Toxicokinetic-Toxicodynamic Model to Assess Thermal Stress.

Temperature is a crucial environmental factor affecting the distribution and performance of ectothermic organisms. This study introduces a new temperature damage model to interpret their thermal stress. Inspired by the ecotoxicological damage model in the General Unified Threshold model for Survival (GUTS) framework, the temperature damage model assumes that damage depends on the balance between temperature-dependent accumulation and constant repair. Mortality due to temperature stress is driven by the damage level exceeding a threshold. Model calibration showed a good agreement with the measured survival of Gammarus pulex exposed to different constant temperatures. Further, model simulations, including constant temperatures, daily temperature fluctuations, and heatwaves, demonstrated the model's ability to predict temperature effects for various environmental scenarios. With this, the present study contributes to the mechanistic understanding of temperature as a single stressor while facilitating the incorporation of temperature as an additional stressor alongside chemicals in mechanistic multistressor effect models.

Toxicokinetics and toxicodynamics of Ag nanomaterials (NM300K) in the soil environment-impact on Enchytraeus crypticus (Oligochaeta).

Silver (Ag) is one of the most used elements in the nanomaterials (NMs) form, which upon release to the environment can be harmful to organisms. We compared the toxicokinetics (TK) and toxicodynamics (TD) of Ag from AgNO3 (0, 15, 45, 135, 405 mg Ag/kg soil) and AgNM300K (0, 75, 150, 300, 600, 1200 mg Ag/kg soil) in the model organism Enchytraeus crypticus. Organisms were exposed in LUFA 2.2 soil, and besides body Ag concentrations, survival and reproduction were determined, in a time series (for 21 days). In the soil, the available (CaCl2 extractable) Ag fraction from Ag NM300K increased from 0 to 21 days but did not consistently change for AgNO3. Internal concentrations reached equilibrium in most exposures to both Ag forms. The organisms were able to internalize and eliminate Ag, but less when exposed to Ag NM300K. The overall uptake rate constants for Ag from AgNO3 and Ag NM300K exposures were 0.05 and 0.06 kg soil/kg organism/day, respectively, the elimination rate constants 0.2 and 0.1 day-1, respectively. For AgNO3 the median lethal concentrations decreased steadily with time, while for Ag NM300K they remained constant during the first 10 days of exposure followed by a 2-fold decline in the last 7 days. The 21-d LC50s for both Ag forms were similar but the LC50inter (based on internal concentrations) were 63 and 121 mg Ag/kg body DW (Dry Weight) for AgNO3 and Ag NM300K, respectively, showing higher toxicity of AgNO3. These results show the importance of assessing time to toxicity, a relevant factor in toxicity assessment, especially for NMs.

Toxicokinetics and toxicodynamics of copper and cadmium in the soil invertebrate Enchytraeus crypticus (Oligochaeta).

The aim of this study was to evaluate the toxicokinetics-toxicodynamics (TKTD) of Cu and Cd in the soil model organism Enchytraeus crypticus, and assess the development of internal effect concentrations over time. Animals were exposed in LUFA 2.2 soil spiked with increasing concentrations of Cu and Cd. Survival, reproduction and internal metal concentrations in the animals were evaluated at different points in time over a period of 21 days. Internal concentrations increased with time, for Cu reaching a steady state after c. 10 days, except for the highest test concentration, and for Cd continuing to increase after 21 days. Applying a one-compartment model to all data together, estimated uptake and elimination rate constants for Cu and Cd were 0.08 and 0.45 kg soil/kg organism/day and 0.4 and 0.04 per day, respectively. Median lethal concentrations, based on total soil concentrations, decreased with time for Cu and did not reach a steady state level, but they did not change with time for Cd. The LC50inter (based on internal concentrations) was 75 mg Cu/kg body DW and > 800 mg Cd/kg body weight. Animals were able to regulate Cu internal concentrations, keeping them low, while for Cd internal concentrations continued to increase showing lack of regulation and also the importance of exposure time. This study highlights the advantages of using a TKTD approach to understand the relation between organism survival and internal Cu or Cd concentrations over time.

Comparing the acute and chronic toxicity of flupyradifurone and imidacloprid to non-target aquatic arthropod species.

