Soil nematode abundance and functional group composition at a global scale.

Soil organisms are a crucial part of the terrestrial biosphere. Despite their importance for ecosystem functioning, few quantitative, spatially explicit models of the active belowground community currently exist. In particular, nematodes are the most abundant animals on Earth, filling all trophic levels in the soil food web. Here we use 6,759 georeferenced samples to generate a mechanistic understanding of the patterns of the global abundance of nematodes in the soil and the composition of their functional groups. The resulting maps show that 4.4 ± 0.64 × 1020 nematodes (with a total biomass of approximately 0.3 gigatonnes) inhabit surface soils across the world, with higher abundances in sub-Arctic regions (38% of total) than in temperate (24%) or tropical (21%) regions. Regional variations in these global trends also provide insights into local patterns of soil fertility and functioning. These high-resolution models provide the first steps towards representing soil ecological processes in global biogeochemical models and will enable the prediction of elemental cycling under current and future climate scenarios.

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.

Flatworm mucus as the base of a food web.

BACKGROUND: By altering their habitats, engineering species can improve their own fitness. However, the effect of this strategy on the fitness of coexisting species or on the structure of the respective food web is poorly understood. In this study, bacteria and bacterivorous nematodes with short (Caenorhabditis elegans) and long (Plectus acuminatus) life cycles were exposed to the mucus secreted by the freshwater flatworm Polycelis tenuis. The growth, reproduction, and feeding preferences of the nematodes in the presence/absence of the mucus were then determined. In addition, confocal laser scanning microscopy (CLSM) was used to examine the structural footprint of the mucus and the mucus colonization dynamics of bacteria and protozoans. RESULTS: Mucus exposure resulted in a greater reproductive output in P. acuminatus than in C. elegans. In a cafeteria experiment, both nematode species were attracted by bacteria-rich patches and were not deterred by mucus. CLSM showed that the flatworms spread a layer of polysaccharide-rich mucus ca. 15 µm thick from their tails. Subsequent colonization of the mucus by bacteria and protozoans resulted in an architecture that progressively resembled a complex biofilm. The presence of protozoans reduced nematode reproduction, presumably due to competition for their bacterial food supply. CONCLUSION: Animal secretions such as mucus may have broader, community-level consequences and contribute to fueling microbial food webs.

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 the water-soluble fraction of a crude oil on freshwater meiofauna and nematode assemblages.

Oil water-soluble fractions (WSFs) compounds have low persistence in water; evaporating in a matter of hours to days. Nonetheless, they pose high toxicity to exposed biota. Their effects may be instantaneous or delayed, affecting, respectively, mainly survival vs growth and reproduction. We investigated the effects of crude oil WSFs on freshwater meiobenthos, with a focus on nematode assemblages, in community microcosm experiments lasting 15 weeks. Treatments consisted of the application of different concentrations of oil WSFs, i.e. high (100%) medium (50%) and low (10%), and effects were assessed one, three, nine and 15 weeks after contamination, allowing us to detect both short-term and lasting effects of oil-WSF. Additionally, we compared the effects of a single contamination event with those of a so-called 'constant' oil-WSF contamination where we replenished evaporated water with water containing the medium concentration of oil WSF. Next to nematodes, the most abundant meiofaunal taxa were rotifers, gastrotrichs, oligochaetes and tardigrades. Total abundance, different diversity indices, the composition of feeding-types and the age structure were investigated in the assessment of direct oil effects on the structure of nematode assemblages. Limited immediate effects were observed, except for a significant decrease of the index of taxonomic distinctness, which already appeared in the first week. Significant impacts on total nematode abundance, diversity and species composition only became apparent after 9-15 weeks of incubation, indicating that delayed effects of a single exposure are far more pronounced than instantaneous effects. Moreover, for most response variables, the strongest impacts were not observed in the highest-concentration treatment, but in a medium-concentration treatment with regular replenishment of oil WSF, suggesting that internal exposure may be important in generating effects. Furthermore, the predictability of the sensitivity of individual species was sometimes poor, which may not only result from these species' sensitivities, but also from alterations in interspecific interactions in polluted communities. Further toxicity tests should be carried out in order to unravel the main modes of action of crude oil WSF which lead to the observed long-term sublethal effects on nematode communities.

Effects of a broad range of experimental temperatures on the population growth and body-size of five species of free-living nematodes.

