Determining the toxicity of organic compounds to the nematode Caenorhabditis elegans based on aqueous concentrations.

Caenorhabditis elegans is used for assessing the toxicity of chemicals in aqueous medium. However, chemicals can absorb to the bacterial food, which reduces the freely dissolved concentrations of the tested compounds. Thus, based on total or nominal concentrations, toxicity is underestimated, resulting in misleading assumptions on toxicity mechanisms or comparisons to other test organisms. As the verification of freely dissolved exposure concentrations (Cfree) is challenging in small test systems, simple partitioning models might by a good option for estimating Cfree. Therefore, C. elegans was exposed to seven differently acting organic chemicals with varying hydrophobicities, thus also different affinities to bind to the food of C. elegans. Measured concentrations of the dissolved aqueous and the bacterial-bound fraction allowed the calculation of binding constants (Kb). Experimental Kb were comparable to literature data of hydrophobic chemicals and correlated well with their hydrophobicity, expressed as log KOW. The chronic toxicity of the various compounds on C. elegans' reproduction, based on their aqueous concentration, was weakly related to their log KOW. Toxicity expressed based on chemical activity and comparisons with a baseline toxicity model, nevertheless, suggested a narcotic mode of action for most hydrophobic compounds (except methylisothiazolinone and trichlorocarbanilide). Although revealing a similar toxicity ranking than Daphnia magna, C. elegans was less sensitive, probably due to its ability to reduce its internal concentrations by means of its very impermeable cuticle or by efficient detoxification mechanisms. It could be shown that measured aqueous concentrations in the nematode test system corresponded well with freely dissolved concentrations that were modeled using simple mass-balance models from nominal concentrations. This offers the possibility to estimate freely dissolved concentrations of chemicals from nominal concentrations, making routine testing of chemicals and their comparison to other species more accurate.

Nematode Community of a Natural Grassland Responds Sensitively to the Broad-Spectrum Fungicide Mancozeb in Soil Microcosms.

Fungicides make up the largest part of total pesticide use, with the dithiocarbamate mancozeb being widely applied as a nonsystemic contact pesticide to protect a wide range of field crops against fungal diseases. Although nematodes are key drivers of soil functioning, data on effects of fungicides, and especially mancozeb, on these nontarget organisms are scarce. Therefore, the effects of mancozeb on a soil nematode community from a natural grassland were assessed in small-scale soil microcosms. Nematodes were exposed to mancozeb-spiked soil in six nominal concentrations (7-133 mg/kg dry soil) and analyzed after 14, 56, and 84 days in terms of densities, genus composition, and functional traits. Because this fungicide is known to quickly degrade in soils (50% degradation time <1 day), mancozeb concentrations were analyzed for all sampling occasions. Chemical analysis revealed considerably lower measured concentrations compared with the aimed nominal soil concentrations at the beginning of the exposure (1-18 mg/kg dry soil), suggesting fast degradation during the spiking process. Nevertheless, the native nematode community responded sensitively to the fungicide mancozeb, revealing lower no-observed-effect concentration and 10% effect concentration (EC10) values than reported for other soil invertebrates such as springtails and earthworms. Using the EC10 for the most sensitive nematode community endpoint (percentage of predators and omnivores: 1.2 mg/kg dry soil), the risk assessment exhibited a toxicity exposure ratio of 0.66 and, thus, a high risk of mancozeb for soil nematodes. Keeping in mind their abundance and their central roles in soil food-web functioning, the demonstrated sensitivity to a widely applied fungicide underscores the relevance of the inclusion of nematodes into routine risk-assessment programs for pesticides. Environ Toxicol Chem 2022;41:2420-2430. © 2022 SETAC.

Long-term exposure of a free-living freshwater micro- and meiobenthos community to microplastic mixtures in microcosms.

