RESUMO
Caffeine (CAF) exposures have been shown to cause several pharmacological and biological effects in target and non-target organisms. Although there are already several ecotoxicological studies with CAF in non-target organisms, they are focused on marine organisms, with relevant concentrations in these ecosystems, therefore, less ecologically relevant to freshwater ecosystems (the main ecoreceptor of this type of anthropogenic contaminant). The present study aimed to assess the chronic effects (28 days) of sub-lethal and environmentally relevant concentrations of CAF (0.16, 0.42, 1.09, 2.84, 7.40, 19.23, and 50 µg/L) in Danio rerio. Biochemical endpoints as biomarkers of antioxidant defense, biotransformation, lipid peroxidation, energy sources, and neurotransmission were assessed. CAF exposure induced alterations in antioxidant defenses (superoxide dismutase and glutathione reductase activities, and glutathione content) preventing lipid peroxidation. Lactate dehydrogenase activity decreased in all the concentrations tested, while acetylcholinesterase activity was only affected by the highest concentrations tested (19.23 and 50 µg/L). We also utilized a multi-biomarker approach (Integrated Biomarker Response version 2, IBRv2) to investigate the effects of CAF in the dispersion scope of individual biochemical responses of D. rerio. IBRv2 showed that the concentration of 50 µg/L promotes the highest stress. However, the results showed that CAF induced disturbances in the metabolic pathways studied in D. rerio. These results demonstrated the toxic effects of CAF on freshwater fish, compromising their physiological functions and evidencing the need for monitoring the residues of CAF released into the inland aquatic environments. Furthermore, this research evidence that phylogenetically and physiologically different species may present different biological responses with concern for ecologically relevant environmental conditions. In this sense, the present study generated ecotoxicologically relevant data, that can be considered by environment regulators, since the here-endpoints evaluated showed sensitivity and consistency in the evaluation of caffeine risks in freshwater environments.
RESUMO
Within the scope of the European Water Framework Directive (WFD), the scientific community recognized clear opportunities to take advantage of the use of ecotoxicological tools in water quality assessments. In this perspective, bioassays and biomarkers were suggested to contribute to the integration of the chemical and biological conditions, and thus to provide an overall insight into the quality of a water body. This study aimed to assess whether current bioassays as feeding rate assays with Daphnia longispina and growth inhibition assays with Lemna minor are suitable to detect potential ecotoxicity, using waters from Portuguese reservoirs. Several sampling sites were defined in reservoirs (Miranda, Pocinho, and Alqueva). The samplings were conducted in autumn of 2018 and spring of 2019. Total chlorophyll, lipid peroxidation, and proline content were also evaluated in L. minor. Results demonstrated that D. longispina showed some sensitivity to water treatments; however, the results were difficult to interpret since no reason or trend can be accurate. All parameters of L. minor did not show sensitivity to detect potential ecotoxicological risks associated with natural water understudy, since no discrimination among the water treatments was recorded. However, biomarkers/bioassays proved to be concordant to each other. Under the conditions evaluated here (reservoirs and sampling periods), the biological responses observed were not consistent, clear, and coherent with the physical-chemical parameters and chemical analyses performed, suggesting that the ecotoxicological tools selected were not sensitive to assess water quality in this type of ecosystems. In this sense, species of different trophic levels are recommended for ecotoxicological analyses due to differences in species sensitivities.