Flupyradifurone (FPF) is a new type of butenolide insecticide. It was launched on the market in 2015 and is considered an alternative to the widely used neonicotinoids, like imidacloprid (IMI), some of which are banned from outdoor use in the European Union. FPF is claimed to be safe for bees, but its safety for aquatic organisms is unknown. Its high water solubility, persistence in the environment, and potential large-scale use make it urgent to evaluate possible impacts on aquatic systems. The current study assessed the acute and chronic toxicity of FPF for aquatic arthropod species and compared these results with those of imidacloprid. Besides, toxicokinetics and toxicokinetic-toxicodynamic models were used to understand the mechanisms of the toxicity of FPF. The present study results showed that organisms take up FPF slower than IMI and eliminate it faster. In addition, the hazardous concentration 5th percentiles (HC05) value of FPF derived from a species sensitivity distribution (SSD) based on acute toxicity was found to be 0.052 µmol/L (corresponding to 15 µg/L), which was 37 times higher than IMI (0.0014 µmol/L, corresponding to 0.36 µg/L). The chronic 28 days EC10 of FPF for Cloeon dipterum and Gammarus pulex were 7.5 µg/L and 2.9 µg/L, respectively. For G. pulex, after 28 days of exposure, the no observed effect concentration (NOEC) of FPF for food consumption was 0.3 µg/L. A toxicokinetic-toxicodynamic (TKTD) model parameterised on the acute toxicity data well predicted the observed chronic effects of FPF on G. pulex, indicating that toxicity mechanisms of FPF did not change with prolonged exposure time, which is not the case for IMI.

Modelling the effects of variability in feeding rate on growth - a vital step for DEB-TKTD modelling.

A major limitation of dietary toxicity studies on rodents is that food consumption often differs between treatments. The control treatment serves as a reference of how animals would have grown if not for the toxicant in their diet, but this comparison unavoidably conflates the effects of toxicity and feeding rate on body weight over time. A key advantage of toxicity models based on dynamic energy budget theory (DEB) is that chemical stress and food consumption are separate model inputs, so their effects on growth rate can be separated. To reduce data requirements, DEB convention is to derive a simplified feeding input, f, from food availability; its value ranges from zero (starvation) to one (food available ad libitum). Observed food consumption in dietary toxicity studies shows that, even in the control treatment, rats limit their food consumption, contradicting DEB assumptions regarding feeding rate. Relatively little work has focused on addressing this mismatch, but accurately modelling the effects of food intake on growth rate is essential for the effects of toxicity to be isolated. This can provide greater insight into the results of chronic toxicity studies and allows accurate extrapolation of toxic effects from laboratory data. Here we trial a new method for calculating f, based on the observed relationships between food consumption and body size in laboratory rats. We compare model results with those of the conventional DEB method and a previous effort to calculate f using observed food consumption data. Our results showed that the new method improved model accuracy while modelled reserve dynamics closely followed observed body fat percentage over time. The new method assumes that digestive efficiency increases with body size. Verifying this relationship through data collection would strengthen the basis of DEB theory and support the case for its use in ecological risk assessment.

An Integrated TK-TD Model for Evaluation of Radix Aconitikusnezoffii (RAK).

OBJECTIVE: An integrated TK-TD model with indirect response to toxicity was established using ADAPT 5 to evaluate abnormal heart rate (HR) and QT interval changes caused by Radix Aconitikusnezoffii (RAK). METHODS: Plasma samples were collected from male SD rats, which were divided into the blank and RAK groups. HR and QT interval indicators were recorded. Four alternative TK models were analyzed, and the best fitting model was determined. An indirect toxicodynamics model was selected, and the relationship of plasma concentration-time-toxicity was linked by Hill's equation. RESULTS: A 1-compartment linear first-order elimination kinetic model with the biophase model - an indirect toxic effect response model - best described the data. The high-dose QT interval was evaluated. Model simulation with the ML method showed that the fitting values of 0-15 h all fell within the confidence interval (95%). AMOS analysis showed that almost all the load factor of the variable was >0.7, and the χ2 value was 4.169 indicating a significant difference. Load factor (correlation coefficient) between the HR and QT intervals was -0.965, indicating negative correlation. CONCLUSIONS: The integrated TK-TD model with linear atrioventricular first-order elimination kinetics and indirect response represents a novel mathematical method to evaluate drug-induced changes in HR and QT.

A biological characteristic extrapolation of compound toxicity for different developmental stage species with toxicokinetic-toxicodynamic model.

