Olivine avoidance behaviour by marine gastropods (Littorina littorea L.) and amphipods (Gammarus locusta L.) within the context of ocean alkalinity enhancement.

Gigaton scale atmospheric carbon dioxide (CO2) removal (CDR) is needed to keep global warming below 1.5 °C. Coastal enhanced olivine weathering is a CDR technique that could be implemented in coastal management programmes, but its CO2 sequestration potential and environmental safety remain uncertain. Large scale olivine spreading would change the surficial sediment characteristics, which could potentially reduce habitat suitability and ultimately result in community composition changes. To test this hypothesis, we investigated the avoidance response of the marine gastropod Littorina littorea (Linnaeus, 1758) and marine amphipod Gammarus locusta (Linnaeus, 1758) to relatively coarse (83 - 332 µm) olivine and olivine-sediment mixtures during short-term choice experiments. Pure olivine was significantly avoided by both species, while no significant avoidance was observed for sediment with 3% or 30% w/w olivine. For L. littorea, aversion of the light green colour of pure olivine (i.e. positive scototaxis) was the main reason for avoidance. Moreover, olivine was not significantly avoided when it was 7.5 cm (45%) closer to a food source/darker microhabitat (Ulva sp.) compared to natural sediment. It is inferred that the amphipod G. locusta avoided pure olivine to reduce Ni and Cr exposure. Yet, a significant increase in whole body Ni concentrations was observed after 79 h of exposure in the 30% and 100% w/w olivine treatments compared to the sediment control, likely as a result of waterborne Ni uptake. Overall, our results are significant for ecological risk assessment of coastal enhanced olivine weathering as they show that L. littorea and G. locusta will not avoid sediments with up to 30% w/w relatively coarse olivine added and that the degree of olivine avoidance is dependent on local environmental factors (e.g. food or shelter availability).

Effect of Polymer Aging on Uptake/Release Kinetics of Metal Ions and Organic Molecules by Micro- and Nanoplastics: Implications for the Bioavailability of the Associated Compounds.

The main driver of the potential toxicity of micro- and nanoplastics toward biota is often the release of compounds initially present in the plastic, i.e., polymer additives, as well as environmentally acquired metals and/or organic contaminants. Plastic particles degrade in the environment via various mechanisms and at different rates depending on the particle size/geometry, polymer type, and the prevailing physical and chemical conditions. The rate and extent of polymer degradation have obvious consequences for the uptake/release kinetics and, thus, the bioavailability of compounds associated with plastic particles. Herein, we develop a theoretical framework to describe the uptake and release kinetics of metal ions and organic compounds by plastic particles and apply it to the analysis of experimental data for pristine and aged micro- and nanoplastics. In particular, we elucidate the contribution of transient processes to the overall kinetics of plastic reactivity toward aquatic contaminants and demonstrate the paramount importance of intraparticulate contaminant diffusion.

Acute bioaccumulation and chronic toxicity of olivine in the marine amphipod Gammarus locusta.

Active atmospheric carbon dioxide removal (CDR) is needed at a gigaton scale in the next decades to keep global warming below 1.5 °C. Coastal enhanced silicate weathering (CESW) aims to increase natural ocean carbon sequestration via chemical weathering of finely ground olivine (MgxFe(1-x)SiO4) rich rock dispersed in dynamic coastal environments. However, the environmental safety of the technique remains in question due to the high Ni and Cr content of olivine. Therefore, we investigated the short term bioaccumulation and chronic toxicity of olivine in the marine amphipod Gammarus locusta. Acute 24-h olivine exposure resulted in significant grain size dependent olivine ingestion and subsequent Ni and Cr accumulation in tissues. Thousands of small (mainly ≤ 10  µm) olivine grains were ingested by G. locusta, but their importance for trace metal bioaccumulation requires additional research. Most olivine grains were egested within 24 h. Chronic 35-day olivine (3-99 µm) exposure reduced amphipod survival, growth, and reproduction, likely as a result of metal induced oxidative stress and disturbance of major cation homeostasis. Amphipod reproduction was significantly reduced at olivine concentrations of 10% w/w and higher. In the context of ecological risk assessment, application of an arbitrary assessment factor of 100 to the highest no observed effect concentration of 1% w/w olivine yields a very low predicted no-effect concentration (PNEC) of 0.01% w/w olivine. This low PNEC value highlights the urgent need for additional marine olivine toxicity data to accurately assess the environmentally safe scale of coastal enhanced weathering for climate change mitigation.

