Search

In the present review, we synthesize information on the mechanisms of chronic copper (Cu) toxicity using an adverse outcome pathway framework and identify three primary pathways for chronic Cu toxicity: disruption of sodium homeostasis, effects on bioenergetics, and oxidative stress. Unlike acute Cu toxicity, disruption of sodium homeostasis is not a driving mechanism of chronic toxicity, but compensatory responses in this pathway contribute to effects on organism bioenergetics. Effects on bioenergetics clearly contribute to chronic Cu toxicity with impacts at multiple lower levels of biological organization. However, quantitatively translating these impacts into effects on apical endpoints such as growth, amphibian metamorphosis, and reproduction remains elusive and requires further study. Copper-induced oxidative stress occurs in most tissues of aquatic vertebrates and is clearly a significant driver of chronic Cu toxicity. Although antioxidant responses and capacities differ among tissues, there is no clear indication that specific tissues are more sensitive than others to oxidative stress. Oxidative stress leads to increased apoptosis and cellular damage in multiple tissues, including some that contribute to bioenergetic effects. This also includes oxidative damage to tissues involved in neuroendocrine axes and this damage likely alters the normal function of these tissues. Importantly, Cu-induced changes in hormone concentrations and gene expression in endocrine-mediated pathways such as reproductive steroidogenesis and amphibian metamorphosis are likely the result of oxidative stress-induced tissue damage and not endocrine disruption. Overall, we conclude that oxidative stress is likely the primary driver of chronic Cu toxicity in aquatic vertebrates, with bioenergetic effects and compensatory response to disruption of sodium homeostasis contributing to some degree to observed effects on apical endpoints. Environ Toxicol Chem 2022;00:1-18. (c) 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Nonpolar narcosis, also known as baseline toxicity, has been described as the minimal toxicity that an organic chemical may elicit based on its lipophilicity. Although lethal effects of narcosis-inducing chemicals (NICs) have been thoroughly investigated, knowledge of sublethal effects is still very limited. We investigated the effects of 3 well-known NICs (phenanthrene, 1,3,5-trichlorobenzene, and pentachlorobenzene) on a variety of organismal endpoints (malformations, swim bladder inflation, respiration, heart rate, swimming activity, and turning angles), which can be plausibly linked to narcosis in zebrafish embryos. Baseline toxicity recorded as mortality is typically observed in similar exposure ranges in a wide variety of species including fish, corresponding to a chemical activity range between 0.01 and 0.1. In the present study, we found that sublethal effects occurred at concentrations approximately 5 times below lethal concentrations. Altered swimming activity and impaired swim bladder inflation were the most sensitive endpoints occurring at exposure levels below the generally accepted threshold for baseline toxicity for 2 out of 3 compounds. Overall, most effective exposure levels across the sublethal endpoints and compounds did fall within the range typically associated with baseline toxicity, and deviations were generally limited to a factor 10. Although there could be benefit in adding sublethal endpoints to toxicity tests, such as the fish embryo acute toxicity (FET) test, based on the present sublethal endpoints and available evidence from our and other studies, the underestimation of toxicity as a result of the sole assessment of mortality as an endpoint in an FET test may be limited for narcosis. Environ Toxicol Chem 2021;40:2802-2812. © 2021 SETAC.

Fish (embryo) toxicity test guidelines are mostly based on aquatic exposures. However, in some cases, other exposure routes can be more practical and relevant. Micro-injection into the yolk of fish embryos could offer a particular advantage for administering hydrophobic compounds, such as many endocrine disruptors. Single-dose micro-injection was compared with continuous aquatic exposure in terms of compound accumulation and biological responses. 17α-Ethinyl estradiol (EE2) was used as a model compound. First, the optimal solvent and droplet size were optimized, and needle variation was assessed. Next, biological endpoints were evaluated. The accumulated internal dose of EE2 decreased over time in both exposure scenarios. Estrogen receptor activation was concentration/injected dose dependent, increased daily, and was related to esr2b transcription. Transcription of vitellogenin 1 (vtg1) and brain aromatase (cyp19a1b) was induced in both scenarios, but the cyp19a1b transcription pattern differed between routes. Injection caused an increase in cyp19a1b transcripts from 48 hours post fertilization (hpf) onward, whereas after aquatic exposure the main increase occurred between 96 and 120 hpf. Some malformations only occurred after injection, whereas others were present for both scenarios. We conclude that responses can differ between exposure routes and therefore micro-injection is not a direct substitute for, but can be complementary to aquatic exposure. Nevertheless, vtg1and cyp19a1b transcription and estrogen receptor activation are suitable biomarkers for endocrine disruptor screening in both scenarios. Environ Toxicol Chem 2019;38:533-547. © 2018 SETAC.

