Matthias Grote

Organic chemicals jeopardize the health of freshwater ecosystems on the continental scale

Significance Protection of freshwater ecosystems from organic pollutants is important to preserve biodiversity and the goods they provide to society, such as clean drinking water and recreation. Organic chemicals have been shown to adversely impact freshwater ecosystems in local and regional studies. Nevertheless, due to paucity of studies on larger spatial scales, it remains unknown how widespread the risk from organic chemicals is. For the first time, to our knowledge, we provide strong evidence that chemicals threaten the ecological integrity and consequently the biodiversity of almost half of the water bodies on a continental scale, based on the analysis of governmental monitoring data from 4,000 European sites. Due to limitations associated with the monitoring programs, our results are likely to underestimate the actual risks. Organic chemicals can contribute to local and regional losses of freshwater biodiversity and ecosystem services. However, their overall relevance regarding larger spatial scales remains unknown. Here, we present, to our knowledge, the first risk assessment of organic chemicals on the continental scale comprising 4,000 European monitoring sites. Organic chemicals were likely to exert acute lethal and chronic long-term effects on sensitive fish, invertebrate, or algae species in 14% and 42% of the sites, respectively. Of the 223 chemicals monitored, pesticides, tributyltin, polycyclic aromatic hydrocarbons, and brominated flame retardants were the major contributors to the chemical risk. Their presence was related to agricultural and urban areas in the upstream catchment. The risk of potential acute lethal and chronic long-term effects increased with the number of ecotoxicologically relevant chemicals analyzed at each site. As most monitoring programs considered in this study only included a subset of these chemicals, our assessment likely underestimates the actual risk. Increasing chemical risk was associated with deterioration in the quality status of fish and invertebrate communities. Our results clearly indicate that chemical pollution is a large-scale environmental problem and requires far-reaching, holistic mitigation measures to preserve and restore ecosystem health.

Confirmation of cause‐effect relationships using effect‐directed analysis for complex environmental samples

The establishment of causal links between chemical contamination and observed ecotoxic effects of environmental samples is a major challenge in ecotoxicology. Effect-directed analysis is an approach combining fractionation with toxicity testing before chemical nontarget analysis to separate and subsequently identify toxic compounds in environmental samples. A final confirmation step in this procedure evaluates how much of the observed toxicity in the environmental sample can be attributed to the mixture of identified toxicants. The aim of the present study is to advance the methodology for the confirmation of mixtures in effect-directed analysis by using tools for the assessment of mixture toxicity accounting for unknown modes of action and heterogeneity of concentration-response curves. For this purpose, toxicants were identified in sediment extracts from two different sites. All identified compounds were tested both individually as pure compounds and in mixtures at ratios equal to those found in the sediment extracts. The observed extract toxicity was then compared with the expected combined effects calculated according to the models of concentration addition and independent action as well as with the observed toxicity of the synthetic mixture. Drawbacks of the established approach using toxic unit summation are demonstrated, and the Index of Confirmation Quality, an easy-to-read representation that allows a quantitative measure of confirmation over a range of different effect levels, is introduced.

Polychlorinated naphthalenes and other dioxin‐like compounds in Elbe River sediments

Contamination of Elbe River (Germany) sediments with dioxin-like toxicants was investigated following the 500-year flood (flood that statistically occurs once in 500 years) of 2002. It was hypothesized that large amounts of particulate matter from river beds and associated dioxin-like toxicants were mobilized and transported during this flood event. The investigation focused on polychlorinated naphthalenes (PCNs) that have not been determined previously in the Elbe River. The in vitro H4IIE-luc assay was used as an overall measure for toxicants capable of binding to the aryl hydrocarbon receptor (AhR). The assay was combined with congener-specific instrumental analyses and fractionation to quantify PCN contributions to total AhR-mediated activity relative to polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) and polychlorinated biphenyls (PCBs). Penta- to octachloronaphthalene concentrations of 30 ng/kg dry weight up to 13 microg/kg dry weight were found in Elbe River sediments downstream of Bitterfeld. Concentrations of penta- to octachloronaphthalenes, however, were only approximately 3 microg/kg dry weight at a site in the vicinity of Bitterfeld, where a level of approximately 3 mg/kg dry weight was reported before the flood. Also, the congener pattern of PCNs at this site changed after the flood, and PCN patterns reported previously for Bitterfeld and assigned to chlor-alkali electrolysis with graphite electrodes could now be observed at the sites from downstream of Bitterfeld and Magdeburg. Whereas PCDD/Fs dominated the dioxin-like activity in the middle and lower Elbe River, PCNs contributed as much as 10% of the total AhR-mediated activity. The contribution of PCBs was less significant (maximum, 0.2%). Thus, in Elbe River sediments, PCNs should be considered as relevant contaminants and be included in future monitoring and risk assessment programs.

