The Water Framework Directive (WFD), 2000/60/EC, requires an integrated approach to the monitoring and assessment of the quality of surface water bodies. The chemical status assessment is based on compliance with legally binding Environmental Quality Standards (EQSs) for selected chemical pollutants (priority substances) of EU-wide concern. In the context of the mandate for the period 2010 to 2012 of the subgroup Chemical Monitoring and Emerging Pollutants (CMEP) under the Common Implementation Strategy (CIS) for the WFD, a specific task was established for the elaboration of a technical report on aquatic effect-based monitoring tools. The activity was chaired by Sweden and co-chaired by Italy and progressively involved several Member States and stakeholders in an EU-wide drafting group. The main aim of this technical report was to identify potential effect-based tools (e.g. biomarkers and bioassays) that could be used in the context of the different monitoring programmes (surveillance, operational and investigative) linking chemical and ecological status assessment. The present paper summarizes the major technical contents and findings of the report.
The European Decision EU 2015/495 included three steroidal estrogens, estrone, 17-estradiol and 17ethinyl estradiol, in the "watch-list" of the Water Framework Directive (WFD). As consequence, these substances have to be chemically monitored at the level of their environmental quality standards, which can be challenging. This project aimed to identify reliable effect-based methods (EBMs) for screening of endocrine disrupting compounds, to harmonise monitoring and data interpretation methods, and to contribute to the current WFD review process. Water and wastewater samples were collected across Europe and analysed using chemical analyses and EBMs. The results showed that 17-estradiol equivalents were comparable among methods, while results can vary between methods based on the relative potencies for individual substances. Further, derived 17-estradiol equivalents were highly correlated with LC-MS/MS analyses. This study shows that the inclusion of effect-based screening methods into monitoring programmes for estrogens in surface waterbodies would be a valuable complement to chemical analysis.
An international round-robin study on the Ames fluctuation test [ISO 11350, 2012], a microplate version of the classic plate-incorporation method for the detection of mutagenicity in water, wastewater and chemicals was performed by 18 laboratories from seven countries. Such a round-robin study is a precondition for both the finalization of the ISO standardization process and a possible regulatory implementation in water legislation. The laboratories tested four water samples (spiked/nonspiked) and two chemical mixtures with and without supplementation of a S9-mix. Validity criteria (acceptable spontaneous and positive control-induced mutation counts) were fulfilled by 92-100%, depending on the test conditions. A two-step method for statistical evaluation of the test results is proposed and assessed in terms of specificity and sensitivity. The data were first subjected to powerful analysis of variance (ANOVA) after an arcsine-square-root transformation to detect significant differences between the test samples and the negative control (NC). A threshold (TH) value based on a pooled NC was then calculated to exclude false positive test results. Statistically, positive effects observed by the William's test were considered negative, if the mean of all replicates of a sample did not exceed the calculated TH. By making use of this approach, the overall test sensitivity was 100%, and the test specificity ranged from 80 to 100%.
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