Current emission and mobilization rates of mercury (Hg) in the environment pose extensive threats to both wildlife and human health. Assessing the exposure risk and effects of Hg contamination in model species such as seabirds is essential to understand Hg risks at the population and ecosystem levels. The lesser black-backed gull (Larus fuscus), a generalist seabird species, is an excellent model species because it forages in both marine and terrestrial habitats, which in turn differ in their Hg exposure risk. To identify possible deleterious effects of Hg exposure on developingL. fuscuschicks, a dietary experiment was carried out and chicks were provided a marine, terrestrial, or mixed diet. The effects of embryonic and dietary Hg exposure on chick body condition and physiological state were assessed at different developmental stages until fledging age (30 d). Overall physiological condition was lower in chicks fed a predominantly marine diet, which coincided with higher Hg loads in blood and primary feathers. However, no effect of dietary uptake of Hg was observed on body condition or in terms of genotoxic damage. Body condition and genotoxic damage correlated instead with Hg exposure during embryonic development, which seems to indicate that embryonic exposure to Hg may result in carry-over effects on later chick development.Environ Toxicol Chem2020;39:2008-2017. (c) 2020 SETAC

It is well known that temperature can affect the ecotoxicity of chemicals (including metals) to aquatic organisms. It was recently reported that nickel (Ni), a priority substance under the European Water Framework directive, showed decreasing chronic toxicity to Daphnia magna with increasing temperature, between 15 and 25 degrees C. We performed a toxicokinetic study to contribute to an increased mechanistic understanding of this effect. More specifically, we investigated the effect of temperature on Ni uptake and elimination in D. magna (in 4 clones) using an experimental design that included Ni exposures with different stable isotopic composition and using a one-compartment model for data analysis. Both Ni uptake and elimination were affected by temperature, and some clear interclonal differences were observed. On average (across all clones), however, a similar pattern of the effect of temperature was observed on both Ni uptake and elimination, that is, the uptake rate constant (k(u)) and elimination rate constant (k(e)) during 72 h of Ni exposure were lower at 25 than at 19 degrees C, by 2.6-fold and 1.6-fold, respectively, and they were similar at 19 and 15 degrees C. This pattern does not correspond to the effects of temperature on chronic Ni toxicity reported previously, suggesting that Ni compartmentalization and/or toxicodynamics may also be affected by temperature. The data gathered with our specific experimental design also allowed us to infer that 1) the k(u) was up-regulated over time, that is, the k(u) after 2 d of Ni exposure was significantly higher than the initial k(u), by 1.5- to 2.3-fold, and 2) the k(e) decreased significantly when the external Ni exposure was stopped, by 1.2- to 1.9-fold. These 2 findings are in contrast with 2 commonly used assumptions in toxicokinetic models, that is, that k(u) is constant during exposure and k(e) is independent of external exposure. We suggest that future toxicokinetic studies consider these factors in their experimental designs and data analyses. Overall, our study contributes to the growing body of evidence that temperature affects toxicokinetics of metals (and chemicals in general), but at the same time we emphasize that knowledge of toxicokinetics alone is not necessarily sufficient to explain or predict temperature effects on (chronic) toxicity. Environ Toxicol Chem 2019;38:784-793. (c) 2019 SETAC