Intraspecific difference in toxicity brings uncertainty to ecological risk assessment (ERA) and water quality criteria (WQC) of chemicals. Here, we compared intraspecies sensitivity to toxicants for Mesocyclops leuckarti of which toxicity data was obtained from published literatures, and zebrafish Danio rerio of which toxicity data was done in this study). Due to the internal concentration of chemicals not measured, simplified toxicokinetic-toxicodynamic (TK-TD) models were used, and we investigated whether TK-TD parameters estimated by Bayesian method might represent the differences in sensitivity between life-stages of 2 species. The results demonstrated that the difference in TK-TD parameters (background mortality m0, no effect concentration NEC, the killing rate ks, and the dominant rate kd) could represent the toxicity difference between life-stages of individual species. The TK-TD model could predict toxicity in individual species (Cyprinus carpio L., Enchytraeus crypticus, Folsomia candida, Hyalella Azteca) exposed to different chemical concentrations and successfully extrapolate toxicity between different life stages of Mesocyclops leuckarti and Danio rerio by scaling several TK-TD parameters. The modified TK-TD model on the extrapolation toxicity of chemicals between life stages for species could be useful for the ERA and for deriving and revising WQC for chemicals.

Gill damage and delayed mortality of Northern shrimp (Pandalus borealis) after short time exposure to anti-parasitic veterinary medicine containing hydrogen peroxide.

Hydrogen peroxide (H2O2) is used as anti-parasitic veterinary medicine in salmon farms worldwide. In the period from 2009 to 2018 a total of 135 million kg of H2O2 was used in Norway, the world's largest producer of Atlantic salmon. Since the treatment water is discharged to the sea, concerns have been raised about effects of H2O2 on the coastal ecosystem. In the present study, Northern shrimp (Pandalus borealis) have been exposed to short pulses of H2O2 in the PARAMOVE® formulation, followed by a recovery period in clean seawater. The exposure concentrations represented 100, 1000 and 10 000 times dilutions of the prescribed treatment concentration for salmon; 15 mg/L, 1.5 mg/L and 0.15 mg/L H2O2. Significantly increased mortality was observed after 2 h exposure to 15 mg/L H2O2 (50%) and after 2 h exposure to 1.5 mg/L H2O2 on 3 consecutive days (33%), but no mortality was observed after 2 h exposure to 0.15 mg/L. The mortality occurred 2-4 days after the first pulse of exposure. The patterns of acute effects (immobility and death) could be captured with a toxicokinetic-toxicodynamic model (GUTS), which allows extrapolations to LC50s for constant exposure, or thresholds for effects given untested exposure profiles. Effects of H2O2 were also detected in shrimp that survived until the end of the recovery period. The feeding rate was 66% lower than in the control after 12 days of recovery for the three-pulse 1.5 mg/L exposure. Furthermore, dose dependent tissue damage was detected in the gills and evidence of lipid peroxidation in the hepatopancreas in shrimp exposed for 1 h to 1.5 mg/L and 15 mg/L and kept in recovery for 8 days. Fluorescence intensity in the hepatopancreas of treated shrimp increased 47% and 157% at 1.5 mg/L and 15 mg/L, respectively, compared to the control. Local hydrodynamic conditions will determine how fast the concentration of H2O2 will be diluted and how far it will be transported horizontally and vertically. Results from dispersion modelling (literature data) together with the current experiments indicate that treatment water with toxic concentrations of H2O2 (1.5 mg/L) could reach P. borealis living more than 1 km from a treated salmon farm.

Combined effect of temperature and ammonia on molecular response and survival of the freshwater crustacean Gammarus pulex.

Freshwater ecosystems are experiencing mounting pressures from agriculture, urbanization, and climate change, which could drastically impair aquatic biodiversity. As nutrient inputs increase and temperatures rise, ammonia (NH3) concentration is likely to be associated with stressful temperatures. To investigate the interaction between NH3 and temperature on aquatic invertebrate survival, we performed a factorial experiment on the survival and molecular response of Gammarus pulex, with temperature (10, 15, 20, and 25°C) and NH3 (0, 0.5, 1, 2, 3, and 4mg NH3/L) treatments. We observed an unexpected antagonistic interaction between temperature and NH3 concentration, meaning survival in the 4mg NH3/L treatment was higher at 25°C than at the control temperature of 10°C. A toxicokinetic-toxicodynamic (TK-TD) model was built to describe this antagonistic interaction. While the No Effect Concentration showed no significant variation across temperatures, the 50% lethal concentration at the end of the experiment increased from 2.7 (2.1-3.6) at 10°C to 5.5 (3.5- 23.4) mg NH3/L at 25°C. Based on qPCR data, we associated these survival patterns to variations in the expression of the hsp70 gene, a generic biomarker of stress. However, though there was a 14-fold increase in hsp70 mRNA expression for gammarids exposed to 25°C compared to controls, NH3 concentration had no effect on hsp70 mRNA synthesis across temperatures. Our results demonstrate that the effects of combined environmental stressors, like temperature and NH3, may strongly differ from simple additive effects, and that stress response to temperature can actually increase resilience to nutrient pollution in some circumstances.

