Effects of naturally sourced bitumen samples from Alberta oil sands region (Canada) on aquatic benthic invertebrates: A case study with Chironomus riparius.

Athabasca oil sands in Alberta, Canada, are large bitumen deposits and are one of the world's largest petroleum reserves. This research contributes to the growing body of knowledge on the influence of this naturally occurring bitumen on freshwaters. Using laboratory-based exposure studies, we examined the life cycle responses of the aquatic midge Chironomus riparius to both naturally formed solid bitumen incorporated in the sediment and its corresponding aqueous extracts, denominated as elutriates. The 28-day partial life cycle assay involved bitumen samples from two distinct geological origins in the Athabasca River Basin (Clearwater and McMurray formations), comprising both weathered and freshly collected bitumen from a total of 4 different rivers. Our results demonstrate a measurable impact of sediment-embedded bitumen on C. riparius life history traits, namely on their growth and emergence patterns. Furthermore, we observed that bitumen samples from the Ells River (McMurray formation), which were freshly collected from exposed river bank soil deposits, exerted the strongest effects on most studied eco-physiological endpoints. Bitumen extracts from the Steepbank River and Athabasca River in the McMurray Formation and Steepbank River in the Clearwater Formation followed, underscoring the geographical variance in bitumen-induced toxicity. Exposure to elutriates, simulating "weathered" bitumen generally did not induce adverse effects in C. riparius life-cycle endpoints compared to elutriates prepared from freshly eroded bank soils. This emphasizes the importance of considering bitumen sources, their age, and the aquatic receiving environment when assessing potential adverse exposure effects. Our study shows that exposure to freshly eroded soils/sediments can potentially affect benthic invertebrates. More research is needed to understand how hydrological changes affect bitumen sediment exposure and the associated risks to aquatic biota.

Toxicity mechanisms of plastic nanoparticles in three terrestrial species: antioxidant system imbalance and neurotoxicity.

The detrimental impacts of plastic nanoparticles (PNPs) are a worldwide concern, although knowledge is still limited, in particular for soil mesofauna. This study investigates the biochemical impact of 44 nm polystyrene PNPs on three soil models-Enchytraeus crypticus (Oligochaeta), Folsomia candida (Collembola) and Porcellionides pruinosus (Isopoda). Exposure durations of 3, 7 and 14 days (d) were implemented at two concentrations (1.5 and 300 mg kg-1 PNPs). Results revealed PNPs impact on the activities of the glutathione-dependent antioxidative enzyme, glutathione S-transferase (GST) and on the neurotransmitter acetylcholinesterase (AChE) for all three species. Catalase (CAT) played a minor role, primarily evident in F. candida at 300 mg kg-1 PNPs (CAT and GST response after 14 d), with no lipid peroxidation (LPO) increase. Even with the antioxidant defence, P. pruinosus was the most sensitive species for lipid oxidative damage (LPO levels increased after 7 d exposure to 300 mg kg-1 PNPs). Significant AChE inhibitions were measured already after 3 d to both PNP concentrations in F. candida and E. crypticus, respectively. Significant AChE inhibitions were also found in P. pruinosus but later (7 d). Overall, the toxicity mechanisms of PNPs involved antioxidant imbalance, being (mostly) the glutathione-associated metabolism part of that defence system. Neurotoxicity, linked to AChE activities, was evident across all species. Sensitivity to PNPs varied: P. pruinosus > F. candida ≅ E. crypticus. This pioneering study on PNPs toxicity in soil invertebrates underscores its environmental relevance, shedding light on altered biochemical responses, that may compromise ecological roles and soil ecosystem fitness.

Responses of soil microbiome to copper-based materials (nano and bulk) for agricultural applications: An indoor-mesocosm experiment.