Pharmaceuticals are essential for human well-being, but their increasing and continuous use pollutes the environment. Although behavioral ecotoxicology is increasingly advocated to assess the effects of pharmaceutical pollution on wildlife and ecosystems, a consensus on the actual environmental risks is lacking for most compounds. The main limitation is the lack of standardized reproducible tests that are based on sensitive behavioral endpoints and that accommodate a high ecological relevance. In the present study, we assessed the impact of a 3-wk exposure to the antidepressant fluoxetine on multiple behavioral traits in the promising new model organism Nothobranchius furzeri (turquoise killifish). Overall, our study shows that fluoxetine can impact feeding behavior, habitat choice in a novel environment, and antipredator response of N. furzeri individuals; effects on spontaneous activity and exploration tendency were less pronounced. However, effects became only apparent when individuals were exposed to fluoxetine concentrations that were 10 times higher than typical concentrations in natural aquatic environments. Ecotoxicologists are challenged to maximize both the reliability and ecological validity of risk assessments of pollutants. Our study contributes to the development of a time- and cost-efficient, standardized ecotoxicological test based on sensitive, ecologically relevant behavioral endpoints in N. furzeri. Environ Toxicol Chem 2019;38:262-270. © 2018 SETAC. ispartof: ENVIRONMENTAL TOXICOLOGY AND CHEMISTRY vol:38 issue:1 pages:262-270 ispartof: location:United States status: published

Recent studies have shown that temperature affects chronic nickel (Ni) toxicity to Daphnia magna at the individual (apical) level. However, the effect of temperature on Ni toxicity to D. magna at the population level is unknown. The present study investigated whether the effect of temperature on chronic Ni toxicity to D. magna assessed on apical endpoints can be extrapolated to the population level. The results of the population experiment showed no consistent Ni effects on total D. magna population abundance at 15, 20, and 25 degrees C, although the Ni concentrations tested were previously reported to significantly reduce reproduction in D. magna individuals. This result supports the idea that ecological risk assessment should not extrapolate as such from apical endpoints to the population level. A dynamic energy budget individual-based model (DEB-IBM) was calibrated using apical Ni toxicity data at 15, 20, and 25 degrees C. The goal was to investigate whether the calibrated DEB-IBM would be able to predict the unexpected absence of effects at the population level and to further investigate the effect of temperature on Ni toxicity to a D. magna population. At the population level, the calibrated DEB-IBM correctly predicted the unexpected absence of an effect of Ni on a D. magna population. Detailed analysis of simulation output suggests that the predicted lower Ni sensitivity at the population level occurs because Ni-induced mortality is compensated by reduced starvation (less intraspecific competition). Extrapolated median effective concentration (EC50) values for population density predicted that the effect of temperature on Ni toxicity to D. magna populations was smaller (1.9-fold higher at 25 degrees C than at 15 degrees C) than on Ni toxicity to D. magna apical reproduction (the EC50 is 6.5-fold higher at 25 degrees C than at 15 degrees C). These results show that the DEB-IBM can help to replace population experiments by in silico simulations and to optimize the experimental design of population studies. Environ Toxicol Chem 2019;38:1423-1433. (c) 2019 SETAC

Freshwater organisms are increasingly exposed to combinations of stressors. However, because it is time-consuming and costly, research on the interaction of stressors, such as compound toxicity and global warming on vertebrates, is scarce. Studies on multigenerational effects of these combined stressors are almost nonexistent. In the present study, we tested the combined effects of 4 °C warming and cadmium (Cd) exposure on life-history traits, biomarkers, bioaccumulation, and multigenerational tolerance in the turquoise killifish, Nothobranchius furzeri. The extremely short life cycle of this vertebrate model allows for assessment of sublethal and multigenerational effects within 4 mo. The applied Cd concentrations had only limited effects on the measured endpoints, which suggests that N. furzeri is more resistant to Cd than fathead minnow and rainbow trout. In contrast, the temperature increase of 4 °C was stressful: it delayed female maturation and lowered adult mass and fecundity. Finally, indications of synergistic effects were found on peak fecundity and embryonic survival. Overall, these results indicate the importance of studying chronic and multigenerational effects of combined stressors. Environ Toxicol Chem 2018;37:2361-2371. © 2018 SETAC. ispartof: ENVIRONMENTAL TOXICOLOGY AND CHEMISTRY vol:37 issue:9 pages:2361-2371 ispartof: location:United States status: published

Ecological risk assessment is commonly based on single-generation ecotoxicological tests that are usually performed at one standard temperature. We investigated the effects of nickel (Ni) on Daphnia magna reproduction at 15, 20, and 25 degrees C over 4 generations. Multigenerational Ni effects on D. magna reproduction depended on the magnitude of the effect in the first generation (F0) and showed very different patterns at different temperatures. At low effect level concentrations (

