Lethal and sublethal toxicity of pesticides and vinasse used in sugarcane cultivation to Ceriodaphnia silvestrii (Crustacea: Cladocera).

With the growing use of agrochemicals in Brazil, there is also a growing need for more realistic toxicity assessments that aid in understanding the potential risks of environmental-realistic agrochemical (mixture) exposures in the natural ecosystems. The aim of the present study was therefore to evaluate the lethal and sublethal effects of environmental realistic (single and mixture) concentrations of the pesticides DMA® 806 BR (active ingredient - a.i. 2,4-D) and Regent® 800 WG (a.i. fipronil) and sugarcane vinasse to the Neotropical cladoceran Ceriodaphnia silvestrii. This evaluation was carried out through lethal (survival), sublethal (reproduction and intrinsic rates of population increase - r) and post-exposure (feeding rate and also reproduction) tests conducted in situ and with water from mesocosms contaminated with the recommended doses of these compounds. The results showed high acute toxicity for treatments containing fipronil and vinasse when acting in isolation, with survival rates only returning to control values on the last sampling day (75 days post application). Reproduction of surviving cladocerans was reduced in all treatments until the end of the experiment and were potentiated effect in the mixture of the three test compounds. The intrinsic rates of population increase were reduced in all treatments except the single 2,4-D treatment. Post-exposure feeding rate and reproduction, however, were not impaired under the conditions analyzed. The results show the high toxicity of recommended doses of fipronil and vinasse (and especially their mixture) and the importance of evaluating the risks of agrochemical mixtures at environmental-realistic concentrations.

Biochemical effects of fipronil and its metabolites on lipid peroxidation and enzymatic antioxidant defense in tadpoles (Eupemphix nattereri: Leiuperidae).

Amphibians are very sensitive to environmental change and pollution because they have both aquatic and terrestrial life cycle stages and high skin permeability. Particularly during the larval stages, when these animals are restricted to small, transient ponds, exposure to high concentrations of pesticides is inevitable in agricultural areas. Given that pesticide application increases during the summer, which coincides with the reproductive season and the occurrence of most neotropical tadpoles in their natural environment, strong indications exist that tadpoles are developing in contaminated ponds. Fipronil is one of the primary insecticides used in sugarcane cultivation in Brazil, and little is known about its toxic effects on non-target organisms such as tadpoles. The purpose of this study was to evaluate the effects of fipronil and its metabolites on oxidative stress in Eupemphix nattereri tadpoles after exposure in water and sediment at concentrations of 35, 120 and 180µgkg-1. We assessed the activities of the antioxidant enzymes glutathione S-transferase (GST), glucose 6-phosphate dehydrogenase (G6PDH) and catalase (CAT) and lipid peroxidation (malondialdehyde, MDA). The results showed that fipronil has an inherent capacity to cause oxidative stress in tadpoles, as evidenced by a decrease in CAT activity and an increase in lipid peroxidation levels at all concentrations tested. Fipronil sulfone also produced elevated MDA levels at two of the tested concentrations and increased G6PDH activity in tadpoles exposed to the highest concentration of this metabolite but did not affect MDA levels. Our data showed that fipronil and its degradation products promoted oxidative stress in Eupemphix nattereri tadpoles exposed to environmentally relevant concentrations and could lead to a decrease in the long-term physiological performance of these animals, leading to detrimental effects at the population level.

Influence of Temperature on the Thyroidogenic Effects of Diuron and Its Metabolite 3,4-DCA in Tadpoles of the American Bullfrog (Lithobates catesbeianus).

Temperature is a key variable affecting the timing of amphibian metamorphosis from tadpoles to tetrapods, through the production and subsequent function of thyroid hormones (TH). Thyroid function can be impaired by environmental contaminants as well as temperature. Tadpoles can experience large temperature fluctuations in their habitats and many species are distributed in areas that may be impacted by agriculture. Diuron is a widely used herbicide detected in freshwater ecosystems and may impact endocrine function in aquatic organisms. We evaluated the influence of temperature (28 and 34 °C) on the action of diuron and its metabolite 3,4-dichloroaniline (3,4-DCA) on thyroid function and metamorphosis in tadpoles of Lithobates catesbeianus. Exposure to both compounds induced more pronounced changes in gene expression and plasma 3,3',5-triiodothyronine (T3) concentrations in tadpoles treated at higher temperature. T3 concentrations were increased in tadpoles exposed to 200 ng/L of diuron at 34 °C and an acceleration of metamorphosis was observed for the same group. Transcriptomic responses included alteration of thyroid hormone induced bZip protein (thibz), deiodinases (dio2, dio3), thyroid receptors (trα, trß) and Krüppel-like factor 9 (klf9), suggesting regulation by temperature on TH-gene expression. These results suggest that environmental temperature should be considered in risk assessments of environmental contaminants for amphibian species.

Antioxidant Defense System of Tadpoles (Eupemphix nattereri) Exposed to Changes in Temperature and pH.

Amphibians are highly susceptible to environmental changes, mainly at the larval stage during which they are restricted to small and ephemeral aquatic habitats, which are subject to large fluctuations of abiotic parameters, such as temperature and pH. Consequently, tadpoles experience changes in biochemical, physiological, and molecular processes related to the maintenance of homeostasis, which may lead them to an oxidative stress state. In the present study, we investigated the effects of stress caused by changes in temperature and pH on the antioxidant enzymes catalase (CAT), glucose-6-phosphate dehydrogenase (G6PDH), glutathione reductase (GR) and glutathione-S-transferase (GST) in tadpoles of Eupemphix nattereri. The results show that changes in temperature and pH conditions induce an antioxidant response in tadpoles. GST and GR showed temperature-dependent activities; GST activity was higher in tadpoles exposed to 28°C, whereas GR exhibited increased activity in response to 28°C and 36°C. At 32°C, both GST and GR had the lowest activity. CAT was induced by treatments with acidic (pH 5.0) and alkaline (pH 8.5) pH. Tadpoles exposed to acidic pH also had increased GR activity. The G6PDH was not changed in either experiment. Our data demonstrate that E. nattereri possesses an efficient antioxidant defense system for coping with the damaging effects of heat and acidity/alkalinity conditions in water. The alterations in antioxidant enzymes are probably a result of immediate physiological adaptation of individuals in response to increased production of ROS under environmental stress conditions.