Individual and combined toxicity of imidacloprid and two seed dressing insecticides on collembolans Folsomia candida.

The aim of this study was to examine the chronic toxicity of imidacloprid (IMI), clothianidin (CLO) and fipronil (FIP) as a single exposure, as well as binary mixtures of IMI with CLO or FIP toward collembolans Folsomia candida, which are fauna present in the soil. Chronic toxicity assays were performed following an ISO guideline in a Tropical Artificial Soil (TAS), and the influence on the number and growth of the juveniles produced were determined. The range of nominal concentrations used in the tests with the individual compounds was 0.08-1.28 mg/kg (IMI), 0.079-1.264 mg/kg (FIP) and 0.007-0.112 mg/kg (CLO), whereas the mixture assays were performed with half the value used in the tests with individual compounds. Based upon single exposures, IMI produced a similar impact of reducing reproduction by 50% (EC50 ranging from 0.74 to 0.85 mg/kg) compared to FIP (EC50 = 0.78 mg/kg), whereas CLO was the most toxic to F. candida (EC50 = 0.08 mg/kg). Their mixtures generally resulted in a diminished effect on reproduction, as evidenced by the higher EC50 values. In contrast, in the case of the IMI+FIP combination at high concentrations at the EC50 level, a synergistic effect on toxicity was observed. The single exposure to the three insecticides and the mixture of IMI-FIP also decreased the size of generated juveniles, which was evidenced by the reduction in the proportion of large juveniles and increased proportion of small juveniles. However, both binary mixtures (IMI-FIP and IMI-CLO) presented antagonistic effects as evidenced by less than expected reductions in growth. Data on the toxic effects of IMI in a mixture with other seed dressing insecticides to collembolans provides useful information to environmental risk assessors by diminishing the uncertainties on the ecological risk of exposure to pesticides, enabling soil management degradation by utilizing multiple insecticides.

Rapid Communication: oxidative stress induced by mixed exposure to glyphosate and silver nanoparticles.

The aim of this study was to examine oxidative stress induced by the binary mixture of silver nanoparticles (AgNP) and glyphosate (Gly) in Daphnia magna by measurement of reactive oxygen species (ROS) production, glutathione (GSH) levels, enzyme activities of catalase (CAT) and superoxide dismutase (SOD) as well as malondialdehyde (MDA) content. Acute exposure of Daphnia magna to binary mixture of AgNP and Gly resulted in significant biochemical responses indicative of oxidative damage. This response seemed to be related to imbalance in enzymatic/non-enzymatic antioxidant enzymes associated with intracellular overproduction of ROS and significant increase in MDA levels, indicating that the integrity and function of the cell membrane was damaged. These changes adversely affected the fitness and survival of Daphnia magna and negatively influenced offspring growth and reproduction.

Application of a phosphonium-based ionic liquid for reactive textile dye removal: Extraction study and toxicological evaluation.

Textile dyeing processes are known for their negative environmental impacts due to the production of aqueous effluents containing toxic dyes. Therefore, new wastewater treatment processes need to be developed to treat such effluents, including Liquid-Liquid Extraction (LLE) process using Ionic Liquids (IL). This work aimed to evaluate the application of the hydrophobic IL trihexyltetradecylphosphonium decanoate to extract black, navy, and royal reactive dyes from water and evaluate the toxicological aspects of the resulting water stream. We investigated the effect of selected parameters, such as pH (2-12), temperature (20-50 °C), salt effects, dye concentration (0.5-50 mg/L), and phase volume ratio (900-9000) on the dye extraction. The results showed extraction yields as high as 97% for the three dyes and an extraction capacity of approximately 300 mg/g for black and navy dyes and 400 mg/g for royal. The toxicity tests involved Lactuca sativa, Triticum aestivium L, and Daphnia magna as bioindicators. The difference between the toxicity of the dye solutions before and after extraction was not statistically significant when L. sativa and Triticum aestivum L were used as bioindicators. However, the extracted solution showed increased toxicity towards D. magna due to traces of IL. Overall, the IL has a high extraction capacity for the black, navy, and royal dyes. Nevertheless, further studies on LLE associated with other processes must be carried out to reduce the risk linked to the toxicity of IL transferred to the water.