In this study toxicity effects of zinc, copper and cadmium, both as single metals and in combination, were examined for the nematode Caenorhabditis elegans. Metal effects on lethality were analysed in a time-dependent manner using different concentrations in K-medium. LC20 concentrations were used to investigate the effects on locomotion and chemosensation. The results showed that copper toxicity was higher compared to cadmium and zinc, resulting in higher mortality rates and a more reduced locomotion. For all metals lethality increased over time. When cadmium concentrations were added to copper, and vice versa, significant increases in toxicity were noted. Different interaction effects were observed for the mixtures ZnCd, ZnCu, CuCd and ZnCuCd. Zinc seemed to have a neutral toxic effect on cadmium, while in combination with copper, a similar additive effect was seen as for the CuCd combination. Binary and tertiary metal mixtures caused a strong decrease in locomotion, except for the ZnCd combination, where zinc seems to have a neutral effect. After LC2024 h exposure, a reduced crawling speed, except for zinc, and a reduced thrashing behaviour, except for zinc and ZnCd mixture were observed. Almost no significant effects were observed on chemosensation. Since the same trend of mixture effects was noted in locomotion and in lethality tests, locomotion could probably be considered as a sensitive endpoint for metal toxicities.

The analysis of metal mixture toxicity to plants is complicated by mutual interactions. In the present study, mixture effects of zinc (Zn), cadmium (Cd), and copper (Cu) on barley (Hordeum vulgare L.) root elongation were analyzed using oxidative stress parameters. The hypothesis was that toxic mixture effects on plant growth are better explained by biochemical parameters than by exposure information, because the former excludes interactions among metals for root uptake. Barley seedlings were exposed for 5 d or 14 d to these metals in nutrient solutions, added in isolation and as mixtures. Root elongation in Cu+Cd mixtures was well predicted from free metal ion concentrations in solution, using concentration addition (CA) or independent action (IA) reference models. In contrast, Zn acted antagonistically when combined with Cu and/or Cd, relative to both CA and IA. This protective effect of Zn correlated with the biomarkers measured in the long-term experiment; oxidative stress (indicated by malondialdehyde level, for example) decreased after addition of Zn. In addition, it was found that some biomarkers were sensitive to both Cu and Cd dosed in isolation, but not to Cu+Cd mixtures. Overall, the exposure explained mixture effects better than most of the 16 measured biomarkers (i.e., the biochemical effects). It is concluded that these biomarkers are not robust indicators for metal mixture toxicity, potentially because different metals have different parallel modes of action on growth that are insufficiently indexed by the biomarkers.

Few studies have considered the effect of temperature on the chronic sensitivity of Daphnia magna to other stressors. The present study investigated the effect of temperature on chronic metal toxicity and whether this effect differed among 4 different D. magna clones. Life table experiments were performed with copper, zinc, and nickel at 15 degrees C, 20 degrees C, and 25 degrees C. General linear modeling indicated that chronic Cu, Zn, and Ni toxicity to D. magna were all significantly affected by temperature. When averaged across clones, our results suggest that chronic metal toxicity to D. magna was higher at 15 degrees C than at 20 degrees C, which is the temperature used in standard toxicity tests. At 15 degrees C, the 21-d median effect concentrations (EC50s) of Cu, Zn, and Ni were 1.4 times, 1.1 times, and 1.3 times lower than at 20 degrees C, respectively. At 25 degrees C, chronic Cu and Zn toxicity did not change in comparison with 20 degrees C, but chronic Ni toxicity was lower (21-d EC50 of nickel at 25 degrees C was 1.6 times higher than at 20 degrees C). The same trends were observed for Cu and Ni when the 21-d 10% and 20% effect concentrations were considered as the effect estimator, but not for Zn, which warns against extrapolating temperature effects on chemical toxicity across effect sizes. Overall, however, chronic metal toxicity was generally highest at the lowest temperature investigated (15 degrees C), which is in contrast with the usually observed higher acute metal toxicity at higher temperatures. Furthermore, the effect of temperature on chronic Ni toxicity depended significantly on the clone. This warns against extrapolating results about effect of temperature on chemical toxicity from single clone studies to the population level. (C) 2016 SETAC

