Mick Hamer

Framework for traits‐based assessment in ecotoxicology

A key challenge in ecotoxicology is to assess the potential risks of chemicals to the wide range of species in the environment on the basis of laboratory toxicity data derived from a limited number of species. These species are then assumed to be suitable surrogates for a wider class of related taxa. For example, Daphnia spp. are used as the indicator species for freshwater aquatic invertebrates. Extrapolation from these datasets to natural communities poses a challenge because the extent to which test species are representative of their various taxonomic groups is often largely unknown, and different taxonomic groups and chemicals are variously represented in the available datasets. Moreover, it has been recognized that physiological and ecological factors can each be powerful determinants of vulnerability to chemical stress, thus differentially influencing toxicant effects at the population and community level. Recently it was proposed that detailed study of species traits might eventually permit better understanding, and thus prediction, of the potential for adverse effects of chemicals to a wider range of organisms than those amenable for study in the laboratory. This line of inquiry stems in part from the ecology literature, in which species traits are being used for improved understanding of how communities are constructed, as well as how communities might respond to, and recover from, disturbance (see other articles in this issue). In the present work, we develop a framework for the application of traits-based assessment. The framework is based on the population vulnerability conceptual model of Van Straalen in which vulnerability is determined by traits that can be grouped into 3 major categories, i.e., external exposure, intrinsic sensitivity, and population sustainability. Within each of these major categories, we evaluate specific traits as well as how they could contribute to the assessment of the potential effects of a toxicant on an organism. We then develop an example considering bioavailability to explore how traits could be used mechanistically to estimate vulnerability. A preliminary inventory of traits for use in ecotoxicology is included; this also identifies the availability of data to quantify those traits, in addition to an indication of the strength of linkage between the trait and the affected process. Finally, we propose a way forward for the further development of traits-based approaches in ecotoxicology.

Genetically modified crops and aquatic ecosystems: considerations for environmental risk assessment and non-target organism testing

Environmental risk assessments (ERA) support regulatory decisions for the commercial cultivation of genetically modified (GM) crops. The ERA for terrestrial agroecosystems is well-developed, whereas guidance for ERA of GM crops in aquatic ecosystems is not as well-defined. The purpose of this document is to demonstrate how comprehensive problem formulation can be used to develop a conceptual model and to identify potential exposure pathways, using Bacillus thuringiensis (Bt) maize as a case study. Within problem formulation, the insecticidal trait, the crop, the receiving environment, and protection goals were characterized, and a conceptual model was developed to identify routes through which aquatic organisms may be exposed to insecticidal proteins in maize tissue. Following a tiered approach for exposure assessment, worst-case exposures were estimated using standardized models, and factors mitigating exposure were described. Based on exposure estimates, shredders were identified as the functional group most likely to be exposed to insecticidal proteins. However, even using worst-case assumptions, the exposure of shredders to Bt maize was low and studies supporting the current risk assessments were deemed adequate. Determining if early tier toxicity studies are necessary to inform the risk assessment for a specific GM crop should be done on a case by case basis, and should be guided by thorough problem formulation and exposure assessment. The processes used to develop the Bt maize case study are intended to serve as a model for performing risk assessments on future traits and crops.

A decision tree for assessing effects from exposures to multiple substances

BackgroundThe Cefic Mixtures Industry Ad-hoc Team (MIAT) has investigated how risks from combined exposures can be effectively identified and managed using concepts proposed in recent regulatory guidance, new advances in risk assessment, and lessons learned from a Cefic-sponsored case study of mixture exposures.ResultsA series of tools were created that include: a decision tree, a system for grouping exposures, and a graphical tool (the MCR-HI plot). The decision tree allows the division of combined exposures into different groups, exposures where one or more individual components are a concern, exposures that are of low concern, and exposures that are a concern for combined effects but not for the effects of individual chemicals. These tools efficiently use available data, identify critical data gaps for combined assessments, and prioritize which chemicals require detailed toxicity information. The tools can be used to address multiple human health endpoints and ecological effects.ConclusionThe tools provide a useful approach for assessing risks associated with combined exposures to multiple chemicals.

A microcosm system to evaluate the toxicity of the triazine herbicide simazine on aquatic macrophytes.

This study evaluates the effects of the triazine herbicide simazine in an outdoor pond microcosm test system that contained two submerged rooted species (Myriophyllum spicatum and Elodea canadensis) and two emergent rooted species (Persicaria amphibia and Glyceria maxima) over a period of 84 days. Simazine was applied to the microcosms at nominal concentrations of 0.05, 0.5 and 5 mg/L. General biological endpoints and physiological endpoints were used to evaluate herbicide toxicity on macrophytes and the algae developing naturally in the system. Concentration-related responses of macrophytes and algae were obtained for the endpoints selected, resulting in a no observed ecologically adverse effect concentration (NOEAEC) at simazine concentrations of 0.05 mg active ingredient/L after 84 days. E. canadensis was the most negatively affected species based on length increase, which was consistently a very sensitive parameter for all macrophytes. The experimental design presented might constitute a suitable alternative to conventional laboratory single-species testing.