Modeling interactions and toxicity of Cu-Zn mixtures to zebrafish larvae.

Quantitative predictions of metal-metal interactions and toxicity in aquatic organisms meet a unique challenge. Accumulation and toxicity of Cu and Zn mixtures in zebrafish larvae has been quantified in binary metal system with variable combinatorial concentrations in order to understand the interactions between essential trace metals and assess availability of the toxicokinetic-toxicodynamic (TK-TD) model which simulated the uptake of metals over time as well as metal toxicity after 24h of exposure. Competitive uptake experiments showed a straightforward antagonistic competition, as would be predicted by Michaelis-Menten competitive equilibrium model. Zn uptake decreased significantly in the presence of Cu2+ concentrations higher than 10-6M. Cu2+ was shown to compete strongly with Zn for uptake, having a higher affinity constant to biotic ligand (BL) sites (KCuBL=105.42M-1) than Zn (KZnBL=104.13M-1). TK-TD model considering potential metal-metal antagonism interactions showed good predictive power in predicting accumulation and toxicity of Cu-Zn mixtures in zebrafish larvae with the high coefficient of determination (r2) and significant level (p). In particular, with the elevated Zn concentrations in mixtures, the TD model showed better predictive power in predicting toxicity of 10-6M Cu concentration in Cu-Zn mixtures. The TK-TD analysis provided some new insights into the interactive mechanism of binary Cu and Zn exposure in aquatic animals and may have important implications for our understanding of quantitative predictions of metal-metal interactions and toxicity in a field where animals are simultaneously exposed to several metals.

Analysis of real-time mixture cytotoxicity data following repeated exposure using BK/TD models.

Cosmetic products generally consist of multiple ingredients. Thus, cosmetic risk assessment has to deal with mixture toxicity on a long-term scale which means it has to be assessed in the context of repeated exposure. Given that animal testing has been banned for cosmetics risk assessment, in vitro assays allowing long-term repeated exposure and adapted for in vitro - in vivo extrapolation need to be developed. However, most in vitro tests only assess short-term effects and consider static endpoints which hinder extrapolation to realistic human exposure scenarios where concentration in target organs is varies over time. Thanks to impedance metrics, real-time cell viability monitoring for repeated exposure has become possible. We recently constructed biokinetic/toxicodynamic models (BK/TD) to analyze such data (Teng et al., 2015) for three hepatotoxic cosmetic ingredients: coumarin, isoeugenol and benzophenone-2. In the present study, we aim to apply these models to analyze the dynamics of mixture impedance data using the concepts of concentration addition and independent action. Metabolic interactions between the mixture components were investigated, characterized and implemented in the models, as they impacted the actual cellular exposure. Indeed, cellular metabolism following mixture exposure induced a quick disappearance of the compounds from the exposure system. We showed that isoeugenol substantially decreased the metabolism of benzophenone-2, reducing the disappearance of this compound and enhancing its in vitro toxicity. Apart from this metabolic interaction, no mixtures showed any interaction, and all binary mixtures were successfully modeled by at least one model based on exposure to the individual compounds.

Reproductive toxicity in birds predicted by physiologically-based kinetics and bioenergetics modelling.