The foreseen increasing application of copper-based nanomaterials (Cu-NMs), replacing or complementing existing Cu-agrochemicals, may negatively impact the soil microbiome. Thus, we studied the effects on soil microbiome function and composition of nano copper oxide (nCuO) or copper hydroxide NMs in a commercial (Kocide®3000) or a lab-synthetized formulation (nCu(OH)2) or bulk copper hydroxide (Cu(OH)2-B), at the commonly recommended Cu dose of 50 mg(Cu)kg-1 soil. Microbial responses were studied over 28 days in a designed indoor mesocosm. On day-28, in comparison to non-treated soil (CT), all Cu-treatments led to a reduction in dehydrogenase (95% to 68%), arylsulfatase (41% to 27%), and urease (40% to 20%) activity. There was a 32% increase in the utilization of carbon substrates in the nCuO-treatment and an increased abundance of viable bacteria in the nCu(OH)2-treatment (75% of heterotrophic and 69% of P-solubilizing bacteria). The relative abundance of Acidobacteria [Kocide®3000, nCuO, and Cu(OH)2-B treatments] and Flavobacteriia [nCu(OH)2-treatment] was negatively affected by Cu exposure. The abundance of Cu-tolerant bacteria increased in soils treated with Kocide®3000 (Clostridia) and nCu(OH)2 (Gemmatimonadetes). All Cu-treated soils exhibited a reduced abundance of denitrification-related genes (0.05% of nosZ gene). The DTPA-extractable pool of ionic Cu(II) varied among treatments: Cu(OH)2-B > Kocide®3000 âˆ¼ nCuO>nCu(OH)2, which may explain changes on the soil microbiome composition, at the genera and OTU levels. Thus, our study revealed that Cu-materials (nano and bulk) influence the soil microbiome with implications on its ecological role. It highlights the importance of assessing the impact of Cu-materials under dynamic and complex exposure scenarios and emphasizes the need for specific regulatory frameworks for NMs.

A Comprehensive Ecotoxicity Study of Molybdenum Disulfide Nanosheets versus Bulk form in Soil Organisms.

The increasing use of molybdenum disulfide (MoS2) nanoparticles (NPs) raises concerns regarding their accumulation in soil ecosystems, with limited studies on their impact on soil organisms. Study aim: To unravel the effects of MoS2 nanosheets (two-dimensional (2D) MoS2 NPs) and bulk MoS2 (156, 313, 625, 1250, 2500 mg/kg) on Enchytraeus crypticus and Folsomia candida. The organisms' survival and avoidance behavior remained unaffected by both forms, while reproduction and DNA integrity were impacted. For E. crypticus, the individual endpoint reproduction was more sensitive, increasing at lower concentrations of bulk MoS2 and decreasing at higher ones and at 625 mg/kg of 2D MoS2 NPs. For F. candida, the molecular endpoint DNA integrity was more impacted: 2500 mg/kg of bulk MoS2 induced DNA damage after 2 days, with all concentrations inducing damage by day 7. 2D MoS2 NPs induced DNA damage at 156 and 2500 mg/kg after 2 days, and at 1250 and 2500 mg/kg after 7 days. Despite affecting the same endpoints, bulk MoS2 induced more effects than 2D MoS2 NPs. Indeed, 2D MoS2 NPs only inhibited E. crypticus reproduction at 625 mg/kg and induced fewer (F. candida) or no effects (E. crypticus) on DNA integrity. This study highlights the different responses of terrestrial organisms to 2D MoS2 NPs versus bulk MoS2, reinforcing the importance of risk assessment when considering both forms.

Ecotoxicological effects of fluvial eroded bitumen sediments from the Alberta oil sands to model aquatic species.