Chemodynamic features of nickel(II) and its complexes: Implications for bioavailability in freshwaters.

A robust description of the bioavailability of Ni(II) in freshwaters is fundamental for the setting of environmental quality standards. Current approaches assume that bioavailability is governed by the equilibrium concentration of the free metal ion in the bulk aqueous medium. Such strategies generally have limited predictive value: a suite of empirical fitting parameters is required to deal with variations in water chemistry. Herein we compile data on Ni(II) speciation under typical freshwater conditions and compute the lability of Ni(II) complexes with typical molecular and nanoparticulate components of dissolved organic carbon. In combination with an analysis of the kinetic setting of Ni(II) biouptake by freshwater organisms, we assess the potential contribution from dissociation of Ni(II) complexes to the diffusive supply flux of free Ni2+. The strategy takes into account the absolute and relative magnitudes of the Michaelis-Menten bioaffinity and bioconversion parameters for a range of freshwater organisms, together with dynamic chemical speciation descriptors under environmentally relevant conditions. The results show that the dissociation kinetics of Ni(II) complexes play a crucial role in buffering the free metal ion concentration at the biointerface. Our results highlight the need to couple the timescales of chemical reactivity with those of biouptake to properly identify the bioavailable fraction of Ni(II) in freshwaters.

Deriving Nickel (Ni(II)) and Chromium (Cr(III)) Based Environmentally Safe Olivine Guidelines for Coastal Enhanced Silicate Weathering.

Enhanced silicate weathering (ESW) by spreading finely ground silicate rock along the coastal zone to remove atmospheric carbon dioxide (CO2) is a proposed climate change mitigation technique. The abundant and fast-dissolving mineral olivine has received the most attention for this application. However, olivine contains nickel (Ni) and chromium (Cr), which may pose a risk to marine biota during a gigaton-scale ESW application. Herein we derive a first guideline for coastal olivine dispersal based on existing marine environmental quality standards (EQS) for Ni and Cr. Results show that benthic biota are at the highest risk when olivine and its associated trace metals are mixed in the surface sediment. Specifically, depending on local sedimentary Ni concentrations, 0.059-1.4 kg of olivine m-2 of seabed could be supplied without posing risks for benthic biota. Accordingly, globally coastal ESW could safely sequester only 0.51-37 Gt of CO2 in the 21st century. On the basis of current EQS, we conclude that adverse environmental impacts from Ni and Cr release could reduce the applicability of olivine in coastal ESW. Our findings call for more in-depth studies on the potential toxicity of olivine toward benthic marine biota, especially in regard to bioavailability and metal mixture toxicity.

Field application of a novel active-passive sampling technique for the simultaneous measurement of a wide range of contaminants in water.

A first test of the field capabilities of a novel in situ sampling technique combining active and passive sampling (APS) was conducted in the sea. The proof-of-concept device uses a pump to draw water into a diffusion cell where dissolved target substances are accumulated onto sorbents which are selective for different classes of contaminants (i.e., metal cations, polar and non-polar organic compounds), simultaneously. A controlled laminar flow established in the diffusion cell enables measurements of contaminant concentrations that are fully independent from the hydrodynamic conditions in the bulk solution. APS measurements were consistent with those obtained using conventional passive sampling techniques such as organic diffusive gradients in thin films (o-DGT) and silicone rubber (SR) samplers (generally < 40% difference), taking into account the prevailing hydrodynamic conditions. The use of performance reference compounds (PRC) for hydrophobic contaminants provided additional information. Field measurements of metal ions in seawater showed large variability due to issues related to the device configuration. An improved field set-up deployed in supplementary freshwater mesocosm experiments provided metal speciation data that was consistent with passive sampling measurements (DGT), taking into account the hydrodynamic conditions. Overall, the results indicate that the APS technique provides a promising approach for the determination of a wide range of contaminants simultaneously, and independently from the hydrodynamic conditions in the bulk solution.

Temperature Effects During a Sublethal Chronic Metal Mixture Exposure on Common Carp (Cyprinus carpio).