Acute and chronic toxicity of amine-functionalized SiO2 nanostructures toward Daphnia magna.

Silicon oxide (SiO2) nanostructures (SiO2NS) are increasingly being incorporated into an array of products, notably in the food, pharmaceutical, medical industries and in water treatment systems. Amorphous SiO2NS have low toxicity, however, due to their great versatility, superficial modifications can be made and these altered structures require toxicological investigation. In this study, SiO2NS were synthetized and amine-functionalized with the molecules (3-aminopropyl)triethoxysilane (APTMS) and 3-[2-(2-aminoethylamino)ethylamino]propyltrimethoxysilane (AEAEAPTMS), named SiO2NS@1 and SiO2NS@3, respectively. The bare SiO2NS, SiO2NS@1 and SiO2NS@3 samples were characterized and the influence of the culture medium used in the toxicological assays was also evaluated. The effect of amine functionalization of SiO2NS was investigated through acute and chronic toxicity assays with Daphnia magna. Modifications to ultrastructures of the intestine and eggs of these organisms were observed in TEM and SEM analysis. The toxicity was influenced by the surface modifications and a possible Trojan horse effect was highlighted, particularly in the case of chronic exposure. Exposure to all NSs promoted alterations in the microvilli and mitochondria of the D. magna intestine and some damage to egg cells was also observed. The results demonstrate the importance of carrying out a full characterization of these materials, since surface modifications can enhance their toxic potential.

Crystalline phase-dependent toxicity of aluminum oxide nanoparticles toward Daphnia magna and ecological risk assessment.

Aluminum oxide nanoparticles (Al2O3 NPs) can be found in different crystalline phases, and with the emergence of nanotechnology there has been a rapid increase in the demand for Al2O3 NPs in different engineering areas and for consumer products. However, a careful evaluation of the potential environmental and human health risks is required to assess the implications of the release of Al2O3 NPs into the environment. Thus, the objective of this study was to investigate the toxicity of two crystalline phases of Al2O3 NPs, alpha (α-Al2O3 NPs) and eta (η-Al2O3 NPs), toward Daphnia magna and evaluate the risk to the aquatic ecology of Al2O3 NPs with different crystalline phases, based on a probabilistic approach. Different techniques were used for the characterization of the Al2O3 NPs. The toxicity toward Daphnia magna was assessed based on multiple toxicological endpoints, and the probabilistic species sensitivity distribution (PSSD) was used to estimate the risk of Al2O3 NPs to the aquatic ecology. The results obtained verify the toxic potential of the NPs toward D. magna even in sublethal concentrations, with a more pronounced effect being observed for η-Al2O3 NPs. The toxicity is associated with an increase in the reactive oxygen species (ROS) content and deregulation of antioxidant enzymatic/non-enzymatic enzymes (CAT, SOD and GSH). In addition, changes in MDA levels were observed, indicating that D. magna was under oxidative stress. The most prominent chronic toxic effects were observed in the organisms exposed to η-Al2O3 NPs, since the lowest LOEC was 3.12 mg/L for all parameters, while for α-Al2O3 NPs the lowest LOEC was 6.25 mg/L for longevity, growth and reproduction. However, the risk assessment results indicate that, based on a probabilistic approach, Al2O3 NPs (alpha, gamma, delta, eta and theta) only a very limited risk to organisms in surface waters.

Toxicity of binary mixtures of Al2O3 and ZnO nanoparticles toward fibroblast and bronchial epithelium cells.

