Bioassay predictive values for chemical health risks in drinking water.

Bioanalytical tools can be used for assessment of the chemical quality of drinking water and its sources. For water managers it is important to know the probability that a bioassay response above an established health-based 'effect-based trigger value' (EBT) indeed implies a harmful chemical (mixture) concentration. This study presents and applies a framework, based on Bayes' theorem, to derive such risk probabilities for bioassay responses. These were evaluated under varying (in silico) chemical mixture concentrations relevant to drinking water (sources), with toxicity data for six in vitro assays from the ToxCast database. For single chemicals and in silico mixtures, the negative predictive value (NPV) was 100 % for all assays. For water managers, this means that when a bioassay response is below the EBT, a chemical risk is reliably absent, and no further action is required. The positive predictive value (PPV) increased with increasing chemical concentrations (2 µg/L) up to 40-80 %, depending on the assay. For in silico mixtures of increasing numbers of chemicals, the PPV did not increase until higher sum concentrations (>2-10 µg/L). Hence, the ability to accurately signal a harmful chemical (mixture) using bioassays will be lowest for highly diverse, low-concentration chemical mixtures. For water managers, this means in practice that further investigations after an EBT exceedance will, in many cases, not reveal chemicals at harmful concentrations. A solution offered is to increase the trigger value for positive responses to achieve a higher PPV and maintain the EBT for negative responses to ensure an optimal NPV.

Application of Effect-Based Methods to Water Quality Monitoring: Answering Frequently Asked Questions by Water Quality Managers, Regulators, and Policy Makers.

Effect-based methods (EBM) have great potential for water quality monitoring as they can detect the mixture effects of all active known and unknown chemicals in a sample, which cannot be addressed by chemical analysis alone. To date, EBM have primarily been applied in a research context, with a lower level of uptake by the water sector and regulators. This is partly due to concerns regarding the reliability and interpretation of EBM. Using evidence from the peer-reviewed literature, this work aims to answer frequently asked questions about EBM. The questions were identified through consultation with the water industry and regulators and cover topics related to the basis for using EBM, practical considerations regarding reliability, sampling for EBM and quality control, and what to do with the information provided by EBM. The information provided in this work aims to give confidence to regulators and the water sector to stimulate the application of EBM for water quality monitoring.

Effect-Based Trigger Values Are Essential for the Uptake of Effect-Based Methods in Water Safety Planning.

Effect-based methods (EBMs) using in vitro bioassays and well plate-based in vivo assays are recommended for water quality monitoring because they can capture the mixture effects of the many chemicals present in water. Many in vitro bioassays are highly sensitive, so an effect in a bioassay does not necessarily indicate poor chemical water quality. Consequently, effect-based trigger values (EBTs) have been introduced to differentiate between acceptable and unacceptable chemical water quality and are required for the wider acceptance of EBMs by the water sector and regulatory bodies. These EBTs have been derived for both drinking water and surface water to protect human and ecological health, respectively, and are available for assays indicative of specific receptor-mediated effects, as well as assays indicative of adaptive stress responses, apical effects, and receptor-mediated effects triggered by many chemicals. An overview of currently available EBTs is provided, and a simple approach is proposed to predict interim EBTs for assays currently without an EBT based on the effect concentration of the assay reference compound. There was good agreement between EBTs predicted using this simplistic approach and EBTs from the literature derived using more robust methods. Finally, an interpretation framework that outlines the steps to take if the effect of a sample exceeds the EBT was developed to help facilitate the uptake of EBMs in routine water quality monitoring and water safety planning for drinking water production. Environ Toxicol Chem 2023;42:714-726. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Application of AOPs to assist regulatory assessment of chemical risks - Case studies, needs and recommendations.

