Female proportion has a stronger influence on dispersal than body size in nematodes of mountain lakes.

Nematodes disperse passively and are amongst the smallest invertebrates on Earth. Free-living nematodes in mountain lakes are highly tolerant of environmental variations and are thus excellent model organisms in dispersal studies, since species-environment relationships are unlikely to interfere. In this study, we investigated how population or organism traits influence the stochastic physical nature of passive dispersal in a topologically complex environment. Specifically, we analyzed the influence of female proportion and body size on the geographical distribution of nematode species in the mountain lakes of the Pyrenees. We hypothesized that dispersal is facilitated by (i) a smaller body size, which would increase the rate of wind transport, and (ii) a higher female proportion within a population, which could increase colonization success because many nematode species are capable of parthenogenetic reproduction. The results showed that nematode species with a low proportion of females tend to have clustered spatial distributions that are not associated with patchy environmental conditions, suggesting greater barriers to dispersal. When all species were pooled, the overall proportion of females tended to increase at the highest elevations, where dispersal between lakes is arguably more difficult. The influence of body size was barely relevant for nematode distributions. Our study highlights the relevance of female proportion as a mechanism that enhances the dispersal success of parthenogenetic species, and that female sex is a determining factor in metacommunity connectivity.

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.

Polystyrene microbeads influence lipid storage distribution in C. elegans as revealed by coherent anti-Stokes Raman scattering (CARS) microscopy.

The exposure of Caenorhabditis elegans to polystyrene (PS) beads of a wide range of sizes impedes feeding, by reducing food consumption, and has been linked to inhibitory effects on the reproductive capacity of this nematode, as determined in standardized toxicity tests. Lipid storage provides energy for longevity, growth, and reproduction and may influence the organismal response to stress, including the food deprivation resulting from microplastics exposure. However, the effects of microplastics on energy storage have not been investigated in detail. In this study, C. elegans was exposed to ingestible sizes of PS beads in a standardized toxicity test (96 h) and in a multigeneration test (∼21 days), after which lipid storage was quantitatively analyzed in individual adults using coherent anti-Stokes Raman scattering (CARS) microscopy. The results showed that lipid storage distribution in C. elegans was altered when worms were exposed to microplastics in form of PS beads. For example, when exposed to 0.1-µm PS beads, the lipid droplet count was 93% higher, the droplets were up to 56% larger, and the area of the nematode body covered by lipids was up to 79% higher than in unexposed nematodes. The measured values tended to increase as PS bead sizes decreased. Cultivating the nematodes for 96 h under restricted food conditions in the absence of beads reproduced the altered lipid storage and suggested that it was triggered by food deprivation, including that induced by the dilutional effects of PS bead exposure. Our study demonstrates the utility of CARS microscopy to comprehensively image the smaller microplastics (<10 µm) ingested by nematodes and possibly other biota in investigations of the effects at the level of the individual organism.

Inventory of the benthic eukaryotic diversity in the oldest European lake.

We have profound knowledge on biodiversity on Earth including plants and animals. In the recent decade, we have also increased our understanding on microorganisms in different hosts and the environment. However, biodiversity is not equally well studied among different biodiversity groups and Earth's systems with eukaryotes in freshwater sediments being among the least known. In this study, we used high-throughput sequencing of the 18S rRNA gene to investigate the entire diversity of benthic eukaryotes in three distinct habitats (littoral sediment and hard substrate, profundal sediment) of Lake Ohrid, the oldest European lake. Eukaryotic sequences were dominated by annelid and arthropod animals (54% of all eukaryotic reads) and protists (Ochrophyta and Ciliophora; together 40% of all reads). Eukaryotic diversity was 15% higher in the deep profundal than on either near-surface hard substrates or littoral sediments. The three habitats differed in their taxonomic and functional community composition. Specifically, heterotrophic organisms accounted for 92% of the reads in the profundal, whereas phototrophs accounted for 43% on the littoral hard substrate. The profundal community was the most homogeneous, and its network was the most complex, suggesting its highest stability among the sampled habitats.

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.

Rapid ingestion and egestion of spherical microplastics by bacteria-feeding nematodes.