The objective of this study was to examine the cytotoxic effects of binary mixtures of Al2O3 and ZnO NPs using mouse fibroblast cells (L929) and human bronchial epithelial cells (BEAS-2B) as biological test systems. The synergistic, additive, or antagonistic behavior of the binary mixture was also investigated. In toxicity experiments, cellular morphology, mitochondrial function (MTT assay), apoptosis, nuclear size and shape, clonogenic assays, and damage based upon oxidative stress parameters were assessed under control and NPs exposure conditions. Although Abbott modeling results provided no clear evidence of the binary mixture of Al2O3 and ZnO NPs exhibiting synergistic toxicity, some specific assays such as apoptosis, nuclear size and shape, clonogenic assay, activities of antioxidant enzymatic enzymes catalase, superoxide dismutase, and levels of glutathione resulted in enhanced toxicity for the mixtures with 1 and 1.75 toxic units (TU) toward both cell types. Data demonstrated that co-presence of Al2O3 and ZnO NPs in the same environment might lead to more realistic environmental conditions. Our findings indicate cytotoxicity of binary mixtures of Al2O3 and ZnO NPs produced greater effects compared to toxicity of either individual compound.

Oxidative stress and hypermethylation induced by exposure of Oreochromis niloticus to complex environmental mixtures of river water from Cubatão do Sul, Brazil.

In this study, we investigated the effects of oxidative stress and hypermethylation through lipid peroxidation and DNA methylation, respectively, in erythrocytes of Oreochromis niloticus exposed to environmental complex mixture of water from Cubatão do Sul River throughout the year. This river is the source of drinking water for the region of Florianópolis, the capital of Santa Catarina State, Brazil. Lipid peroxidation was quantified by the rate of malondialdehyde (MDA) formation, and DNA methylation was quantified by the rate of 5-methyldeoxycytosine (m(5)dC) formation. In all studied sites, the river water samples caused metabolic changes in O. niloticus. MDA formation rates were significantly different when compared to the negative control (except for samples from Site 1 during spring 2010, summer 2011 and fall 2011). All samples (except Site 1, spring 2010) induced increases in the m(5)dC formation rates, and at the end of the study, the values were near the values found in the positive control (potassium dichromate 2.5mg/L). The results showed that samples of environmental complex mixtures of water from Cubatão do Sul River are capable of inducing high levels of oxidative damage and hypermethylation in O. niloticus.

Toxicity of PAMAM-coated gold nanoparticles in different unicellular models.

Polyamidoamine (PAMAM) dendrimers are used for many pharmaceutical and biomedical applications. However, the toxicological risks of several PAMAM-based compounds are still not fully evaluated, despite evidences of PAMAM deleterious effects on biological membranes, leading to toxicity. In this report, we investigated the toxicity of generation 0 PAMAM-coated gold nanoparticles (AuG0 NPs) in four different models to determine how different cellular systems are affected by PAMAM-coated NPs. Toxicity was evaluated in two mammalian cell lines, Neuro 2A and Vero, in the green alga Chlamydomonas reinhardtii and the bacteria Vibrio fischeri. AuG0 NP treatments reduced cell metabolic activity in algal and bacterial cells, measured by esterase enzymatic activity (C. reinhardtii) and luminescence emission (V. fischeri). EC50 value after 30 min of treatment was similar in both organisms, with 0.114 and 0.167 mg mL(-1) for C. reinhardtii and V. fischeri, respectively. On the other hand, AuG0 NPs induced no change of mitochondrial activity in mammalian cells after 24 h of treatment to up to 0.4 mg mL(-1) AuG0 NPs. Change in the absorption spectra of AuG0 NP in the mammalian cell culture media may indicate an alteration of NP properties that contributed to the low toxicity of AuG0 NPs in mammalian cells. For a safe development of PAMAM-based nanomaterials, the difference of sensitivity between mammalian and microbial cells, as well as the modulation of NPs toxicity by medium properties, should be taken into account when designing PAMAM NPs for applications that may lead to their introduction in the environment.

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.

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.

Effects of exposure to soluble fraction of industrial solid waste on lipid peroxidation and DNA methylation in erythrocytes of Oreochromis niloticus, as assessed by quantification of MDA and m5dC rates.