Future needs and recommendations in the development of species sensitivity distributions: Estimating toxicity thresholds for aquatic ecological communities and assessing impacts of chemical exposures.

A species sensitivity distribution (SSD) is a probability model of the variation of species sensitivities to a stressor, in particular chemical exposure. The SSD approach has been used as a decision support tool in environmental protection and management since the 1980s, and the ecotoxicological, statistical, and regulatory basis and applications continue to evolve. This article summarizes the findings of a 2014 workshop held by the European Centre for Toxicology and Ecotoxicology of Chemicals and the UK Environment Agency in Amsterdam, The Netherlands, on the ecological relevance, statistical basis, and regulatory applications of SSDs. An array of research recommendations categorized under the topical areas of use of SSDs, ecological considerations, guideline considerations, method development and validation, toxicity data, mechanistic understanding, and uncertainty were identified and prioritized. A rationale for the most critical research needs identified in the workshop is provided. The workshop reviewed the technical basis and historical development and application of SSDs, described approaches to estimating generic and scenario-specific SSD-based thresholds, evaluated utility and application of SSDs as diagnostic tools, and presented new statistical approaches to formulate SSDs. Collectively, these address many of the research needs to expand and improve their application. The highest priority work, from a pragmatic regulatory point of view, is to develop a guidance of best practices that could act as a basis for global harmonization and discussions regarding the SSD methodology and tools. Integr Environ Assess Manag 2017;13:664-674.

Response of the mayfly (Cloeon dipterum) to chronic exposure to thiamethoxam in outdoor mesocosms

Thiamethoxam is a widely used neonicotinoid insecticide that has been detected in surface water monitoring programs in North America and Europe. This has led to questions about its toxicity to nontarget insects, specifically those with an aquatic life stage. To address the uncertainty associated with possible impacts from environmental exposures, a chronic (35-d) outdoor mesocosm study with a formulated product containing thiamethoxam was conducted. The specific focus of the study was the response of mayflies (Ephemeroptera), which have been reported to be particularly sensitive in laboratory studies. A range of concentrations (nominally 0.1, 0.3, 1.0, 3.0, and 10.0 µg/L thiamethoxam), plus untreated controls were tested, and the abundance and emergence of mayflies (Cloeon dipterum) were assessed weekly for 35 d. Mean measured time-weighted average exposures were within 6% of nominal over the duration of the study, with the mean half-life of thiamethoxam in each treatment ranging from 7 to 13 d. Statistically significant reductions in both larval abundance and adult emergence were observed at 10.0, 3.0, and 1.0 μg/L following 1, 2, and 3 wk of exposure, respectively. Exposure to 0.1 and 0.3 µg/L thiamethoxam had no statistically significant effect on larval mayfly abundance or adult emergence at any point in the study. These findings support a 35-d no-observed-effect concentration (NOEC) of 0.3 µg thiamethoxam/L for mayflies (C. dipterum) under chronic conditions. Furthermore, because the 95th percentile of environmental concentrations has been reported to be 0.054 µg/L, these results indicate that populations of C. dipterum and similarly sensitive aquatic insects are unlikely to be significantly impacted by thiamethoxam exposure in natural systems represented by the conditions in our study. Environ Toxicol Chem 2018;37:1040-1050.

Pairing behavior and reproduction in Hyalella azteca as sensitive endpoints for detecting long-term consequences of pesticide pulses

The aim of the present study was to examine acute and delayed effects of pulse exposure of the pyrethroid pesticide, permethrin, on precopulatory pairs of Hyalella azteca. Pairs of H. azteca were exposed to a single 1h pulse of different nominal concentrations of permethrin: 0, 0.3, 0.9 or 2.7 μg/L. During exposure, pairing behavior was observed, and during a 56 day post-exposure period the treatments were monitored for pairing behavior, survival and reproductive output. All permethrin-exposed pairs separated within minutes during exposure and shortly thereafter became immobile; however they regained mobility after transfer to clean water. The time to re-form pairs was significantly longer in all tested concentrations compared to the control, although all surviving pairs re-formed within the 56 day test period. Nevertheless not all pairs exposed to 0.9 and 2.7 μg/L reproduced. Furthermore the numbers of juveniles produced by pairs exposed to 0.9 and 2.7 μg/L, but not 0.3 μg/L, were lower throughout the entire post-exposure period compared to the control groups, and the total numbers of juveniles produced during 56 days were significantly lower in organisms exposed to 0.9 and 2.7 μg/L, but not 0.3 μg/L, compared to the control groups. The long-term effects of short-term exposure on reproductive behavior of pairs could potentially have consequences for the population dynamics of H. azteca. However, since individual-level responses can both overestimate and underestimate effects at the population level, appropriate population models are needed to reduce the uncertainty in extrapolating between these levels of biological organization.

