Isolated and mixed effects of pure and formulated abamectin and difenoconazole on biochemical biomarkers of the gills of Danio rerio.

Pesticides are released into the environment daily, and their effects on nontarget species in aquatic ecosystems have been widely reported. To evaluate the adverse effects caused in adults of Danio rerio species exposed to the pesticides abamectin, difenoconazole, and their commercial formulations (Kraft 36EC® and Score 250EC®), both isolated and in mixtures, biochemical biomarkers were analyzed in the gills of organisms exposed to sublethal concentrations. To this end, the activities of the enzymes 7-ethoxyresorufin-O-deethylase (EROD), glucuronosyltransferase (UDPGT), glutathione-S-transferase (GST), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), lipid hydroperoxide (LH), and malondialdehyde (MDA), which are indicative of oxidative stress, were measured after 48 h of exposure to the different pesticide treatments. The results showed a significant increase in EROD activity and MDA levels in the gills of fish exposed to the commercial formulation of abamectin. When the fish were exposed to difenoconazole and its commercial formulation, an increase in GST, GPx, and MDA levels and a decrease in GR activity were observed in the gills. Furthermore, the responses of the biomarkers were more pronounced in organisms exposed to mixtures of both active ingredients and commercial formulations. It is concluded that the commercial formulations Kraft 36EC® and Score 250EC® and their mixtures cause significant alterations in the detoxification metabolism of exposed organisms and induce oxidative stress in fish.

Proposal for a tiered regulatory framework for the aquatic risk assessment of pesticides in Brazil.

Aquatic risk assessment is essential to guarantee the sustainable use of pesticides and the conservation of water resources near agricultural fields. This article discusses a proposal for a tiered regulatory framework for the aquatic risk assessment of pesticides in Brazil. The first step is problem formulation, which includes establishing general and specific protection goals. In the exposure assessment, the Estimated Environmental Concentrations in water should be calculated based on realistic worst-case assumptions regarding application rate and frequency, the entry into the edge-of-field water body, and fate in the water body, using scenario-dependent models suggested by the Brazilian Environmental Agency. These calculations can be refined by including Efate studies with variable exposures to reflect realistic environmental conditions accurately and include mitigation measures that impact the modeling. In the hazard assessment, ecotoxicological data for toxicity to fish, aquatic invertebrates, algae, and aquatic plants should be required for all pesticides based on standardized protocols and species. Tier 2 has several refinement options, including incorporating toxicity data from additional test species and effect modeling. In Tier 3, population- and community-level effects are evaluated using semi-field studies. Considering the case study in Brazil, Tier 1 demonstrated that, from the 12 pesticides that were assessed, seven (58%) failed based on the value of the Risk Quotient. In Tier 2, when exposure refinement options and mitigation measures such as buffer zones are considered, all seven pesticides, for which Tier 1 indicated risk, still failed the assessment. The risk for four of these seven pesticides could be refined by considering toxicity information from additional species. Refinement options and mitigation measures that could be applied to the agricultural scenario in Brazil were discussed. In conclusion, the proposed tiered risk assessment is a feasible way to evaluate whether a pesticide will pose an unacceptable risk to aquatic organisms. Integr Environ Assess Manag 2024;00:1-15. © 2024 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Residual determination and acute toxicity of the neonicotinoid clothianidin in the neotropical stingless bee Tetragonisca angustula Latreille, 1811 (Apidae: Meliponini).

Bees play a crucial role as natural pollinators, ensuring the maintenance and stability of the world's biodiversity and agricultural crops. Native bees in neotropical regions belong to the Meliponini tribe, a larger group that differs significantly in behavior and biology from honeybees (e.g., Apis mellifera) and solitary bees (e.g., Osmia spp.). Hence, the exposure and effects of pesticides is also likely to vary among these different species. The aim of this study was to develop an analytical method to determine the presence of the neonicotinoid clothianidin in the Brazilian native stingless bee Tetragonisca angustula (local common name: Jataí). The method used for the chemical analysis involved a QuEChERS technique combined with UHPLC-MS/MS analysis. The developed method was subsequently used to analyze collected field samples. In addition, the acute toxicity of the pesticide to T. angustula was evaluated in a laboratory bioassay evaluating both lethal and sublethal endpoints. The analytical method was successfully developed with detection and quantification limits of 1.55 and 5 µg L-1, respectively, along with a linear range of 1-5 ng mL-1. Clothianidin was detected in environmental samples (9.2-32.9 ng g-1), and the exposure experiments demonstrated acute oral toxicity to adults of T. angustula, (24 h-LD50 of 0.16 ng a.i./bee), as well as no significative interference in acetylcholinesterase activity. Considering the obtained toxicity endpoints for T. angustula and those reported in the literature for other bee species, this study revealed that T. angustula is more (lethally) sensitive to clothianidin than other bee species, including those commonly used in environmental risk assessment studies. This thus also supports the call for using native test species in (regional) risk assessment evaluations.

