Modeling Full Life-Cycle Effects of Copper on Brook Trout (Salvelinus fontinalis) Populations.

Population models are increasingly used to predict population-level effects of chemicals. For trout, most toxicity data are available on early-life stages, but this may cause population models to miss true population-level effects. We predicted population-level effects of copper (Cu) on a brook trout (Salvelinus fontinalis) population based on individual-level effects observed in either a life-cycle study or an early-life stage study. We assessed the effect of Cu on predicted trout densities (both total and different age classes) and the importance of accounting for effects on the full life cycle compared with only early-life stage effects. Additionally, uncertainty about the death mechanism and growth effects was evaluated by comparing the effect of different implementation methods: individual tolerance (IT) versus stochastic death (SD) and continuous versus temporary growth effects. For the life-cycle study, the same population-level no-observed-effect concentration (NOECpop) was predicted as the lowest reported individual-level NOEC (NOECind; 9.5 µg/L) using IT. For SD, the NOECpop was predicted to be lower than the NOECind for young-of-the-year and 1-year-old trout (3.4 µg/L), but similar for older trout (9.5 µg/L). The implementation method for growth effects did not affect the NOECpop of the life-cycle study. Simulations based solely on the early-life stage effects within the life-cycle study predicted unbounded NOECpop values (≥32.5 µg/L), that is, >3.4 times higher than the NOECpop based on all life-cycle effects. For the early-life stage study, the NOECpop for both IT and SD were predicted to be >2.6 times higher than the lowest reported NOECind. Overall, we demonstrate that effects on trout populations can be underestimated if predictions are solely based on toxicity data with early-life stages. Environ Toxicol Chem 2024;00:1-15. © 2024 SETAC.

Contribution of combined stressors on density and gene expression dynamics of the copepod Temora longicornis in the North Sea.

The impact of multiple environmental and anthropogenic stressors on the marine environment remains poorly understood. Therefore, we studied the contribution of environmental variables to the densities and gene expression of the dominant zooplankton species in the Belgian part of the North Sea, the calanoid copepod Temora longicornis. We observed a reduced density of copepods, which were also smaller in size, in samples taken from nearshore locations when compared to those obtained from offshore stations. To assess the factors influencing the population dynamics of this species, we applied generalised additive models. These models allowed us to quantify the relative contribution of temperature, nutrient levels, salinity, turbidity, concentrations of photosynthetic pigments, as well as chemical pollutants such as polychlorinated biphenyls and polycyclic aromatic hydrocarbons (PAHs), on copepod density. Temperature and Secchi depth, a proxy for turbidity, were the most important environmental variables predicting the densities of T. longicornis, followed by summed PAH and chlorophyll concentrations. Analysing gene expression in field-collected adults, we observed significant variation in metabolic and stress-response genes. Temperature correlated significantly with genes involved in proteolytic activities, and encoding heat shock proteins. Yet, concentrations of anthropogenic chemicals did not induce significant differences in the gene expression of genes involved in the copepod's fatty acid metabolism or well-known stress-related genes, such as glutathione transferases or cytochrome P450. Our study highlights the potential of gene expression biomonitoring and underscores the significance of a changing environment in future studies.

Extrapolation of Metal Toxicity Data for the Rotifer Brachionus calyciflorus Using an Individual-Based Population Model.

Ecological risk assessment (ERA) of metals typically starts from standardized toxicity tests, the data from which are then extrapolated to derive safe concentrations for the envisioned protection goals. Because such extrapolation in conventional ERA lacks ecological realism, ecological modeling is considered as a promising new approach for extrapolation. Many published population models are complex, that is, they include many processes and parameters, and thus require an extensive dataset to calibrate. In the present study, we investigated how individual-based models based on a reduced version of the Dynamic Energy Budget theory (DEBkiss IBM) could be applied for metal effects on the rotifer Brachionus calyciflorus. Data on survival over time and reproduction at different temperatures and food conditions were used to calibrate and evaluate the model for copper effects. While population growth and decline were well predicted, the underprediction of population density and the mismatch in the onset of copper effects were attributed to the simplicity of the approach. The DEBkiss IBM was applied to toxicity datasets for copper, nickel, and zinc. Predicted effect concentrations for these metals based on the maximum population growth rate were between 0.7 and 3 times higher in all but one case (10 times higher) than effect concentrations based on the toxicity data. The size of the difference depended on certain characteristics of the toxicity data: both the steepness of the concentration-effect curve and the relative sensitivity of lethal and sublethal effects played a role. Overall, the present study is an example of how a population model with reduced complexity can be useful for metal ERA. Environ Toxicol Chem 2024;43:324-337. © 2023 SETAC.

