Toxicity of fipronil and 2,4-D pesticides in Daphnia similis: a multiple endpoint approach.

In Brazil, among the pesticides widely applied simultaneously in sugarcane monocultures are the Regent® 800 WG insecticide (active ingredient (a.i.) fipronil) and the DMA® 806 BR herbicide (a.i. 2,4-D). Thus, this study aimed to investigate, through different endpoints, the effects of the fipronil and 2,4-D pesticides, isolated and as mixtures, on the cladoceran Daphnia similis. To do this, acute toxicity tests were carried out with the compounds acting in isolation and in mixture, where the survival of the organisms was evaluated, and chronic toxicity tests with the isolated compounds, where reproduction and maternal and neonatal body length were evaluated. In this study, the physiological endpoints of D. similis were also analyzed, through the analysis of feeding rates (filtration and ingestion) in exposure and post-exposure scenarios, in order to verify the cladoceran food recovery capacity. In addition, D. similis data were compared with other species when exposed to the studied pesticides, using species sensitivity distribution curves. Acute toxicity tests of the fipronil and 2,4-D showed an average EC50-48 h of 66.68 µg a.i./L and 327.07 mg a.i./L, respectively. In both cases, D. similis showed lower sensitivity compared to other species. For the mixture test, the evaluation by the IA model (independent action) and deviation DR (dose ratio dependent) indicated the occurrence of mostly antagonistic effects. The chronic test with fipronil showed a decrease in the fecundity of the organism at a concentration of 16 µg a.i./L, a concentration already found in aquatic environments. For 2,4-D, no significant differences were observed for reproduction at the concentrations tested. Regarding the maternal body length, there were no significant changes when D. similis were exposed to both fipronil and 2,4-D, but these differences were observed in the body length of the neonates only for 2,4-D. There were no significant changes in the feeding rates of the organisms when exposed to both pesticides.

Short and long-term exposure to the pesticides fipronil and 2,4-D: Effects on behavior and life history of Daphnia magna.

The high levels of contamination in aquatic ecosystems caused by pesticides and the organisms' consequent continuous exposure to it has made them vulnerable to damage. However, mobile organisms can avoid this continued exposure to contaminants by moving to less disturbed habitats. Therefore, through the use of the Heterogenous Multi-Habitat Assay System (HeMHAS), our objective was to evaluate the ability of Daphnia magna to detect and avoid habitats contaminated by fipronil and 2,4-D, in a spatially connected landscape. Further, the role of contamination by these pesticides, isolated and in mixtures, concerning the colonization of habitats by daphnids was also evaluated. Given that not all organisms successfully escape contamination, the chronic toxicity of the same pesticides using different parameters for D. magna (maternal survival, fecundity and maternal body length) was also evaluated. When evaluating the avoidance response by D. magna exposed to pesticides, there was no preference for the less contaminated areas for both compounds. However, organisms did not move to contaminated zones in the colonization experiments, with no immigration of daphnids to the zones with intermediate and the highest levels of fipronil, nor to the highest concentration of 2,4-D. Finally, the colonization by daphnids was significantly prevented when exposed to a mixture of the pesticides, in which the areas with the highest combinations of pesticide concentrations were not colonized by D. magna. Regarding the long-term chronic effects, negative consequences were observed, particularly for maternal body length, fecundity and maternal survival, due to the exposure to fipronil. Considering that pesticides can limit the areas colonized by organisms by making them unattractive, the risk of local population extinction may be underestimated if only standard endpoints involving forced exposure are studied.

Avoidance responses by Danio rerio reveal interactive effects of warming, pesticides and their mixtures.

Temperature variations and thermal extremes events caused by climate change can have profound implications for the toxicity of pesticides in aquatic organisms. Using an innovative system (Heterogeneous Multi-Habitat Test System - HeMHAS) that allows the simulation of different scenarios within a spatially heterogeneous landscape, the effects on the habitat selection of Danio rerio fish caused by the pesticides fipronil and 2,4-D were studied as single compounds and in mixture and integrated with air temperature variation (20, 24 and 28 °C). As a result, D. rerio detected and avoided both pesticides at air temperatures of 20 and 24 °C; however, at 28 °C no significant difference was observed in habitat choice by fish. Additionally, when pesticides were mixed in a heterogeneously contaminated landscape, it was observed that D. rerio detected contamination and preferred the clean zone at 20 and 24 °C; however, at 28 °C the potential to escape from the most contaminated areas was impaired. Thus, contamination by both pesticides made the habitat selection behavior of fish at 20 and 24 °C more noticeable. In addition, the association between pesticides and temperature showed negative effects on the response of fish to detect and escape from contaminated environments, suggesting the influence of temperature in altering the ability of the organism to provide an efficient response to stress.

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.

Effects of diuron and carbofuran pesticides in their pure and commercial forms on Paramecium caudatum: The use of protozoan in ecotoxicology.

Toxic effects of diuron and carbofuran on Paramecium caudatum were evaluated. Acute and chronic tests were conducted with diuron and carbofuran active ingredients and their commercial formulations, Diuron Nortox(®) 500 SC and Furadan(®) 350 SC, respectively. The sensitivity range of P. caudatum to reference substance sodium chloride was established. A preliminary risk assessment of diuron and carbofuran for Brazilian water bodies was performed. The tests indicated that toxicity of pure diuron and its commercial formulation was similar, while the commercial product carbofuran was more toxic than its pure form. In acute tests, readings were carried out at 2, 3, 4 and 6 h and showed an increase of mortality with increasing exposure time. The sensitivity of P. caudatum to NaCl ranged from 3.31 to 4.44 g L(-1), averaging 3.88 g L(-1). For diuron, the 6 h LC50 was 64.6 ± 3.3 mg L(-1) for its pure form and 62.4 ± 2.5 mg L(-1) for its commercial formulation. Carbofuran active ingredient was less toxic than that of diuron, presenting a 6 h LC50 of 142.0 ± 2.4 mg L(-1) for its pure form and 70.4 ± 2.2 mg L(-1) for its commercial product. Chronic tests showed that these pesticides cause significant decrease on population growth, generation number and biomass of P. caudatum. The 24 h IC50 was 7.10 ± 0.58 mg L(-1) for pure diuron, 6.78 ± 0.92 mg L(-1) for commercial diuron, 22.95 ± 3.57 mg L(-1) for pure carbofuran and 4.98 ± 0.62 mg L(-1) for commercial carbofuran. Preliminary risk assessment indicated that diuron and carbofuran present potential ecological risks for Brazilian water bodies. P. caudatum was a suitable and sensitive test organism to evaluate diuron and carbofuran toxicity to freshwater protozooplankton and, taking into account the relevant role of protozoans in aquatic environments, we strongly recommend its inclusion in ecotoxicological studies.