Microplastics in a wide range of shapes and polymer types (MPs; <5 mm) accumulate in freshwater sediments, where they may pose an environmental threat to sediment-dwelling micro- and meiobenthos. To date, the effects of MPs on those organisms have mostly been studied in single-species experiments exposed to high particle concentrations. By contrast, there have been few investigations of the effects resulting from the long-term exposure of natural communities to environmental relevant MPs. This research gap was addressed in the present study. A microcosm experiment was conducted to examine the impact of a mixture of MPs of varying polymer composition, shape, and size (50% polystyrene (PS) beads: 1-µm diameter; 37% polyethylene terephthalate (PET) fragments: 32 × 21 µm in size, and 13% polyamide (PA) fibers 104 × 15 µm in size; % based on the total particle number) provided at two concentrations (low: 4.11 × 105 MPs/kg sediment dw and high: 4.11 × 107 MPs/kg sediment dw) and two exposure durations (4 and 12 weeks) on a micro- and meiobenthic community collected from a freshwater sediment. MPs exposure did not alter the abundance of protozoa (ciliates and flagellates) as well as the abundance and biomass of meiobenthic organisms (nematodes, rotifers, oligochaetes, gastrotrichs, nauplii), whereas the abundance and biomass of harpacticoid copepods was affected. Neither nematode species diversity (species richness, Shannon-Wiener index, and evenness) nor the NemaSPEAR[%]-index (pollution-sensitive index based on freshwater nematodes) changed in response to the MPs. However, changes in the structure of the meiobenthic and nematode community in the presence of environmentally relevant MPs mixtures cannot be excluded, such that microcosms experiments may be of value in detecting subtle, indirect effects of MPs.

Food availability is crucial for effects of 1-µm polystyrene beads on the nematode Caenorhabditis elegans in freshwater sediments.

Freshwater sediments represent a sink for microplastic (<5 mm) through various processes. Thus, benthic organisms can be exposed to relatively high concentrations of microplastics. Surprisingly, studies on benthic organisms are still underrepresented in the field of ecotoxicological effect assessment of microplastics. Therefore, we studied the effects of 1-µm polystyrene (PS) beads on the reproduction of the nematode Caenorhabditis elegans using a standardized protocol for toxicity testing in freshwater sediments (96 h; ISO 10872:2020), combined with ingestion experiments using fluorescent PS beads. To investigate the role of sediment properties (e.g., textures, organic contents) for ingestion and effects of PS beads, five different artificial and field-collected sediments were used. Body burdens of 1-µm PS beads in the intestinal tract of the nematodes after 96 h differed between the sediments, however, differences were not significant over the whole course of the experiment. EC10 and EC50-values of 1-µm PS beads for C. elegans' reproduction in the various sediments ranged from 0.9 to 2.0 and 4.8 to 11.3 mg PS/g dry sediment, respectively. The ECx-values showed to be considerably higher than values reported for water exposure (EC10/50: 0.2 and 0.6 mg PS/ml, respectively), which was probably due to higher food densities in sediment compared to water exposure. Based on the PS beads/bacteria ratio, ECx-values were comparable between sediment and water exposure, suggesting that also in sediments microplastic reduces the food availability for C. elegans causing lower reproduction. This indirect effect mechanism was confirmed by experiments with varying food densities. Thus, the nutritious conditions might play a crucial role for the overall ecological risk of microplastics in benthic ecosystems.

The Uptake of Sporopollenin Exine Capsules and Associated Bioavailability of Adsorbed Oestradiol in Selected Aquatic Invertebrates.

Lycopodium clavatum sporopollenin exine capsules (SpECs) are known to both adsorb and absorb chemicals. The aim of the present work was to determine whether oestradiol (E2) is 'bioavailable' to bioindicator species, either pre-adsorbed to, or in the presence of, SpECs. SpEC uptake was confirmed for Daphnia magna and Dreissena bugensis. E2 levels varied among treatments for Caenorhabditis elegans though there was no relationship to SpEC load. E2 was not detected in D. bugensis tissues. Expression changes of general stress and E2-specific genes were measured. For C. elegans, NHR-14 expression suggested that SpECs modulate E2 impacts, but not general health responses. For D. magna, SpECs alone and with E2 changed Vtg1 and general stress responses. For D. bugensis, SpECS were taken up but no E2 or change in gene expression was detected after exposure to E2 and/or SpECs. The present study is the first to investigate SpECs and bound chemical dynamics.

On the balance between practical relevance and standardization - Testing the effects of zinc and pyrene on native nematode communities in soil microcosms.

