoform 1A in fish under these particular experimental conditions. PROD activity was significantly altered in both concentrations, by means of 5 and 15 µg L-1, when compared to control. This result can indicate an important role of PROD on the metabolism of E2 present in water. Regarding to the BROD activity, it could be observed differences statistically significant between control and both groups of treatments. Two or more CYP isoforms can contribute to the metabolism of the same compound, what makes BROD a candidate as a next bioindicator of the exposure to E2 in aquatic ecosystems. Analysis of variance could confirm the effect of E2 statistically significant on the GST activity in liver tissues with >90% of significance (Prob>F = 0.0753). Furthermore, it was possible to observe that the values of GSTs activities in liver and gills in both, control and treatments, follow a tendency, that means, enzymatic activity in gills increase as the increasing of the activity in the liver tissues. In this study, the 17β–estradiol was found in measurable concentrations in three sampled points, and these values were similar to the findings of other authors at different locations in Brazil. In addition, those values are much higher than the minimum concentration that presented observable effects (10 ng L-1).
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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In the aquatic environment, biotransformation enzymes are established biomarkers for assessing PAH exposure in fish, but little is known about the effect of 17β-estradiol (E2) on these enzymes during exposure to benzo(a)pyrene (BaP). In this study, Nile tilapia (Oreochromis niloticus) were exposed for 3, 5, and 10 days to BaP (300 μg L(-1)) and E2 (5 μg L(-1)). These substances were applied isolated or mixed. In the mixture experiment, fish were analyzed pre- and postexposure in order to better understand whether preexposure to the hormone masks the responses activated by PAH or vice versa. Phase I enzymes ethoxyresorufin-O-deethylase (EROD), pentoxyresorufin-O-depenthylase (PROD), and benzyloxyresorufin-O-debenzylase (BROD) activities as well as the phase II enzyme glutathione S-transferase (GST) were analyzed. Isolated E2 treatment decreased EROD activity after 3 days, but this enzyme activity returned to control values after 5 and 10 days of exposure. Isolated BaP treatment significantly induced EROD activity after 3 and 5 days, and the activity returned to control levels after ten exposure days. Combined treatment (E2 + Bap) significantly increased EROD activity, both in the pre- and postexposure. This increase was even higher than in the isolated BaP treatment, suggesting a synergism between these two compounds. When E2 and BaP were used singly, they did not change BROD and PROD activities. However, combined treatment (E2 + Bap) significantly increased PROD activity. Isolated BaP treatment increased GST activity after 10 days. However, this response was not observed in the mixture treatment, suggesting that E2 suppressed the GST induction modulated by BaP. The results put together indicated that E2 altered the biotransformation pathway regarding enzymes activated by BaP in Nile tilapia.
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Oil refinery effluents contain many chemicals at variable concentrations. Therefore, it is difficult to predict potential effects on the environment. The Atibaia River (SP, Brazil), which serves as a source of water supply for many municipalities, receives the effluents of one of the biggest oil refinery of this country. The aim of this study was to identify the (eco)toxicity of fresh water sediments under the influence of this oil refinery through neutral red (cytotoxicity) and ethoxyresorufin-O-deethylase (EROD) assays (AhR-mediated toxicity) in RTL-W1 cells (derived from fish liver). Once the refinery captures the waters of Jaguarí River for the development of its activities and discharges its effluents after treatment into the Atibaia River, which then flows into Piracicaba River, sediments from both river systems were also investigated. The samples showed a high cytotoxic potential, even when compared to well-known pollution sites. However, the cytotoxicity of samples collected downstream the effluent was not higher than that of sediments collected upstream, which suggested that the refinery discharges are not the main source of pollution in those areas. No EROD activity could be recorded, which could be confirmed by chemical analyses of polycyclic aromatic hydrocarbons (PAHs) that revealed a high concentration of phenanthrene, anthracene, fluoranthene, and pyrene, which are not EROD inducers in RTL-W1 cells. In contrast, high concentrations of PAHs were found upstream the refinery effluent, corroborating cytotoxicity results from the neutral red assay. A decrease of PAHs was recorded from upstream to downstream the refinery effluent, probably due to dilution of compounds following water discharges. On the other hand, these discharges apparently contribute specifically to the amount of anthracene in the river, since an increase of anthracene concentrations could be recorded downstream the effluent. Since the extrapolation of results from acute toxicity to specific toxic effects with different modes of action is a complex task, complementary bioassays covering additional specific effects should be applied in future studies for better understanding of the overall ecotoxicity of those environments.