The influence of pH and dissolved organic carbon on the ecotoxicity of ampicillin and clarithromycin.

The impacts of water chemistry properties including pH and dissolved organic carbon (DOC) on the ecotoxicity of active pharmaceutical ingredients (APIs) are increasingly evident. These impacts are a result of alterations in API bioavailability: pH regulates the bioavailability of many ionizable APIs via chemical speciation, whereas DOC interacts with several APIs to inhibit the APIs from traversing the membrane system of organisms. In this study, we examined the influences of pH and DOC on the bioavailability of ampicillin (AMP) and clarithromycin (CLA) with the help of a bioavailability model. The effects on bioavailability were quantified by ecotoxicity observed in cyanobacteria growth inhibition tests with Microcystis aeruginosa PCC7806. The median effect concentration (96 h-EC50total) of AMP increased by 5-fold when pH raised from 7.4 to 9.0, suggesting the zwitterionic AMP+/- species being higher in bioavailability than the negatively charged AMP- species. CLA ecotoxicity showed no significant pH-dependency, suggesting CLA+ and CLA0 species to be equally bioavailable, albeit it correlated significantly with M. aeruginosa growth rate in negative controls. In addition, DOC demonstrated no significant effects on the ecotoxicity of AMP or CLA. Overall, together with earlier results on ciprofloxacin, our data show that bioavailability relations with pH and DOC are variable among different antibiotics. Factors other than chemical speciation alone could play a role in their bioavailability, such as their molecular size and polarity.

Ecotoxicity of Lead to a Phytoplankton Community: Effects of pH and Phosphorus Addition and Implications for Risk Assessment.

Ecological risk assessment and water quality criteria for lead (Pb) are increasingly making use of bioavailability-based approaches to account for the impact of toxicity-modifying factors, such as pH and dissolved organic carbon. For phytoplankton, which are among the most Pb-sensitive freshwater species, a Pb bioavailability model has previously been developed based on standard single-species exposures at a high phosphorus (P) concentration and pH range of 6.0 to 8.0. It is well known that P can affect metal toxicity to phytoplankton and that the pH of many surface waters can be above 8.0. We aimed to test whether the single-species bioavailability model for Pb could predict the influence of pH on Pb toxicity to a phytoplankton community at both low and high P supply. A 10-species phytoplankton community was exposed to Pb for 28 days at two different pH levels (7.2 and 8.4) and two different P supply levels (low and high, i.e., total P input 10 and 100 µg/L, respectively) in a full factorial 2 × 2 test design. We found that the effects of total Pb on three community-level endpoints (biodiversity, community functioning, and community structure) were highly dependent on both pH and P supply. Consistent lowest-observed-effect concentrations (LOECs) ranged between 21 and >196 µg total Pb/L and between 10 and >69 µg filtered Pb/L. Long-term LOECs were generally higher, that is, 69 µg total Pb/L (42 µg filtered Pb/L) or greater, across all endpoints and conditions, indicating recovery near the end of the exposure period, and suggesting the occurrence of acclimation to Pb and/or functional redundancy. The highest toxicity of Pb for all endpoints was observed in the pH 7.2 × low P treatment, whereas the pH 8.4 × low P and pH 8.4 × high P treatment were the least sensitive treatments. At the pH 7.2 × high P treatment, the algal community showed an intermediate Pb sensitivity. The effect of pH on the toxicity of filtered Pb could not be precisely quantified because for many endpoints no effect was observed at the highest Pb concentration tested. However, the long-term LOECs (filtered Pb) at low P supply suggest a decrease in Pb toxicity of at least 1.6-fold from pH 7.2 to 8.4, whereas the single-species algal bioavailability model predicted a 2.5-fold increase. This finding suggests that bioavailability effects of pH on Pb toxicity cannot be extrapolated as such from the single species to the community level. Overall, our data indicate that, although the single-species algal Pb bioavailability model may not capture pH effects on Pb ecotoxicity in multispecies systems, the bioavailability-based hazardous concentration for 5% of the species was protective of long-term Pb effects on the structure, function, and diversity of a phytoplankton community in a relevant range of pH and P conditions. Environ Toxicol Chem 2023;42:2684-2700. © 2023 SETAC.

