Ecotoxicological effects of chemical contaminants adsorbed to microplastics in the clam <i>Scrobicularia plana</i>

Although microplastics (MPs) are distributed globally in the marine environment, a great deal of unknowns relating to their ecotoxicological effects on the marine biota remains. Due to their lipophilic nature, microplastics have the potential to adsorb persistent organic pollutants present in contaminated regions, which may increase their detrimental impact once assimilated by organisms. This study investigates the ecotoxicological effects of exposure to low-density polyethylene (LDPE) microplastics (11–13 μm), with and without adsorbed contaminants (benzo[a]pyrene—BaP and perfluorooctane sulfonic acid—PFOS), in the peppery furrow shell clam, Scrobicularia plana. Environmentally relevant concentrations of contaminants (BaP−16.87 ± 0.22 μg g−1 and PFOS−70.22 ± 12.41 μg g−1) were adsorbed to microplastics to evaluate the potential role of plastic particles as a source of chemical contamination once ingested. S. plana were exposed to microplastics, at a concentration of 1 mg L−1, in a water-sediment exposure setup for 14 days. Clams were sampled at the beginning of the experiment (day 0) and after 3, 7, and 14 days. BaP accumulation, in whole clam tissues, was analyzed. A multi-biomarker assessment was conducted in the gills, digestive gland, and haemolymph of clams to clarify the effects of exposure. This included the quantification of antioxidant (superoxide dismutase, catalase, glutathione peroxidase) and biotransformation (glutathione-S-transferases) enzyme activities, oxidative damage (lipid peroxidation levels), genotoxicity (single and double strand DNA breaks), and neurotoxicity (acetylcholinesterase activity). Results suggest a potential mechanical injury of gills caused by ingestion of microplastics that may also affect the analyzed biomarkers. The digestive gland seems less affected by mechanical damage caused by virgin microplastic exposure, with the MPs-adsorbed BaP and PFOS exerting a negative influence over the assessed biomarkers in this tissue

The NORMAN interlaboratory study on biotesting of spiked water extracts

The NORMAN network is a permanent self-sustaining network for the monitoring and biomonitoring of emerging environmental contaminants. The NORMAN working group on Bioassays (Bio WG) focuses on the application of bioanalytical tools for environmental quality monitoring. A main objective is to provide recommendations for the implementation of effect-based tools into regulatory frameworks. In this context, a blind interlaboratory study (ILS) was performed. The aim was to verify if a bioassay battery conducted in different laboratories following own protocols would produce comparable results when testing spiked surface water extracts. The lead in planning and organizing was taken by the Department of Ecosystem Analysis (ESA), RWTH Aachen University (DE). The ILS bioassay battery included acute-toxicity assays with organisms from different trophic levels (Algae, Daphnia, Zebrafish embryos); and mechanism-specific bioassays for estrogenicity (YES, ER-Luc cell lines) and mutagenicity (Ames fluctuation) assessment. Three to four participants performed each bioassay, including: BfG (DE), Waternet (NL), Waterproef (NL), INERIS (FR), IFREMER (FR), RECETOX (CZ), ISSeP (BE), ITM (SE), IVM-VU (NL), Entox/University of Queensland (AU), Ecotox Centre (CH), ESA-RWTH (DE). Clean water from a reference site was concentrated 10.000 times with large volume solid-phase extraction. Four emerging pollutants were used for spiking: triclosan, acridine, 3-nitrobezanthrone and 17-alpha-ethinylestradiol. Extracts were spiked with either single chemicals or a mixture, in concentrations aimed to produce full dose-response curves in bioassays. The spiked extracts were prepared, separated in aliquots, identified with codes, and sent to the participating laboratories. Standardized bioassay methods (OECD, ISO) were recommended but not mandatory, and biotesters could use their own protocols. Results were sent to RWTH, and a summary of the full ILS was provided to the ILS participants. In October 2014, a workshop was held at RWTH Aachen to present and discuss the ILS results. Bioassays produced mostly highly comparable results, even when protocols differed significantly. Suggestions for future improvements include harmonization of methods for data analysis and results evaluation. An important expected outcome of the ILS is the promotion of biotesting for water quality monitoring at the level of European policy-makers

An ecotoxicological view on neurotoxicity assessment

The numbers of potential neurotoxicants in the environment are raising and pose a great risk for humans and the environment. Currently neurotoxicity assessment is mostly performed to predict and prevent harm to human populations. Despite all the efforts invested in the last years in developing novel in vitro or in silico test systems, in vivo tests with rodents are still the only accepted test for neurotoxicity risk assessment in Europe. Despite an increasing number of reports of species showing altered behaviour, neurotoxicity assessment for species in the environment is not required and therefore mostly not performed. Considering the increasing numbers of environmental contaminants with potential neurotoxic potential, eco-neurotoxicity should be also considered in risk assessment. In order to do so novel test systems are needed that can cope with species differences within ecosystems. In the field, online-biomonitoring systems using behavioural information could be used to detect neurotoxic effects and effect-directed analyses could be applied to identify the neurotoxicants causing the effect. Additionally, toxic pressure calculations in combination with mixture modelling could use environmental chemical monitoring data to predict adverse effects and prioritize pollutants for laboratory testing. Cheminformatics based on computational toxicological data from in vitro and in vivo studies could help to identify potential neurotoxicants. An array of in vitro assays covering different modes of action could be applied to screen compounds for neurotoxicity. The selection of in vitro assays could be guided by AOPs relevant for eco-neurotoxicity. In order to be able to perform risk assessment for eco-neurotoxicity, methods need to focus on the most sensitive species in an ecosystem. A test battery using species from different trophic levels might be the best approach. To implement eco-neurotoxicity assessment into European risk assessment, cheminformatics and in vitro screening tests could be used as first approach to identify eco-neurotoxic pollutants. In a second step, a small species test battery could be applied to assess the risks of ecosystems