Exploration of Daphnia behavioral effect profiles induced by a broad range of toxicants with different modes of action

Behavior is increasingly reported as a sensitive and early indicator of toxicant stress in aquatic organisms. However, the systematic understanding of behavioral effects and comparisons between effect profiles is hampered because the available studies are limited to few chemicals and differ in the exposure conditions and effect parameters examined. The aims of the present study were 1) to explore behavioral responses of Daphnia magna exposed to different toxicants, 2) to compare behavioral effect profiles with regard to chemical modes of action, and 3) to determine the sensitivity and response time of behavioral parameters in a new multi-cell exposure system named Multi-DaphTrack compared with currently utilized tests. Twelve compounds covering different modes of toxic action were selected to sample a wide range of potential effect profiles. Acute standard immobilization tests and 48 h of behavioral tracking were performed in the customized Multi-DaphTrack system and a single-cell commercialized biological early warning system. Contrasting behavioral profiles were observed for average speed (i.e., intensity, time of effect onset, effect duration), but no distinct behavioral profiles could be drawn from the chemical mode of action. Most compounds tested in the Multi-DaphTrack system induced an early and significant average speed increase at concentrations near or below the 10% effective concentration (48 h) of the acute immobilization test, demonstrating that the Multi-DaphTrack system is fast and sensitive. To conclude, behavior endpoints could be used as an alternative or complement to the current acute standard test or chemical analysis for the predictive evaluation of ecotoxic effects of effluents or water bodies.

Physiological sensitivity of freshwater macroinvertebrates to heavy metals

Macroinvertebrate species traits, such as physiological sensitivity, have successfully been introduced in trait-based bioassessment approaches and are important predictors of species sensitivity in the field. The authors ranked macroinvertebrate species according to their physiological sensitivity to heavy metals using toxicity data from acute laboratory assays. Rankings for each of the heavy metals, Cd, Cu, Cr, Ni, Pb, Zn, and Hg, were standardized based on all available species data. Rankings for different heavy metals on the species level showed no significant difference between compounds and were reasonably well correlated pairwise (0.50<r<0.73). Thus, an aggregated heavy metal ranking was developed, which assigns a single physiological sensitivity value (S(metal) ) to macroinvertebrate taxa. Considering the high variation, especially for higher taxonomic levels, that is, in the order level, it is recommended to use S values of the genus or species level for meaningful analyses. In terms of taxonomic ranking, crustaceans were overall the most sensitive taxonomic group, whereas insects were generally the most tolerant group. Species in the order of Cladocera were three orders of magnitude more sensitive than insects of the order of Trichoptera. By contrast, mollusks covered a wide range of sensitivities, with bivalves being on average one order of magnitude more sensitive than gastropods. The authors concluded that physiological sensitivity represents a promising trait for trait-based risk assessment that together with other demographic and recolonization traits may help to identify the effects of heavy metal pollution in aquatic ecosystems.