The aquatic environment is the final sink of various pollutants including metals, which can pose a threat for aquatic organisms. Waterborne metal mixture toxicity might be influenced by environmental parameters such as the temperature. In the present study, common carp were exposed for 27 days to a ternary metal mixture of Cu, Zn, and Cd at two different temperatures, 10 and 20°C. The exposure concentrations represent 10% of the 96 h-LC50 (concentration lethal for the 50% of the population in 96 h) for each metal (nominal metal concentrations of Cu: 0.08 µM; Cd: 0.02 µM and Zn: 3 µM). Metal bioaccumulation and toxicity as well as changes in the gene expression of enzymes responsible for ionoregulation and induction of defensive responses were investigated. Furthermore the hepatosomatic index and condition factor were measured as crude indication of overall health and energy reserves. The obtained results showed a rapid Cu and Cd increase in the gills at both temperatures. Cadmium accumulation was higher at 20°C compared to 10°C, whereas Cu and Zn accumulation was not, suggesting that at 20°C, fish had more efficient depuration processes for Cu and Zn. Electrolyte (Ca, Mg, Na, and K) levels were analyzed in different tissues (gills, liver, brain, muscle) and in the remaining carcasses. However, no major electrolyte losses were observed. The toxic effect of the trace metal ion mixture on major ion uptake mechanisms may have been compensated by ion uptake from the food. Finally, the metal exposure triggered the upregulation of the metallothionein gene in the gills as defensive response for the organism. These results, show the ability of common carp to cope with these metal levels, at least under the condition used in this experiment.

Effect of Water pH on the Uptake of Acidic (Ibuprofen) and Basic (Propranolol) Drugs in a Fish Gill Cell Culture Model.

Water pH is predicted to affect the uptake of ionizable pharmaceuticals in fish. The current study used an in vitro primary fish gill cell culture system to assess the effect of pH values in the range of 4.5-8.75 on the uptake rates of the base propranolol (pKa 9.42) and the acid ibuprofen (pKa 4.59). The rate-limiting step in the uptake was the diffusive supply flux of the unionized form from the water to the apical membrane, with subsequent rapid transfer across the epithelium. Computed uptake rate based on the unionized fraction best described the uptake of propranolol and ibuprofen over the range of pH values 5-8 and 6-8.75, respectively. For ibuprofen, the computed uptake rate overestimated the uptake below pH 6 where the unionized fraction increased from 4% at pH 6 to 55% at pH 4.5. As the unionized fraction increased, the uptake rate plateaued suggesting a saturation of the transport process. For both drugs, large variations in the uptake occur with only small fluctuations in pH values. This occurs between pH values 6 and 8, which is the pH range acceptable in regulatory test guidelines and seen in most of our freshwaters.

Chemodynamic features of nanoparticles: Application to understanding the dynamic life cycle of SARS-CoV-2 in aerosols and aqueous biointerfacial zones.

We review concepts involved in describing the chemodynamic features of nanoparticles and apply the framework to gain physicochemical insights into interactions between SARS-CoV-2 virions and airborne particulate matter (PM). Our analysis is highly pertinent given that the World Health Organisation acknowledges that SARS-CoV-2 may be transmitted by respiratory droplets, and the US Center for Disease Control and Prevention recognises that airborne transmission of SARS-CoV-2 can occur. In our theoretical treatment, the virion is assimilated to a core-shell nanoparticle, and contributions of various interaction energies to the virion-PM association (electrostatic, hydrophobic, London-van der Waals, etc.) are generically included. We review the limited available literature on the physicochemical features of the SARS-CoV-2 virion and identify knowledge gaps. Despite the lack of quantitative data, our conceptual framework qualitatively predicts that virion-PM entities are largely able to maintain equilibrium on the timescale of their diffusion towards the host cell surface. Comparison of the relevant mass transport coefficients reveals that virion biointernalization demand by alveolar host cells may be greater than the diffusive supply. Under such conditions both the free and PM-sorbed virions may contribute to the transmitted dose. This result points to the potential for PM to serve as a shuttle for delivery of virions to host cell targets. Thus, our critical review reveals that the chemodynamics of virion-PM interactions may play a crucial role in the transmission of COVID-19, and provides a sound basis for explaining reported correlations between episodes of air pollution and outbreaks of COVID-19.

A comparative study on the effects of three different metals (Cu, Zn and Cd) at similar toxicity levels in common carp, Cyprinus carpio.