To fully understand the ecological and cumulative effects of mining activities on the surrounding aquatic systems of the Canadian oil sands region, it is essential to understand the consequences of exposure to bitumen-containing soils/sediment from natural geomorphological processes. Both physical and chemical stress on aquatic biota can potentially result from exposure to natural bitumen, resulting from hillslope erosional processes and slumping of bankside soils into the rivers, affecting both riverbed habitat and water quality. The magnitude and duration of bitumen-containing soil's fluvial soils/erosional input into receiving watersheds depends on the interannual variability in the regional hydroclimatology and related seasonal and extreme flow events. The primary objective of this study was to evaluate the potential ecotoxicological effects associated with erosional input of riverbank bitumen soils using controlled exposures. A series of integrated, laboratory-based ecotoxicological bioassays were conducted using organisms with different ecological and functional traits (Daphnia magna (Cladocera), Physa acuta (Gastropoda), and Aliivibrio fischeri (Bacteria)). All model organisms were exposed to elutriates produced from natural bitumen from four different regional rivers: fresh bitumen from shoreline outcrops at the Steepbank River upper and lower reaches (STB-CF and STB-MF, respectively) and the lower-Ells River (EL-MF) and aged, fluvially processed/weathered bitumen from the shoreline of the Athabasca River (ATB-MF). All tested organisms responded negatively to STB-MF and EL-MF elutriates. Low toxicity was also observed in the STB-CF and ATB-MF samples. These results follow the chemical analysis of the parental material and elutriates, where higher levels of metals, polycyclic aromatic compounds and naphthenic acids were detected in the EL-MF sample. In summary, this study shows that eroded and transported bitumen-containing soils and sediments could be a natural source of contaminant exposure to aquatic biota. This fluvial pathway should be considered when assessing background toxicity and the toxicological and ecological effects of oil sands mining activities.

The effects of different temperatures in mercury toxicity to the terrestrial isopod Porcellionides pruinosus.

Climate changes and metal contamination are pervasive stressors for soil ecosystems. Mercury (Hg), one of the most toxic metals, has been reported to interact with temperature. However, compared to aquatic biota, little is known about how temperature affects Hg toxicity and bioaccumulation to soil organisms. Here, toxicity and bioaccumulation experiments were replicated at 15 °C, 20 °C, and 25 °C to understand how sub-optimal temperatures affect the toxicokinetics and toxicodynamics of Hg via soil. Genotoxicity and energy reserves were also assessed to disclose potential trade-offs in life-history traits. Results underpin the complexity of temperature-Hg interactions. Survival was determined mainly by toxicokinetics, but toxicodynamics also played a significant role in defining survival probability during early stages. The processes determining survival probability were faster at 25 °C: General Unified Threshold of Survival (GUTS) model identified an earlier/steeper decline in survival, compared to 20 °C or 15 °C, but it also approached the threshold faster. Despite potentiation of Hg genotoxicity, temperature promoted faster detoxification, either increasing toxicokinetics rates or damage repair mechanisms. This metabolism-driven increase in detoxification led to higher depletion of energy reserves and likely triggered stress response pathways. This work emphasized the need for comprehensive experimental approaches that can integrate the multiple processes involved in temperature-metal interactions.

Susceptibility of Folsomia candida to Agrochemicals after Multigenerational Exposure to Human Pharmaceuticals.

In realistic environmental scenarios, soil organisms can be exposed to a combination of pharmaceuticals and agriproducts or within different time frames. Therefore, it is necessary to increase knowledge on soil organism susceptibility under a complex mixture exposure scenario. The present study aimed to assess the susceptibility of the collembolan Folsomia candida to copper and dimethoate on a pre-exposure for 3 generations to human pharmaceuticals (fluoxetine and carbamazepine). Carryover effects on reproductive output and survival were observed after a multigenerational pre-exposure to carbamazepine or fluoxetine, considerably increasing the sensitivity of collembolans to both copper and dimethoate. This was more evident for collembolans pre-exposed to the highest concentrations of both pharmaceuticals (40 mg/kg soil), as demonstrated by a significant reduction in the number of juveniles and increased mortality. In addition, pre-exposure to carbamazepine and fluoxetine induced varying effects on subsequent exposure to the same chemical. Although pre-exposure to carbamazepine led to a decrease in collembolan reproduction, even when transferred to a clean medium, fluoxetine induced severe effects but only when collembolans were exposed to other contaminants (i.e., not when transferred to clean soil). The present study highlighted the need to consider carryover effects and possible interactions between pharmaceuticals and other contaminants under simultaneous exposure. Environ Toxicol Chem 2022;41:592-600. © 2021 SETAC.