Effects on the growth and reproduction of birds are important endpoints in the environmental risk assessment (ERA) of pesticides. Toxicokinetic-toxicodynamic models based on dynamic energy budget theory (DEB) are promising tools to predict these effects mechanistically and make extrapolations relevant to ERA. However, before DEB-TKTD models are accepted as part of ERA for birds, ecotoxicological case studies are required so that stakeholders can assess their capabilities. We present such a case-study, modelling the effects of the fluopyram metabolite benzamide on the northern bobwhite quail (Colinus virginianus). We parametrised a DEB-TKTD model for the embryo stage on the basis of an egg injection study, designed to provide data for model development. We found that information on various endpoints, such as survival, growth, and yolk utilisation were needed to clearly distinguish between the performance of model variants with different TKTD assumptions. The calibration data were best explained when it was assumed that chemical uptake occurs via the yolk and that benzamide places stress on energy assimilation and mobilisation. To be able to bridge from the in vitro tests to real-life exposure, we developed a physiologically-based toxicokinetic (PBK) model for the quail and used it to predict benzamide exposure inside the eggs based on dietary exposure in a standard reproductive toxicity study. We then combined the standard DEB model with the TKTD module calibrated to the egg injection studies and used it to predict effects on hatchling and 14-day chick weight based on the exposure predicted by the PBK model. Observed weight reductions, relative to controls, were accurately predicted. Thus, we demonstrate that DEB-TKTD models, in combination with suitable experimental data and, if necessary, with an exposure model, can be used in bird ERA to predict chemical effects on reproduction.

Comparing Sensitivity of Different Bee Species to Pesticides: A TKTD modeling approach.

Risk assessment for bees is mainly based on data for honey bees; however, risk assessment is intended to protect all bee species. This raises the question of whether data for honey bees are a good proxy for other bee species. This issue is not new and has resulted in several publications in which the sensitivity of bee species is compared based on the values of the 48-h median lethal dose (LD50) from acute test results. When this approach is used, observed differences in sensitivity may result both from differences in kinetics and from inherent differences in species sensitivity. In addition, the physiology of the bee, like its overall size, the size of the honey stomach (for acute oral tests), and the physical appearance (for acute contact tests) also influences the sensitivity of the bee. The recently introduced Toxicokinetic-Toxicodynamic (TKTD) model that was developed for the interpretation of honey bee tests (Bee General Uniform Threshold Model for Survival [BeeGUTS]) could integrate the results of acute oral tests, acute contact tests, and chronic tests within one consistent framework. We show that the BeeGUTS model can be calibrated and validated for other bee species and also that the honey bee is among the more sensitive bee species. In addition, we found that differences in sensitivity between species are smaller than previously published comparisons based on 48-h LD50 values. The time-dependency of the LD50 and the specifics of the bee physiology are the main causes of the wider variation found in the published literature. Environ Toxicol Chem 2024;43:1431-1441. © 2024 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Extrapolating Metal (Cu, Ni, Zn) Toxicity from Individuals to Populations Across Daphnia Species Using Mechanistic Models: The Roles of Uncertainty Propagation and Combined Physiological Modes of Action.

Mechanistic effect modeling is a promising tool to improve the ecological realism of environmental risk assessment. An open question for the mechanistic modeling of metal toxicity is whether the same physiological mode of action (PMoA) could be assumed for closely related species. The implications of various modeling choices, such as the use of parameter point estimates and assumption of simplistic toxicodynamic models, are largely unexplored. We conducted life-table experiments with Daphnia longispina, Daphnia magna, and Daphnia pulex exposed to the single metals Cu, Ni, and Zn, and calibrated toxicokinetic-toxicodynamic (TKTD) models based on dynamic energy budget theory. We developed TKTD models with single and combined PMoAs to compare their goodness-of-fit and predicted population-level sensitivity. We identified the PMoA reproduction efficiency as most probable in all species for Ni and Zn, but not for Cu, and found that combined-PMoA models predicted higher population-level sensitivity than single-PMoA models, which was related to the predicted individual-level sensitivity, rather than to mechanistic differences between models. Using point estimates of parameters, instead of sampling from the probability distributions of parameters, could also lead to differences in the predicted population-level sensitivity. According to model predictions, apical chronic endpoints (cumulative reproduction, survival) are conservative for single-metal population effects across metals and species. We conclude that the assumption of an identical PMoA for different species of Daphnia could be justified for Ni and Zn, but not for Cu. Single-PMoA models are more appropriate than combined-PMoA models from a model selection perspective, but propagation of the associated uncertainty should be considered. More accurate predictions of effects at low concentrations may nevertheless motivate the use of combined-PMoA models. Environ Toxicol Chem 2024;43:338-358. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Identifying and Predicting Delayed Mortality with Toxicokinetic-Toxicodynamic Models.