Ecological risk assessment for metals in sediment and waters from the Brazilian Amazon region.

Pollution by metals is a matter of concern around the world. In recent decades, the high population growth in urban centers has significantly magnified the entry of these pollutants into aquatic ecosystems. The Amazon region, intense migratory flow, gold mining, and industrialization have been considered the main driving forces for increasing metal pollution. Thus, the main aim of this study is to conduct, for the first time, an Ecological Risk Assessment (ERA) based on metal concentrations measured in the sediment and water of several aquatic environments from the Amazon basin, based on the risk quotient values (RQ = measured environmental concentration - MEC/predicted no effect concentration - PNEC). In addition, the metal contamination factor (CF) was estimated. Although metal concentrations in water were generally low, these values were far above the limits established by current national legislation in many areas, showing higher concentrations for the metals Co, Pb, Cr, Cu, and Ni. Concentrations of Mn, Cu, Ba, Pb, Co, Ni, Cr, Zn, Cd, and As were especially high in the sediment for several evaluated environments. The ERA for the water compartment revealed that 56% of the studied areas presented high risk (RQ > 1) for aquatic biota. In the sediment, 66% of the sites presented a high risk and 40% medium risk (RQ = 0.1-1). The CF indicated that 49% of the sampling points had high contamination and only 24%, had low contamination. These results reveal that monitoring studies in the Amazon region, provides important information so that public policies for the preservation of water resources can be strengthened in the Amazon.

Abamectin and difenoconazole monitoring in strawberry flowers and pollen sampled from Tetragonisca angustula (Latreille) (Hymenoptera: Apidae) hives located in crop vicinities.

The increase in agricultural productivity associated with the emergence and the extensive use of pesticides is undeniable. However, strong evidence indicates that this continuous demand is causing serious environmental impacts and bringing toxic effects to associated biota as pollinating insects. The present work aims the determination of the insecticide abamectin (ABA) and the fungicide difenoconazole (DIF) in strawberry flowers (Fragaria x ananassa DUCH.) and pollen sampled from beehives of the stingless bee Tetragonisca angustula Latreille (Hymenoptera: Apidae) located nearby strawberry fields. For analysis, QuEChERS method was optimized, and the analytical performance of those two pesticides was verified. Then, the method was applied to strawberry flowers and the pollen was sampled during three field campaigns. While abamectin was not detected, the systemic fungicide difenoconazole was determined in almost all flowers and pollen samples, demonstrating the major persistence of this pesticide in investigated matrices. The results were then discussed about the difenoconazole application rate and transport to colonies to estimate a preliminary environmental risk assessment for stingless native bees. All calculations were proceeded considering exposure rates and toxicity data from the literature, adapted from Apis mellifera studies. In this sense, the determination, application, and discussion about risk assessment figure out as an important tool to the knowledge about the preliminary risks of native bees exposed to pesticides.

Toxicity of Binary Metal Mixtures to the Tropical Ostracod Strandesia Trispinosa.

The ecotoxicity of metals is generally assessed individually, in part because current knowledge does not allow for the accurate prediction of the toxicity of metal mixtures to aquatic organisms. The objective of this study was to investigate the toxic effects of binary combinations of metal salts (copper sulphate-CuSO4, cadmium chloride-CdCl2, mercury chloride-HgCl2 and manganese sulphate-MnSO4) on the tropical ostracod Strandesia trispinosa through acute toxicity tests. To this end, ostracods were exposed to each individual metal salt as well as to their combinations by applying a full factorial design. The model that best explained the effects of the mixtures CuSO4 x CdCl2, CuSO4 x HgCl2 and CuSO4 x MnSO4 on the survival of S. trispinosa was Concentration Addition, whereas this was Independent Action for the CdCl2 x HgCl2 mixture. The observed synergistic interactions are likely to result in unacceptable risks to aquatic ecosystems under real field conditions. This is especially the case if CuSO4 predominates the metal mixture, as observed for its combination with mercury and manganese.