Soils are among the most densely inhabited and biodiverse habitats on our planet, and many important soil ecosystem services depend on the health condition of the native soil fauna. Anthropogenic stress such as chemical pollution acting on the native soil fauna might jeopardize these functions. Laboratory microcosm tests are an appropriate tool for assessing the risk of chemicals on the native soil fauna and can be regarded as intermediate tier tests, bridging the gap between single species toxicity tests and field testing. Nematodes are one of the most abundant and divers soil invertebrates, and as such native nematode communities might be suitable for ecotoxicological assessments in laboratory microcosm set ups. In order to test such a small-scale (30 g soil) microcosm system, two different chemicals (zinc and pyrene) were assessed in various soil types for their effects on the respective native nematode communities. Various community parameters such as total nematode density, genus richness and genus composition, as well as trait-related indices (e.g. maturity index) were monitored over a period of 8-10 weeks. The response of the nematode communities strongly varied between soil types, and these differences were more pronounced for Zn than for pyrene. Interestingly, the structure of the respective native nematode communities was shown to play a larger role for explaining the varying toxic effects than soil properties governing the bioavailability of the spiked chemicals. We demonstrated that exposure of natural nematode communities in their original soil matrix to the metal zinc and to pyrene under climatically highly controlled conditions resulted in quantitatively and qualitatively distinct responses. Upon comparison of various community indices, the maturity index was shown to be the most sensitive toxicity endpoint for all tested soils and chemicals.

Food bacteria and synthetic microparticles of similar size influence pharyngeal pumping of Caenorhabditis elegans.

Toxicity tests using the model organism Caenorhabditis elegans have shown that exposure to small microplastics such as polystyrene (PS) beads lead to high body burdens and dietary restrictions that in turn inhibit reproduction. Pharyngeal pumping is the key mechanism of C. elegans for governing the uptake of food and other particles and can be easily monitored by determining the pumping rates. In this study, pharyngeal pumping of C. elegans was examined in response to increasing quantities of food bacteria (E. coli: 106-1010 cells ml-1) and synthetic particles (107-109 beads ml-1) of similar size (1 µm). While the average pumping rate of C. elegans exposed to E. coli depended on the density of the bacterial cells, this was not the case for the synthetic beads. At 107 items ml-1, bacterial cells and synthetic beads triggered a basic stimulation of the pumping rate, independent of the nutritional value of the particle. At quantities >107 items ml-1, however, the nutritional value was essential to maximize the pumping rate, as it was upregulated only by E. coli cells, which can be chemosensorially recognized by C. elegans. Given the unselective uptake of all particles in the size range of bacteria, restricting the pumping rates for particles with low nutritional value to a basic rate, prevents the nematodes from wasting energy by high-frequency pumping, but still allows a food-quality screening at low food levels.

Bacterial consumption by nematodes is disturbed by the presence of polystyrene beads: The roles of food dilution and pharyngeal pumping.

Microplastics (MPs; <5 mm) released into freshwaters from anthropogenic sources accumulate in sediments, where they may pose an environmental threat to benthic organisms, such as nematodes. Several studies have examined the effects of nano- and microplastics on the nematode Caenorhabditis elegans, whereas reduced food availability was suggested as a possible explanation for the observed inhibitory effects. Therefore, this study should clarify whether micro-beads of different sizes (1.0 and 6.0 µm in diameter) and materials (polystyrene PS, silica) are able to interfere with the feeding of C. elegans on its bacterial diet (Escherichia coli), and, by this, lowering its consumption rate within 7 h of exposure. Moreover, it was examined whether an inhibited bacterial consumption was caused by a reduction of the nematode's pumping rate, as a primary indicator of food ingestion. Bacterial consumption by C. elegans was significantly decreased in the presence of 1.0- and 6.0-µm PS beads (49-67% lower bacterial consumption compared to control), whereas in the presence of 1.0-µm silica beads feeding was not impeded. Interestingly, the pumping rate was significantly lower in the presence of non-ingestible 6.0-µm PS beads with 161 ± 16 pumps min-1, while it was largely unchanged for nematodes exposed to ingestible 1.0-µm PS beads with 205 ± 12 pumps min-1, compared to control conditions with 210 ± 18 pumps min-1, respectively. As reduced bacterial consumption leads to generally lower energy reserves in C. elegans, these results allow to link observed inhibitory effects of MPs on the nematodes to a lower food availability. Such indirect, food-web related, effects of MPs should raise concern of ecological consequences in natural habitats, where temporal food deficiencies can occur. Consequently, disturbances in food availability and feeding efficiency should be regarded as important parameters in environmental risk assessments focusing on MPs.