Microplastics, anthropogenically released into freshwaters, settle in sediments, where they are directly ingested by benthic organisms. However, to the best of our knowledge, fine-scale studies of microplastic ingestion and egestion by nematodes, one of the most abundant meiofaunal taxa, are lacking. We therefore conducted a time series of the ingestion and egestion by adult Caenorhabditis elegans and Pristionchus pacificus of 0.5- and 1.0-µm fluorescent polystyrene (PS) beads along with bacteria. The nematodes were exposed to 107 beads ml-1 in aqueous medium for 5 min-24 h and pumping rates of C. elegans were determined. In the egestion study, PS bead egestion was monitored in nematodes with high microplastic body burdens for 5 min-24 h in microplastic-free medium. Ingested beads were detected already within 5 min and up to 203 ± 15 PS beads (1.0 µm; C. elegans) were found after 30 min. Overall, significantly more 1.0-µm than 0.5-µm PS beads were taken up. The distinct feeding behaviors of the two species influenced their PS bead body burdens. Ingested PS beads were almost completely egested within the first 20-40 min in the presence of sufficient food. In C. elegans, 1.0-µm beads were egested less rapidly than 0.5-µm PS beads. Given the rapid ingestion and egestion of the beads, our study demonstrates that the actual amount of ingested and egested microplastics by nematodes in the environment may be several times higher than the microplastic body burdens may imply. However, spherical PS beads did not bioconcentrate in nematodes.

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.

Ingestion of microplastics by meiobenthic communities in small-scale microcosm experiments.

Microplastics have been detected in many different environments. Nematodes are a rife meiofaunal taxon and occupy an important trophic position in benthic food webs. Laboratory-based ingestion experiments have demonstrated the susceptibility of single nematode species to microplastic uptake. However, the determinants of ingestion by meiofaunal assemblages, especially those of nematodes, have yet to be fully examined. We therefore conducted a microcosm study in which field-collected freshwater sediment was spiked with fluorescent polystyrene (PS) beads (1.0, 3.0 and 6.0 µm) in concentrations of 103 and 107 PS beads ml-1 and the ingestion by the most dominant indigenous meiofaunal taxa (nematodes, rotifers, chironomids, copepods) was investigated after 2, 4 and 8 days using fluorescence microscopy. In additional small-scale microcosms, PS bead ingestion by nematode assemblages was quantified as a function of feeding type, exposure time (1-10 days), concentration (103, 105, 107 PS beads ml-1) and bead size (0.5, 1.0, 3.0, 6.0 µm). PS beads at 107 beads ml-1 were largely ingested by chironomids and copepods. Exposure time and concentration correlated positively with PS bead ingestion for all taxa. The most relevant size class for ingestion for the majority of meiofaunal taxa was PS beads of 1.0 µm. Nematode communities, especially deposit-feeding species, effectively ingested micropastics from sediment, as >30% of the exposed individuals and 56% of the species ingested 1.0-µm PS beads in <24 h. Ingestion rates were mainly influenced by PS bead size and nematode feeding type/habit, with the exception of a bead concentration of 103 beads ml-1, at which exposure time was also an important factor. Sediment particles reduced microplastic ingestion considerably for all investigated meiobenthic organisms. Our study demonstrates the ability of free-living nematodes communities to readily ingest PS beads of various sizes. If the feeding-type distribution is known, the potential exposure of nematode communities may be predicted.

Preface: Patterns and processes of meiofauna in freshwater ecosystems.

Never heard of harpacticoids, ostracods, gastrotrichs or microturbellarians? This is no surprise, they are so tiny! Yet these taxa and many others more famous (nematodes, rotifers, or tardigrades) show complex behaviours and extraordinary physiologies that allow them to colonize inland waters worldwide. This exuberant fauna is better known as the meiofauna (or meiobenthos). Meiofaunal organisms have been fascinating study objects for zoologists since the seventeenth century and recent research has demonstrated their intermediate role in benthic food webs. This special issue highlights how meiofauna can help freshwater ecologists to describe and predict species distribution patterns, to assess production of biomass and trait functions relationships, as well as to examine the trophic links between microscopic and macroscopic worlds and to better understand species' resilience to environmental extremes. Overall, meiofaunal organisms are bridging scales, and as such they deserve better integration to develop more comprehensive concepts and theories in ecology.

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.

Selective grazing behaviour of chironomids on microalgae under pesticide pressure.