A Scientometric Study on Industrial Effluent and Sludge Toxicity.

The growth of industrialization has led to an increase in the production of highly contaminated wastewater. Industrial wastewater contains highly complex compounds varying in characteristics and required to be treated before its discharge into a water medium from various industries. However, the efficiency of the treated wastewater from the toxicity reduction perspective is unclear. In order to overcome this barrier, toxicity assessment of the industrial wastewater before and after treatment is crucial. Thus, in this study, a scientometric analysis has been performed on the toxicity assessment of industrial wastewater and sludges, which have been reported in the literature. Web of Science (WoS) core collection database has been considered the main database to execute this analysis. Via the search of pre-researched keywords, a total number of 1038 documents were collected, which have been published from 1951 to 2020. Via CiteSpace software and WoS analyser, these documents went under analysis regarding some of the scientometry criteria, and the detailed results obtained are provided in this study. The total number of published documents on this topic is relatively low during such a long period of time. In conclusion, the need for more detailed contributions among the scientific and industrial communities has been felt.

Biochar in soil mitigates dimethoate hazard to soil pore water exposed biota.

Soil contamination is a worldwide problem urging for mitigation. Biochar is a carbonaceous material used as soil amendment that can immobilize chemical compounds, potentially turning them unavailable for soil biota. The aim of our study was to evaluate biochar's capacity to immobilize dimethoate in soil and, therefore, decreasing the toxicity to soil organisms. Two biochar application rates (2.5% and 5% w/w) were chosen to assess dimethoate potential immobilization, looking at changes in its toxicity to the collembolan Folsomia candida and the plant Brassica rapa upon soil amendment. Complementarily, chemical analyses were performed on soil pore water. Results showed that biochar may sorb and decrease dimethoate concentrations in soil pore water, influencing dimethoate bioavailability and consequent toxicity. Contrary to dimethoate solo impact on collembolans (LC50 0.69 mg kg-1, EC50 0.46 mg kg-1), their survival rate and offspring production were not affected by dimethoate when biochar was applied, regardless of application rate (LC50 and EC50 > 1.6 mg kg-1). Shoot length, fresh and dry weights of B. rapa were less affected by dimethoate upon biochar addition (EC50 values increase for all endpoints). Our study shows that biochar may contribute to decrease dimethoate bioavailability and toxicity to soil porewater exposed organisms.

Effects of climate conditions on the avoidance behavior of Folsomia candida and Enchytraeus crypticus towards metal(loid)-contaminated soils.

Global climate changes are predicted for the 21st century. Alterations in soil contaminants' availability and soil invertebrates' behavior are expected, which may interfere with the avoidance capacity that invertebrates may have towards contaminated soils and, therefore, compromise their role in soil functioning. This study aimed to assess the individual effects of air temperature, ultraviolet (UV) radiation and atmospheric CO2 concentration on the avoidance behavior of the arthropod Folsomia candida and the soft-bodied oligochaete Enchytraeus crypticus towards metal(loid)-contaminated soils. Avoidance behavior was evaluated under distinct climate treatments (simulating increases in air temperature, UV radiation exposure or atmospheric CO2 concentration) and compared to the response obtained at the standard conditions recommended by ISO guidelines. Both soil invertebrate species behave differently under standard conditions, with F. candida not avoiding the contaminated soils while E. crypticus did. Increases in air temperature and exposure to UV radiation did not change F. candida behavior towards contaminated soils. However, high atmospheric CO2 concentration modified this pattern and induced avoidance towards contaminated soils. As for E. crypticus, contaminated soils were also avoided under the different climate treatments simulated. Thus, our study shows that, depending on the species and the climate factor, changes in climate conditions may alter soil invertebrates' behavioral pattern towards meta(loid)-contaminated soils.