Dataset supporting the use of nematodes as bioindicators of polluted sediments.

We provide the dataset supporting the research article "Nematodes as bioindicators of polluted sediments using metabarcoding and microscopic taxonomy" [1]. Nematodes are frequently used as bioindicators and the NemaSPEAR[%] is an validated index that is originally based on morphological data. The index was compared to molecular sequence data for the 28S rDNA, 18S rDNA and COI gene for 7 locations. This dataset includes chemical analyses of the sediments for 33 different substances. The sequence data for OTU-based analyses for the 28S rDNA, 18S rDNA and COI gene is given, together with the read distribution during bioinformatics processing. We furthermore include alternative ASV data, based on a cluster-independent approach. The morphological data is presented, including the biomass for each species, as well as an overview about whether the species is represented in the NCBI database. Furthermore, rarefaction analysis is given for the morphological data, and furthermore NMDS plots for the species and genus level based on morphological and molecular data. The correlation between the mean PEC-Q and the NemaSPEAR[%] values is given in order to compare the efficiency of the index, based on morphological and molecular data.

Nematodes as bioindicators of polluted sediments using metabarcoding and microscopic taxonomy.

The use of bioindicator species is a widely applied approach to evaluate ecological conditions, and several indices have been designed for this purpose. To assess the impact of pollution, especially in sediments, a pollution-sensitive index based on nematodes, one of the most abundant and species-rich groups of metazoa, was developed. The NemaSPEAR[%] index in its original form relies on the morphological inspection of nematode species. The application of a morphologically based NemaSPEAR[%] at the genus-level was previously validated. The present study evaluated a NemaSPEAR[%] index based on metabarcoding of nematode communities and tested the potential of fragments from the 28S rDNA, 18S rDNA and cytochrome c oxidase subunit I (COI) genes. In general, molecular-based results tended to show a poorer condition than morphology-based results for the investigated sites. At the genus level, NemaSPEAR[%] values based on morphological data strongly correlated with those based on molecular data for both the 28S rDNA and the 18S rDNA gene fragments (R2 = 0.86 and R2 = 0.74, respectively). Within the dominant genera (>3%) identified by morphology, 68% were detected by at least one of the two ribosomal markers. At the species level, however, concordance was less pronounced, as there were several deviations of the molecular from the morphological data. These differences could mostly be attributed to shortcomings in the reference database used in the molecular-based assignments. Our pilot study shows that a molecularly based, genus-level NemaSPEAR[%] can be successfully applied to evaluate polluted sediment. Future studies need to validate this approach further, e.g. with bulk extractions of whole meiofaunal communities in order to circumvent time-consuming nematode isolation. Further database curation with abundant NemaSPEAR[%] species will also increase the applicability of this approach.

Limited effects of pesticides on stream macroinvertebrates, biofilm nematodes, and algae in intensive agricultural landscapes in Sweden.

Pesticides are frequently detected in surface waters, sometimes at levels exceeding ecotoxicological guidelines. We screened for almost 100 pesticides in 32 streams from intense agricultural areas in Southern Sweden, in concert with water chemistry parameters. In addition, we investigated the communities of benthic macroinvertebrates, biofilm nematodes and algae and calculated multiple bioassessment metrics. The number of pesticides found in each stream ranged between 2 and 52, but the sum of Toxic Units (ΣTU) for the mixtures was generally low, and exceeded the European Uniform Principles only in a single sample for algae and in 2% of the samples for Daphnia. Only nematode communities were significantly correlated with the ΣTU, potentially due to their higher pesticide exposure in biofilms. Diatom metrics showed that most streams were impacted by eutrophication and macroinvertebrate metrics showed good status in most streams, whereas the SPEARpesticides (SPEcies At Risk) index, specifically designed to indicate pesticide effects, showed that about half of the samples were at risk. Interestingly, SPEARpesticides was not correlated to ΣTUDaphnia, and this discrepancy suggests that redefining the boundaries for quality classes might be necessary for this index. Moreover, SPEARpesticides was positively correlated with the commonly used macroinvertebrate index ASPT, although disparate results were found for several streams. We argue that this questions the scaling of both metrics and the specificity of their responses. We discuss that the overall good/moderate status of the streams, despite the intense agriculture in the catchments, can be due to the fact that i) a sampling strategy with repeated grab samples did not capture peak pesticide concentrations, thus underestimating acute exposure, ii) pesticide run-off indeed was low, due to measures such as buffer strips, and iii) the nutrient-rich conditions and high sediment loads counteracted pesticide toxicity. We conclude that agricultural land use was the overriding stressor in the investigated streams, including strong effects of nutrients, less apparent effects of pesticides and likely impact of hydromorphological alterations (not specifically addressed in this study).

