ABSTRACT
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.
Subject(s)
Ecotoxicology , Mortality , Pharmacokinetics , Toxicity Tests, Acute , Animals , Humans , Daphnia magna/drug effects , Datasets as Topic , Death , Ecotoxicology/methods , Fluoroquinolones/toxicity , Pesticides/toxicity , Risk Assessment , Stochastic Processes , Time Factors , Toxicity Tests, Acute/methods , Toxicity Tests, Acute/standardsABSTRACT
The extrapolation of effects from controlled standard laboratory tests to real environmental conditions is a major challenge facing ecological risk assessment (ERA) of chemicals. Toxicokinetic-toxicodynamic (TKTD) models, such as those based on dynamic energy budget (DEB) theory, can play an important role in filling this gap. Through the years, different practical TKTD models have been derived from DEB theory, ranging from the full "standard" DEB animal model to simplified "DEBtox" models. It is currently unclear what impact a different level of model complexity can have on the regulatory risk assessment. In the present study, we compare the performance of two DEB-TKTD models with different levels of complexity, focusing on model calibration on standard test data and on forward predictions for untested time-variable exposure profiles. The first model is based on the standard DEB model with primary parameters, whereas the second is a reduced version with compound parameters, based on DEBkiss. After harmonization of the modeling choices, we demonstrate that these two models can achieve very similar performances both in the calibration step and in the forward prediction step. With the data presented in the present study, selection of the most suitable TKTD model for ERA therefore cannot be based alone on goodness-of-fit or on the precision of model predictions (within current ERA procedures for pesticides) but would likely be based on the trade-off between ease of use and model flexibility. We also stress the importance of modeling choices, such as how to fill gaps in the information content of experimental toxicity data and how to accommodate differences in growth and reproduction between different data sets for the same chemical-species combination. Environ Toxicol Chem 2024;43:440-449. © 2023 ibacon GmbH. Bayer AG and The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.
Subject(s)
Pesticides , Animals , Risk Assessment , Pesticides/toxicity , EcotoxicologyABSTRACT
The demand for mineral resources is increasing mining activities worldwide. In Norway, marine tailing disposal (MTD) is practiced, introducing mineral particles into fjord ecosystems. We investigated the effects of two concentrations (high and low) of fine tailings from a CaCO3 processing plant on early life stages of the marine copepod Calanus finmarchicus. Results show that the exposure did not significantly impact hatching success or development in non- and early feeding life stages. However, feeding stage nauplii ingested tailings, which caused a significantly slower development in later nauplii stages in high exposure groups, with most individuals being two stages behind the control group. Further, high mortality occurred in late nauplii and early copepodite stages in low exposure groups, which could be caused by insufficient energy accumulation and depleted energy reserves during development. Individuals exposed to high exposure concentrations seemed to survive by arresting development and potentially by reduced activity, thereby conserving energy reserves. In nature, slower development could affect lipid storage buildup and reproduction.
ABSTRACT
The Dynamic Energy Budget theory (DEB) enables ecotoxicologists to model the effects of chemical stressors on organism life cycles through the coupling of toxicokinetic-toxicodynamic (TK-TD) models. While good progress has been made in the application of DEB-TKTD models for aquatic organisms, applications for soil fauna are scarce, due to the lack of dedicated experimental designs suitable for collecting the required time series effect data. Enchytraeids (Annelida: Clitellata) are model organisms in soil ecology and ecotoxicology. They are recognised as indicators of biological activity in soil, and chemical stress in terrestrial ecosystems. Despite this, the application of DEB-TKTD models to investigate the impact of chemicals has not yet been tested on this family. Here we assessed the impact of the pyrethroid insecticide cypermethrin on the life cycle of Enchytraeus crypticus. We developed an original experimental design to collect the data required for the calibration of a DEB-TKTD model for this species. E. crypticus presented a slow initial growth phase that has been successfully simulated with the addition of a size-dependent food limitation for juveniles in the DEB model. The DEB-TKTD model simulations successfully agreed with the data for all endpoints and treatments over time. The highlighted physiological mode of action (pMoA) for cypermethrin was an increase of the growth energy cost. The threshold for effects on survival was estimated at 73.14 mg kg- 1, and the threshold for effects on energy budget (i.e., sublethal effects) at 19.21 mg kg- 1. This study demonstrates that DEB-TKTD models can be successfully applied to E. crypticus as a representative soil species, and may improve the ecological risk assessment for terrestrial ecosystems, and our mechanistic understanding of chemical effects on non-target species.
