Locomotion behavior testing as a complementary tool in Collembola avoidance assays with neurotoxic insecticides.

This study aimed (1) to assess the ability of collembolans Folsomia candida to avoid soils contaminated with three seed dressing insecticides imidacloprid, clothianidin, and fipronil; (2) to assess the effects of the insecticides on collembolans' locomotion behavior; (3) to check if changes in the locomotion behavior would explain the avoidance/preference responses; and (4) to evaluate the possibility to use locomotion behavior as toxicity biomarker of the tested insecticides. Avoidance and locomotion behavior assays with collembolans F. candida were performed with commercial seed dressing formulations of three insecticides (imidacloprid, clothianidin, and fipronil). Results showed no avoidance behavior at any concentration, while a "preference" was observed with increasing concentrations of the three tested insecticides. Significant reductions in the locomotion of exposed collembolans were observed at ≥ 1 mg kg-1 for imidacloprid (18-38%) and fipronil (29-58%) and ≥ 4 mg kg-1 for clothianidin (10-47%). At the higher insecticide concentrations, the collembolans had their trajectories restricted to smaller areas, with a tendency for circular movements. Our results confirm that the "preference" for contaminated soils with neurotoxic substances is likely due to locomotion inhibition impairing the ability of organisms to escape. This effect highlights that only avoidance assays may be not sufficient to assure the safety of some substances and confirm the potential of locomotion behavior as a sensitive toxicity biomarker for neurotoxic insecticides.

A systematic review of the toxic effects of a nanopesticide on non-target organisms: Estimation of protective concentrations using a species sensitivity distribution (SSD) approach - The case of atrazine.

Nanopesticides, such as nanoencapsulated atrazine (nATZ), have been studied and developed as eco-friendly alternatives to control weeds in fields, requiring lower doses. This review contains a historical and systematic literature review about the toxicity of nATZ to non-target species. In addition, the study establishes protective concentrations for non-target organisms through a species sensitivity distribution (SSD) approach. Through the systematic search, we identified 3197 publications. Of these, 14 studies addressed "(nano)atrazine's toxicity to non-target organisms". Chronological and geographic data on the publication of articles, characterization of nATZ (type of nanocarrier, size, polydispersity index, zeta potential), experimental design (test species, exposure time, measurements, methodology, tested concentrations), and toxic effects are summarized and discussed. The data indicate that cell and algal models do not show sensitivity to nATZ, while many terrestrial and aquatic invertebrates, aquatic vertebrates, microorganisms, and plants have high sensitivity to nAZT. The SSD results indicated that D. similis is the most sensitive species to nATZ, followed by C. elegans, E. crypticus, and P. subcapitata. However, the limitations in terms of the number of species and endpoints available to elaborate the SSD reflect gaps in knowledge of the effects of nATZ on different ecosystems.

Influence of temperature and soil moisture on the toxic potential of clothianidin to collembolan Folsomia candida in a tropical field soil.

Climate change can alter the toxic effects of pesticides on soil invertebrates. However, the nature and magnitude of the influence of climatic factors on clothianidin impacts in tropical soils are still unknown. The influence of increasing atmospheric temperature and the reduction in soil moisture on the toxicity and risk of clothianidin (seed dressing formulation Inside FS®) were assessed through chronic toxicity tests with collembolans Folsomia candida in a tropical field soil (Entisol). The risk of clothianidin for collembolans was estimated using the Toxicity-Exposure Ratio (TER) approach. Organisms were exposed to increasing clothianidin concentrations at 20, 25 and 27 °C in combination with two soil moisture conditions (30 and 60% of the maximum water holding capacity-WHC). The effect of temperature and soil moisture content on clothianidin toxicity was verified through the number of F. candida juveniles generated after 28 days of exposure to the spiked soil. The toxicities estimated at 25 °C (EC50_30%WHC = 0.014 mg kg-1; EC50_60%WHC = 0.010 mg kg-1) and 27 °C (EC50_30%WHC = 0.006 mg kg-1; EC50_60%WHC = 0.007 mg kg-1) were 2.9-3.0-fold (25 °C) and 4.3-6.7-fold (27 °C) higher than those found at 20 °C (EC50_30%WHC = 0.040 mg kg-1; EC50_60%WHC = 0.030 mg kg-1), indicating that clothianidin toxicity increases with temperature. No clear influence of soil moisture content on clothianidin toxicity could be observed once the EC50 values estimated at 30% and 60% WHC, within the same temperature, did not significantly differ. A significant risk was detected in all temperatures and soil moisture scenarios studied, and the TER values indicate that the risk can increase with increasing temperatures. Our results revealed that temperature could overlap with soil moisture in regulating clothianidin toxicity and reinforce the importance of including climatic factors in the prospective risk assessment of pesticides.

The use of sewage sludge as remediation for imidacloprid toxicity in soils.