Surface-Related Toxicity of Polystyrene Beads to Nematodes and the Role of Food Availability.

Microplastics released into freshwaters from anthropogenic sources settle in the sediments, where they may pose an environmental threat to benthic organisms. However, few studies have considered the ecotoxicological hazard of microplastic particles for nematodes, one of the most abundant taxa of the benthic meiofauna. This study investigated the toxic effects of polystyrene (PS) beads (0.1-10.0 µm) and the underlying mechanisms thereof on the reproduction of the nematode Caenorhabditis elegans. The observed effect of the PS beads on the nematodes correlated well with the total surface area of the beads per volume, with a 50% inhibition of reproduction at 55.4 ± 12.9 cm2/mL, independent of the bead size. The adverse effects were not explained by styrene monomers leaching from the beads because chemical activities of styrene in PS suspensions were well below the toxic levels. However, the observed effects could be related to the bead material because the same-sized silica (SiO2) beads had considerably less impact, probably due to their higher specific density. PS and SiO2 beads affected the food availability of C. elegans, with greater effects by the PS beads. Our results demonstrate the importance of including indirect food web effects in studies of the ecological risks posed by microplastics.

Ingestion of microplastics by nematodes depends on feeding strategy and buccal cavity size.

Microplastics are hardly biodegradable and thus accumulate rather than decompose in the environment. Due to sedimentation processes, meiobenthic fauna is exposed to microplastics. Within the meiofauna, nematodes are a very abundant taxon and occupy an important position in benthic food webs by connecting lower and higher trophic levels. However, the key determinants of the uptake of microplastics by freshwater nematodes are still unknown. To investigate the bioaccessibility of microplastics for nematodes, we performed single- and multi-species ingestion experiments in which the ability of seven nematode species (six bacterial and one fungal feeder), diverse in their buccal cavity morphology (1.3-10.5 µm), to ingest fluorescence-labelled polystyrene (PS) beads along with their natural diet was examined. Applied beads sizes (0.5, 1.0, 3.0 and 6.0 µm), exposure time (4, 24 and 72 h) and concentration (3 × 106 PS beads ml-1 and 107 PS beads ml-1) were varied. Ingested beads were localized and quantified via fluorescence microscopy in the nematodes. In contrast to fungal-feeding nematode species with a stylet, bacterial-feeding species ingested 0.5- and 1.0-µm PS beads with up to 249 and 255 beads after 24 h, respectively. Microplastics ≥0.5 µm could only be ingested and transported into the gastrointestinal tract, if the buccal cavities were considerably (>1.3 times) larger than the beads. At concentrations of 107 PS beads ml-1 ingestion rates were influenced by exposure time and PS bead concentration. In case of a known microplastic size distribution in the environment, predictions on the potential ingestion for nematode communities can be made based on the feeding type composition and the size of their buccal cavities.

Comparing the effects of fludioxonil on non-target soil invertebrates using ecotoxicological methods from single-species bioassays to model ecosystems.