Assessing the acute and chronic toxicity of exposure to naturally occurring oil sands deposits to aquatic organisms using Daphnia magna.

In the Athabasca region, the oil sands are located at or near the surface making open-pit mining viable. In addition, the Athabasca River and its tributaries flow through these oil sands deposits, thereby receiving bitumen-associated contaminants through natural fluvial erosional and weathering processes. A key knowledge gap has been related to understanding both the magnitude and significance of the toxicological and ecological effects on aquatic organisms exposed to naturally occurring bitumen entering fluvial systems. Using the Daphnia magna model system, this study assessed the ecotoxicological effects of exposure to bitumen-elutriate treatments that simulated the early stages of fluvial/erosional exposure conditions. No significant among-site differences were observed in the survival of D. magna after 48 h exposure to elutriates produced from a 24 h extraction cycle, and chemical analysis indicated low concentration of a complex mixture of hydrocarbon and metal contaminants. In contrast, the same elutriates impaired reproduction and growth after a 21-day chronic exposure. F1 neonates from the chronic tests were tested for sensitivity to the reference substance potassium dichromate, revealing a decrease in their sensitivity. Inter-generational effects were also observed, with a significant decrease in subsequent neonate production, when daphnids were moved to a clean medium. Supplemental acute toxicity assays using 48 and 72 h bitumen extraction cycles progressively increased daphnid mortality after a 48-h exposure to the respective elutriates. This indicates that bitumen-associated contaminants are being liberated after initial input and fluvial washing (24 h), highlighting the need for future work to assess toxicity responses and associated elutriate water chemistry of a longer fluvial exposure time-series. This work contributes to our understanding of the possible effects of natural bitumen exposure on riverine aquatic ecosystems, providing new information to inform the delineation of baseline conditions to assess environmental change and the design of future regional effects-based monitoring programs.

Effects of the organic UV-filter, 3-(4-methylbenzylidene) camphor, on benthic invertebrates and ecosystem function in artificial streams.

In the last decades, the use of organic ultraviolet-filters (UV-filters) has increased worldwide, and these compounds are now considered emerging contaminants of many freshwater ecosystems. The present study aimed to assess the effects of 3-(4-methylbenzylidene) camphor (4-MBC) on a freshwater invertebrate community and on associated ecological functions. For that, artificial streams were used, and a natural invertebrate benthic community was exposed to sediments contaminated with two concentrations of 4-MBC. Effects were evaluated regarding macroinvertebrate abundance and community structure, as well as leaf decomposition and primary production. Results showed that the macroinvertebrate community parameters and leaf decomposition rates were not affected by 4-MBC exposure. On the other hand, primary production was strongly reduced. This study highlights the importance of higher tier ecotoxicity experiments for the assessment of the effects of low concentrations of organic UV-filters on freshwater invertebrate community structure and ecosystem functioning.

Long-term exposure of Daphnia magna to carbendazim: how it affects toxicity to another chemical or mixture.