To improve our understanding of underlying toxic mechanisms, it is important to evaluate differences in effects that a variety of metals exert at concentrations representing the same toxic level to the organism. Therefore, the main goal of the present study was to compare the effects of waterborne copper (Cu(II)), zinc (Zn(II)) and cadmium (Cd (II)) on a freshwater fish, the common carp (Cyprinus carpio), at concentrations being 0%, 25%, 50% and 100% of the 96 h LC50 (the concentration which is lethal to 50% of the population in 96 h). All the exposures were performed for a period of 1 week at 20°C. Our results show a rapid increase in the amount of copper and cadmium accumulated in the gills, while zinc only started to increase by the end of the experiment. All three metal ions increased metallothionein gene expression in both gills and liver. However, clear adverse effects were mainly observed for the Cu exposed group. Cu caused a decrease in Na level in gill tissue; it altered the expression of genes involved in ionoregulation such as Na+ /K+ -ATPase and H+ -ATPase as well as the expression of oxidative stress-related genes, such as catalase, glutathione reductase and glutathione S-transferase. Zinc and cadmium exposure did not alter the ion levels in the gills. In addition, no obvious effect of oxidative stress was observed, except for a transient increase in glutathione reductase at the highest cadmium concentration.

Uptake and Release Kinetics of Organic Contaminants Associated with Micro- and Nanoplastic Particles.

A generic theoretical framework is presented for describing the kinetics of uptake and release of organic compounds that associate with plastic particles. The underlying concepts account for the physicochemical features of the target organic compounds and the plastic particles. The developed framework builds on concepts established for dynamic speciation analysis by solid-phase microextraction and the size-dependent reactivity features of particulate complexants. The theoretical framework is applied to interpretation of literature data, thereby providing more rigorous insights into previous observations. The presented concepts enable predictions of the sink/source functioning of plastic particles and their impact on the dynamic chemical speciation of organic compounds in aqueous environmental media and within biota. Our results highlight the fundamental influence of particle size on the uptake and release kinetics. The findings call for a comprehensive description of the physicochemical features of plastic particles to be provided in experimental studies on micro- and nanoplastics in different types of aquatic environmental media.

The synergistic toxicity of Cd(II) and Cu(II) to zebrafish (Danio rerio): Effect of water hardness.

We have evaluated the interactive toxicity of Cu(II) and Cd(II) in water with different hardness levels using adult zebrafish (Danio rerio). Zebrafish were exposed to Cd(II) (0.2-22 µM) or Cu(II) (0.1-8 µM) in single or binary exposures in very soft, moderately hard or very hard water. The whole body burdens of Cd(II) and Cu(II) reflect the net effect of biouptake and elimination, mortality was the indicator of toxicity, and whole body major ion content was measured to assess ion regulatory functions. Cu(II) was found to be more toxic than Cd(II) for zebrafish, and Cu(II) and Cd(II) exhibited a significant synergistic effect. The toxicity of metal ions increased upon decreasing the ionic strength of the exposure medium, probably due to elevated competition between metal ions with other cations in hard water and increased activity of Ca2+ pathways in soft water treatments. Whole body metal accumulation and the accumulation rate of both Cu and Cd increased as the metal ion concentration in the exposure medium increased. Nevertheless, neither parameter explained the observed synergistic effect on mortality. Finally, we observed a significant loss of whole body Na+ in fish which died during the metal exposure compared to surviving fish, irrespective of exposure conditions. Such an effect was not observed for other major cations (K+, Ca2+ and Mg2+). This observation suggests that, under the applied exposure conditions, survival was correlated to the capacity of the organism to maintain Na+ homeostasis.

The effect of copper on behaviour, memory, and associative learning ability of zebrafish (Danio rerio).

Copper is an essential element in many biological processes, but may exert toxic effects at levels surplus to metabolic requirements. Herein we assess the effect of copper on zebrafish behaviour using two assays, namely the novel tank diving test and a T-maze test with food reward. Novel tank diving tests were conducted on days 0, 4, and 10 of a 10 day Cu exposure (at concentrations of 0.77 µM (25% of the 240 h LC50) and 1.52 µM (50% of the 240 h LC50) to assess the alterations of behavioural responses in repeating novel tank diving assays and the effect of Cu on these patterns. Results demonstrate habituation to novelty, which is an indicator of spatial memory. Copper exposure had no effect on the latency of entry into the upper zones of the tank, nor on the total time spent therein, but did cause a greater number of freezing bouts in comparison to the control group. Additionally, Cu exposure had no effect on the habituation responses of zebrafish. Using the T-maze assay, we tested the effect of prior exposure to Cu for 10 days on subsequent behavioural trainings. The T-maze protocol was based on associative learning, where a visual stimulus (colour) was linked with a natural stimulus (food). Results of the control group showed that zebrafish are able to perform associative learning tasks. Moreover, Cu was found to negatively affect the associative learning capabilities. Specifically, while zebrafish in the control group achieved a significant number of correct choices (leading to food reward) throughout the T-maze training, such a trend was not observed for Cu exposed fish. Thus at the exposure concentrations and exposure times considered herein, Cu has no determinative impact on instinctual behavioural responses of zebrafish in repeated novel tank diving assays but does limit the associative learning capabilities.