The lower tier toxicity tests used for risk assessment of plant protection products are conducted with single species, only regarding direct effects of the tested substances. However, it is not clear, if lower tier tests are able to protect in situ soil communities, as these tests are not able to account for direct and indirect effects of chemicals on multi-species systems in natural soil communities. This knowledge gap between single-species tests and field studies can be bridged using model ecosystems (microcosms), which allow for the assessment of direct and indirect effects of the compounds under evaluation. In the present study, single-species toxicity tests and soil-spiked microcosms were used to comparatively investigate the toxicity of the non-systemic fungicide fludioxonil (FDO) on non-target soil organisms, with nematodes being the test organisms of choice. The potential effects of FDO on nematodes were investigated in two different test systems: (i) standardized toxicity tests using Caenorhabditis elegans exposed to FDO-spiked soil (FDO concentrations 50-1207 mg/kg soil dry weight) and (ii) in situ nematode communities sampled from microcosms containing FDO-spiked soil (FDO concentrations 75-600 mg/kg soil dry weight). FDO dose-dependently inhibited the reproduction of C. elegans, with an effect concentration (EC50) of 209.9 mg FDO/kg soil dry weight and a no observed effect concentration (NOEC) of 63.0 mg FDO/kg soil dry weight. In the microcosms, FDO significantly affected trait-based indices, such as the Maturity Index (MI25) and the Enrichment Index (EI), which responded already at FDO concentrations of 14.3 and 62.4 mg/kg dry soil. Overall, this study provides new insights into the impact of the non-systemic fungicide FDO on non-target soil organisms and demonstrates the suitability of nematode-based tools, that allow for a quick and cost-effective lower and higher tier risk assessment of plant protection products.

Effects of waste materials on Caenorhabditis elegans (Nematoda) using the ISO standard soil toxicity test.

The ecotoxicological characterization of waste according to the European Waste List (HP14) is part of its hazard classification, which is based on 15 different hazardous properties and should include toxicity tests representing the aquatic and terrestrial compartment. Besides established soil toxicity tests with bacteria, plants, and earthworms, the standardized test with the nematode Caenorhabditis elegans (ISO 10872:2010) is suitable for testing soils and wastes, however, has never been validated for the purpose of waste toxicity assessment. Therefore, 23 different waste samples were tested for their toxicity on growth and reproduction of C. elegans to validate the suitability of ISO 10872 (ISO 2010) for assessing the ecotoxicological hazard of waste and to compare the results with those of other ecotoxicological test systems. C. elegans showed a comparable response to the waste samples as the earthworm avoidance test, however, with single samples where the nematodes indicated a higher toxicity than the E. fetida, thus, providing non-redundant information to the ecotoxicological hazard assessment of wastes. Also due to the short duration (4 days) and small-scale test set-up, the soil toxicity test with C. elegans turned out to be a valuable addition to already existing test batteries, for assessing the ecotoxicity of wastes in the soil compartment.

Pro-Angiogenic Effects of Latent Heparanase and Thrombin Receptor-Mediated Pathways-Do They Share a Common Ground in Melanoma Cells?

Heparanase (HPSE) is an endo-ß-D-glucuronidase that cleaves heparan sulphate (HS) chains of proteoglycans (HSPGs). Besides a remodelling of the extracellular matrix, HPSE increases the bioavailability of pro-angiogenic mediators, such as HS-associated vascular endothelial growth factor (VEGF), thereby contributing to metastatic niche formation. Notably, HPSE also induces release of VEGF from tumour cells independent of its enzymatic activity, but the underlying molecular mechanisms remain unresolved. We found that exogenous addition of latent HPSE stimulates VEGF release from human MV3 melanoma cells. The same effect was noted upon direct stimulation of thrombin receptor (protease-activated receptor 1 [PAR-1]) by Thrombin Receptor Activator Peptide 6 (TRAP-6). The matricellular ligand cysteine-rich 61 protein (Cyr61) was identified as pathway component since Cyr61 knockdown in MV3 cells abolished the VEGF release by TRAP-6 and HPSE. Since both TRAP-6 and HPSE mediated an up-regulation of phosphorylated focal adhesion kinase, which could be blocked by antagonizing PAR-1, we postulated a crosstalk between latent HPSE and PAR-1 in promoting pro-angiogenic pathways. To test this hypothesis at a molecular level, we applied dynamic mass redistribution (DMR) technique measuring intracellular mass relocation as consequence of direct receptor activation. Indeed, latent HPSE evoked a concentration-dependent DMR signal in MV3 cells as TRAP-6 did. Both could be modulated by targeting G-protein receptor signalling in general or by the PAR-1 inhibitor RWJ 56110. Using cells devoid of cell surface HS synthesis, we could confirm HPSE effects on PAR-1, independent of HSPG involvement. These data indicate, for the first time, a crosstalk between latent HPSE, thrombin receptor activation and G-protein signalling in general.