Subject(s)
Insecticides , Oligochaeta , Pyrethrins , Animals , Insecticides/toxicity , Research Design , Soil , Ecosystem , Pyrethrins/toxicity , Life Cycle StagesABSTRACT
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.
Subject(s)
Chlorpyrifos , Insecticides , Oligochaeta , Animals , Chlorpyrifos/toxicity , Daphnia , Insecticides/chemistry , ToxicokineticsABSTRACT
Thanks to growing interest and research in the field, toxicokinetic-toxicodynamic (TKTD) models are close to realising their potential in environmental risk assessment (ERA) of chemicals such as plant protection products. A fundamental application is to find a multiplicative scale factor which-when applied to an exposure profile-results in some specified effect relative to a control. The approach is similar to applying assessment factors to experimental results, common in regulatory frameworks. It also relies on the same core assumption: that increasing the scaling always produces more extreme effects. Unlike experimental approaches, TKTD models offer an opportunity to interrogate this assumption in a mathematically rigorous manner. For four well-known TKTD models we seek to prove that the approach guarantees a unique scale factor for any percentage effect. Somewhat surprisingly, certain model configurations may have multiple scale factors which result in the same percentage effect. These cases require a more cautious regulatory approach and generate open biological and mathematical questions. We provide examples of the violations and suggest how to deal with them. Mathematical proofs provide the strongest possible backing for TKTD modelling approaches in ERA, since the applicability of the models can be determined exactly.
Subject(s)
Models, Theoretical , Risk Assessment/methodsABSTRACT
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.
Subject(s)
Pesticides , Humans , Infant, Newborn , Models, Biological , Pesticides/toxicity , Reproduction , Risk Assessment , ToxicokineticsABSTRACT
In this study we investigated potential impacts of Cu exposure at low, environmentally relevant, concentrations on early live stages of Atlantic cod (Gadus morhua). Cod embryos and larvae were exposed to 0.5 µg/L (low), 2 µg/L (medium), and 6 µg/L (high) Cu from 4 to 17 days post fertilisation (dpf). Hatching success, mortality, oxygen consumption, biometric traits, and malformations were determined. A dynamic energy budget (DEB) model was applied to identify potential impacts on bioenergetics. A positive correlation was found between Cu exposure concentrations and Cu body burden in eggs, but not in larvae. The tested concentrations did not increase mortality in neither embryos nor larvae, or larvae deformations. Further, the DEB model did not indicate effects of the tested Cu concentrations.
ABSTRACT
Marine tailing disposal (MTD) is sometimes practiced as an alternative to traditional mine tailing deposition on land. Environmental challenges connected to MTD include spreading of fine particulate matter in the water column and the potential release of metals and processing chemicals. This study investigated if tailing exposure affects the marine copepod Calanus finmarchicus, and whether effects are related to exposure to mineral particles or the presence of metals and/or processing chemicals in the tailings. We investigated the impacts of three different tailing compositions: calcium carbonate particles with and without processing chemicals and fine-grained tailings from a copper ore. Early life stages of C. finmarchicus were exposed over several developmental stages to low and high suspension concentrations for 15 days, and their development, oxygen consumption and biometry determined. The data was fitted in a dynamic energy budget (DEB) model to determine mechanisms underlying responses and to understand the primary modes of action related to mine tailing exposure. Results show that copepods exposed to tailings generally exhibited slower growth and accumulated less lipids. The presence of metals and processing chemicals did not influence these responses, suggesting that uptake of mineral particles was responsible for the observed effects. This was further supported by the applied DEB model, confirming that ingestion of tailing particles while feeding can result in less energy being available for growth and development.