Our ability to conduct whole-organism toxicity tests to understand chemical safety has been outpaced by the synthesis of new chemicals for a wide variety of commercial applications. As a result, scientists and risk assessors are turning to mechanistically based studies to increase efficiencies in chemical risk assessment and making greater use of in vitro and in silico methods to evaluate potential environmental and human health hazards. In this context, the adverse outcome pathway (AOP) framework has gained traction in regulatory science because it offers an efficient and effective means for capturing available knowledge describing the linkage between mechanistic data and the apical toxicity end points required for regulatory assessments. A number of international activities have focused on AOP development and various applications to regulatory decision-making. These initiatives have prompted dialogue between research scientists and regulatory communities to consider how best to use the AOP framework. Although expert-facilitated discussions and AOP development have been critical in moving the science of AOPs forward, it was recognized that a survey of the broader scientific and regulatory communities would aid in identifying current limitations while guiding future initiatives for the AOP framework. To that end, a global horizon scanning exercise was conducted to solicit questions concerning the challenges or limitations that must be addressed to realize the full potential of the AOP framework in research and regulatory decision-making. The questions received fell into several broad topical areas: AOP networks, quantitative AOPs, collaboration on and communication of AOP knowledge, AOP discovery and development, chemical and cross-species extrapolation, exposure/toxicokinetics considerations, and AOP applications. Expert ranking was then used to prioritize questions for each category, where 4 broad themes emerged that could help inform and guide future AOP research and regulatory initiatives. In addition, frequently asked questions were identified and addressed by experts in the field. Answers to frequently asked questions will aid in addressing common misperceptions and will allow for clarification of AOP topics. The need for this type of clarification was highlighted with surprising frequency by our question submitters, indicating that improvements are needed in communicating the AOP framework among the scientific and regulatory communities. Overall, horizon scanning engaged the global scientific community to help identify key questions surrounding the AOP framework and guide the direction of future initiatives. Environ Toxicol Chem 2017;36:1411-1421. © 2017 SETAC.

Inflation of the posterior and/or anterior swim bladder is a process previously demonstrated to be regulated by thyroid hormones. We investigated whether inhibition of deiodinases, which convert thyroxine (T4) to the more biologically active form, 3,5,3′-triiodothyronine (T3), would impact swim bladder inflation. Two experiments were conducted using a model deiodinase inhibitor, iopanoic acid (IOP). First, fathead minnow embryos were exposed to 0.6, 1.9, or 6.0 mg/L or control water until 6 d postfertilization (dpf), at which time posterior swim bladder inflation was assessed. To examine anterior swim bladder inflation, a second study was conducted with 6-dpf larvae exposed to the same IOP concentrations until 21 dpf. Fish from both studies were sampled for T4/T3 measurements and gene transcription analyses. Incidence and length of inflated posterior swim bladders were significantly reduced in the 6.0 mg/L treatment at 6 dpf. Incidence of inflation and length of anterior swim bladder were significantly reduced in all IOP treatments at 14 dpf, but inflation recovered by 18 dpf. Throughout the larval study, whole-body T4 concentrations increased and T3 concentrations decreased in all IOP treatments. Consistent with hypothesized compensatory responses, deiodinase-2 messenger ribonucleic acid (mRNA) was up-regulated in the larval study, and thyroperoxidase mRNA was down-regulated in all IOP treatments in both studies. These results support the hypothesized adverse outcome pathways linking inhibition of deiodinase activity to impaired swim bladder inflation. Environ Toxicol Chem 2017;36:29422952. Published 2017 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.

To improve the ecological relevance of environmental risk assessment, an improved understanding is needed of 1) the influence of environmental conditions on the toxicity of pollutants, and 2) the effect of these factors in combination with possible interpopulation variability. The influences of salinity and dissolved organic carbon (DOC) on the accumulation and effect of copper (Cu) to settled mussels were investigated with mussels from a North Sea and a Baltic Sea population. We found that both populations were equally Cu-sensitive, even though the Baltic Sea population lives in suboptimal conditions. Baltic Sea mussels, however, accumulated more Cu. This suggests that these populations may have different ways of coping with excess Cu. The influence of salinity on Cu toxicity to settled mussels was limited for both populations. An increase in DOC did not decrease the Cu accumulation or effect in either population. This suggests that DOC-Cu complexes are bioavailable for settled mussels. These findings are in contrast with previous research which indicated that DOC decreased the toxicity and accumulation of Cu in the D-larvae life stage. As a consequence, the mussel larval stage is not the most Cu-sensitive life stage at high DOC concentrations. Furthermore, a DOC correction factor for Cu toxicity cannot be used for settled mussels. This should be accounted for in future marine Cu environmental risk assessment. (C) 2017 SETAC