Water brownness regulates the bioavailability of a fluoroquinolone antibiotic: UV-absorbance as a predictor of ciprofloxacin ecotoxicity.

Dissolved organic carbon (DOC) is a powerful regulator of the ecotoxicity of ciprofloxacin (CIP), a widely applied fluoroquinolone antibiotic. In this study, we investigated the impact of DOC from a variety of sources on CIP bioavailability, using a cyanobacteria growth inhibition test with Microcystis aeruginosa. We analyzed the impact from two perspectives: (1) DOC concentration, and (2) water brownness, defined in this work as the light absorbance of DOC solutions. The toxicity tests were conducted with (1) unprocessed freshwater DOC in the naturally occurring state, (2) DOC extracted from a freshwater stream (Schwarzbach stream, Küchelscheid, Belgium), and (3) the commercial DOC product Suwannee River organic matter. Across all DOC sources investigated, a strong negative correlation was observed between CIP ecotoxicity and light absorbance at four wavelengths across the ultraviolet-visible range (e.g., A350), whereas CIP ecotoxicity correlated poorly with the DOC concentration. In addition, the interactions between CIP and DOC were modelled as a CIP-DOC binding process to allow the quantification of the inhibitory effects of DOC on CIP toxicity via binding constants (Kd,CIPx, with x being the ionic charge + or +/-, L g-1). Processed DOC sources showed higher binding potency than most of the unprocessed DOC sources, suggesting that toxicity tests employing only processed DOC potentially overestimates the impact of DOC in natural environments. Nonetheless, the light absorption coefficient (i.e., ε350) appeared a reliable predictor of the Kd,CIP+/- (and thus of the potential of the DOC source to reduce ecotoxicity of CIP) of both processed and unprocessed DOC. The relationship can be further incorporated into model simulations to estimate CIP bioavailability in dynamic environments. It is concluded that the brownness of water is a better predictor of the impact of DOC on CIP bioavailability than the DOC concentration itself.

Decadal decline of dominant copepod species in the North Sea is associated with ocean warming: Importance of marine heatwaves.

Time-series are crucial to understand the status of zooplankton communities and to anticipate changes that might affect the entire food web. Long-term time series allow us to understand impacts of multiple environmental and anthropogenic stressors, such as chemical pollution and ocean warming, on the marine ecosystems. Here, a recent time series (2018-2022) of abundance data of four dominant calanoid and one harpacticoid copepod species from the Belgian Part of the North Sea was combined with previously collected (2009-2010, 2015-2016) datasets for the same study area. The time series reveals a significant decrease (up to two orders of magnitude) in calanoid copepod abundance (Temora longicornis, Acartia clausi, Centropages spp., Calanus helgolandicus), while this was not the case for the harpacticoid Euterpina acutifrons. We applied generalized additive models to quantify the relative contribution of temperature, nutrients, salinity, primary production, turbidity and pollution (anthropogenic chemicals, i.e., polychlorinated biphenyls and polycyclic aromatic hydrocarbons) to the population dynamics of these species. Temperature, turbidity and chlorophyll a concentrations were the only variables consistently showing a relative high contribution in all models predicting the abundances of the selected species. The observed heat waves which occurred during the summer periods of the investigated years coincided with population collapses (versus population densities in non-heatwave years) and are considered the most likely cause for the observed copepod abundance decreases. Moreover, the recorded water temperatures during these heatwaves correspond to the physiological thermal limit of some of the studied species. As far as we know, this is the first study to observe ocean warming and marine heat waves having such a dramatic impact (population collapse) on the dominant zooplankton species in shallow coastal areas.

Investigating Population-Level Toxicity of the Antidepressant Citalopram in Harpacticoid Copepods Using In Vivo Methods and Bioenergetics-Based Population Modeling.