The temperature-size rule postulates that the growth rates of ectotherms increase under rising temperatures, while the sizes of these organisms at maturity decrease. However, the upper temperature-tolerance range is also typically represented by a metabolic tipping point after which growth suddenly ceases. Free-living nematodes are important members of ecosystems, but little is known about their thermal tolerance. In the present study we measured the population growth rates and body-size distributions of five species of free-living bacterivorous nematodes exposed in the laboratory to a broad range of temperatures. This allowed a determination of their different thermal tolerance ranges, even of closely related species, including Plectus acuminatus (thermal optimum of 20-25 °C) and P. cf. velox (10-15 °C). With the exception of Acrobeloides nanus, which had the broadest thermal tolerance range, the population growth of the other species declined between 25 and 30 °C. Our results were consistent with the temperature-size rule, as the body-size of the tested species at maturity decreased with increasing temperature. This reduction was accompanied by a smaller number of eggs carried by mature females. Although our study was purely experimental, it suggests that heat waves or other alterations in the thermal regime affect the population dynamics and body-size structure of nematode communities in the field.

Interactions among competing nematode species affect population growth rates.

Investigations of the interplay of organisms in an ecological community are a prerequisite to understanding the processes that shape the structures of those communities. Among several types of interactions, interest in the positive interactions of species that compete for the same resource has grown, as they may provide a mechanism enabling coexistence. In the laboratory experiment described herein, the effects of interspecific interaction on the population growth of two bacterial-feeding nematode species, Panagrolaimus cf. thienemanni and Poikilolaimus cf. regenfussi, were investigated. Specifically, we asked: (1) whether there is an interspecific interaction between organisms competing for a mutual resource and (2) whether these interactions are altered by the competitors' initial densities and (3) their variable growth rates (induced by different food supplies). Each treatment initially contained 48 nematode individuals, but at different species ratios (48:0; 32:16; 24:24; 16:32; 0:48). The populations were provided with three different bacterial densities (108, 109, and 1010 cells ml-1) as food. The data were analyzed using a generalized linear mixed model. The best-fitting model revealed a significant decline in population growth rates with an increasing species ratio, but depending on the food density and species. These results provide strong evidence for positive interspecific interactions that vary with both species density and food-supply level. They also suggest important roles for positive interspecific interactions in habitat colonization and in maintaining the coexistence of species in the same trophic group.

Is stemflow a vector for the transport of small metazoans from tree surfaces down to soil?

BACKGROUND: Stemflow is an essential hydrologic process shaping the soil of forests by providing a concentrated input of rainwater and solutions. However, the transport of metazoans by stemflow has yet to be investigated. This 8-week study documented the organisms (< 2 mm) present in the stemflow of different tree species. Because the texture of the tree bark is a crucial determination of stemflow, trees with smooth bark (Carpinus betulus and Fagus sylvatica) and rough bark (Quercus robur) were examined. RESULTS: Up to 1170 individuals per liter of stemflow were collected. For rotifers and nematodes, a highly positive correlation between abundance and stemflow yield was determined. Both taxa were predominant (rotifers: up to 70%, nematodes: up to 13.5%) in the stemflow of smooth-barked trees whereas in that of the oak trees collembolans were the most abundant organisms (77.3%). The mean number of organisms collected per liter of stemflow from the two species of smooth-barked trees was very similar. A higher number of nematode species was found in the stemflow of these trees than in the stemflow of rough-barked oak and all were typical colonizers of soil- and bark-associated habitats. CONCLUSION: This pilot study showed for the first time that stemflow is a transport vector for numerous small metazoans. By connecting tree habitats (e.g., bark, moss, lichens or water-filled tree holes) with soil, stemflow may influence the composition of soil fauna by mediating intensive organismal dispersal.

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.

Analyzing life-history traits and lipid storage using CARS microscopy for assessing effects of copper on the fitness of Caenorhabditis elegans.

Lipid storage provides energy for cell survival, growth, and reproduction and is closely related to the organismal response to stress imposed by toxic chemicals. However, the effects of toxicants on energy storage as it impacts certain life-history traits have rarely been investigated. Here, we used the nematode Caenorhabditis elegans as a test species for a chronic exposure to copper (Cu) at EC20 (0.50 mg Cu/l). Effects on the fatty acid distribution in C. elegans body were determined using coherent anti-Stokes Raman spectroscopy (CARS) to link population fitness responses with individual ecophysiological responses. Cu inhibited nematode reproductive capacity and offspring growth in addition to shortening the lifespan of exposed individuals. In adult nematodes, Cu exposure led to significant reduction of lipid storage compared to the Cu-free control: Under Cu, lipids filled only 0.5% of the nematode body volume vs. 7.5% in control nematodes, lipid droplets were on average 74% smaller and the number of tiny lipids (0-10 µm2) was increased. These results suggest that (1) Cu has an important effect on the life-history traits of nematodes; (2) the quantification of lipid storage can provide important information on the response of organisms to toxic stress; and (3) CARS microscopy is a promising tool for non-invasive quantitative and qualitative analyses of lipids as a measure of nematode fitness.