Subject(s)
Copepoda , Animals , Calcium Carbonate , Copper/toxicity , Metals , Particulate MatterABSTRACT
Current methods to assess the impact of chemical mixtures on organisms ignore the temporal dimension. The General Unified Threshold model for Survival (GUTS) provides a framework for deriving toxicokinetic-toxicodynamic (TKTD) models, which account for effects of toxicant exposure on survival in time. Starting from the classic assumptions of independent action and concentration addition, we derive equations for the GUTS reduced (GUTS-RED) model corresponding to these mixture toxicity concepts and go on to demonstrate their application. Using experimental binary mixture studies with Enchytraeus crypticus and previously published data for Daphnia magna and Apis mellifera, we assessed the predictive power of the extended GUTS-RED framework for mixture assessment. The extended models accurately predicted the mixture effect. The GUTS parameters on single exposure data, mixture model calibration, and predictive power analyses on mixture exposure data offer novel diagnostic tools to inform on the chemical mode of action, specifically whether a similar or dissimilar form of damage is caused by mixture components. Finally, observed deviations from model predictions can identify interactions, e.g., synergism or antagonism, between chemicals in the mixture, which are not accounted for by the models. TKTD models, such as GUTS-RED, thus offer a framework to implement new mechanistic knowledge in mixture hazard assessments.
Subject(s)
Daphnia , Models, Biological , Animals , Bees , Calibration , Risk Assessment , ToxicokineticsABSTRACT
Toxicokinetic-toxicodynamic (TKTD) models offer a mechanistic understanding of individual-level toxicity over time and allow for meaningful extrapolations from laboratory tests to exposure conditions in the field. Thereby, they hold great potential for ecotoxicological studies, both in a regulatory context as well as for basic research. In contrast to mechanistic effect models at higher levels of biological organization, TKTD models can be, and generally are, parameterized by fitting them to data (results from toxicity tests). Fitting models comes with a range of statistical and numerical challenges, which may hamper the application of TKTD models in a practical setting. Especially in the context of environmental risk assessment, there is a need for robust and user-friendly software tools to automatically extract the best-fitting model parameters and quantify their uncertainty from any data set. The study presents a general outline for TKTD model analysis, rooted in likelihood-based ("frequentist") inference. The general outline is followed by a presentation of the specific algorithm that has been implemented into software for the robust and automated analysis of toxicity data for survival. However, the presented approach is more broadly applicable to low-dimensional problems. Integr Environ Assess Manag 2021;17:388-397. © 2020 SETAC.
Subject(s)
Ecotoxicology , Toxicity Tests , Likelihood Functions , Models, Biological , Risk Assessment , Toxicokinetics , UncertaintyABSTRACT
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 10â¯000 times dilutions of the prescribed treatment concentration for salmon; 15â¯mg/L, 1.5â¯mg/L and 0.15â¯mg/L H2O2. Significantly increased mortality was observed after 2â¯h exposure to 15â¯mg/L H2O2 (50%) and after 2â¯h exposure to 1.5â¯mg/L H2O2 on 3 consecutive days (33%), but no mortality was observed after 2â¯h exposure to 0.15â¯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 1.5â¯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 1â¯h to 1.5â¯mg/L and 15â¯mg/L and kept in recovery for 8 days. Fluorescence intensity in the hepatopancreas of treated shrimp increased 47% and 157% at 1.5â¯mg/L and 15â¯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 (1.5â¯mg/L) could reach P. borealis living more than 1â¯km from a treated salmon farm.
Subject(s)
Antiparasitic Agents/toxicity , Gills/drug effects , Hydrogen Peroxide/toxicity , Pandalidae/drug effects , Veterinary Drugs/toxicity , Water Pollutants, Chemical/toxicity , Animals , Dose-Response Relationship, Drug , Ecosystem , Hepatopancreas/drug effects , Lethal Dose 50 , Models, Biological , Norway , Seawater/chemistry , Survival Analysis , Time FactorsABSTRACT
Organism size and growth curves are important biological characteristics. Current methods to measure organism size, and in particular growth curves, are often resource intensive because they involve many manual steps. Here we demonstrate a method for automated, high-throughput measurements of size and growth in individual aquatic invertebrates kept in microtiter well-plates. We use a spheroid counter (Cell3iMager, cc-5000) to automatically measure size of seven different freshwater invertebrate species. Further, we generated calibration curves (linear regressions, all p < 0.0001, r2 >=0.9 for Ceriodaphnoa dubia, Asellus aquaticus, Daphnia magna, Daphnia pulex; r2 >=0.8 for Hyalella azteca, Chironomus spec. larvae and Culex spec. larvae) to convert size measured on the spheroid counter to traditional, microscope based, length measurements, which follow the longest orientation of the body. Finally, we demonstrate semi-automated measurement of growth curves of individual daphnids (C. dubia and D. magna) over time and find that the quality of individual growth curves varies, partly due to methodological reasons. Nevertheless, this novel method could be adopted to other species and represents a step change in experimental throughput for measuring organisms' shape, size and growth curves. It is also a significant qualitative improvement by enabling high-throughput assessment of inter-individual variation of growth.