While human regulatory risk assessment (RA) still largely relies on animal studies, new approach methodologies (NAMs) based on in vitro, in silico or non-mammalian alternative models are increasingly used to evaluate chemical hazards. Moreover, human epidemiological studies with biomarkers of effect (BoE) also play an invaluable role in identifying health effects associated with chemical exposures. To move towards the next generation risk assessment (NGRA), it is therefore crucial to establish bridges between NAMs and standard approaches, and to establish processes for increasing mechanistically-based biological plausibility in human studies. The Adverse Outcome Pathway (AOP) framework constitutes an important tool to address these needs but, despite a significant increase in knowledge and awareness, the use of AOPs in chemical RA remains limited. The objective of this paper is to address issues related to using AOPs in a regulatory context from various perspectives as it was discussed in a workshop organized within the European Union partnerships HBM4EU and PARC in spring 2022. The paper presents examples where the AOP framework has been proven useful for the human RA process, particularly in hazard prioritization and characterization, in integrated approaches to testing and assessment (IATA), and in the identification and validation of BoE in epidemiological studies. Nevertheless, several limitations were identified that hinder the optimal usability and acceptance of AOPs by the regulatory community including the lack of quantitative information on response-response relationships and of efficient ways to map chemical data (exposure and toxicity) onto AOPs. The paper summarizes suggestions, ongoing initiatives and third-party tools that may help to overcome these obstacles and thus assure better implementation of AOPs in the NGRA.

Effect-based monitoring to integrate the mixture hazards of chemicals into water safety plans.

Water safety plans (WSPs) are intended to assure safe drinking water (DW). WSPs involve assessing and managing risks associated with microbial, chemical, physical and radiological hazards from the catchment to the consumer. Currently, chemical hazards in WSPs are assessed by targeted chemical analysis, but this approach fails to account for the mixture effects of the many chemicals potentially present in water supplies and omits the possible effects of non-targeted chemicals. Consequently, effect-based monitoring (EBM) using in vitro bioassays and well plate-based in vivo assays are proposed as a complementary tool to targeted chemical analysis to support risk analysis, risk management and water quality verification within the WSP framework. EBM is frequently applied to DW and surface water and can be utilised in all defined monitoring categories within the WSP framework (including 'system assessment', 'validation', 'operational' and 'verification'). Examples of how EBM can be applied within the different WSP modules are provided, along with guidance on where to apply EBM and how frequently. Since this is a new area, guidance documents, standard operating procedures (SOPs) and decision-making frameworks are required for both bioassay operators and WSP teams to facilitate the integration of EBM into WSPs, with these resources being developed currently.

Sensitivity of phytoplankton, zooplankton and macroinvertebrates to hydrogen peroxide treatments of cyanobacterial blooms.

Addition of hydrogen peroxide (H2O2) is a promising method to acutely suppress cyanobacterial blooms in lakes. However, a reliable H2O2 risk assessment to identify potential effects on non-target species is currently hampered by a lack of appropriate ecotoxicity data. The aim of the present study was therefore to quantify the responses of a wide diversity of freshwater phytoplankton, zooplankton and macroinvertebrates to H2O2 treatments of cyanobacterial blooms. To this end, we applied a multifaceted approach. First, we investigated the 24-h toxicity of H2O2 to three cyanobacteria (Planktothrix agardhii, Microcystis aeruginosa, Anabaena sp.) and 23 non-target species (six green algae, eight zooplankton and nine macroinvertebrate taxa), using EC50 values based on photosynthetic yield for phytoplankton and LC50 values based on mortality for the other organisms. The most sensitive species included all three cyanobacterial taxa, but also the rotifer Brachionus calyciflores and the cladocerans Ceriodaphnia dubia and Daphnia pulex. Next, the EC50 and LC50 values obtained from the laboratory toxicity tests were used to construct a species sensitivity distribution (SSD) for H2O2. Finally, the species predicted to be at risk by the SSD were compared with the responses of phytoplankton, zooplankton and macroinvertebrates to two whole-lake treatments with H2O2. The predictions of the laboratory-based SSD matched well with the responses of the different taxa to H2O2 in the lake. The first lake treatment, with a relatively low H2O2 concentration and short residence time, successfully suppressed cyanobacteria without major effects on non-target species. The second lake treatment had a higher H2O2 concentration with a longer residence time, which resulted in partial suppression of cyanobacteria, but also in a major collapse of rotifers and decreased abundance of small cladocerans. Our results thus revealed a trade-off between the successful suppression of cyanobacteria at the expense of adverse effects on part of the zooplankton community. This delicate balance strongly depends on the applied H2O2 dosage and may affect the decision whether to treat a lake or not.