What´s in the tank? Nematodes and other major components of the meiofauna of bromeliad phytotelms in lowland Panama.

BACKGROUND: Nematodes are a very diverse and extremely abundant group of animals, but their occurrence in the tropics is surprisingly little understood. We investigated the meiofauna of epiphytic tank bromeliads in the lowlands of Panama with particular emphasis on nematodes. RESULTS: We encountered 89 morphospecies of nematodes in 54 bromeliad tanks, which were sampled in the wet and the dry season. Rotifers were by far the most abundant group in both the dry and the wet season (with up to 960 individual ml(-1)), followed by nematodes, annelids and harpacticoid copepods. Individual plants hosted up to 25 nematode species. These nematodes represented a diversity of feeding guilds, suction-feeders and deposit-feeders being most abundant. The relative abundances of feeding-types of nematodes differed considerably in the wet and dry season. Both species richness and abundance were strongly correlated with the size of the phytotelms and the season, while species diversity assessed with the Shannon-index was affected by neither of the two. CONCLUSION: This is the first study with a particular focus on the diversity of nematodes in tank bromeliads. We document a meiofauna of considerable abundance and diversity, which suggests important functional roles in ecological processes such as decomposition, which in turn warrants further study.

Phenanthrene bioaccumulation in the nematode Caenorhabditis elegans.

The contribution of food to the bioaccumulation of xenobiotics and hence toxicity is still an ambiguous issue. It is becoming more and more evident that universal statements cannot be made, but that the relative contribution of food-associated xenobiotics in bioaccumulation depends on species, substance, and environmental conditions. Yet, small-sized benthic or soil animals such as nematodes have largely been disregarded so far. Bioaccumulation of the polycyclic aromatic hydrocarbon phenanthrene in the absence and presence of bacterial food was measured in the nematode Caenorhabditis elegans. Elimination of phenanthrene in the nematodes was biphasic, suggesting that there was a slowly exchanging pool within the nematodes or that biotransformation of phenanthrene took place. Even with food present, dissolved phenanthrene was still the major contributor to bioaccumulated compound in nematode tissues, whereas the diet only contributed about 9%. Toxicokinetic parameters in the treatment without food were different from the ones of the treatment with bacteria, possibly because nematodes depleted their lipid reserves during starvation.

Free-living nematodes in the freshwater food web: a review.

Free-living nematodes are well-recognized as an abundant and ubiquitous component of benthic communities in inland waters. Compelling evidence from soil and marine ecosystems has highlighted the importance of nematodes as trophic intermediaries between microbial production and higher trophic levels. However, the paucity of empirical evidence of their role in freshwater ecosystems has hampered their inclusion in our understanding of freshwater food web functioning. This literature survey provides an overview of research efforts in the field of freshwater nematode ecology and of the complex trophic interactions between free-living nematodes and microbes, other meiofauna, macro-invertebrates, and fishes. Based on an analysis of the relevant literature and an appreciation of the potential of emerging approaches for the evaluation of nematode trophic ecology, we point out research gaps and recommend relevant directions for further research. The latter include (i) interactions of nematodes with protozoans and fungi; (ii) nonconsumptive effects of nematodes on microbial activity and the effects of nematodes on associated key ecosystem processes (decomposition, primary production); and (iii) the feeding selectivity and intraspecific feeding variability of nematodes and their potential impacts on the structure of benthic communities.

Experimental studies with nematodes in ecotoxicology: an overview.

With respect to their high abundances, their role as intermediaries between microorganisms and higher trophic levels, and their ubiquitous occurrence in all habitats, nematodes are of strong potential interest as environmental indicators. Ecotoxicological methods to evaluate the risk of anthropogenic pollutants on ecosystems require both in vitro and in vivo toxicity tests to investigate either mechanisms or pathways of toxicity and to set accurate toxicity thresholds. For this, the interest in nematodes as model organisms in ecotoxicology increased over the past few decades and existing appropriate experimental methods are reviewed in this manuscript. An overview of the various existing ecotoxicological tools for nematodes, ranging from molecular laboratory methods to experimental model ecosystem approaches, and their role as indicator organisms is given. The reviewed studies, approaches that range from species-based to community-based methods, reveal exciting possibilities for the future use of nematodes in ecotoxicological studies. Suitable ecotoxicological tools and ecological indices for nematodes should be integrated in weight-of-evidence approaches for assessing the ecological risk of contamination.

Predator effects on a detritus-based food web are primarily mediated by non-trophic interactions.