Aquatic organisms might be exposed episodically or continuously to chemicals for long-term periods throughout their life span. Pesticides are one example of widely used chemicals and thus represent a potential hazard to aquatic organisms. In addition, these chemicals may be present simultaneously in the environment or as pulses, being difficult to predict accurately how their joint effects will take place. Therefore, the aim of the present study was to investigate how Daphnia magna (clone k6) exposed throughout generations to a model pesticide (the fungicide carbendazim) would react upon an exposure to another chemical compound (triclosan) and to a mixture of both chemicals (carbendazim and triclosan). Responses of daphnids continuously exposed to carbendazim and kept in clean medium will be compared using immobilization tests and the comet assay (DNA integrity). The results showed that triclosan presented similar toxicity to daphnids exposed for 12 generations (F12) to carbendazim (similar 48-h-LC50 values for immobilization data), when compared with daphnids kept in clean medium. However, at subcellular level, daphnids previously exposed to carbendazim for 12 generations (F12) showed different responses than those from clean medium, presenting a higher toxicity; a general higher percentage of DNA damage was observed, after exposure to a range of concentrations of triclosan and to the binary combination of triclosan + carbendazim. The patterns of toxicity observed for the binary mixture triclosan + carbendazim were generally similar for daphnids in clean medium and daphnids exposed to carbendazim, with a dose level deviation with antagonism observed at low doses of the chemical mixture for the immobilization data and a dose ratio deviation with synergism mainly caused by triclosan for DNA damage. With this study, we contributed to the knowledge on long-term induced effects of carbendazim exposure, while looking at the organismal sensitivity to another chemical (triclosan) and to a mixture of carbendazim and triclosan using lethality as an endpoint at the individual level and DNA damage as a subcellular endpoint.

Soil moisture influences the avoidance behavior of invertebrate species in anthropogenic metal(loid)-contaminated soils.

Water availability is paramount in the response of soil invertebrates towards stress situations. This study aimed to evaluate the effects of forecasted soil moisture scenarios on the avoidance behavior of two invertebrate species (the arthropod Folsomia candida and the soft-bodied oligochaete Enchytraeus crypticus) in soils degraded by different types of anthropogenic metal(loid) contamination (mining soil and agricultural soil affected by industrial chemical wastes). Different soil moisture contents (expressed as % of the soil water holding capacity, WHC) were evaluated: 50% (standard soil moisture conditions for soil invertebrates' tests); 75% (to simulate increasing soil water availability after intense rainfalls and/or floods); 40%, 30%, 25% and 20% (to simulate decreasing soil water availability during droughts). Invertebrates' avoidance behavior and changes in soil porewater major ions and metal(loid)s were assessed after 48 h exposure. Soil incubations induced a general solubilization/mobilization of porewater major ions, while higher soil acidity favored the solubilization/mobilization of porewater metal(loid)s, especially at 75% WHC. Folsomia candida preferred soils moistened at 50% WHC, regardless the soils were contaminated or not and the changing soil porewater characteristics. Enchytraeus crypticus avoided metal(loid) contamination, but this depended on the soil moisture conditions and the corresponding changes in porewater characteristics: enchytraeids lost their capacity to avoid contaminated soils under water stress situations (75% and 20-25% WHC), but also when contaminated soils had greater water availability than control soils. Therefore, forecasted soil moisture scenarios induced by global warming changed soil porewater composition and invertebrates capacity to avoid metal(loid)-contaminated soils.

Mercury accumulation from food decreases collembolans' growth.

In the terrestrial environment, mercury (Hg) contamination can be originated from different inorganic and metal-organic sources, redistributed and transformed in soils. In the present study, the effects of contaminated food with environmentally relevant concentrations of Hg were evaluated in the soil-dwelling invertebrate Folsomia candida. Changes in growth rate and Hg bioaccumulation levels were observed at different concentrations of Hg in food, which can be complementary for data already available for reproduction and survival from standardized protocols. Collembolan growth was recorded every two days, and their growth rate along with a Von Bertalanffy's growth curve were derived showing that growth was dependent on Hg food concentration. Also, the final length of animals reflected the Hg concentration in food, with differences in all treatments comparing to non-exposed organisms. Toxicokinetic patterns from different Hg concentrations in food were not significantly different during the uptake phase, but differences were found in the depuration phase. Combining the two approaches, collembolans seem to invest their energy for depuration processes, neglecting other vital processes, such as growth. Also, contaminated food avoidance possibly occurred, thus decreasing their feeding and contaminant intake. Therefore, growth tests in collembolans can act as complementary tools to bioaccumulation and reproductive assays, towards a mechanistic understanding of how organisms use their energy upon contamination. Changes in growth rate, even at low and environmentally relevant concentrations, could be a warning signal when occurring in species with key roles in ecosystems. Also, this study highlights the importance of these complementary tests for a better and complete approach to risk assessment studies.

