Monitoring estrogenic activities of waste and surface waters using a novel in vivo zebrafish embryonic (EASZY) assay: Comparison with in vitro cell-based assays and determination of effect-based trigger values

Brion, François; Gussem, Valentin De; Buchinger, Sebastian; Hollert, Henner; Carere, Mario; Porcher, Jean‐Marc; Piccini, Benjamin; Féray, Christine; Dulio, Valeria; Könemann, Sarah; Simon, Eszter; Werner, Inge; Kaše, Robert; Aı̈t-Aı̈ssa, Sélim

doi:10.1016/j.envint.2019.06.006

Cited by 54 publications

(39 citation statements)

References 47 publications

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“…It is noticeable that in vivo estrogenic activity at the three NS sites was correctly predicted by the in vitro bioassays in the present study. Since the EBT value of 0.56 ng/L was built based on a comparison of MELN and EASZY data (Brion et al 2019), the present dataset confirms the suitability of this EBT value in another environmental context. Furthermore, the use of the in vivo EASZY assay enabled us to confirm the estrogenic activity observed in vitro for NS2…”

Section: Identification Of Hot Spots and Comparison With Effect-based Trigger Valuessupporting

confidence: 70%

“…The estrogenic activity of Novi Sad samples was assessed in vivo using the EASZY assay based on transgenic cyp19a1b-GFP zebrafish line (Brion et al 2012). The assay procedure for sample testing has been described in Sonavane et al (2016) and Brion et al (2019). In brief, 15 newly fertilized transgenic eggs (< 4h post fertilization) were selected for each experimental group and exposed for h in 15 mL of acclimated water in glass crystallizers.…”

Section: Zebrafish Embryo-based Bioassay (Easzy Assay)mentioning

confidence: 99%

“…For instance, Kase et al (2018) and Escher et al (2018) (Escher et al 2018) or assuming all possible mixture effects based on empirical response of fish embryos to environmental samples (Brion et al 2019). With the exception of the EBT proposed by Kunz et al (2017) which is a direct translation of the EQS of E2 into EEQ (0.4 ng/L EEQ), all other approaches consider the bioassay-specific sensitivity (REP) to estrogenic chemicals in deriving the EBT.…”

Section: Identification Of Hot Spots and Comparison With Effect-based Trigger Valuesmentioning

confidence: 99%

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Estrogenic activity of surface waters using zebrafish- and human-based in vitro assays: The Danube as a case-study

Serra

Brion

Chardon

et al. 2020

Environmental Toxicology and Pharmacology

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Most in vitro reporter gene assays used to assess estrogenic contamination are based on human estrogen receptor α (hERα) activation. However, fish bioassays can have distinct response to estrogenic chemicals and mixtures, questioning the relevance of human-based bioassays for assessing risk to this species. In this study, zebrafish liver cells stably expressing zebrafish ERβ2 (ZELHβ2) and human breast cancer cells expressing hERα (MELN) were used to quantify the estrogenic activity of 25 surface water samples of the Danube River, for which chemicals have been previously quantified. Most samples had a low estrogenic activity below 0.1 ng/L 17β-estradiol-equivalents that was more often detected by MELN cells, while ZELHβ2 response tend to be lower than predicted based on the chemicals identified.Nevertheless, both bioassays quantified well a higher estrogenic activity at two sites, which was confirmed in vivo using a transgenic zebrafish assay. The results are discussed considering the effect-based trigger values proposed for water quality monitoring.

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Section: Identification Of Hot Spots and Comparison With Effect-based Trigger Valuessupporting

confidence: 70%

Section: Zebrafish Embryo-based Bioassay (Easzy Assay)mentioning

confidence: 99%

Section: Identification Of Hot Spots and Comparison With Effect-based Trigger Valuesmentioning

confidence: 99%

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Estrogenic activity of surface waters using zebrafish- and human-based in vitro assays: The Danube as a case-study

Serra

Brion

Chardon

et al. 2020

Environmental Toxicology and Pharmacology

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“…This approach is scientifically sound, but also renders the interpretation of the results more difficult, especially if, as in the current study, two bioassays with the same endpoint are resulting in a differing number of exceedances. Taking into account the relevance of in vitro responses in the ERα-CALUX for in vivo effects in zebrafish embryos, Brion, De Gussem [74] recently suggested an EBT value of 0.28 ng/L.…”