Predator effects on ecosystems can extend far beyond their prey and are often not solely lethally transmitted. Change in prey traits in response to predation risk can have important repercussions on community assembly and key ecosystem processes (i.e. trait-mediated indirect effects). In addition, some predators themselves alter habitat structure or nutrient cycling through ecological engineering effects. Tracking these non-trophic pathways is thus an important, yet challenging task to gain a better grasp of the functional role of predators. Multiple lines of evidence suggest that, in detritus-based food webs, non-trophic interactions may prevail over purely trophic interactions in determining predator effects on plant litter decomposition. This hypothesis was tested in a headwater stream by modulating the density of a flatworm predator (Polycelis felina) in enclosures containing oak (Quercus robur) leaf litter exposed to natural colonization by small invertebrates and microbial decomposers. Causal path modelling was used to infer how predator effects propagated through the food web. Flatworms accelerated litter decomposition through positive effects on microbial decomposers. The biomass of prey and non-prey invertebrates was not negatively affected by flatworms, suggesting that net predator effect on litter decomposition was primarily determined by non-trophic interactions. Flatworms enhanced the deposition and retention of fine sediments on leaf surface, thereby improving leaf colonization by invertebrates - most of which having strong affinities with interstitial habitats. This predator-induced improvement of habitat availability was attributed to the sticky nature of the mucus that flatworms secrete in copious amount while foraging. Results of path analyses further indicated that this bottom-up ecological engineering effect was as powerful as the top-down effect on invertebrate prey. Our findings suggest that predators have the potential to affect substantially carbon flow and nutrient cycling in detritus-based ecosystems and that this impact cannot be fully appreciated without considering non-trophic effects.

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.

First evidence of cryptic species diversity and significant population structure in a widespread freshwater nematode morphospecies (Tobrilus gracilis).

Free-living nematodes are ubiquitous and highly abundant in terrestrial and aquatic environments, where they sustain ecosystem functioning by mineralization processes and nutrient cycling. Nevertheless, very little is known about their true diversity and intraspecific population structure. Recent molecular studies on marine nematodes indicated cryptic diversity and strong genetic differentiation of distinct populations, but for freshwater nematode species, analogous studies are lacking. Here, we present the first extensive molecular study exploring cryptic species diversity and genetic population structure of a widespread freshwater nematode morphospecies, Tobrilus gracilis, from nine postglacially formed European lakes. Taxonomic species status of individuals, analysed for fragments of the mitochondrial COI gene and for the large (LSU) and small (SSU) ribosomal subunits, were determined by morphological characteristics. Mitochondrial and nuclear markers strongly supported the existence of three distinct genetic lineages (Tg I-III) within Tobrilus gracilis, suggesting that this morphospecies indeed represents a complex of highly differentiated biological species. High genetic diversity was also observed at the population level. Across the nine lakes, 19 mitochondrial, and seven (LSU) and four (SSU) nuclear haplotypes were determined. A phylogeographical analysis revealed remarkable genetic differentiation even among neighbouring lake populations for one cryptic lineage. Priority and persistent founder effects are possible explanations for the observed population structure in the postglacially colonized lakes, but ask for future studies providing direct estimates of freshwater nematode dispersal rates. Our study suggests therefore that overall diversity of limnetic nematodes has been so far drastically underestimated and challenges the assumed ubiquitous distribution of other, single freshwater nematode morphospecies.

Effects of a bioassay-derived ivermectin lowest observed effect concentration on life-cycle traits of the nematode Caenorhabditis elegans.

The pharmaceutical ivermectin is used to treat parasitic infections, such as those caused by nematodes. While several studies have demonstrated the severe effects of ivermectin on non-target organisms, little is known about the drug's impact on free-living nematodes. In the present work, a full life-cycle experiment was conducted to estimate how an ivermectin lowest observed effect concentration derived from a Caenorhabditis elegans bioassay (endpoint reproduction) might translate into effects at the population level of this free-living nematode. The results showed that fecundity decreased to levels similar to those determined in the bioassay after a time of corresponding duration (18.6 % inhibition compared to the control), but the impact then rather weakened until the end of the experiment, at which point the net reproductive rate (R(0)) was still, but not significantly, reduced by 12.4 %. Moreover, the average lifespan, length of the reproductive period, maximum daily reproduction rate, and intrinsic rate of increase (r(m)) were significantly reduced by 30.0, 25.9, 11.2, and 3.5 %, respectively. The experiment revealed that a 4-day bioassay is protective enough for C. elegans with respect to ivermectin's effects on fecundity. However, the pronounced effects of a low drug concentration on survival, a highly elastic trait, may better account for the observed population-level response, i.e., a decrease of r(m), than the effects on fecundity. These results emphasize that full life-cycle experiments are valuable for assessment of pollutants, because the effects on several life-cycle traits can be simultaneously measured and integrated into an ecologically relevant parameter, the population growth rate, that reflects a population's response to a specific pollutant.

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.