Toward a tiered regulatory framework for the prospective aquatic effect assessment of pesticides in (Neo)tropical areas.

Research and regulatory interest into (Neo)tropical aquatic pesticide ecotoxicology has increased considerably over the past few years. However, pesticide effect assessment frameworks in (Neo)tropical areas remain largely undeveloped and as such not implemented. The present study provides an overview of studies into the comparison of pesticide sensitivity between (Neo)tropical and temperate aquatic taxa. Recommended assessment factors to be applied to temperate toxicity data for use in tropical effect assessments differ significantly between these studies. Shortcomings and bottlenecks of available pesticide sensitivity comparison studies based on results from bioassays, species sensitivity distributions, and model ecosystem studies are scrutinized. Current data lacks and (subsequent) indications for future research are also covered. Ultimately, implications for procedures aimed at the derivation of regulatory acceptable concentrations for pesticides in (Neo)tropical edge-of-field surface waters are discussed. Integr Environ Assess Manag 2023;19:638-647. © 2022 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Responses of Chironomus sancticaroli to the simulation of environmental contamination by sugarcane management practices: Water and sediment toxicity.

Sugarcane management practices include the application of pesticides, including the herbicide 2,4-D and the insecticide fipronil. In addition, a by-product from the ethanol industry, called vinasse, is commonly applied to fertilize sugarcane areas. The potential risks of these practices to the edge-of-field aquatic ecosystems were assessed in the present study. This was done by contaminating mesocosms with (single and mixtures of) both pesticides and vinasse and evaluating the effects on the midge Chironomus sancticaroli through in-situ and laboratory bioassays. To this end, outdoor mesocosms were treated with fipronil (F), 2,4-D (D), and vinasse (V) alone and with the mixture of fipronil and 2,4-D (M), as well as with both pesticides and vinasse (MV). C. sancticaroli was deployed in mesocosms before contamination in cages, which were taken out 4- and 8-days-post-contamination. Water and sediment samples were also taken for laboratory bioassays on the first day of contamination, as well as 7-, 14-, 21-, 30-, 45-, and 75-days post-contamination. The responses assessed in subchronic assays (8-day) were survival, growth, head capsule width, development, and mentum deformities. Low survival occurred in the in-situ experiments of all treatments due to the low oxygen levels. In the laboratory tests, effects on survival occurred for F, V, and M over time after exposure to both water and sediment. All organisms died post-exposure to water samples from the MV treatment, even 75-days-post-contamination. Impairments in body length and head capsule width occurred for F, V, and M for water and F, V, M, and MV for sediment samples over time. All treatments increased mentum deformities in exposed larvae for any of the sampling periods. The negative effects observed were more significant in the mixture mesocosms (M and MV), thus indicating increased risks from management practices applying these compounds together or with a short time interval in crops.

Influence of temperature on the toxicity of the elutriate from a pesticide contaminated soil to two cladoceran species.

Brazil has become one of the largest consumers of pesticides in the world. However, there are still few studies evaluating pesticide toxicity integrating local aquatic and terrestrial environments. In addition, there is growing concern about the influence of temperature conditions related with climate change on contaminants toxicity. The aim of the present study was to evaluate the elutriate toxicity of the insecticide Kraft® 36 EC (a.i. abamectin), the fungicide Score® 250 EC (a.i. difenoconazole) and their mixture to the cladocerans Ceriodaphnia silvestrii and Daphnia similis, using model ecosystems (mesocosms). To this end, mesocosms were filled with natural soil and subjected to the following treatments: Control (Milli-Q water), Kraft (10.8 g abamectin ha-1), Score (20 g difenoconazole ha-1), and Kraft + Score (10.8 g abamectin ha-1 + 20 g difenoconazole ha-1). The experiment lasted 18 days, and the applications were made on days 1, 8, and 15; the occurrence of rainfall was simulated on days 1, 8, and 15 after applications and only rainfall simulation on days 4, 11, and 18. The experiment was conducted under two different temperatures: 23 °C and 33 °C. At 23 °C, single Kraft treatment and in combination with Score showed high toxicity to both cladocerans. At 33 °C, elutriate of the Kraft® and mixture treatments were highly toxic to D. similis but not to C. silvestrii. The results indicate that while Kraft had higher toxicity than Score to both cladocerans, this toxicity was counteracted at 33 °C only for the exotic species, D. similis. The results portray the complexity of pesticide toxicity when considering realistic experimental settings including different organisms and temperature treatments.