Subject(s)
Arthropods/growth & development , Fresh Water , Larva/growth & development , AnimalsABSTRACT
The risk of accidental oil spills in the Arctic is on the rise due to increased shipping and oil exploration activities, making it essential to calibrate parameters for risk assessment of oil spills to Arctic conditions. The toxicokinetics of crude oil components were assessed by exposing one lipid-poor (CIII) and one lipid-rich (CV) stage of the Arctic copepod Calanus hyperboreus to crude oil WSF (water-soluble fraction). Water concentrations and total body residues (BR), as well as lipid volume fractions, were measured at regular intervals during exposure and recovery. Bioconcentration factors (BCFs) and elimination rates ( ke) for 26 petrogenic oil components were estimated from one-compartment models fitted to the BR data. Our parameters were compared to estimations made by the OMEGA bioaccumulation model, which uses the octanol-water partitioning coefficient ( KOW) in QSAR (quantitative structure-activity relationship) predictions. Our parameters for the lipid-poor CIIIs generally agreed with the OMEGA predictions, while neither the BCFs nor the kes for the lipid-rich CVs fitted within the realistic range of the OMEGA parameters. Both the uptake and elimination rates for the CVs were in general half an order of magnitude lower than the OMEGA predictions, showing an overestimation of these parameters by the OMEGA model.
Subject(s)
Copepoda , Petroleum , Water Pollutants, Chemical , Animals , Arctic Regions , ToxicokineticsABSTRACT
Environmental risk assessment (ERA) of chemicals relies on the combination of exposure and effects assessment. Exposure concentrations are commonly estimated using mechanistic fate models, but the effects side is restricted to descriptive statistical treatment of toxicity data. Mechanistic effect models are gaining interest in a regulatory context, which has also sparked discussions on model quality and good modeling practice. Proposals for good modeling practice of effect models currently focus very much on population and community models, whereas effects models also exist at the individual level, falling into the category of toxicokinetic-toxicodynamic (TKTD) models. In contrast to the higher-level models, TKTD models are usually completely parameterized by fitting them to experimental data. In fact, one of their explicit aims is to replace descriptive methods for data analysis. Furthermore, the construction of these models does not fit into an orderly modeling cycle, given that most TKTD models have been under continuous development for decades and are being applied by many different research groups, for many different purposes. These aspects have considerable consequences for the application of frameworks for model evaluation. For example, classical sensitivity analysis becomes rather meaningless when all model parameters are fitted to a data set. We illustrate these issues with the General Unified Threshold model for Survival (GUTS), relate them to the quality issues for currently used models in ERA, and provide recommendations for the evaluation of TKTD models and their analyses. Integr Environ Assess Manag 2018;14:604-614. ©2018 SETAC.
Subject(s)
Environmental Monitoring/methods , Environmental Pollutants/toxicity , Models, Chemical , Toxicokinetics , Ecotoxicology , Risk AssessmentABSTRACT
As ecotoxicologists we strive for a better understanding of how chemicals affect our environment. Humanity needs tools to identify those combinations of man-made chemicals and organisms most likely to cause problems. In other words: which of the millions of species are at risk from pollution? And which of the tens of thousands of chemicals contribute most to the risk? We identified our poor knowledge on physiological modes of action (how a chemical affects the energy allocation in an organism), and how they vary across species and toxicants, as a major knowledge gap. We also find that the key to predictive ecotoxicology is the systematic, rigorous characterization of physiological modes of action because that will enable more powerful in vitro to in vivo toxicity extrapolation and in silico ecotoxicology. In the near future, we expect a step change in our ability to study physiological modes of action by improved, and partially automated, experimental methods. Once we have populated the matrix of species and toxicants with sufficient physiological mode of action data we can look for patterns, and from those patterns infer general rules, theory and models.