Recent research has revealed various lethal and sublethal effects of the selective serotonin reuptake inhibitor citalopram hydrobromide on the harpacticoid copepod Nitocra spinipes. In the present study, an individual-based model (IBM) grounded in the dynamic energy budget (DEB) theory was developed to extrapolate said effects to the population level. Using a generic DEB-IBM as a template, the model was designed to be as simple as possible, keeping model components that are outside the scope of the core DEB theory to a minimum. To test the model, a 56-day population experiment was performed at 0, 100, and 1000 µg citalopram hydrobromide L-1 . In the experiment, the populations quickly reached a plateau in the control and at 100 µg L-1 , which was correctly reproduced by the model and could be explained by food limitations hindering further population growth. At 1000 µg L-1 , a clear mismatch occurred: Whereas in the experiment the population size increased beyond the supposed (food competition-induced) capacity, the model predicted a suppression of the population size. It is assumed that the IBM still misses important components addressing population density-regulating processes. Particularly crowding effects may have played an important role in the population experiment and should be further investigated to improve the model. Overall, the current DEB IBM for N. spinipes should be seen as a promising starting point for bioenergetics-based copepod population modeling, which-with further improvements-may become a valuable individual-to-population extrapolation tool in the future. Environ Toxicol Chem 2023;42:1094-1108. © 2023 SETAC.

A Bioavailability Model to Predict the Impact of pH and Dissolved Organic Carbon on Ciprofloxacin Ecotoxicity to the Cyanobacterium Microcystis aeruginosa.

Ciprofloxacin (CIP) is a pseudopersistent antibiotic detected in freshwater worldwide. As an ionizable chemical, its fate in freshwater is influenced by water chemistry factors such as pH, hardness, and dissolved organic carbon (DOC) content. We investigated the effect of pH, DOC, and Ca2+ levels on the toxicity of CIP to Microcystis aeruginosa and developed a bioavailability model on the basis of these experimental results. We found that the zwitterion (CIP+/- ) is the most bioavailable species of CIP to M. aeruginosa, whereas DOC is the most dominant factor reducing CIP toxicity, possibly via binding of both CIP+/- and CIP+ to DOC. pH likely also regulates CIP-DOC binding indirectly through its influence on CIP speciation. In addition, higher tolerance to CIP by M. aeruginosa was observed at pH < 7.2, but the underlying mechanism is yet unclear. Calcium was identified as an insignificant factor in CIP bioavailability. When parameterized with the data obtained from toxicity experiments, our bioavailability model is able to provide accurate predictions of CIP toxicity because the observed and predicted total median effective concentrations deviated by <28% from each other. Our model predicts that changes in pH and DOC conditions can affect CIP toxicity by up to 10-fold, suggesting that CIP in many natural environments is likely less toxic than in standard laboratory toxicity experiments. Environ Toxicol Chem 2022;41:2835-2847. © 2022 SETAC.

Predicting Combined Effects of Chemical Stressors: Population-Level Effects of Organic Chemical Mixtures with a Dynamic Energy Budget Individual-Based Model.

Most regulatory ecological risk-assessment frameworks largely disregard discrepancies between the laboratory, where effects of single substances are assessed on individual organisms, and the real environment, where organisms live together in populations and are often exposed to multiple simultaneously occurring substances. We assessed the capability of individual-based models (IBMs) with a foundation in the dynamic energy budget (DEB) theory to predict combined effects of chemical mixtures on populations when they are calibrated on toxicity data of single substances at the individual level only. We calibrated a DEB-IBM for Daphnia magna for four compounds (pyrene, dicofol, α-hexachlorocyclohexane, and endosulfan), covering different physiological modes of action. We then performed a 17-week population experiment with D. magna (designed using the DEB-IBM), in which we tested mixture combinations of these chemicals at relevant concentrations, in a constant exposure phase (7-week exposure and recovery), followed by a pulsed exposure phase (3-day pulse exposure and recovery). The DEB-IBM was validated by comparing blind predictions of mixture toxicity effects with the population data. The DEB-IBM accurately predicted mixture toxicity effects on population abundance in both phases when assuming independent action at the effect mechanism level. The population recovery after the constant exposure was well predicted, but recovery after the pulse was not. The latter could be related to insufficient consideration of stochasticity in experimental design, model implementation, or both. Importantly, the mechanistic DEB-IBM performed better than conventional statistical mixture assessment methods. We conclude that the DEB-IBM, calibrated using only single-substance individual-level toxicity data, produces accurate predictions of population-level mixture effects and can therefore provide meaningful contributions to ecological risk assessment of environmentally realistic mixture exposure scenarios. Environ Toxicol Chem 2022;41:2240-2258. © 2022 SETAC.