This study investigated the influence of the sewage sludge (SS) soil amendment on the chronic toxicity of imidacloprid (through the seed dressing formulation MUCH 600 FS®-600 g active ingredient L-1) to collembolans Folsomia candida. Individuals 10-12 days old were exposed to two contrasting tropical soils (Oxisol and Entisol) amended with SS doses (0, 20, 40, 80, 160, and 320 g SS kg-1 soil; the SS doses have low intrinsic toxicity, which was checked before its application) in a full factorial combination with five imidacloprid concentrations (varying from 0.25 to 4 mg kg-1 in Oxisol and 0.03-0.5 mg kg-1 in Entisol) plus a control. None of the SS doses (without imidacloprid) in both soils reduced the number of generated juvenile collembolans. The imidacloprid concentrations reducing the collembolan reproduction in 50% (EC50) in Oxisol and Entisol without SS were 0.49 and 0.08 mg kg-1, respectively. However, the EC50 values generally increased with increasing SS doses in soils, varying from 1.03 to 1.41 in Oxisol and 0.07 to 0.21 in Entisol. The SS-amended soils showed 2.1- to 2.9-fold lower imidacloprid toxicity (EC50-based) in Oxisol and 1.8- to 2.7-fold lower toxicity in Entisol. Our results suggest the most effective SS doses alleviating the imidacloprid toxicity (EC50-based) to collembolans are 20 g kg-1 in Oxisol and 80 g kg-1 in Entisol. These results indicate that the tested SS has the potential to be employed as a soil amendment agent by reducing the toxicity of imidacloprid to the reproduction of F. candida.

Role of climatic factors in the toxicity of fipronil toward earthworms in two tropical soils: effects of increased temperature and reduced soil moisture content.

The aim of this study was to assess the effect of temperature on the toxicity of fipronil toward earthworms (Eisenia andrei) in two Brazilian soils (Entisol and Oxisol) with contrasting textures. In the case of Entisol, the influence of soil moisture content on toxicity was also investigated. Earthworms were exposed for 56 days to soils spiked with increasing concentrations of fipronil (8.95, 19.48, 38.22, 155.61, and 237.81 mg kg-1 for Entisol; 12.99, 27.94, 48.42, 204.67, and 374.29 mg kg-1 for Oxisol) under scenarios with different combinations of temperature (20, 25 and 27 °C) and soil moisture content (60 and 30% of water holding capacity (WHC) for Entisol and 60% WHC for Oxisol). The number of juveniles produced was taken as the endpoint, and a risk assessment was performed based on the hazard quotient (HQ). In Entisol, at 60% WHC the fipronil toxicity decreased at 27 °C compared with the other temperatures tested (EC50 = 52.58, 48.48, and 110 mg kg-1 for 20, 25, and 27 °C, respectively). In the case of Oxisol at 60% WHC, the fipronil toxicity increased at 27 °C compared with other temperatures (EC50 = 277.57, 312.87, and 39.89 mg kg-1 at 20, 25, and 27 °C, respectively). An increase in fipronil toxicity was also observed with a decrease in soil moisture content in Entisol at 27 °C (EC50 = 27.95 and 110 mg kg-1 for 30% and 60% WHC, respectively). The risk of fipronil was only significant at 27 °C in Entisol and Oxisol with water contents of 30% and 60% WHC, respectively, revealing that higher temperatures are able to increase the risk of fipronil toxicity toward earthworms depending on soil type and soil moisture content. The results reported herein show that soil properties associated with climatic shifts could enhance the ecotoxicological effects and risk of fipronil for earthworms, depending on the type of soil.

Toxicity of fipronil to Folsomia candida in contrasting tropical soils and soil moisture contents: effects on the reproduction and growth.

This study assessed the influence of three tropical soil types and soil moisture content on the toxicity and risk of the insecticide fipronil to collembolans Folsomia candida. Chronic toxicity tests were performed in a Tropical Artificial Soil (TAS), an Oxisol and an Entisol spiked with increasing concentrations of fipronil to assess the effects on the reproduction and growth of the species. The soil moisture contents were kept at 60% (standard condition) and 30 or 45% (water restriction) of their water holding capacity (WHC). The toxicity of fipronil on collembolans reproduction was about three times higher in Entisol compared to TAS or Oxisol. Higher toxicities were also found in the drier TAS (EC50 30%WHC = 0.20 vs EC50 60%WHC = 0.70 mg kg-1) and Oxisol (EC50 45%WHC = 0.27 vs EC50 60%WHC = 0.54 mg kg-1), while in Entisol lower impacts were found in the drier samples (EC50 30%WHC = 0.41 vs EC50 60%WHC = 0.24 mg kg-1). For all tested soils, the size of generated collembolans was reduced by the fipronil concentrations, regardless of soil moisture. However, the drier condition increased the effect on the growth in TAS and Entisol for some concentrations. A significant risk of exposure was found in TAS and Oxisol at drier conditions and, for Entisol, regardless of the soil moisture. The toxic effects and risk of fipronil on collembolans were higher in the natural sandy soil. The soil moisture content increase or decrease the toxicity of the insecticide for collembolans, depending on soil type.