Modified QuEChERS Method for Extracting Thiamethoxam and Imidacloprid from Stingless Bees: Development, Application, and Green Metrics.

In the present study, a method for the determination of residues of the neonicotinoid insecticides thiamethoxam and imidacloprid in the stingless bee Melipona scutellaris Latreille (1811) was optimized through a factorial design, tested using green metrics, and then applied to exposed bees. It combines the extraction with a modified quick, easy, cheap, effective, rugged, and safe method and the determination by liquid chromatography-tandem mass spectrometry analysis. Different parameters such as the mass of the sample, dispersive sorbents, and elution solvents were assessed. Method validation parameters were checked and include sensitivity, specificity, and linearity. The limit of quantification of 0.0025 µg g-1 was obtained for both insecticides, where accuracy was 94%-100% with satisfactory intraday and interday precisions (relative standard deviation <10%). The qualified method was applied to orally and topically exposed bee samples, and the results indicated that it is suitable for the determination and quantification of neonicotinoid pesticide residues in this species. Moreover, green analytical metrics like the National Environmental Methods Index, Eco Scale score, high-performance liquid chromatography with an environmental assessment tool (HPLC-EAT), waste generation, and amount of sample were compared with methods described in the literature involving neonicotinoid analysis in honeybees. As a result, the present study displayed the highest Eco Scale score and HPLC-EAT score and the second smallest amount of sample and waste generated. Thus, the method meets green analytical metrics more than other methods. In this sense, besides the application, the multicriteria decision analysis tool employed suggests that this is a good option as a green analytical method. Environ Toxicol Chem 2022;41:2365-2374. © 2022 SETAC.

Chronic environmentally relevant levels of pesticides disrupt energy reserves, feeding rates, and life-cycle responses in the amphipod Hyalella meinerti.

When pesticides reach the aquatic environment, they can distribute in water and sediment, increasing the risks to benthic organisms, such as amphipods that play a key role in the aquatic food webs. Thus, the present study assessed the consequences of exposure to the insecticide fipronil and herbicide 2,4-D (alone and in mixture) on biochemical markers, feeding rates and the partial life-cycle of Hyalella meinerti. Three concentrations of fipronil (0.1, 0.3, and 0.7 µg L-1) and 2,4-D (19, 124, and 654 µg L-1), and six mixture combinations were assessed. The first experiment was carried out with males and females separately assessing the feeding rates, total carbohydrate content, and lipid profile. The second (partial life-cycle) lasted 49 days, and the survival, growth, and reproductive endpoints were assessed. Both pesticides and their mixture caused decreases in feeding rates, mainly in females. Females also suffered a change in the total carbohydrate content. In addition, there were changes in the percentage of triacylglycerol and phospholipids in males and females. Furthermore, alterations occurred in the percentual of triacylglycerol and phospholipids to both sexes. In the second experiment, fipronil and the mixtures caused decreases in the survival of H. meinerti over time. Exposure to 2,4-D, fipronil, and their mixture impaired the 28-day growth leading to biomass loss ranging from 17-23%, 54-60%, and 22-49%, respectively. The insecticide and mixture caused increases in time to sexual maturation of up to 10 and 6 days, respectively, and reduced the number of formed couples. Furthermore, fipronil decreased reproduction up to 36 times and no juveniles were produced in some mixture combinations. In addition, the pesticides on isolation decreased the juvenile size. Finally, exposure to both pesticides, alone or in a mixture, decreased the intrinsic rate of population growth. The results were observed in concentrations already quantified in water bodies, with risks for ecosystems functioning due to the importance of amphipods in aquatic ecosystems.

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.

Bioaccumulation and chronic toxicity of arsenic and zinc in the aquatic oligochaetes Branchiura sowerbyi and Tubifex tubifex (Annelida, Clitellata).