Multigenerational effects of carbendazim in Daphnia magna: From a subcellular to a population level.

Anthropogenic activities such as the use of pesticides may affect aquatic biota populations, due to potential agricultural runoffs or disposals. Carbendazim is one example of a widely used fungicide with a high potential to end up in aquatic ecosystems through runoff. Deleterious effects observed at the individual level are possibly explained by changes in homeostasis at the cellular level, and both factors can then be used to predict effects at the population level. In the present study, an isoclonal population of Daphnia magna (clone K6) was exposed to a concentration that mimics relevant levels of carbendazim in the environment over 12 generations. The effects of carbendazim were assessed in some generations using the following endpoints: biochemical biomarkers (cholinesterase, catalase, and glutathione-S-transferase), lipid peroxidation and energy-related parameters (carbohydrates, lipids, and proteins along with available energy and energy consumption), parental longevity, and population growth (r). Long-term exposure to carbendazim had no effect on the intrinsic rate of natural increase (r) of adult D. magna, but longevity was decreased at the F12 generation compared to that of control. Differences between the exposed and nonexposed populations were found for cholinesterase, glutathione-S-transferase, and lipid peroxidation. However, for catalase and energy-related parameters, no differences were observed between these 2 populations. Natural variability was seen throughout the test period, under control conditions, within the 12 generations. Overall, carbendazim induced some effects at the subcellular level that translated into changes in longevity but these later vanished in terms of population effects. Environ Toxicol Chem 2019;38:412-422. © 2018 SETAC.

Joint effects of chlorpyrifos and mancozeb on the terrestrial isopod Porcellionides pruinosus: A multiple biomarker approach.

The exposure to pesticides by nontarget soil biota has long been regarded as a serious downside of modern agricultural regimes and the subject of heated debate. Of utmost relevance is the exposure to pesticide mixtures because their effects have been shown to not necessarily reflect the individual toxicity of their components, and even the simple addition of effects may lead to consequences not clearly anticipated. In the present study, a multiple biomarker approach was employed to identify the mechanistic and time effects underlying several single and mixture treatments of chlorpyrifos (CPF) and mancozeb (MCZ) in juveniles and adults of the terrestrial isopod Porcellionides pruinosus. The effects of the individual pesticides and the mixtures at recommended doses were mostly transitory under these controlled conditions and one-pulse exposure. Whereas imbalances were identified on detoxification and oxidative stress-related enzymes, the isopods generally showed the ability to recover through the end of the experiment. However, juveniles displayed greater vulnerability than adults. Most of the differences between life stages occurred in energy-related parameters where distinct performances and stress-handling behaviors were observed, suggesting higher metabolic costs to juveniles. Our results stress that understanding the time dependence of the underlying mechanisms governing the joint effects of the pesticides can help in assessing and anticipating the effects of the pesticide mixtures. Moreover, we emphasize the importance of taking life stage-related differences into consideration when evaluating the environmental risks of pesticides and pesticide mixtures. Environ Toxicol Chem 2018;37:1446-1457. © 2018 SETAC.

The comet assay in Folsomia candida: A suitable approach to assess genotoxicity in collembolans.