Metal pollution is a serious threat to environmental health. While the aquatic isopod Asellus aquaticus L. (Isopoda) is an important decomposer of freshwater ecosystems, very little research has reported its long-term or incipient lethal concentrations for metals. Moreover, the lethal concentrations at a certain percentage (LCxs) that can be found in the literature are often based on unmeasured concentrations, which could lead to a severe underestimation of the actual toxicity. In the present exploratory study, the 1-, 4-, 7-, and 14-d LC10, LC20, and LC50 values of copper, cadmium, and lead for adult A. aquaticus were determined. The LCx values were calculated using the nominal concentrations, the effective concentrations, and the free ion activities. Incipient lethal values were determined as well. In general, surprisingly lower lethal concentrations were found than in other studies. Furthermore, the present study shows that lethal concentrations based on free ion activities were generally much lower than nominal and effective concentrations. Although almost all LC values were higher than the environmental quality standards (EQS), the Cu 14-d LC10 and several (incipient) Pb LC10 and LC20 values, all calculated with free ion activities, were lower than the EQS. We conclude that lethal or effective concentrations based on free ion activities provide a more realistic view, which is vital for environmental policy making

Although aquatic organisms in the environment are exposed to mixtures of metals, risk assessment for metals is most commonly performed on a metal-by-metal basis. To increase the knowledge about chronic mixture effects, the authors investigated whether metal mixture effects are dependent on the biological species, mixture composition, and metal concentration ratio. The authors evaluated the effects of quaternary Ni-Zn-Cu-Cd and ternary Ni-Zn-Cu mixtures on 48-h algal growth rate (Pseudokirchneriella subcapitata) and 7-d daphnid reproduction (Ceriodaphnia dubia) using a ray design. Single metals were 3-fold to 42-fold more toxic for C. dubia than for P. subcapitata, based on the 50% effective concentration expressed as free metal activity, the range representing different metals. Statistical analysis of mixture effects showed that the ternary and quaternary mixture effects were antagonistic on algal growth relative to the concentration addition (CA) model, when the analysis was based on dissolved concentrations and on free metal ion activities. Using the independent action (IA) model, mixture effects in both rays were statistically noninteractive for algal growth when the analysis was based on dissolved concentrations; however, the interactions shifted toward antagonism when based on free ion activities. The ternary Ni-Zn-Cu mixture acted antagonistically on daphnid reproduction relative to both reference models, either expressed as free ion activities or dissolved concentrations. When Cd was added to the mixture, however, the mixture effects shifted toward noninteractivity for daphnids. The metal concentration ratio did not significantly influence the magnitude of observed antagonistic effects. Regardless of statistical interactions observed, based on the present study, CA and in most instances also IA can serve as a protective model for ternary Ni-Zn-Cu and quaternary Ni-Zn-Cu-Cd toxicity to both species. Environ Toxicol Chem 2017;36:1056-1066. © 2016 SETAC.

Metal contamination is mostly a mixture of different metals, and these multicomponent mixtures can produce significant mixture effects. The present study was set up to investigate the toxicity of multiple metal mixtures of Cu, Ni, Cd, and Zn to plants at metal doses individually causing low-level phytotoxic effects. Barley (Hordeum vulgare L.) root elongation toxicity tests were performed in resin-buffered nutrient solutions to control metal speciation. Treatments included single-metal concentrations and binary, ternary, and quaternary mixtures. Mixtures of different metals at free ion concentrations, each causing