Mixture Toxicity of Herbicides with Dissimilar Modes of Action to Myriophyllum spicatum.

Considering the vital role of rooted macrophytes in the aquatic ecosystem, validating assumptions on the interactive effects of herbicides with different modes of action at an environmentally relevant mixture ratio is necessary. We investigated the effects of diflufenican (a carotenoid biosynthesis inhibitor) and iodosulfuron-methyl-sodium (IMS; an acetolactate synthase inhibitor) in a 14-day growth inhibition experiment with Myriophyllum spicatum, wherein single compounds and their combination were tested in parallel (n = 84). The assessment was done using three different methods: significance testing, model deviation ratio (MDR), and mixture interaction factor (MIF). Interactions relative to both concentration addition and independent action were assessed via significance testing. This revealed that diflufenican and IMS acted antagonistically relative to both models for fresh weight and total shoot length (p < 0.05) and that there was slight synergism for the number of side shoots (p < 0.001) relative to concentration addition. The MDR and MIF can only assess interactions relative to the concentration addition model. According to MDR, the mixture appeared to show no interaction (neither antagonistic nor synergistic), whereas the MIF method revealed that the compounds acted antagonistically for fresh weight and that there was a slight synergism for total shoot length and number of side shoots. We conclude that inferences about mixture toxicity interactions are method- and endpoint-dependent, which can have implications for regulatory mixtures assessment. Environ Toxicol Chem 2022;41:2209-2220. © 2022 SETAC.

Sea Spray Aerosols Contain the Major Component of Human Lung Surfactant.

Marine phytoplankton influence the composition of sea spray aerosols (SSAs) by releasing various compounds. The biogenic surfactant dipalmitoylphosphatidylcholine (DPPC) is known to accumulate in the sea surface microlayer, but its aerosolization has never been confirmed. We conducted a 1 year SSA sampling campaign at the Belgian coast and analyzed the SSA composition. We quantified DPPC at a median and maximum air concentration of 7.1 and 33 pg m-3, respectively. This discovery may be of great importance for the field linking ocean processes to human health as DPPC is the major component of human lung surfactant and is used as excipient in medical aerosol therapy. The natural airborne exposure to DPPC seems too low to induce direct human health effects but may facilitate the effects of other marine bioactive compounds. By analyzing various environmental variables in relation to the DPPC air concentration, using a generalized linear model, we established that wave height is a key environmental predictor and that it has an inverse relationship. We also demonstrated that DPPC content in SSAs is positively correlated with enriched aerosolization of Mg2+ and Ca2+. In conclusion, our findings are not only important from a human health perspective but they also advance our understanding of the production and composition of SSAs.

Integrating Bioavailability of Metals in Fish Population Models.

Population models are increasingly being used to extrapolate individual-level effects of chemicals, including metals, to population-level effects. For metals, it is also important to take into account their bioavailability to correctly predict metal toxicity in natural waters. However, to our knowledge, no models exist that integrate metal bioavailability into population modeling. Therefore, our main aims were to 1) incorporate the bioavailability of copper (Cu) and zinc (Zn) into an individual-based model (IBM) of rainbow trout (Oncorhynchus mykiss), and 2) predict how survival-time concentration data translate to population-level effects. For each test water, reduced versions of the general unified threshold model of survival (GUTS-RED) were calibrated using the complete survival-time concentration data. The GUTS-RED individual tolerance (IT) showed the best fit in the different test waters. Little variation between the different test waters was found for 2 GUTS-RED-IT parameters. The GUTS-RED-IT parameter "median of distribution of thresholds" (mw ) showed a strong positive relation with the Ca2+ , Mg2+ , Na+ , and H+ ion activities. Therefore, mw formed the base of the calibrated GUTS bioavailability model (GUTS-BLM), which predicted 30-d x% lethal concentration (LCx) values within a 2-fold error. The GUTS-BLM was combined with an IBM, inSTREAM-Gen, into a GUTS-BLM-IBM. Assuming that juvenile survival was the only effect of Cu and Zn exposure, population-level effect concentrations were predicted to be 1.3 to 6.2 times higher than 30-d laboratory LCx values, with the larger differences being associated with higher interindividual variation of metal sensitivity. The proposed GUTS-BLM-IBM model can provide insight into metal bioavailability and effects at the population level and could be further improved by incorporating sublethal effects of Cu and Zn. Environ Toxicol Chem 2021;40:2764-2780. © 2021 SETAC.