The prevalence of standardized toxicity testing in ecotoxicology has largely obscured the notion that toxicity is a function of time as well. The necessity of considering time is vividly demonstrated by observations of delayed mortality, that is, deaths continue to occur even when animals are no longer exposed to a toxicant. In this contribution, I explore to what extent toxicokinetic-toxicodynamic (TKTD) models from the framework of the General Unified Threshold model for Survival (GUTS) can capture delayed mortality, and to what extent this phenomenon can be predicted from short-term standard tests. I use a previously published data set for fluoroquinolones in Daphnia magna that shows strongly delayed mortality (using immobilization as a proxy for death). The model analysis shows that the GUTS stochastic death models can capture delayed mortality in the complete data set with a long recovery phase, but that the delayed effects would not have been predicted from a 2-day standard test. The study underlines the limited information content of standard acute test designs. Toxicokinetic-toxicodynamic modeling offers a handle on the time aspects of toxicity but cannot always be relied on to provide accurate extrapolations based on severely limited standard tests. The phenomenon of delayed toxicity requires more structured study to clarify its prevalence and impact; I discuss several avenues for further investigation. Environ Toxicol Chem 2024;43:1030-1035. © 2024 SETAC.

Toxicokinetics and toxicodynamics of chromium in the soil invertebrate Enchytraeus crypticus (Oligochaeta).

Chromium emissions led to increased concentrations in soil, where it can affect soil organisms to relevant levels. With the aim of better understanding the effects of Cr throughout time, its toxicokinetics-toxicodynamics (TKTD) were evaluated in the soil model organism Enchytraeus crypticus to assess the development of internal concentrations and consequent toxic effects. To achieve this goal, organisms were exposed in LUFA 2.2 soil spiked with increasing CrCl3 concentrations. During the 21-day exposure period, survival, internal concentrations, and reproduction were evaluated at several time points up to 21 days. Uptake and elimination rate constants were 0.0044 kg soil/kg organism/day and 0.023 per day, respectively. Internal Cr concentrations increased with time, generally reaching equilibrium within 14 days with an estimated LC50inter (based on internal metal concentrations) of 57.7 mg Cr/kg body DW. Internal Cr concentrations were regulated by the organisms up to exposure to 360 mg Cr/kg soil DW, where the elimination rate was highest, but at 546 mg Cr/kg soil DW the animals were no longer able to eliminate Cr, and the internal concentrations were well above the estimated LC50inter. At day 21, exposure to 546 mg Cr/kg soil DW significantly reduced survival by 23 %, while reproduction EC50 was 344 mg Cr/kg soil DW. This study highlights the advantages of using a TKTD approach to understand the development of internal metal concentrations in time and relate it to the phenotypical effects observed. Toxicity is better understood when also taking into account time and not just exposure concentration alone.

A Framework for Algae Modeling in Regulatory Risk Assessment.

The use of toxicokinetic-toxicodynamic (TKTD) modeling in regulatory risk assessment of plant protection products is increasingly popular, especially since the 2018 European Food Safety Authority (EFSA) opinion on TKTD modeling announced that several established models are ready for use in risk assessment. With careful adherence to the guidelines laid out by EFSA, we present a stepwise approach to validation and use of the Simple Algae Model Extended (SAM-X) for regulatory submission in Tier 2C. We demonstrate how the use of moving time windows across time-variable exposure profiles can generate thousands of virtual laboratory mimic simulations that seamlessly predict the effects of time-variable exposures across a full exposure profile while maintaining the laboratory conditions of the standard Organisation for Economic Co-operation and Development (OECD) growth inhibition test. Thus, every virtual laboratory test has a duration of 72 h, with OECD medium and constant light and temperature conditions. The only deviation from the standard test setup is the replacement of constant exposure conditions for time-variable concentrations. The present study demonstrates that for simulation of 72-h toxicity tests, the nutrient dynamics in the SAM-X model are not required, and we propose the alternative use of a simplified model version. For risk assessment, in accordance with the EFSA guidelines we use a median exposure profile of 10 as a threshold, meaning that if a time window within the exposure profile causes 50% growth inhibition when magnified by a factor of 10, the threshold will have been exceeded. We present a simplified example for chlorotoluron and isoproturon. The present case study brings to life our proposed framework for TKTD modeling of algae to establish whether a given exposure can be considered to be of low risk. Environ Toxicol Chem 2023;42:1823-1838. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Impact of the antidepressant Bupropion on the Dynamic Energy Budget of Daphnia magna.