Prospective aquatic risk assessment for chemical mixtures in agricultural landscapes

Abstract Environmental risk assessment of chemical mixtures is challenging because of the multitude of possible combinations that may occur. Aquatic risk from chemical mixtures in an agricultural landscape was evaluated prospectively in 2 exposure scenario case studies: at field scale for a program of 13 plant‐protection products applied annually for 20 yr and at a watershed scale for a mixed land‐use scenario over 30 yr with 12 plant‐protection products and 2 veterinary pharmaceuticals used for beef cattle. Risk quotients were calculated from regulatory exposure models with typical real‐world use patterns and regulatory acceptable concentrations for individual chemicals. The results could differentiate situations when there was concern associated with single chemicals from those when concern was associated with a mixture (based on concentration addition) with no single chemical triggering concern. Potential mixture risk was identified on 0.02 to 7.07% of the total days modeled, depending on the scenario, the taxa, and whether considering acute or chronic risk. Taxa at risk were influenced by receiving water body characteristics along with chemical use profiles and associated properties. The present study demonstrates that a scenario‐based approach can be used to determine whether mixtures of chemicals pose risks over and above any identified using existing approaches for single chemicals, how often and to what magnitude, and ultimately which mixtures (and dominant chemicals) cause greatest concern. Environ Toxicol Chem 2018;37:674–689.

Is an ecosystem services-based approach developed for setting specific protection goals for plant protection products applicable to other chemicals?

Clearly defined protection goals specifying what to protect, where and when, are required for designing scientifically sound risk assessments and effective risk management of chemicals. Environmental protection goals specified in EU legislation are defined in general terms, resulting in uncertainty in how to achieve them. In 2010, the European Food Safety Authority (EFSA) published a framework to identify more specific protection goals based on ecosystem services potentially affected by plant protection products. But how applicable is this framework to chemicals with different emission scenarios and receptor ecosystems? Four case studies used to address this question were: (i) oil refinery waste water exposure in estuarine environments; (ii) oil dispersant exposure in aquatic environments; (iii) down the drain chemicals exposure in a wide range of ecosystems (terrestrial and aquatic); (iv) persistent organic pollutant exposure in remote (pristine) Arctic environments. A four-step process was followed to identify ecosystems and services potentially impacted by chemical emissions and to define specific protection goals. Case studies demonstrated that, in principle, the ecosystem services concept and the EFSA framework can be applied to derive specific protection goals for a broad range of chemical exposure scenarios. By identifying key habitats and ecosystem services of concern, the approach offers the potential for greater spatial and temporal resolution, together with increased environmental relevance, in chemical risk assessments. With modifications including improved clarity on terminology/definitions and further development/refinement of the key concepts, we believe the principles of the EFSA framework could provide a methodical approach to the identification and prioritization of ecosystems, ecosystem services and the service providing units that are most at risk from chemical exposure.

High concentrations of protein test substances may have non-toxic effects on Daphnia magna: implications for regulatory study designs and ecological risk assessments for GM crops.

Laboratory testing for possible adverse effects of insecticidal proteins on non-target organisms (NTOs) is an important part of many ecological risk assessments for regulatory decision-making about the cultivation of insect-resistant genetically modified (IRGM) crops. To increase confidence in the risk assessments, regulatory guidelines for effects testing specify that representative surrogate species for NTOs are exposed to concentrations of insecticidal proteins that are in excess of worst-case predicted exposures in the field. High concentrations in effects tests are achieved by using protein test substances produced in microbes, such as Escherichia coli. In a study that exposed Daphnia magna to a single high concentration of a microbial test substance containing Vip3Aa20, the insecticidal protein in MIR162 maize, small reductions in growth were observed. These effects were surprising as many other studies strongly suggest that the activity of Vip3Aa20 is limited to Lepidoptera. A plausible explanation for the effect on growth is that high concentrations of test substance have a non-toxic effect on Daphnia, perhaps by reducing its feeding rate. A follow-up study tested that hypothesis by exposing D. magna to several concentrations of Vip3Aa20, and a high concentration of a non-toxic protein, bovine serum albumin (BSA). Vip3Aa20 and BSA had sporadic effects on the reproduction and growth of D. magna. The pattern of the effects suggests that they result from non-toxic effects of high concentrations of protein, and not from toxicity. The implications of these results for regulatory NTO effects testing and ERA of IRGM crops are discussed.