In this study, lipid peroxidation and DNA methylation were observed in erythrocytes of Oreochromis niloticus exposed to soluble fractions of textile, metal-mechanic and pulp and paper industrial waste, after a period of 48 h. Lipid peroxidation was quantified by the rate of malondialdehyde (MDA) and DNA methylation was quantified by the rate of 5-methyldeoxycytosine (m5dC). Soluble fractions of textile industrial waste caused metabolic changes for all studied samples. In organisms exposed to samples TX1 and TX2 (textile waste) MDA rates were 132.36 and 140.28 nM MDA/mg protein, respectively, while in control organism the MDA rates were 27.5 nM MDA/mg protein. All samples from soluble fractions of textile industrial waste induced increases in m5dC rates, increases varied between 300 percent and 700 percent when compared to the control organism. All the organisms exposed to soluble fractions of metal-mechanic industrial waste presented increases between 360 percent and 600 percent in the rates of MDA, and one of them (sample MM3) induced an increase of 180 percent in the rate of m5dC, when compared to control. Although a significant increase was not observed in the MDA rate of fish exposed to the soluble fractions of pulp and paper industrial waste, there was an increase of 460 percent in the rate of m5dC in one of the samples (sample PP2), when compared to control. The results showed that the soluble fractions of these industrial wastes are capable of inducing oxidative damage and altering the DNA methylation of O. niloticus. Thus, the MDA and m5dC rates demonstrated to be effective biomarkers of exposure, which could be used to evaluate the toxicity of soluble fractions of industrial solid waste.

Acute and chronic toxicity of soluble fractions of industrial solid wastes on Daphnia magna and Vibrio fischeri.

Industrial wastes may produce leachates that can contaminate the aquatic ecosystem. Toxicity testing in acute and chronic levels is essential to assess environmental risks from the soluble fractions of these wastes, since only chemical analysis may not be adequate to classify the hazard of an industrial waste. In this study, ten samples of solid wastes from textile, metal-mechanic, and pulp and paper industries were analyzed by acute and chronic toxicity tests with Daphnia magna and Vibrio fischeri. A metal-mechanic waste (sample MM3) induced the highest toxicity level to Daphnia magna(CE(50,48 h) = 2.21%). A textile waste induced the highest toxicity level to Vibrio fischeri (sample TX2, CE(50,30 min) = 12.08%). All samples of pulp and paper wastes, and a textile waste (sample TX2) induced chronic effects on reproduction, length, and longevity of Daphnia magna. These results could serve as an alert about the environmental risks of an inadequate waste classification method.

Individual and combined multigenerational effects induced by polystyrene nanoplastic and glyphosate in Daphnia magna (Strauss, 1820).

In this study, the acute and multigenerational effects of the individual and combined toxicity of polystyrene nanoplastic (PSNP - 15.6, 31.2 62.5, 125, 250 and 500 mg/L) and glyphosate (Gly - 6.2, 12.5, 25, 50, 100 and 200 mg/L) on the freshwater crustacean Daphnia magna were investigated. The acute toxicity interactions were predicted mathematically using Abbott's model and multiple toxicological endpoints. In the multigenerational tests, we evaluated the effects in filial (F1 and F2) generations of daphnids after parental (F0) exposure to Gly and PSNP, as individual compounds and as a mixture, during their life history. Based on Abbott's model, the combined individual toxicities of Gly and PSNP are increased when they are present as a mixture. This indicates synergy between the components of the mixture, especially in the case of co-exposure to Gly and PSNP in higher equitoxic proportions. The mixture of PSNP and Gly caused an increase in immobility and ROS production and decrease in swimming activity. Multigenerational responses indicated that the exposure of F0 daphnids to Gly and PSNP as a mixture induced effects in the F1 and F2 reproduction parameters in the recovery tests. Thus, the results reported herein provide important information on the interaction of hydrophilic organic and nanoplastic pollutants in aqueous ecosystems. This will be useful in future studies on the toxicity of mixtures and multigenerational effects and provide a basis for management decisions aimed at the protection of environmental health.

A simple method to evaluate the toxic effects of Prorocentrum lima extracts to fish (sea bass) kidney cells.