Oligochaetes feed on bulk sediment and penetrate the sediment through the construction of burrows, making them especially vulnerable to sediment metal contamination. However, the few oligochaete species that have been tested to date are almost exclusively temperate test species. Although the warmwater adapted species Branchiura sowerbyi has been indicated as a promising candidate for tropical sediment toxicity testing, few (especially chronic) studies have been conducted so far to confirm this. Therefore, the aim of the present study was to evaluate the bioaccumulation and chronic 28d lethal and sublethal toxicity of arsenic (As) and zinc (Zn) to both the warmwater-adapted B. sowerbyi and the coldwater-adapted oligochaete Tubifex tubifex for comparison. Arsenic was more toxic to both oligochaete species than Zn. Inter- and intra-species variability in toxicity values of the two test species and other benthic invertebrates was within an order of magnitude. However, B. sowerbyi was the most sensitive species to As even for sediment concentration (EC50: 36.6 ± 2.1 µg/g and 147.1 ± 21.7 µg/g, for B. sowerbyi and T. tubifex, respectively) and for tissue concentration (ER50: 9.2 ± 0.9 µg/g and 887.0 ± 35.0 µg/g, for B. sowerbyi and T. tubifex, respectively). Finally, the Tissue Residue-effects Approach (TRA) using Effective Tissue Residues appears to be a promising way forward in advancing in this since it considers internal body concentrations.

Acute toxicity of the insecticide abamectin and the fungicide difenoconazole (individually and in mixture) to the tropical stingless bee Melipona scutellaris.

Stingless bees have been recognized as essential plant pollinators and producers of various natural products in neotropical areas. Research into the potential risks of pesticides they may be exposed to in agricultural fields, however, remains meagre. Especially the toxicity of pesticide mixtures likely to occur under real-world conditions and that are likely to exert synergetic effects has been poorly studied. The aim of the present study was therefore to evaluate the single and mixture acute contact and oral toxicity of commercial products containing the insecticide abamectin and the fungicide difenoconazole in laboratory bioassays with the Brazilian native stingless bee Melipona scutellaris. In addition, a comparison of the insecticide sensitivity of stingless bees relative to the honeybee Apis mellifera was made based on previously published toxicity data. Except for oral exposure to abamectin, M. scutellaris appeared to be more sensitive that A. mellifera in the single compound toxicity tests. A difenoconazole concentration at the NOEC (no observed effect concentration) level indicated a synergetic toxic interaction with abamectin. A sensitivity comparison based on published toxicity data for A. mellifera and stingless bees indicated several insecticidal modes of action having a high relative sensitivity to stingless bees that need especial consideration in future studies. The research findings highlight the need for testing native bee species and environmentally relevant pesticide mixtures in risk assessments to avoid underestimation of potential risks to bee populations and the subsequent loss of pollination ecosystem services.

Multi-generational exposure to fipronil, 2,4-D, and their mixtures in Chironomus sancticaroli: Biochemical, individual, and population endpoints.

Conventional farming delivers a range of pesticides to aquatic ecosystems leading to implications for the indigenous species. Due to the multiple applications and persistence of molecules, organisms may be exposed for a prolonged period over multiple generations. The present study outlines a full life-cycle design over three generations of Chironomus sancticaroli exposed to the insecticide fipronil, the herbicide 2,4-D, and their mixtures. The experiment started with newly hatched larvae from the parental generation and lasted with the emerged adults from the second generation. Five nominal concentrations of fipronil and 2,4-D were tested, as well as six combinations of both pesticides. As additional responses, the total carbohydrates and the lipid classes were evaluated in the parental generation. The first and second generations were more susceptible to the tested compounds compared with the parental ones. Survival of larvae and pupae was decreased by both pesticides and their mixtures along with the generations. Only fipronil impaired the survival of emerged adults. Both pesticides (isolated and in the mixture) altered the emergence and the fraction of males and females. Moreover, the number of eggs produced, and their hatchability decreased. Only one combination of the pesticides increased the content of carbohydrates. Fipronil, 2,4-D, and its mixture altered the profile of the lipid classes. All mixture treatments and the three highest concentrations of fipronil extinguished the population of C. sancticaroli at the end of the first generation. In the remaining treatments with the insecticide, the population did not survive the second generation. Only three concentrations of 2,4-D and the control persisted until the end of the experiment. The results indicate that a prolonged exposition to these pesticides may disrupt the natural populations of exposed organisms with consequences to ecosystems' functioning, considering the importance of chironomids to aquatic and terrestrial environments.