Metals occur as mixtures in the environment. Risk assessment procedures for metals currently lack a framework to incorporate chronic metal mixture toxicity. In the present study, the toxicity of binary and ternary mixture combinations of Ni, Zn, and Pb was investigated in 3 large-scale experiments using the standard chronic (7-d) Ceriodaphnia dubia reproductive toxicity test. These metals were selected because of anticipated differences in mode of action. The toxicity of the metals in most mixtures, expressed as either free metal ion activities or as dissolved metal concentrations, were antagonistic relative to the concentration addition model, whereas no significant (p < 0.05) interactive effects were observed relative to the independent action model. The only exception was the binary Pb-Zn mixture, for which mixture effects were noninteractive based on the dissolved concentrations, but antagonistic based on free ion activities all relative to the independent action model. Overall, the independent action model fitted the observed toxicity better than the concentration addition model, which is consistent with the different modes of action of these metals. The concentration addition model mostly overestimated toxicity. Finally, the present study warns against extrapolation of the type of interactive effects between species, even when they are closely related. Environ Toxicol Chem 2016;35:1796-1805. (c) 2015 SETAC

Metal ecotoxicity to soil organisms (for example, in enchytraeids) has been addressed mainly by assessing effects on survival and reproduction, but very little is known about the underlying molecular mechanisms of responses. The main purpose of the present study was to assess and compare the transcriptional responses of Enchytraeus albidus to an essential (Zn) and a nonessential (Cd) metal. Exposure was performed with two concentrations with a known effect on reproduction (effective concentration for 50% [EC50] and 90% [EC90]) at three time points (2, 4, and 8 d). Results showed that transcriptional responses were influenced by exposure duration but, independently of that, the mechanisms of response to Cd and Zn were consistently different. Both metals affected pathways related to the regulation of gene expression, calcium homeostasis, and cellular respiration. Mechanisms of toxicity that were exclusively associated with Cd exposures were the inhibition of DNA repair and the impairment of ubiquitin-mediated proteolysis. The microarray for E. albidus was a useful tool for detecting molecular pathways affected by metal exposures. Transcriptional responses strongly correlated with known mechanisms of Cd and Zn responses in other organisms, suggesting cross-species conserved mechanisms of action. It should be highlighted not only that the authors could retrieve mechanistic information but also that genes responded within 2 to 8 d of exposure. This represents an additional advantage of using such molecular endpoints as a complement to the traditional, more time-consuming endpoints. published

To incorporate metal mixture toxicity effects into risk-assessment procedures, more information is needed about combined and interactive effects of metal mixtures during chronic exposure. The authors investigated the toxicity of binary Ni-Zn mixtures in 2 independent full-factorial experiments using standard chronic (21-d) Daphnia magna reproduction toxicity tests. Global statistical analysis (i.e., when considering all investigated mixture treatments simultaneously) showed noninteractive effects according to the concentration addition model and significant synergistic effects according to the independent action model. However, treatment-specific statistical analysis revealed that both occurrence and type of interactive effect were dependent on the effect size at which Ni and Zn were combined in the mixture. Only noninteractive or weakly antagonistic effects occurred in mixture treatments in which each of the individual metals produced only weak adverse effects on its own (i.e., 20% reduction of reproductive performance). On the other side of the spectrum, synergistic mixture effects occurred in all mixture treatments where both metals already caused a >20% (for independent action) and a >40% (for concentration addition) effect on reproduction on their own. Because low effect sizes are the most relevant in most regulatory frameworks, the authors' data suggest that the concentration addition and independent action mixture toxicity models can both serve as conservative models for predicting effects of Ni-Zn mixtures. The present study highlights the importance of investigating metal mixture toxicity at low effect sizes and warns against extrapolating conclusions about metal mixture interactions from high to low effect sizes. Environ Toxicol Chem 2015;34:1091-1102. (c) 2015 SETAC

Predicting copper (Cu) toxicity in marine and estuarine environments is challenging because of the influence of anions on Cu speciation, competition between Cu2+ and other cations at the biotic ligand and the effect of salinity on the physiology of the organism. In the present study the combined effect of salinity and dissolved organic carbon (DOC) on Cu toxicity to larvae of Mytilus galloprovincialis was assessed. Two statistical models were developed and used to elucidate the relationship between Cu toxicity, salinity, and DOC. All models based on dissolved Cu indicate a decrease in Cu toxicity with increasing DOC concentrations, which can partly be explained by complexation of Cu2+ ions with DOC. These models also indicate an increase in Cu toxicity (modeled with dissolved Cu or Cu2+ activity) with increasing salinity, suggesting a salinity-induced alteration in the physiology of the mussel larvae. When based on Cu body burdens, neither of the models indicates an effect of salinity or DOC. This shows that the Cu body burden is a more constant predictor of Cu toxicity, regardless of the water chemistry influencing Cu speciation or competition and possible physiological alterations or changes in Cu speciation or competition. Environ Toxicol Chem 2015;34:1330-1336. (c) 2015 SETAC