The diarrhetic shellfish toxins (DSTs) okadaic acid (OA) and its analogues - the dinophysistoxins (DTXs) - are produced by dinoflagellates such as Prorocentrum lima and can bioaccumulate in filter-feeding organisms as they are transferred through the food web. Although there is no assessment of the harmful effects of these toxins on the fish's immune system, this study developed a primary culture protocol for kidney cells from marine fish Centropomus parallelus and evaluated the immunotoxic effects to P. lima extracts containing DSTs. The cells were obtained by mechanical dissociation, segregated with Percoll gradient, and incubated for 24 h at 28 °C in a Leibovitz culture medium supplemented with 2% fetal bovine serum and antibiotics. The exposed cells were evaluated in flow cytometry using the CD54 PE antibody. We obtained >5.0 × 106 viable cells per 1.0 g of tissue that exhibited no cell differentiation. Exposure to 1.2 or 12 ng DST mL-1 stimulated the immune system activation and increased the proportion of activated macrophages and monocytes in 48 to 52% and in 127 to 146%, respectively. The protocol proved to be an alternative tool to assess the immunotoxic effects of DST exposure on fish's anterior kidney cells.

Membrane adsorption with polyacrylonitrile prepared with superfine powder-activated carbon, case study: separation process applied in water treatment containing diclofenac.

Polyacrylonitrile membranes (PAN) have high stability against chemical agents, making them suitable for a wide range of applications as such Ultrafiltration processes. Ultrafiltration membranes composed of PAN/Superfine powder activated carbon (S-PAC) mixtures can be a good research route, aiming the development of a new separation processes for water treatment. The association of materials to form a single product can have technological and economic advantages in separation processes. In this study, S-PAC impregnated into PAN membranes were prepared, characterized and used, as a case study, to remove diclofenac (DCF) from water. The membranes (PAN/S-PAC) were synthesized with different concentrations of S-PAC (0.2, 0.6, 1.0, 3.0 and 5.0 wt%) by a phase inversion process. The results of the TEM characterizations of the S-PAC indicated the presence of micro and nanoparticles (∼10 nm) and tending to form micrometric clusters. The infrared spectra of the membranes were characteristic of PAN; however, vibrational bands attributed to the S-PAC spectrum were also observed, which indicated an interaction between the materials. The case study showed an increase in the water flux and in the DCF rejection efficiency, for composite membranes (PAN/S-PAC) with higher concentration of S-PAC. The results of static adsorption tests indicated that the mechanism of DCF rejection occurred predominantly by adsorption. There were indications that the PAN/S-PAC membranes formed a composite material and the PAN/S-PAC (3.0) presented the best study composition given the results. Although the research is in its initial phase, the results indicated that the composition can improve many water treatment systems.

Investigation of animal and algal bioassays for reliable saxitoxin ecotoxicity and cytotoxicity risk evaluation.

Contamination of water bodies by saxitoxin can result in various toxic effects in aquatic organisms. Saxitoxin contamination has also been shown to be a threat to human health in several reported cases, even resulting in death. In this study, we evaluated the sensitivity of animal (Neuro-2A) and algal (Chlamydomonas reinhardtii) bioassays to saxitoxin effect. Neuro-2A cells were found to be sensitive to saxitoxin, as shown by a 24 h EC50 value of 1.5 nM, which was obtained using a cell viability assay. Conversely, no saxitoxin effect was found in any of the algal biomarkers evaluated, for the concentration range tested (2-128 nM). These results indicate that saxitoxin may induce toxic effects in animal and human populations at concentrations where phytoplankton communities are not affected. Therefore, when evaluating STX risk of toxicity, algal bioassays do not appear to be reliable indicators and should always be conducted in combination with animal bioassays.

Removal of coloured compounds from textile industry effluents by UV/H2O2 advanced oxidation and toxicity evaluation.