Phycotoxin-Enriched Sea Spray Aerosols: Methods, Mechanisms, and Human Exposure.

To date, few studies have examined the role of sea spray aerosols (SSAs) in human exposure to harmful and beneficial marine compounds. Two groups of phycotoxins (brevetoxins and ovatoxins) have been reported to induce respiratory syndromes during harmful algal blooms. The aerosolization and coastal air concentrations of other common marine phycotoxins have, however, never been examined. This study provides the first (experimental) evidence and characterization of the aerosolization of okadaic acid (OA), homoyessotoxin, and dinophysistoxin-1 using seawater spiked with toxic algae combined with the realistic SSA production in a marine aerosol reference tank (MART). The potential for aerosolization of these phycotoxins was highlighted by their 78- to 1769-fold enrichment in SSAs relative to the subsurface water. To obtain and support these results, we first developed an analytical method for the determination of phycotoxin concentrations in SSAs, which showed good linearity (R2 > 0.99), recovery (85.3-101.8%), and precision (RSDs ≤ 17.2%). We also investigated natural phycotoxin air concentrations by means of in situ SSA sampling with concurrent aerosolization experiments using natural seawater in the MART. This approach allowed us to indirectly quantify the (harmless) magnitude of OA concentrations (0.6-51 pg m-3) in Belgium's coastal air. Overall, this study provides new insights into the enriched aerosolization of marine compounds and proposes a framework to assess their airborne exposure and effects on human health.

Making Sense of Life-History Effects of the Antidepressant Citalopram in the Copepod Nitocra spinipes Using a Bioenergetics Model.

The global consumption of human antidepressants has steadily increased over the last years. The most widely prescribed antidepressants are the selective serotonin reuptake inhibitors (SSRIs), which have been linked to various life-history effects in nontarget organisms. We investigated the effects of the SSRI citalopram hydrobromide on the life history of the copepod Nitocra spinipes. Slight but significant developmental delay effects were observed at nominal concentrations of 0.1 and 1 µg/L, with stronger effects occurring at measured concentrations of 178 µg/L and above. At 77 µg/L and above, a significant increase in adult body length and offspring production/brood was found, although the time between brood releases remained unaffected. The pre-adult surviving fraction was significantly reduced (by 44%) at 765 µg/L. For a mechanistic evaluation of these observations, we used a bioenergetics model for N. spinipes based on the dynamic energy budget theory. Toxicokinetic and toxicodynamic submodels were used to dynamically simulate the chemical uptake and elimination, as well as dose-response relationships for hypothetical physiological modes of action and survival over time. Although none of the commonly invoked physiological modes of action, acting on assimilation, maintenance, growth, or offspring production, could explain the observed combination of effects, a newly proposed physiological mode of action acting on the process of maturation delivered correct predictions in terms of each effect's direction. The model fits could be further improved by allowing for a gentler concentration-effect slope and by adding an auxiliary physiological mode of action acting on the reproduction efficiency. The quantitative explanations provided in the present study offer a starting point for exploratory simulation studies investigating the effects of SSRIs at higher ecological levels. Environ Toxicol Chem 2021;40:1928-1939. © 2021 SETAC.

Neonicotinoid Insecticides from a Marine Perspective: Acute and Chronic Copepod Testing and Derivation of Environmental Quality Standards.