Section: Discussionmentioning

confidence: 99%

Effects of treated wastewater on the ecotoxicity of small streams – Unravelling the contribution of chemicals causing effects

et al. 2019

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Wastewater treatment plant effluents are important point sources of micropollutants. To assess how the discharge of treated wastewater affects the ecotoxicity of small to medium-sized streams we collected water samples up- and downstream of 24 wastewater treatment plants across the Swiss Plateau and the Jura regions of Switzerland. We investigated estrogenicity, inhibition of algal photosynthetic activity (photosystem II, PSII) and growth, and acetylcholinesterase (AChE) inhibition. At four sites, we measured feeding activity of amphipods (Gammarus fossarum) in situ as well as water flea (Ceriodaphnia dubia) reproduction in water samples. Ecotoxicological endpoints were compared with results from analyses of general water quality parameters as well as a target screening of a wide range of organic micropollutants with a focus on pesticides and pharmaceuticals using liquid chromatography high-resolution tandem mass spectrometry. Measured ecotoxicological effects in stream water varied substantially among sites: 17β-estradiol equivalent concentrations (EEQbio, indicating the degree of estrogenicity) were relatively low and ranged from 0.04 to 0.85 ng/L, never exceeding a proposed effect-based trigger (EBT) value of 0.88 ng/L. Diuron equivalent (DEQbio) concentrations (indicating the degree of photosystem II inhibition in algae) ranged from 2.4 to 1576 ng/L and exceeded the EBT value (70 ng/L) in one third of the rivers studied, sometimes even upstream of the WWTP. Parathion equivalent (PtEQbio) concentrations (indicating the degree of AChE inhibition) reached relatively high values (37 to 1278 ng/L) mostly exceeding the corresponding EBT (196 ng/L PtEQbio). Decreased feeding activity by amphipods or decreased water flea reproduction downstream compared to the upstream site was observed at one of four investigated sites only. Results of the combined algae assay (PSII inhibition) correlated best with results of chemical analysis for PSII inhibiting herbicides. Estrogenicity was partly and AChE inhibition strongly underestimated based on measured steroidal estrogens respectively organophosphate and carbamate insecticides. An impact of dissolved organic carbon on results of the AChE inhibition assay was obvious. For this assay more work is required to further explore the missing correlation of bioassay data with chemical analytical data. Overall, the discharge of WWTP effluent led to increased estrogenicity, PSII and AChE inhibition downstream, irrespective of upstream land use.

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“…An additional approach specific for drinking water has been proposed that converts the concentrations of reference compounds considered safe in vivo to concentrations that can be detected using in vitro assays using differences in the toxicokinetics of different compounds to correct the EBT (Brand et al 2013). Brion et al (2019) derived EBTs for estrogenicity by comparing the effects detected in vitro to effects detected in vivo. In addition, other studies have derived EBTs for surface water using a combination of approaches, including converting from in vivo toxicity data and field investigations, often following multiple lines of evidence in a more qualitative manner (van der Oost et al 2017; de Baat et al 2020), or using distributions of a large set of water samples (Besselink et al 2017).…”

Section: Introductionmentioning

confidence: 99%

Effect-Based Trigger Values for Mixtures of Chemicals in Surface Water Detected with In Vitro Bioassays

Escher

Neale

2020

Environmental Toxicology and Chemistry

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Effect-based trigger (EBT) values for in vitro bioassays are important for surface water quality monitoring because they define the threshold between acceptable and poor water quality. EBTs have been derived for highly specific This article is protected by copyright. All rights reserved. Accepted Article bioassays, such as hormone-receptor activation in reporter gene bioassays, by reading across from existing chemical guideline values. This read-across method is not easily applicable to bioassays indicative of adaptive stress responses, which are triggered by many different chemicals, and activation of nuclear receptors for xenobiotic metabolism, to which many chemicals bind with rather low specificity. We propose an alternative approach to define the EBT from the distribution of specificity ratios of all active chemicals. Specificity ratios are the ratio between the predicted baseline toxicity of a chemical in a given bioassay and its measured specific endpoint. Unlike many previous read-across methods to derive EBTs, the proposed method accounts for mixture effects and includes all chemicals, not only high-potency chemicals. The EBTs were derived from a cytotoxicity EBT that was defined as equivalent to 1% of cytotoxicity in a native surface water sample. The cytotoxicity EBT was scaled by the median of the log-normal distribution of specificity ratios to derive the EBT for effects specific for each bioassay. We illustrate the new approach using the example of the AREc32 assay indicative of the oxidative stress response and two nuclear receptor assays targeting the peroxisome proliferator activated receptor PPAR and the arylhydrocarbon receptor AhR. The EBTs were less conservative than previously proposed but were able to differentiate untreated and insufficiently treated wastewater from wastewater treatment plant effluent with secondary or tertiary treatment and surface water.

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Monitoring estrogenic activities of waste and surface waters using a novel in vivo zebrafish embryonic (EASZY) assay: Comparison with in vitro cell-based assays and determination of effect-based trigger values

Cited by 54 publications

References 47 publications

Estrogenic activity of surface waters using zebrafish- and human-based in vitro assays: The Danube as a case-study

Estrogenic activity of surface waters using zebrafish- and human-based in vitro assays: The Danube as a case-study

Effects of treated wastewater on the ecotoxicity of small streams – Unravelling the contribution of chemicals causing effects

Effect-Based Trigger Values for Mixtures of Chemicals in Surface Water Detected with In Vitro Bioassays

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