Estudo de variações bioquímicas e genotoxicidade induzidas por mistura de contaminantes em tilápia (Oreochromis niloticus), como biomarcadores de contaminação ambiental

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Enzimas de biotransformação em tilápia (Oreochromis niloticus) exposta ao hormônio natural 17b-estradiol

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).

PROD and BROD modulation in Nile tilapia after exposure to 17 beta-estradiol

The activities of 7-ethoxyresorufin-O-deetylase (EROD), 7-benzyloxyresorufin-O-debenzylase (BROD), 7-pentoxyresorufin-O-depentilase (PROD), and glutathione S-transferase (GST) were measured in Nile tilapias exposed for 7 days of 5 and 15 μg/L 17 β-estradiol. EROD and GST activities were unchanged. PROD activity increased in animals exposed to the higher dose of the hormone, while BROD was increased after 7 exposure days to both doses of the compound. These results indicate the usefulness of these enzymes as biomarkers for 17 β-estradiol exposure.