Neonicotinoid insecticides have become of global concern for the aquatic environment. Harpacticoid copepods are among the organisms most sensitive to neonicotinoids. We exposed the brackish copepod Nitocra spinipes to 4 neonicotinoid insecticides (clothianidin, imidacloprid, thiacloprid, and thiamethoxam) to investigate acute toxicity on adults (96-h exposure) and effects on larval development (7-d exposure). We used these results in combination with publicly available ecotoxicity data to derive environmental quality standards (EQS). These EQS were ultimately used in a single-substance and mixture risk assessment for the Belgian part of the North Sea. Acute toxicity testing revealed that immobilization is a more sensitive endpoint than mortality, with 96-h median effect concentration (EC50) values of 6.9, 7.2, 25, and 120 µg L-1 for clothianidin, thiacloprid, imidacloprid, and thiamethoxam, respectively. In addition, the larval development tests resulted in 7-d no-observed-effect concentrations (NOECs) of 2.5, 2.7, 4.2, and >99 µg L-1 for clothianidin, thiacloprid, imidacloprid, and thiamethoxam, respectively. The derived saltwater annual average (AA-)EQS were 0.05, 0.0048, 0.002, and 0.016 µg L-1 for clothianidin, thiacloprid, imidacloprid, and thiamethoxam, respectively. Finally, the risk characterization revealed some exceedances of the AA-EQS in Belgian harbors for imidacloprid (number of exceedances, n = 2/4), for thiacloprid (n = 1/4), for thiamethoxam (n = 1/4), and for the mixture of the 4 neonicotinoids (n = 4/4), but not at the open sea. At the open sea site, the toxic unit sums relative to the AA-EQS were 0.72 and 0.22, suggesting no mixture risk, albeit with a relatively small margin of safety. Including short-term EC10 (96-h) values of N. spinipes for the AA-EQS derivation led to a refinement of the AA-EQS for clothianidin and thiamethoxam, suggesting their use for the AA-EQS derivation because one of the overarching goals of the definition of EQS is to protect species at the population level. Environ Toxicol Chem 2021;40:1353-1367. © 2021 SETAC.

Development and Validation of a Mixture Toxicity Implementation in the Dynamic Energy Budget-Individual-Based Model: Effects of Copper and Zinc on Daphnia magna Populations.

Mechanistic population models are gaining considerable interest in ecological risk assessment. The dynamic energy budget approach for toxicity (DEBtox) and the general unified threshold model for survival (GUTS) are well-established theoretical frameworks that describe sublethal and lethal effects of a chemical stressor, respectively. However, there have been limited applications of these models for mixtures of chemicals, especially to predict long-term effects on populations. We used DEBtox and GUTS in an individual-based model (IBM) framework to predict both single and combined effects of copper and zinc on Daphnia magna populations. The model was calibrated based on standard chronic toxicity test results with the single substances. A mixture toxicity implementation based on the general independent action model for mixtures was developed and validated with data from a population experiment with copper and zinc mixtures. Population-level effects of exposure to individual metals were accurately predicted by DEB-IBM. The DEB-IBM framework also allowed us to identify the potential mechanisms underlying these observations. Under independent action the DEB-IBM was able to predict the population dynamics observed in populations exposed to the single metals and their mixtures (R2 > 65% in all treatments). Our modeling shows that it is possible to extrapolate from single-substance effects at the individual level to mixture toxicity effects at the population level, without the need for mixture toxicity data at the individual level from standard mixture toxicity tests. The application of such modeling techniques can increase the ecological realism in risk assessment. Environ Toxicol Chem 2021;40:513-527. © 2020 SETAC.

Interplay between dietary lipids and cadmium exposure in rainbow trout liver: Influence on fatty acid metabolism, metal accumulation and stress response.