Subject(s)
Ecotoxicology/methods , Environmental Pollutants/toxicity , Hazardous Substances/toxicity , Models, Biological , Animals , Environmental Pollutants/chemistry , Environmental Pollutants/pharmacokinetics , Hazardous Substances/chemistry , Hazardous Substances/pharmacokinetics , Quantitative Structure-Activity Relationship , Risk Assessment/methods , Species SpecificityABSTRACT
Efficiently assessing and managing the risks of pollution in the marine environment requires mechanistic models for toxic effects. The General Unified Threshold model for Survival (GUTS) provides a framework for deriving toxicokinetic-toxicodynamic (TKTD) models for the end point survival. Two recurring questions in the application of GUTS concern the most appropriate death mechanism, and whether the total body residue is a proper dose metric for toxic effects. We address these questions with a case study for dimethylnaphthalene in the marine copepod Calanus finmarchicus. A detailed analysis revealed that body residues were best explained by representing copepods with two toxicokinetic compartments: separating structural biomass and lipid storage. Toxicity is most likely related to the concentration in structure, which led to identification of "stochastic death" as the most appropriate death mechanism. Interestingly, the parametrized model predicts that lipid content will have only minor influence on short-term toxicity. However, the toxicants stored in lipids may have more substantial impacts in situations not included in our experiments (e.g., during diapause and gonad maturation), and for contaminant transfer to eggs and copepod predators.
Subject(s)
Copepoda , Lipid Metabolism/drug effects , Naphthalenes/toxicity , Water Pollutants, Chemical/toxicity , Animals , Biological Transport , Lipids , Metamorphosis, Biological , ToxicokineticsABSTRACT
Mechanistic models are essential tools for interpreting and predicting the consequences of a changing environment and stressors such as pollution on the life histories of marine organisms. Here, we apply the simple and generic energy-budget model DEBkiss to the life history of the marine copepod Calanus finmarchicus. Model modifications were needed to accommodate the copepod life cycle, which deviates in several respects from most other animals (e.g., a sudden stop of growth after the final moult). We identified an acceleration of growth in the early copepodite stages, which could be linked to an increase in the specific feeding rate of the animals. Lipid storage, an essential element of C. finmarchicus biology, was successfully captured with the reproduction buffer of the DEBkiss model. The resulting model was fitted to a detailed data set from the literature and was able to explain growth, development and lipid storage from egg to adult, at different temperatures and food availabilities, within a single consistent framework. The parameterised model could subsequently be used to elucidate the energetic constraints on gonad maturation and reproduction. Interestingly, the overhead costs for egg production seem to be substantially higher than the default value applied in DEB-based studies. The current model provides a solid basis for applications in stress ecology, although our model analysis also identified several knowledge gaps. Specifically, further research is needed to cover the dynamics of diapause and gonad maturation, to explain the dependence of maximum body size on food and temperature, and to verify the predicted high costs for maturity maintenance.
ABSTRACT
In this investigation, acute toxicity data were used from two previously reported studies where cold-water copepods were exposed to mechanically dispersed (MD) and chemically (CD) dispersed oil. In one of these studies, concentration-dependent mortality was observed, whereas no apparent relationship between exposure concentration and mortality was found in the other. The only marked difference between the studies is that copepods in the first experiment displayed a lower lipid sac volume (on average) than in the second one. In this study additional biometric data on lipid content were utilized and observed effects and toxicokinetics modeling applied in order to investigate whether differences in sensitivity between copepod cohorts might be explained by differences in lipid content. Results suggest that although a considerable lipid sac might retard toxicokinetics, the observed differences in lipid volume are not sufficient to explain differences in toxicity. Further, there are no apparent indications that acute toxic stress leads to lipid depletion, or that acute increased mortality rate selectively affects lipid-poor individuals. It is conceivable that other potential explanations exist, but the causal relationship between lipid content and increased mortality frequency remains elusive.