The herbicide diuron and the insecticide imidacloprid are amongst the most frequently detected pesticides in French rivers, and each is known to affect many aquatic organisms. However, the question of whether and how both pesticides together might induce multi-stress conditions, which could induce indirect effects such as the modification of biological interactions within freshwater microbial communities has not received much attention. This study was undertaken to determine the effect of diuron and imidacloprid alone and in combination on the feeding behaviour of chironomid larvae. An initial experiment measured the impact of the different contamination conditions at environmental concentrations (5 µg L-1 for each pesticide) on the grazing rate of chironomids on three microalgae species, independently. Two diatom species, Gomphonema gracile (two different morphotypes: normal and teratogen) and Planothidium lanceolatum, and one green alga Desmodesmus sp. were offered as food, during 24 h. Chironomids grazing rates varied according to the pesticide and algae species. Indeed, diuron impacted algae more strongly and probably affected their palatability, leading chironomids to increase grazing pressure on less nutritionally interesting algae. Imidacloprid, by targeting insect larvae, increased or inhibited their grazing capacity depending on the food source. In a second experiment (cafeteria design), the food selectivity of chironomids on previous algae was determined under similar contamination conditions during 4 h: under diuron, larvae switched equally between the microalgae and were as mobile as in the control without pesticide. However, imidacloprid and the pesticide mixture condition altered chironomid movements and grazing behaviour. By investigating the impact of an herbicide and an insecticide, alone and in combination, on the responses of food (algae growth rate) and biological (mortality) and behavioural (mobility, food selection) responses of chironomid larvae, this study provided new insights on the direct and indirect effects of pesticide contamination on a simplified trophic web.

A global database of soil nematode abundance and functional group composition.

As the most abundant animals on earth, nematodes are a dominant component of the soil community. They play critical roles in regulating biogeochemical cycles and vegetation dynamics within and across landscapes and are an indicator of soil biological activity. Here, we present a comprehensive global dataset of soil nematode abundance and functional group composition. This dataset includes 6,825 georeferenced soil samples from all continents and biomes. For geospatial mapping purposes these samples are aggregated into 1,933 unique 1-km pixels, each of which is linked to 73 global environmental covariate data layers. Altogether, this dataset can help to gain insight into the spatial distribution patterns of soil nematode abundance and community composition, and the environmental drivers shaping these patterns.

Comparison of morphological, DNA barcoding, and metabarcoding characterizations of freshwater nematode communities.

Biomonitoring approaches and investigations of many ecological questions require assessments of the biodiversity of a given habitat. Small organisms, ranging from protozoans to metazoans, are of great ecological importance and comprise a major share of the planet's biodiversity but they are extremely difficult to identify, due to their minute body sizes and indistinct structures. Thus, most biodiversity studies that include small organisms draw on several methods for species delimitation, ranging from traditional microscopy to molecular techniques. In this study, we compared the efficiency of these methods by analyzing a community of nematodes. Specifically, we evaluated the performances of traditional morphological identification, single-specimen barcoding (Sanger sequencing), and metabarcoding in the identification of 1500 nematodes from sediment samples. The molecular approaches were based on the analysis of the 28S ribosomal large and 18S small subunits (LSU and SSU). The morphological analysis resulted in the determination of 22 nematode species. Barcoding identified a comparable number of operational taxonomic units (OTUs) based on 28S rDNA (n = 20) and fewer OTUs based on 18S rDNA (n = 12). Metabarcoding identified a higher OTU number but fewer amplicon sequence variants (AVSs) (n = 48 OTUs, n = 17 ASVs for 28S rDNA, and n = 31 OTUs, n = 6 ASVs for 18S rDNA). Between the three approaches (morphology, barcoding, and metabarcoding), only three species (13.6%) were shared. This lack of taxonomic resolution hinders reliable community identifications to the species level. Further database curation will ensure the effective use of molecular species identification.

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.

Environmental factors and river network position allow prediction of benthic community assemblies: A model of nematode metacommunities.

The field of metacommunity studies is growing rapidly, including recent applications to river networks. Most of these studies have targeted a single river network but whether their findings are relevant to other river systems is unknown. This study investigated the influence of environmental, spatial and temporal parameters on the community structure of nematodes in the river networks of the Elbe and Rhine. We asked whether the variance in community structure was better explained by spatial variables representing the watercourse than by overland distances. After determining the patterns in the Elbe river network, we tested whether they also explained the Rhine data. The Elbe data were evaluated using a boosted regression tree analysis. The predictive ability of the model was then assessed using the Rhine data. In addition to strong temporal dynamics, environmental factors were more important than spatial factors in structuring riverine nematode communities. Community structure was more strongly influenced by watercourse than by Euclidean distances. Application of the model's predictions to the Rhine data correlated significantly with field observations. Our model shows that the consequences of changes in environmental factors or habitat connectivity for aquatic communities across different river networks are quantifiable.