The interactive effect of copper(II) and conspecific alarm substances on behavioural responses of zebrafish (Danio rerio).

Environmental contaminants such as metal ions can have detrimental effects on aquatic organisms at the molecular, organismal and population levels. In the present work, we examined the interactive effect of Cu(II) and conspecific alarm substance on zebrafish behavioural responses utilizing the novel tank diving assay. To this end, 3 novel tank diving tests (on day 0, 3 and 10 of the experimental phase) were conducted on zebrafish in 4 experimental groups: (1) control: no Cu(II) and no alarm substance, (2) Cu(II) only: exposed to 0.78 µM Cu(II) (25 % of the 240 h LC50) in the home tank for 10 days, (3) alarm substance only: exposed to alarm substance for 6 min concomitant with behavioural testing, and (4) Cu(II) + alarm substance: exposed to 0.78 µM Cu(II) in the home tank for 10 days and treated with alarm substance for 6 min during the behavioural testing. Results showed robust habituation response of zebrafish. Exposure to Cu(II) did not affect the behavioural phenotypes of zebrafish in the novel tank diving test or habituation responses. Alarm substance treatment evoked strong anxiety-like behaviour. Finally, zebrafish in the Cu(II) + alarm substance group lost their sensitivity to alarm substance in repeated novel tank assays throughout the concomitant Cu(II) exposure; this observation is tentatively ascribed to Cu(II)-induced olfactory impairment.

Characterization of the accumulation of metals and organic contaminants on a novel active-passive sampling device under controlled water flow conditions.

Recently, a new sampling device combining active and passive sampling (APS) was developed for the measurement of time-averaged concentrations of metal species and both polar and non-polar organic contaminants in water. By coupling a diffusion cell (loaded with a set of sorbents selective for different substances) with a small pump and a flow meter, the APS device is able to perform in situ measurements that are independent of the hydrodynamic conditions in the exposure medium. In the present study, the diffusion layer thickness (δ) at the sorbent/solution interface within the diffusion cell was characterised under controlled flow conditions. Laboratory tests indicated that, in the range of flow rates investigated, the average diffusion layer thickness (δ¯) varied from ∼60 to ∼110 µm, depending on the type of substance measured and the position of the sorbent with respect to the flow direction. Due to its ability to maintain an approximately constant δ¯, good to excellent agreement was found between measurements performed with the APS device in non-complexing media and concentrations measured in discrete water samples for all the substances investigated. These results suggest that the APS device could overcome issues affecting the quantitative interpretation of measurements by conventional passive sampling devices and serve as a useful tool for simultaneously monitoring a wide range of contaminants in water.

Rigorous Physicochemical Framework for Metal Ion Binding by Aqueous Nanoparticulate Humic Substances: Implications for Speciation Modeling by the NICA-Donnan and WHAM Codes.

Latest knowledge on the reactivity of charged nanoparticulate complexants toward aqueous metal ions is discussed in mechanistic detail. We present a rigorous generic description of electrostatic and chemical contributions to metal ion binding by nanoparticulate complexants, and their dependence on particle size, particle type (i.e., reactive sites distributed within the particle body or confined to the surface), ionic strength of the aqueous medium, and the nature of the metal ion. For the example case of soft environmental particles such as fulvic and humic acids, practical strategies are delineated for determining intraparticulate metal ion speciation, and for evaluating intrinsic chemical binding affinities and heterogeneity. The results are compared with those obtained by popular codes for equilibrium speciation modeling (namely NICA-Donnan and WHAM). Physicochemical analysis of the discrepancies generated by these codes reveals the a priori hypotheses adopted therein and the inappropriateness of some of their key parameters. The significance of the characteristic time scales governing the formation and dissociation rates of metal-nanoparticle complexes in defining the relaxation properties and the complete equilibration of the metal-nanoparticulate complex dispersion is described. The dynamic features of nanoparticulate complexes are also discussed in the context of predictions of the labilities and bioavailabilities of the metal species.

The effect of metal mixture composition on toxicity to C. elegans at individual and population levels.