Inter‐laboratory trial of a standardized sediment contact test with the aquatic plant Myriophyllum aquaticum (ISO 16191)

A whole-sediment toxicity test with Myriophyllum aquaticum has been developed by the German Federal Institute of Hydrology and standardized within the International Organization for Standardization (ISO; ISO 16191). An international ring-test was performed to evaluate the precision of the test method. Four sediments (artificial, natural) were tested. Test duration was 10 d, and test endpoint was inhibition of growth rate (r) based on fresh weight data. Eighteen of 21 laboratories met the validity criterion of r ≥ 0.09 d(-1) in the control. Results from 4 tests that did not conform to test-performance criteria were excluded from statistical evaluation. The inter-laboratory variability of growth rates (20.6%-25.0%) and inhibition (26.6%-39.9%) was comparable with the variability of other standardized bioassays. The mean test-internal variability of the controls was low (7% [control], 9.7% [solvent control]), yielding a high discriminatory power of the given test design (median minimum detectable differences [MDD] 13% to 15%). To ensure these MDDs, an additional validity criterion of CV ≤ 15% of the growth rate in the controls was recommended. As a positive control, 90 mg 3,5-dichlorophenol/kg sediment dry mass was tested. The range of the expected growth inhibition was proposed to be 35 ± 15%. The ring test results demonstrated the reliability of the ISO 16191 toxicity test and its suitability as a tool to assess the toxicity of sediment and dredged material.

A New Multi-cell Exposure System for Continuous Tracking of Daphnia Behavior for Toxicity Assessments

For several years, video tracking systems have been developed to analyze alterations in the swimming behavior of daphnia to provide early signals of chemical stress. However, these systems have limited testing abilities that do not allow for a systematic analysis of the robustness of behavioral endpoints. With recent advances in behavior tracking technology, we were able to develop a new behavioral analysis multi-cell exposure system named “Multi-DaphTrack” with a high-throughput testing capacity for assessing the behavioral response of Daphnia magna. The insecticide esfenvalerate was chosen as chemical model and tested on daphnid neonates at several concentrations for 48 h to (i) evaluate the performance of this new system and (ii) compare the sensitivity of our new multi-cell system with the standard immobilization assay and the Bbe®Daphnia Toximeter. Overall, the results demonstrated that our new “Multi- DaphTrack” system can detect significant behavioral effects of esfenvalerate at concentrations as low as 0.14 μg/L from a minimum of 1 h of exposure. Similar rapid behavioral effect trends were observed with the Bbe®Daphnia Toximeter. The behavior proved to be more sensitive than the standard immobilization endpoint. Significant behavioral changes were observed at the esfenvalerate concentrations that occur in contaminated rivers from agricultural areas in Europe and North America. Our results indicate that the “Multi-DaphTrack” system represents a powerful and convenient tool for the assessment of c and water quality

Bioavailability of copper in contaminated sediments assessed by a DGT approach and the uptake of copper by the aquatic plant Myriophyllum aquaticum

The assessment of the potentially harmful effects of metals on biota depends on the speciation and bioavailability of the metals. In the present study, the authors investigated Cu accumulation and toxicity in the aquatic plant Myriophyllum aquaticum after exposure to artificial sediments varying in peat or ferric hydroxide content and spiked with Cu (5-200 mg kg(-1)). Modeling of the kinetic diffusive gradient in thin film (DGT) measurements revealed fast and slow Cu resupply from the solid phase for sediment formulated with and without peat, respectively. Myriophyllum aquaticum proved to be sensitive to Cu, as the Cu accumulation and growth differed depending on the sediment composition and Cu concentration. Comparing the Cu accumulation in M. aquaticum with total dissolved concentration, free concentration, and concentration in solution derived from DGT measurements (CDGT), Cu concentrations revealed that CDGT concentrations were a better predictor of accumulation than the others. However, the relatively weak correlation observed (r(2)  = 0.6) and the fact that plant uptake does not increase proportionally to DGT fluxes suggest that Cu uptake in plants was not diffusion limited. Thus, the free Cu concentrations near the root surface were sufficient to meet the plants demand during the experiment. Furthermore, labile complexes that continuously resupply the Cu(2+) pool may also contribute to the concentrations available for plant uptake. In the range of Cu concentrations investigated in the present study, saturation of uptake processes as well as toxicity are considered responsible for the poor DGT prediction of plant uptake.