Psychiatric drugs are considered among the emerging contaminants of concern in ecological risk assessment, due to their potential to disrupt homeostasis in aquatic organisms. Bupropion is an antidepressant that acts by selective reuptake inhibition of norepinephrine and dopamine. Little is known about this compound's effects on aquatic organisms, despite being detected in significant concentrations in both water and biota close to waste-water treatment plants and densely populated areas. Dynamic Energy Budget (DEB) models are flexible mechanistic tools that can be applied to understand toxic effects and extrapolate individual responses to higher biological levels and under untested environmental conditions. In this work, we used the stdDEB-TKTD (an application of the DEB theory to ecotoxicology) approach to investigate the possible physiological mode of action of Bupropion on the model organism Daphnia magna. Next, Dynamic Energy Budget Individual-Based Models (DEB-IBM) were used to extrapolate the results to the population level and to predict the combined effects of Bupropion exposure and food availability on the daphnids. Our results revealed an increasing negative effect of this antidepressant on the reproduction and survival of the animals with increasing concentration (0.004, 0.058, 0.58 and 58 µM). At the population level, we found that even the lowest used doses of Bupropion could reduce the population density and its reproductive output. The impacts are predicted to be stronger under limited food conditions.

Considerations for using reproduction data in toxicokinetic-toxicodynamic modeling.

Toxicokinetic-toxicodynamic (TKTD) modeling is essential to make sense of the time dependence of toxic effects, and to interpret and predict consequences of time-varying exposure. These advantages have been recognized in the regulatory arena, especially for environmental risk assessment of pesticides, where time-varying exposure is the norm. We critically evaluate the link between the modeled variables in TKTD models and the observations from laboratory ecotoxicity tests. For the endpoint reproduction, this link is far from trivial. The relevant TKTD models for sublethal effects are based on dynamic energy budget (DEB) theory, which specifies a continuous investment flux into reproduction. In contrast, experimental tests score egg or offspring release by the mother. The link between model and data is particularly troublesome when a species reproduces in discrete clutches and, even more so, when eggs are incubated in the mother's brood pouch (and release of neonates is scored in the test). This situation is quite common among aquatic invertebrates (e.g., cladocerans, amphipods, mysids), including many popular test species. In this discussion paper, we treat these and other issues with reproduction data, reflect on their potential impact on DEB-TKTD analysis, and provide preliminary recommendations to correct them. Both modelers and users of model results need to be aware of these complications, as ignoring them could easily lead to unnecessary failure of DEB-TKTD models during calibration, or when validating them against independent data for other exposure scenarios. Integr Environ Assess Manag 2022;18:479-487. © 2021 SETAC.

How to analyse and account for interactions in mixture toxicity with toxicokinetic-toxicodynamic models.

The assessment of chemical mixture toxicity is one of the major challenges in ecotoxicology. Chemicals can interact, leading to more or less effects than expected, commonly named synergism and antagonism respectively. The classic ad hoc approach for the assessment of mixture effects is based on dose-response curves at a single time point, and is limited to identifying a mixture interaction but cannot provide predictions for untested exposure durations, nor for scenarios where exposure varies in time. We here propose a new approach using toxicokinetic-toxicodynamic modelling: The General Unified Threshold model of Survival (GUTS) framework, recently extended for mixture toxicity assessment. We designed a dedicated mechanistic interaction module coupled with the GUTS mixture model to i) identify interactions, ii) test hypotheses to identify which chemical is likely responsible for the interaction, and finally iii) simulate and predict the effect of synergistic and antagonistic mixtures. We tested the modelling approach experimentally with two species (Enchytraeus crypticus and Mamestra brassicae) exposed to different potentially synergistic mixtures (composed of: prochloraz, imidacloprid, cypermethrin, azoxystrobin, chlorothalonil, and chlorpyrifos). Furthermore, we also tested the model with previously published experimental data on two other species (Bombus terrestris and Daphnia magna) exposed to pesticide mixtures (clothianidin, propiconazole, dimethoate, imidacloprid and thiacloprid) found to be synergistic or antagonistic with the classic approach. The results showed an accurate simulation of synergistic and antagonistic effects for the different tested species and mixtures. This modelling approach can identify interactions accounting for the entire time of exposure, and not only at one time point as in the classic approach, and provides predictions of the mixture effect for untested mixture exposure scenarios, including those with time-variable mixture composition.