Mouthpart deformities in chironomids have been reported to indicate adverse effects of environmental pollutants. The authors assessed rates of mouthpart deformities in tributyltin-exposed, inbred, and outcrossed Chironomus riparius larvae over multiple generations. The authros found that the occurrence of mouthpart deformities was significantly correlated with inbreeding, whereas no correlation was found with the tributyltin exposure. The present study confirms the strong effect of high inbreeding rates on developmental deformities in chironomids.

Copper uptake and accumulation by the marine mussel Mytilus edulis were studied at different Cu concentrations in chemically defined artificial seawater in the presence and absence of fulvic acids. Both short-term uptake of Cu by excised mussel gills and Cu accumulation in whole mussels after 24 h of exposure decreased in the presence of fulvic acids compared with their absence at similar dissolved Cu concentrations. Calculations of Cu speciation based on previous measurements of labile Cu by anodic stripping voltammetry demonstrated that Cu uptake and accumulation depended on the concentration of labile Cu, in agreement with the free ion activity model. No evidence of a significant uptake of Cufulvic acid complexes was observed. Environ. Toxicol. Chem.

Environmental pollution with natural or synthetic estrogens may pose a serious threat to reproduction of wildlife species. This study describes the effects of 17-alpha-ethynylestradiol (EE2) on fish reproductive organs in a laboratory model. Adult zebrafish were semistatically exposed to nominal concentrations of 0, 10, and 25 ng/L EE2 for 24 d and then transferred to EE2-free medium. Gonadosomatic index (GSI), plasma vitellogenin concentration (VTG), and histology of the gonads (control and 10 ng/L only) were examined as a function of time. It was found that EE2 has an adverse impact on both male and female reproductive organs. Notably in females, gonadal changes were observed through histological evaluation after 3 d of exposure to 10 ng/L EE2, and this was followed by a reduction of GSI at day 6 of exposure. In males, a reduction of GSI and altered testis histology was found after 24 d of exposure to 10 ng/L. The observed effects on the ovary after EE2 exposure, combined with complete recovery after 24 d is considered to be triggered by feedback at the level of the pituitary. In both males and females, VTG was induced in response to EE2 and normalized during the recovery period. The observed correlation between VTG and ovarian somatic index (OSI) demonstrates that excessive VTG induction may be predictive for adverse effects of EE2 on ovarian function in female zebrafish. These results indicate that long-term stimulation by synthetic estrogens such as EE2 might impair reproductive function in zebrafish in a reversible manner.

First-stage larvae of the midge Chironomus riparius were exposed in small enclosures at 19 sites located in three different river basins in Flanders (Belgium). Sediments were sampled and sieved at 200 mum at all exposure sites. A layer of approximately 2 cm of sediment was placed in each cage and 100 midge larvae were added. Cages were placed in watercourses where resident midge larvae were present. Accumulation of Cd, Cr, Cu, Pb, Ni, and Zn was determined after four weeks of exposure when larvae had reached the fourth stage. Comparing metal levels between caged and resident larvae revealed no significant differences. A significant correlation between metal levels in caged and resident larvae was found when all sites were considered. However, such correlation was low (r(2) = 0.28) for Pb. The highly significant r(2) values found for Cu and Ni probably were due to only one site. Metal levels in tissue were related to levels in water and sediment, taking into account some sediment characteristics (particle size distribution and organic carbon) and oxygen level in the water. To determine the relative importance of these different sediment factors contributing to the variation in metal accumulation by the chironomids, nonlinear regression models were constructed. With the models used, 56.1, 32.2, and 57.4% of the variation for Cd, Pb, and Zn, respectively, could be described. None and 26.9% of the variation could be described for Cu and Ni, respectively. Among the environmental factors, organic carbon and oxygen levels in water were important in describing the accumulation of metals.