Linking ecological health to co-occurring organic and inorganic chemical stressors in a groundwater-fed stream system.

Freshwaters are among the most endangered ecosystems worldwide, due predominantly to excessive anthropogenic practices compromising the future provisioning of ecosystem services. Despite increased awareness of the role of multiple stressors in accounting for ecological degradation in mixed land-use stream systems, risk assessment approaches applicable in field settings are still required. This study provides a first indication for ecological consequences of the interaction of organic and inorganic chemical stressors, not typically evaluated together, which may provide a missing link enabling the reconnection of chemical and ecological findings. Specifically, impaired ecological conditions - represented by lower abundance of meiobenthic individuals - were observed in the hyporheic zone where a contaminant groundwater plume discharged to the stream. These zones were characterized by high xenobiotic organic concentrations, and strongly reduced groundwater (e.g. elevated dissolved iron and arsenic) linked to the dissolution of iron hydroxides (iron reduction) caused by the degradation of xenobiotic compounds in the plume. Further research is still needed to separate whether impact is driven by a combined effect of organic and inorganic stressors impacting the ecological communities, or whether the conditions - when present simultaneously - are responsible for enabling a specific chemical stressor's availability (e.g. trace metals), and thus toxicity, along the study stream. Regardless, these findings suggest that benthic meioinvertebrates are promising indicators for supporting biological assessments of stream systems to sufficiently represent impacts resulting from the co-occurrence of stressors in different stream compartments. Importantly, identification of the governing circumstances is crucial for revealing key patterns and impact drivers that may be needed in correctly prioritizing stressor impacts in these systems. This study further highlights the importance of stream-aquifer interfaces for investigating chemical stressor effects in multiple stressor systems. This will require holistic approaches for linking contaminant hydrogeology and eco(toxico)logy in order to positively influence the sustainable management of water resources globally.

Glyphosate, a chelating agent-relevant for ecological risk assessment?

Glyphosate-based herbicides (GBHs), consisting of glyphosate and formulants, are the most frequently applied herbicides worldwide. The declared active ingredient glyphosate does not only inhibit the EPSPS but is also a chelating agent that binds macro- and micronutrients, essential for many plant processes and pathogen resistance. GBH treatment may thus impede uptake and availability of macro- and micronutrients in plants. The present study investigated whether this characteristic of glyphosate could contribute to adverse effects of GBH application in the environment and to human health. According to the results, it has not been fully elucidated whether the chelating activity of glyphosate contributes to the toxic effects on plants and potentially on plant-microorganism interactions, e.g., nitrogen fixation of leguminous plants. It is also still open whether the chelating property of glyphosate is involved in the toxic effects on organisms other than plants, described in many papers. By changing the availability of essential as well as toxic metals that are bound to soil particles, the herbicide might also impact soil life, although the occurrence of natural chelators with considerably higher chelating potentials makes an additional impact of glyphosate for most metals less likely. Further research should elucidate the role of glyphosate (and GBH) as a chelator, in particular, as this is a non-specific property potentially affecting many organisms and processes. In the process of reevaluation of glyphosate its chelating activity has hardly been discussed.

Response of nematode communities to metals and PAHs in freshwater microcosms.

Freshwater microcosms were used to investigate the effects of various metals and polycyclic aromatic hydrocarbons (PAHs), applied as single substances and in mixtures, on sediment-inhabiting nematode assemblages. Several community measures, including nematode abundance, biomass, species richness, species composition and the NemaSPEAR[%]-index, were assessed and their changes in response to the chemical treatments were compared with the results of single-species toxicity testing using the nematode Caenorhabditis elegans. The diagnostic power of endpoints revealing toxic effects was then evaluated to assess the general suitability of nematodes as bioindicators of sediment contamination by a set of relevant chemicals. Overall, community measures based on species-level (detected with principle response curves) responded most sensitively to chemical stress in the microcosms, especially in terms of biomass, with the C. elegans toxicity test was for the most part as sensitive to the chemicals as the nematode species composition. Generally, this study justified the suitability of nematodes in assessments of the risk of chemicals frequently associated with freshwater sediments and underlined the value of C. elegans in the prospective risk assessment of chemicals, as this species was able to indicate the negative effects of comparatively low sediment concentrations of the contaminants.