The toxicity of zinc (Zn), copper (Cu), and cadmium (Cd) to the nematode Caenorhabditis elegans was characterised under single metal and mixture scenarios at different organisational levels. The effects on population size and body length were investigated at two concentrations corresponding to the 24 h LC5 and LC20 levels. Metal toxicity was dependent on metal concentration, exposure time and mixture composition. Populations exposed to LC20 levels of Cd, ZnCu, CuCd and ZnCuCd plummeted, while for all LC5 concentrations, population size continued to increase, albeit that single metals were less harmful than mixtures. Combinations of the LC20 concentration of Cd with a range of Zn concentrations showed concentration dependent mitigating effects on population size and antagonistic effects on mortality. By combining effects at different organisational levels, more insight into metal toxicity was obtained. Metal effects were more evident on population size than on body length or mortality, suggesting that population size could be considered as a sensitive endpoint. Furthermore, our observations of ZnCd mixture effects at the individual and population levels are consistent with literature data on the dose-dependent expression of the cdf-2 gene, which is involved in mediation of Zn and Cd toxicity.

The effect of thermal pre-incubation and exposure on sensitivity of zebrafish (Danio rerio) to copper and cadmium single and binary exposures.

Zebrafish (Danio rerio) is a prominent model organism in a wide range of biological studies including toxicology. However, toxicological studies are often performed at species specific optimum temperature, and knowledge on the effect of different temperature regimes on the toxicity of metal ions is rather limited. To address this knowledge gap, present study investigates the effect of various thermal scenarios (simultaneous and sequential; acute and chronic) on the toxicity of Cu and Cd in zebrafish. For this purpose we assessed mortality and whole body metal burdens as indicators of toxicity and bioavailability, respectively, and whole body electrolyte concentrations and body condition as the indicators of physiological condition. Thermal pre-incubations (for 12 or 96 h or 28 days) and subsequent metal ion exposures (for 10 days) were conducted at 17, 22, 25, 28, 32 and 34 °C. The metal exposures were performed at Cu concentrations of 1.2 µM and Cd concentrations of 0.2 µM, both singly and in binary mixtures. Irrespective of thermal treatments, Cu exposures resulted in greater mortality than Cd exposures at the given concentrations. Moreover, the Cu and Cd mixture indicated a synergistic effect. While acute pre-incubation for 12 or 96 h at elevated temperatures increased mortality in the subsequent metal exposure at the optimum temperature (28 °C), pre-incubation at cold temperatures in this scenario appeared to increase tolerance towards the subsequent metal exposure. Chronic thermal pre-incubation of zebrafish to a range of temperatures for 28 days moderated the effect of temperature fluctuations on subsequent metal toxicity at the optimum temperature. Chronic thermal pre-incubation at a range of temperatures followed by metal exposure at the same temperature showed that environmental temperature variations (higher or lower than optimal temperature) coupled with metal exposure, led to increased mortality, furthermore, the highest whole body metal burdens were measured in this scenario. Nevertheless, neither the whole body burden of metals, nor the metal accumulation rate, were predictors of mortality, i.e. these two values were not higher in dead fish in comparison to those that survived the exposures. Finally, we observed a significant decrease in the whole body Na+ level of dead fish in comparison to fish which survived the exposure conditions, suggesting that survival depends on maintaining Na+ homeostasis under the applied multi-stress conditions. Overall, our results show that thermal pre-history and ambient temperature play an important role in determining the tolerance of zebrafish towards metal ion stress.

Advancing the Zebrafish embryo test for endocrine disruptor screening using micro-injection: Ethinyl estradiol as a case study.

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.

The interplay between chemical speciation and physiology determines the bioaccumulation and toxicity of Cu(II) and Cd(II) to Caenorhabditis elegans.

Using the well-documented model organism Caenorhabditis elegans, a combined analysis of metal speciation in the exposure medium and body burdens of metals (Zn, Cu and Cd) was performed, and factors that are predictive of toxicological endpoints in single metal and mixed metal exposures were identified. Cu, and to a lesser extent Cd, is found to associate with Escherichia coli in the exposure medium (the food source for C. elegans) as evidenced by the observed decrease in both their dissolved and free metal ion concentrations. Together with a critical analysis of literature data, our results suggest that free metal ion concentrations and thus aqueous uptake routes are the best predictor of internal concentrations under all conditions considered, and of metal toxicity in single metal exposures. Additional factors are involved in determining the toxicity of metal mixtures. In general, the eventual adverse effects of metals on biota are expected to be a consequence of the interplay between chemical speciation in the exposure medium, timescale of exposure, exposure route as well as the nature and timescale of the biotic handling pathways.