The present study aimed at investigating interactive effects between dietary lipids and both short- and long-term exposures to a low, environmentally realistic, cadmium (Cd) concentration. Juvenile rainbow trout were fed four isolipidic diets (31.7 g/kg) enriched in either linoleic acid (LA, 18:2n-6), alpha-linolenic acid (ALA, 18:3n-3), eicosapentaenoic acid (EPA, 20:5n-3) or docosahexaenoic acid (DHA, 22:6n-3). From the 4th week of this 10-week experiment, the lipid level of the diet was increased (120.0 g/kg) and half of the fish fed each diet were aqueously exposed to Cd (0.3 µg/L) while the other half were not exposed to Cd (control). Fish were sampled and their liver was harvested for fatty acid profile, hepatic Cd and calcium concentrations, total glutathione level and gene expression assessment, either (i) after 4 weeks of feeding and 24 h of Cd contamination (day 29) (short-term Cd exposure) or (ii) after 10 weeks of feeding and 6 weeks of Cd contamination (day 70) (long-term Cd exposure). We found that both dietary lipids and Cd exposure influenced fatty acid homeostasis and metabolism. The hepatic fatty acid profile mostly reflected that of the diet (e.g. n-3/n-6 ratio) with some differences, including selective retention of specific long chain polyunsaturated fatty acids (LC-PUFAs) like DHA and active biotransformation of dietary LA and ALA into LC-PUFAs. Cd effects on hepatic fatty acid profiles were influenced by the duration of the exposure and the nutritional status of the fish. The effects of diet and Cd exposure on the fatty acid profiles were only sparsely explained by variation of the expression pattern of genes involved in fatty acid metabolism. The biological responses to Cd were also influenced by dietary lipids. Fish fed the ALA-enriched diet seemed to be the least affected by the Cd exposure, as they showed a higher detoxifying ability against Cd with an early upregulation of protective metallothionein a (MTa) and apoptosis regulator BCL2-Like1 (BCLx) genes, an increased long-term phospholipid synthesis and turnover and fatty acid bioconversion efficiency, as well as a lower long-term accumulation of Cd in their liver. In contrast, fish fed the EPA-enriched diet seemed to be the most sensitive to a long-term Cd exposure, with an impaired growth performance and a decreased antioxidant capacity (lower glutathione level). Our results highlight that low, environmentally realistic aqueous concentrations of Cd can affect biological response in fish and that these effects are influenced by the dietary fatty acid composition.

A margin of safety approach for the assessment of environmentally realistic chemical mixtures in the marine environment based on combined passive sampling and ecotoxicity testing.

Organisms in the marine environment are being exposed to an increasing variety of chemicals. This research presents an effect-based monitoring method for the derivation of a margin of safety for environmentally realistic chemical mixtures. The method is based on a combination of passive sampling and ecotoxicity testing. First, passive sampling was performed using H2O-philic divinylbenzene Speedisks during 3 sampling campaigns between 2016 and 2018 at 4 sampling locations in the Belgian part of the North Sea. Next, we exposed the marine diatom Phaeodactylum tricornutum to Speedisk extracts that were reconstituted in HPLC-grade water and defined the MoS of each sample as the highest no-observed effect concentration, expressed as relative enrichment factor (REF). A REF was defined by comparing the concentrations of 89 personal care products, pesticides and pharmaceuticals in the biotest medium with those measured in water grab samples to relate exposure concentrations in the tests to environmental concentrations. Across eight marine samples, diatom growth inhibition was observed at REF ≥ 3.2 and margins of safety were found between REF 1.1-11.0. In addition, we found that reconstitution of extracts in HPLC-water was suitable to overcome the solvent-related challenges in biotesting that are usually associated with passive sampler extract spiking, whilst it still allowed REFs up to 44 in the biotest medium to be achieved. This method, however, likely covers mainly the polar fraction of environmentally realistic chemical mixtures and less the non-polar fraction. Nevertheless, for 5 out of 8 samples, the Margin of Safety (MoS) was found to be lower than 10, which represents the typically lowest possible assessment factor applied to no effects ecotoxicological data in conventional environmental risk assessments, suggesting ecological risks for these samples.

Spatio-temporal patterns in the gene expression of the calanoid copepod Temora longicornis in the Belgian part of the North Sea.

Marine zooplankton are increasingly being affected by recent environmental changes, such as climate change, and respond with profound spatial relocations and shifts in phenology and physiology. In order to predict whether populations are able to persist or adapt to such new conditions, it is essential to understand the molecular basis of such adaptations, which ultimately get translated into these physiological responses. To explore variation in population gene expression across time and space, we investigated transcriptome-level profiles of the calanoid copepod Temora longicornis, that were collected at four different locations in the Belgian Part of the North Sea (BPNS) on three different time points (April, June, October) in 2018. RNA-seq analysis of field collected adults identified large seasonal differences in gene expression, mainly between spring-summer and autumn samples. The largest log-fold changes occurred in a set of genes encoding for ribosomal and myosin (heavy chain) transcripts. Enrichment analysis revealed a strong seasonal pattern in vitellogenin, cuticle and glycolytic gene expression as well. We also found a positive correlation between vitellogenin expression and densities of T. longicornis. No clear spatial variation in expression patterns was found in the BPNS. This study underlines the potential of field gene expression studies for biomonitoring purposes and the significance of considering seasonal variation in future studies.