Chronic effects of clothianidin to non-target soil invertebrates: Ecological risk assessment using the species sensitivity distribution (SSD) approach.

This study aimed to assess the chronic toxicity and risk of clothianidin in a seed dressing formulation to non-target soil invertebrates. The toxicity assays were performed with two oligochaetes (earthworms Eisenia andrei and enchytraeids Enchytraeus crypticus) and three collembolans (Folsomia candida, Proisotoma minuta and Sinella curviseta) species following ISO protocols. Risk assessment (via Hazard Quotient approach - HQ) was based on the hazardous concentrations for 95% of the species (HC5), derived from chronic Species Sensitivity Distributions (SSD) for clothianidin, and on its predicted environmental concentrations (PEC). Four SSD scenarios were generated with literature and/or this study data, following different data selection criteria (i.e., general, only data from tests using similar formulations, similar soils, or identical soil/formulation). In our experiments, a higher clothianidin toxicity (EC50-based) was found for collembolans (varying from 0.11 to 0.28 mg kg-1 between species) followed by the earthworms (4.35 mg kg-1), while the enchytraeids were the least sensitive (33.5 mg kg-1). HQ indicated a significant risk of clothianidin to soil invertebrates because the estimated PEC were at least 16.6 times higher than HC5 and are expected to affect the whole group of collembolans. Despite the criteria for data inclusion have influenced the HC5 values, no substantial changes were observed for the risk outcomes. To our knowledge, this is the first study assessing the chronic ecological risk of clothianidin to beneficial soil fauna based on a probabilistic SSD approach. Data from this study can help to derive more reliable protection thresholds for clothianidin in soils.

Effect of temperature on the toxicity of imidacloprid to Eisenia andrei and Folsomia candida in tropical soils.

The influence of temperature on the chronic toxicity and risk of imidacloprid to soil non-target species was assessed in tropical soils. Earthworms Eisenia andrei and collembolans Folsomia candida were exposed to a tropical artificial soil (TAS) and two natural tropical soils from Brazil (Entisol and Oxisol) with increasing concentrations of imidacloprid under atmospheric temperatures of 20, 25 and 28 °C. The effect of temperature on the reproduction of both species was assessed through the number of juveniles and earthworm's growth, and the risk associated was estimated through the Toxicity-Exposure Ratio (TER). Toxicity of imidacloprid increased with temperature in all tested soils, being generally lower in TAS soil (EC50s of 1.48, 0.66 and 0.40 mg kg-1 for E. andrei and 0.3, 0.2 and 0.06 mg kg-1 for F. candida at 20, 25 and 28 °C, respectively) compared to Entisol (EC50s of 0.19, 0.03 and 0.14 mg kg-1 for E. andrei and 0.04, 0.02, 0.01 mg kg-1 for F. candida at 20, 25 and 28 °C, respectively) and Oxisol (EC50s of 0.21, 0.07, 0.06 mg kg-1 for E. andrei and 0.16, 0.09, 0.06 mg kg-1 for F. candida at 20, 25 and 28 °C, respectively) within each temperature for both species. These values indicate that properties of TAS may not be representative of natural/local soils to adequately estimate the toxicity of pesticides to non-target soil species. At higher temperatures, the variability of imidacloprid toxicity between soils was lower, which suggests that the influence of soil properties on imidacloprid toxicity was overshadowed by temperature. TER values revealed that risk is also greater at higher temperatures. Data reported enforce the need for the inclusion of more realistic conditions in single-species tests in prospective risk assessment of pesticides to avoid underestimation of risk to non-target species.

Ecotoxicological assessment of Fluazuron: effects on Folsomia candida and Eisenia andrei.

The cattle production in Brazil has increased considerably in the last years, mainly due to the control of parasite infestation of the animals, which cause loss of productivity to the sector. Fluazuron is an active ingredient (a.i.) of the benzoylurea class used to control ticks in cattle. As this a.i. has been found unchanged in animal feces, which may present a risk to edaphic organisms, this study aimed to assess the effects of fluazuron on survival, reproduction, and behavior of the soil invertebrates Folsomia candida and Eisenia andrei, through ecotoxicological assays. We carried out bioassays in a tropical artificial soil (TAS) spiked with increasing doses of the insecticide. Earthworm mortality was found only at the highest tested fluazuron concentration (LOEC = 160 mg a.i. kg-1 dry soil and NOEC = 80 mg kg-1), while the reproduction of F. candida and E. andrei was reduced at lower fluazuron concentrations (EC50 = 4.48 mg kg-1 and EC50 = 20.8 mg kg-1, respectively). Avoidance behavior was detected for both species at lower concentrations than those that caused impacts on reproduction, indicating that the substance may affect the soil habitat function. Since the possible adverse effects of fluazuron on edaphic fauna are still unknown or neglected, this study also warns about the possible harmful effect of veterinary pharmaceutical products on edaphic fauna.