Comparative field study on bioassay responses and micropollutant uptake of POCIS, Speedisk and SorbiCell polar passive samplers.

Routine water quality monitoring is generally performed with chemical analyses of grab samples, which has major limitations. First, snapshot samples will not give a good representation of the water quality. Second, it is not sufficient to analyze only a limited number of (priority) pollutants. These limitations can be circumvented by an alternative environmental risk assessment that combines time-integrated passive sampling (PS) with effect-based methods. This study aimed to select which of three polar PS devices was best suited for effect-based monitoring strategies. In the first part of this study, Speedisk, SorbiCell and POCIS polar PS devices were compared by simultaneous deployment at five sites. Chemical analyses of 108 moderately polar compounds (-1.82 < log D < 6.28) revealed that highest number of compounds, with the widest range of log KOW, log D and pKa, were detected in extracts of POCIS, followed by Speedisk. SorbiCell samplers accumulated the lowest numbers and concentrations of compounds, so they were not further investigated. In a follow-up study, bioassay responses were compared in extracts of POCIS and Speedisk devices deployed at eight sites. The passive sampler extracts were subjected to bioassays for non-specific toxicity, endocrine disruption, and antibiotics activities. More frequent and higher responses were induced by POCIS extracts, leading to more exceedances of effect-based trigger values for environmental risks. As POCIS outperformed Speedisk, it is better suited as PS device targeting polar compounds for semi-quantitative effect-based water quality monitoring.

Smarter Sediment Screening: Effect-Based Quality Assessment, Chemical Profiling, and Risk Identification.

Sediments play an essential role in the functioning of aquatic ecosystems but simultaneously retain harmful compounds. However, sediment quality assessment methods that consider the risks caused by the combined action of all sediment-associated contaminants to benthic biota are still underrepresented in water quality assessment strategies. Significant advancements have been made in the application of effect-based methods, but methodological improvements can still advance sediment risk assessment. The present study aimed to explore such improvements by integrating effect-monitoring and chemical profiling of sediment contamination. To this end, 28 day life cycle bioassays with Chironomus riparius using intact whole sediment cores from contaminated sites were performed in tandem with explorative chemical profiling of bioavailable concentrations of groups of legacy and emerging sediment contaminants to investigate ecotoxicological risks to benthic biota. All contaminated sediments caused effects on the resilient midge C. riparius, stressing that sediment contamination is ubiquitous and potentially harmful to aquatic ecosystems. However, bioassay responses were not in line with any of the calculated toxicity indices, suggesting that toxicity was caused by unmeasured compounds. Hence, this study underlines the relevance of effect-based sediment quality assessment and provides smarter ways to do so.

Persistence of chironomids in metal polluted andean high altitude streams: does melanin play a role?

In high altitude Andean streams an intense solar radiation and coinciding metal pollution allow the persistence of only a few specialized taxa, including chironomids. The aim of the present study was therefore to determine the mechanisms underlying the persistence of chironomids under these multiple stress conditions, hypothesizing that melanin counteracts both the adverse effects of solar radiation and of metals. Melanin was determined in chironomids from reference and metal polluted streams at 3000 and 4000 m altitude, being 2-fold higher at 4000 m compared to 3000 m, and 2-fold higher in polluted streams than in reference streams at both altitudes. The field observations were experimentally verified by assessing the combined effects of Cu and UV-B on the survival and melanin concentration in larvae of the model species Chironomus riparius (Chironomidae, Diptera). In laboratory exposures, the highest melanin concentrations were found in larvae surviving toxic Cu concentrations, but not in those exposed to the highest UV-B radiation. Pre-exposure to UV-B decreased the sensitivity of the larvae to UV-B and to Cu+UV-B. It is concluded that in the field, melanin may protect chironomids partially against both elevated metal concentrations and solar radiation, allowing them to persist under the harshest conditions in high altitude streams.