The present study shows the comet assay technique being successfully applied for the first time to one of the most widely used soil organisms in standardized ecotoxicological tests, Folsomia candida, providing a step forward in assessing the genotoxicity induced by xenobiotics. Because collembolans have a high content of chitin, a new methodology was developed in which the heads of the collembolans were separated from the rest of the body, allowing the hemolymph to leak out. This procedure allows the cells to be released, and after lysis the genetic material is available for the comet assay. Among other key procedures, the use of 30 organisms (20- to 22-d-old adults) per replicate and the correct amount of cells with genetic material (translated as 10 µL of suspension) applied on the agarose gel were determinants for the success of the results obtained. The methodology was validated by exposing F. candida to a representative metallic element (cadmium) and a representative of organophosphates, the insecticide dimethoate, for a shorter time period of 10 d, compared with the 28 d for the International Organization for Standardization 11267 method. Within this method, the relatively low percentage of DNA damage (30%) observed in controls and the significant increase in terms of percentage of DNA damage for almost all the concentrations of dimethoate and Cd (reaching 52% and 56% of damage in the highest concentrations, respectively) confirmed the genotoxic effect of both compounds and validated this technique. The comet assay proved to be a sensitive technique to detect DNA strand breaks in collembolans' cells. Environ Toxicol Chem 2017;36:2514-2520. © 2017 SETAC.

Multigenerational effects of carbendazim in Daphnia magna.

Carbendazim is a fungicide largely used in agriculture as a plant protection product. As a result of agricultural runoffs, drainage, and leaching, it reaches surface waters at concentrations possibly hazardous to aquatic communities. Because of potential and continuous release of carbendazim to aquatic systems, long-term exposure to aquatic organisms should be addressed. To fill the knowledge gap, the present study evaluated the responses of multiple generations of Daphnia magna (clone K6) to an environmentally relevant concentration of carbendazim (5 µg/L). Twelve successive generations were evaluated, and the effects in these offspring were compared with those from a control population. Neonates' fitness was assessed through immobilization, reproduction, and feeding activity tests, along with the comet assay for in vivo DNA damage evaluation. Recovery from long-term exposure was also assessed. In the F5 generation, the results revealed that when daphnids were re-exposed to carbendazim, DNA damage was higher in daphnids continuously exposed to carbendazim than those from clean medium. After daphnids were moved to a clean medium, a low recovery potential was observed for DNA damage. Daphnids exposed continuously for 6 generations (F6) to carbendazim displayed an increase in feeding rates when re-exposed to carbendazim compared with F6 daphnids reared in clean medium. Continuous exposure of daphnids to carbendazim induced a significant increase in DNA damage from the F0 to the F12 generation. Deleterious effects of the multigenerational exposure to carbendazim were more prominent at a subcellular level (DNA damage) compared with the individual level. Environ Toxicol Chem 2017;36:383-394. © 2016 SETAC.

Ecotoxicity and genotoxicity of cadmium in different marine trophic levels.

Cadmium ecotoxicity and genotoxicity was assessed in three representative species of different trophic levels of marine ecosystems - the calanoid copepod Acartia tonsa, the decapod shrimp, Palaemon varians and the pleuronectiform fish Solea senegalensis. Ecotoxicity endpoints assessed in this study were adult survival, hatching success and larval development ratio (LDR) for A. tonsa, survival of the first larval stage (zoea I) and post-larvae of P. varians, egg and larvae survival, as well as the presence of malformations in the larval stage of S. senegalensis. In vivo genotoxicity was assessed on adult A. tonsa, the larval and postlarval stage of P. varians and newly hatched larvae of S. senegalensis using the comet assay. Results showed that the highest sensitivity to cadmium is displayed by A. tonsa, with the most sensitive endpoint being the LDR of nauplii to copepodites. Sole eggs displayed the highest tolerance to cadmium compared to the other endpoints evaluated for all tested species. Recorded cadmium toxicity was (by increasing order): S. senegalensis eggs < P. varians post-larvae < P. varians zoea I < S. senegalensis larvae < A. tonsa eggs < A. tonsa LDR. DNA damage to all species exposed to cadmium increased with increasing concentrations. Overall, understanding cadmium chemical speciation is paramount to reliably evaluate the effects of this metal in marine ecosystems. Cadmium is genotoxic to all three species tested and therefore may differentially impact individuals and populations of marine taxa. As A. tonsa was the most sensitive species and occupies a lower trophic level, it is likely that cadmium contamination may trigger bottom-up cascading effects in marine trophic interactions.