RESUMO
The aim of this study was to evaluate and compare the effects on biochemical parameters and organosomatic indices in the freshwater bivalve Diplodon chilensis exposed to a glyphosate-based formulation under direct and dietary exposures (4 mg a.p./L). After 1, 7, and 14 days of exposure, reduced glutathione (GSH) and thiobarbituric acid reactive substances (TBARS) levels and the activities of glutathione-S- transferase (GST), superoxide dismutase (SOD), and catalase (CAT) were evaluated in the gills and digestive gland. The hepatosomatic (HSI) and branchiosomatic (BSI) indices were also analyzed. Direct and dietary glyphosate-based formulation exposure altered the redox homeostasis in the gills and digestive gland throughout the experimental time, inducing the detoxification response (GST), the antioxidant defenses (SOD, CAT, GSH), and causing lipid peroxidation. After 14 days of exposure, the HSI and BSI increased significantly (43% and 157%, respectively) only in the bivalves under direct exposure. Greater changes in the biochemical parameters were induced by the dietary exposure than by the direct exposure. Furthermore, the gills presented an earlier response compared to the digestive gland. These results suggested that direct and dietary exposure to a glyphosate-based formulation induced oxidative stress in the gills and digestive glands of D. chilensis. Thus, the presence of glyphosate-based formulations in aquatic ecosystems could represent a risk for filter-feeding organisms like bivalves.
Assuntos
Bivalves , Poluentes Químicos da Água , Animais , Glifosato , Exposição Dietética , Ecossistema , Estresse Oxidativo , Catalase/metabolismo , Superóxido Dismutase/metabolismo , Peroxidação de Lipídeos , Brânquias/metabolismo , Glutationa Transferase/metabolismo , Poluentes Químicos da Água/metabolismo , Biomarcadores/metabolismoRESUMO
In the Upper Valley of Río Negro and Río Neuquén in Argentina, agriculture represents the second most important economic activity. Azinphos-methyl has been found in water from this region throughout the year at a maximum concentration of 22.48 µg L(-1) during the application period. Toxicological studies on local non-target species have been performed mostly on vertebrates, while mollusks, which could be more sensitive, have not been studied so far. This work aims to characterize cholinesterase (ChE) and carboxilesterase (CE) activities of Chilina gibbosa, a freshwater gastropod native to southern Argentina and Chile. These enzymes, together with neurotoxicity signals, are evaluated herein after as sensitive biomarkers of exposure to azinphos-methyl at environmentally relevant concentrations. Effects of azinphos-methyl on antioxidant defenses: glutathione (GSH), catalase (CAT), superoxide dismutase (SOD) and glutathione S-transferase (GST) are also studied in order to complete a set of biomarkers with different sensitivity and specificity, to propose C. gibbosa as a sentinel species. The highest specific activity was obtained with acetylthiocholine as substrate, followed by propionylthiocholine (83% in comparison to acetylthiocholine) and butyrylthiocholine (19%).The lowest Km and the highest efficiency for ChE were obtained with acetylthiocholine. Regarding CEs activities, a higher efficiency was obtained with p-nitrophenyl butyrate than with p-nitrophenyl acetate. Eserine produced significant inhibition of ChE activity (81% with 0.001 mM and 98% with 1mM) while iso-OMPA did not produce any significant effect on ChE. Our results show that C. gibbosa ChE is very sensitive to azinphos-methyl (CI50 0.02 µg L(-1)) while CEs are inhibited at higher concentrations (CI50 1,000 µg L(-1)). CEs have been reported to be more sensitive to OPs than ChEs in most of the aquatic invertebrates protecting the organisms from neurotoxic effects. In contrast, C. gibbosa, has ChE which are much more sensitive to azinphos-methyl than CEs and shows marked signs of neurotoxicity. Regarding antioxidant defenses, GSH levels were significantly increased by 0.02 and 20 µg L(-1) azinphos-methyl (80 and 103%, respectively), CAT activity was increased 85% only at 0.02 µg L(-1) and SOD and GST did not show any significant response. Since ChE activity, neurotoxicity signs, GSH and CAT are sensitive biomarkers of acute exposure to azinphos-methyl at environmental concentrations C. gibbosa could be included as sentinel species in monitoring programs of pesticide hazard in regions of Argentina and Chile.