Study of the Potential of Water Treatment Sludges in the Removal of Emerging Pollutants.

Presently, water quantity and quality problems persist both in developed and developing countries, and concerns have been raised about the presence of emerging pollutants (EPs) in water. The circular economy provides ways of achieving sustainable resource management that can be implemented in the water sector, such as the reuse of drinking water treatment sludges (WTSs). This study evaluated the potential of WTS containing a high concentration of activated carbon for the removal of two EPs: the steroid hormones 17ß-estradiol (E2) and 17α-ethinylestradiol (EE2). To this end, WTSs from two Portuguese water treatment plants (WTPs) were characterised and tested for their hormone adsorbance potential. Both WTSs showed a promising adsorption potential for the two hormones studied due to their textural and chemical properties. For WTS1, the final concentration for both hormones was lower than the limit of quantification (LOQ). As for WTS2, the results for E2 removal were similar to WTS1, although for EE2, the removal efficiency was lower (around 50%). The overall results indicate that this method may lead to new ways of using this erstwhile residue as a possible adsorbent material for the removal of several EPs present in wastewaters or other matrixes, and as such contributing to the achievement of Sustainable Development Goals (SDG) targets.

Toxicity of fipronil and 2,4-D formulations (alone and in a mixture) to the tropical amphipod Hyalella meinerti.

Conventional farming uses a large volume of pesticides that may reach aquatic ecosystems. This is also the case for the insecticide fipronil and the herbicide 2,4-D, which are widely used in many crops. This study aimed at evaluating the individual and mixture toxicity of these pesticides to the tropical amphipod Hyalella meinerti. To this end, acute toxicity tests (96 h) were conducted. Chronic bioassays (10 days) were also carried out, in which the body length and dry biomass were evaluated as endpoints. In addition, a complete factorial mixture chronic toxicity test was carried out. H. meinerti was sensitive to fipronil in the acute toxicity tests, with a LC50-96-h of 0.86 µg L-1 (95% CI 0.26-0.46), and no acute effects were observed after 2,4-D exposure even at the highest test concentration of 100 mg L-1. In the chronic toxicity tests, all tested concentrations of both pesticides decreased the growth of H. meinerti, in which losses on biomass reached 45% and 65% for 2,4-D and fipronil, respectively. The pesticide mixture indicated antagonism although it still significantly decreased the body growth. The results obtained indicate a high sensitivity of H. meinerti exposed to environmentally realistic concentrations, demonstrating that there are risks for the species in real field conditions.

Integrated ecosystem models (soil-water) to analyze pesticide toxicity to aquatic organisms at two different temperature conditions.

In order to increase the knowledge about pesticides considering the soil-water interaction, ecosystem models (mesoscosms) were used to analyze the of leachate on the immobility and feeding rate of the cladocerans, Ceriodaphnia silvestrii and D. similis and algae Raphidocelis subcapitata, at two different temperatures. Mesocosm were filled with natural soil (latosolo) that were contaminated with insecticide/acaricide Kraft 36 EC® and fungicide Score 250 EC®, using the recommended concentration for strawberry crops (10.8 g abamectin/ha and 20 g difenoconazole/ha). Pesticides were applied once (hand sprayers) and the precipitation was simulated twice a week (Days 1, 4, 8, 11, 15 and 18). The mesocosm were kept in a room with a controlled temperature (23 and 33 °C) and photoperiod (12h light/12h dark). The Kraft 36 EC® insecticide showed toxicity for both species of cladocerans tested, with effects on immobility and feeding rate, both at 23 and 33 °C. Score 250 EC® showed to be toxic only for the experiments that analyzed the immobility of C. silvestrii at 23 °C and the feeding of D. smilis at 33 °C, demonstrating that the effects are species-specific and related to the temperature at which they are tested. While for species R. subcapitata there was an effect only for mixture treatments of the pesticides analyzed at both temperatures. Thereby, zooplanktonic organisms may be at risk when exposed to this compound even after percolating in a soil column, which could lead to effects on the entire aquatic trophic chain and that temperature can influence the organism response to the contaminant.