This study has investigated the reduction in coloured substances and toxic compounds present in textile industry effluent by the use of an advanced oxidation process using hydrogen peroxide (H2O2) as oxidant, activated by ultraviolet radiation. The investigation was carried out on industrial effluents, both raw and after biological treatment, using different concentrations of H2O2 in a photochemical reactor equipped with a 250 W high-pressure mercury vapour lamp. The results showed that after 60 minutes of ultraviolet irradiation a H2O2 concentration of 500 mg L(-1) was able to remove approximately 73% of the coloured compounds present in raw effluent and 96% of those present in biologically treated effluent. Additionally, post-treatment toxicity tests performed using the microcrustacean Daphnia magna showed a significant effective reduction in the acute toxicity of the raw effluent. In tests carried out with treatment at a concentration of 750 and 1000 mg L(-1) H2O2, analysis of the frequency ofmicronuclei in erythrocytes of Tilapia cf rendalli exposed to treated effluent samples confirmed that there were no mutagenic effects on the fish. Together, these results indicate that the oxidation process offers a good alternative for the removal of colour and toxicity from textile industry effluent.

Interaction Effect of EDTA, Salinity, and Oxide Nanoparticles on Alga Chlamydomonas reinhardtii and Chlamydomonas euryale.

The interaction effects of organic ligand ethylene diamine tetra-acetic acid (EDTA) and oxide nanoparticles (magnetite Fe3O4-NPs and copper CuO-NPs) were investigated during a 72 h period on two green algal species-Chlamydomonas reinhardtii under freshwater conditions and Chlamydomonas euryale under saltwater conditions. Fe3O4-NPs had larger agglomerates and very low solubility. CuO-NPs, having smaller agglomerates and higher solubility, were more toxic than Fe3O4-NPs in freshwater conditions for similar mass-based concentrations, especially at 72 h under 100 mg L-1. Furthermore, the effect of EDTA increased nanoparticle solubility, and the salinity caused a decrease in their solubility. Our results on C. euryale showed that the increase in salinity to 32 g L-1 caused the formation of larger nanoparticle agglomerates, leading to a decrease in the toxicity impact on algal cells. In addition, EDTA treatments induced a toxicity effect on both freshwater and saltwater Chlamydomonas species, by altering the nutrient uptake of algal cells. However, C. euryale was more resistant to EDTA toxicity than C. reinhardtii. Moreover, nanoparticle treatments caused a reduction in EDTA toxicity, especially for CuO-NPs. Therefore, the toxicity impact caused by these environmental factors should be considered in risk assessment for metallic nanoparticles.

Toxicological effects of leachate extracts from asphalt mixtures nanomodified under Daphnia magna and Landoltia punctata test organisms.

The incorporation of nanomaterials in binders in the paving area has been studied to improve the mechanical behavior of asphalt mixtures. However, asphalt mixture compounds are susceptible to leaching and deposition in the environment. In this context, this research aimed to investigate the toxic effect of two leachate extracts from asphalt mixtures nanomodified with 2% carbon nanotube and 3% organophilic nanoclay, compared to conventional mixture, using Daphnia magna and Landoltia punctata as test organisms. The nanomaterials were characterized to confirm morphology, stability and effective diameter. Extracts were chemically characterized using the Fourier-Transform Infrared Spectroscopy (FTIR) technique, which indicated presence of functional groups of the asphalt binder in greater intensity in the leachate from conventional mixture. Acute toxicity with D. magna indicated EC50,48h of 83.5 ± 6.2 mL/L for leachate extract from conventional mixture, 306.0 ± 87.6 mL/L for leachate extract from mixture with nanoclay and 464.8 ± 32.1 mL/L for leachate extract from mixture with carbon nanotube. No leachate caused significant chronic toxicity. As for L.punctata, concentrations that caused 50% growth inhibition were 127.5 mL/L for the leachate extract from mixture with carbon nanotube, 196.9 mL/L for the leachate extract from mixture with nanoclay and 205 mL/L for the leachate extract from conventional mixture. For these test organisms, there is no evidence of negative impacts directly associated with the use of the present nanomaterials in asphalt mixtures. The incorporation of these nanos may also reduce the acute toxicity of the mixtures.

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.