Impact of 2,4-D and fipronil on the tropical midge Chironomus sancticaroli (Diptera: Chironomidae).

Increased use of pesticides in conventional agriculture implies potential risks to the environment. In aquatic ecosystems, benthic organisms may be exposed to pesticides via contaminated water and sediment, leading to several potential cascading effects on the food web. The aim of this study was to assess the functional implications of environmental realistic concentrations of the herbicide 2,4-D and the insecticide fipronil (alone and in combination) to the native tropical chironomid Chironomus sancticaroli. These two pesticides are widely applied to different crops and have frequently been detected (together) in surface water bodies in Brazil and elsewhere. Commercial products containing fipronil (Regent® 800WG) and 2,4-D (DMA® 806BR) were evaluated in 8-day toxicity tests for their effects on larval survival, growth (body length and biomass), head capsule width, development, and mentum deformities. Fipronil decreased the larval survival at the highest test concentration and the effective concentrations (EC) after eight days of exposure were: EC10 = 0.48 µg L-1 (0.395-0.565), EC20 = 1.06 µg L-1 (0.607-1.513), and EC50 = 3.70 µg L-1 (1.664-5.736). All sublethal test concentrations of fipronil decreased the larval growth, causing reductions in biomass up to 72%. The two highest test concentrations of fipronil decreased the head capsule width and after exposure to 3.7 µg fipronil L-1, only half of the larvae reached the fourth instar. The incidence of deformities was increased by fipronil in a concentration dependent manner with an increase ranging from 23% to 75%. The highest test concentration of 2.4-D (426 µg L-1) decreased the head capsule width, but larval development was unaffected at all concentrations evaluated. In the mixture tests, antagonism was observed at lower fipronil concentrations and synergism at higher fipronil concentrations for growth. The incidence of deformities rose with increasing fipronil concentrations. The results showed that environmental realistic concentrations of fipronil may have serious ecological implications for C. sancticaroli populations and that a mixture with the herbicide 2,4-D can have synergistic effects, potentiating the risks to the aquatic ecosystem.

Functional responses of Hyalella meinerti after exposure to environmentally realistic concentrations of 2,4-D, fipronil, and vinasse (individually and in mixture).

Sugarcane crops management in Brazil includes the use of pesticides, as well as alternative organic fertilizers such as vinasse obtained from waste of the ethanol industry. In order to assess the effects of the environmental contamination generated by such sugarcane practices, this study was aimed to investigate the effects of the pesticides 2,4-Dichlorophenoxyacetic acid (2,4-D) and fipronil, as well as vinasse, on the survival, behavior, and reproduction of the native epibenthic macroinvertebrate Hyalella meinerti through in situ and laboratory experiments. In situ assays were conducted in mesocosms with six treatments, i.e. untreated control, 2,4-D, fipronil, and vinasse, the mixture of the two pesticides, and both pesticides mixed with vinasse. Survival, swimming behavior, and reproduction were evaluated over time post contamination, from 0-96 h (T1) and 7-14 days (T2) through in situ experiments and 30-44 days (T3) and 75-89 days (T4) post contamination by laboratory bioassays with mesocosm water. In the T1 period, survival of H. meinerti was registered only in controls and mesocosms treated with 2,4-D. In the T2 period, treatments containing fipronil and vinasse (isolated or in both mixture treatments) still caused 100 % of mortality. Survival was recorded only in 2,4-D and control treatments, whereas reproduction only occurred in the control. In the T3 period, no survival occurred to fipronil and both mixture treatments. Vinasse and 2,4-D decreased total reproduction in comparison to control. In the T4 period, amphipods survival was detected when exposed to fipronil and its mixture with 2,4-D. However, these same treatments decreased the amplexus rates and total reproduction, with synergism denoted for the pesticide mixture. The swimming activity of males, females, and couples was decreased in surviving organisms exposed to 2,4-D, fipronil, vinasse, and the mixture of pesticides along all experimental periods. Our study showed that the application of fipronil, 2,4-D, and vinasse isolated or mixed at realistic concentrations of actual sugarcane management practices may negatively impact functional responses of indigenous amphipods in natural aquatic systems.