Kai Ristau

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.

Using meiofauna to assess pollutants in freshwater sediments: A microcosm study with cadmium

The direct and indirect effects of Cd on benthic communities were assessed in a freshwater microcosm study over a period of seven months (218 d). Cadmium was regarded as a model substance to evaluate the usefulness of small-scale laboratory microcosm with microscopic fauna. In particular, effects on the meiofauna community, an ecologically important but rather neglected benthic component, were investigated. In addition, some microfaunal parameters (protozoan abundance and microbial activity) were determined. The sediment was spiked with nominal Cd concentrations of 10, 100, and 1,000 mg/kg dry weight. Because of the strong binding of Cd to sediment particles, measured Cd pore-water concentrations never exceeded 129.5 ± 40.7 µg/L. At 1,000 mg/kg dry weight, the abundances of the two dominant meiofauna taxa, nematodes and oligochaetes, were significantly reduced throughout the present study. Regarding nematodes, species of bacterivorous taxa (Daptonema, Eumonhystera) decreased, whereas species of predacious and omnivorous taxa (Mononchus, Dorylaimus, and Ironus) increased in dominance in microcosms of the highest Cd concentration. Transient effects on microfauna were observed, especially in the first half of the present study, with a reduction in microbial activity and protozoan abundance. However, in microcosms receiving the highest Cd concentration, the abundance of the flagellate Euglena mutabilis increased significantly toward the end of the present study. The results of the present study support the use of small-scale microcosms with natural meiofauna communities as a suitable tool to assess the impact of pollutants in freshwater sediments.

Relation between nematode communities and trophic state in southern Swedish lakes

The aim of this study was to examine whether littoral nematode community patterns are shaped by lake trophic state. It was hypothesized that trophic level is associated negatively with the proportion of omnivores and positively with the percentages of bacterial feeders, but not at all with the diversity, abundance, and biomass of freshwater nematodes. Sediment samples were taken at littoral sites of eight southern Swedish lakes of different trophy in spring and autumn 2007. Trophic level was found to strongly influence species richness, as oligotrophic and mesotrophic lakes supported the greatest species numbers, whereas nematode abundance, biomass, and Shannon index were unaffected. Furthermore, our results indicated effects on the nematode community’s trophic structure, with a larger proportion of predatory nematodes in oligotrophic and mesotrophic lakes but no differences in the other feeding types (bacteria, algae and suction feeders, omnivorous species). Multivariate analysis indicated a shift in species compositions along the threshold from mesotrophic to eutrophic conditions, with the presence of Tobrilus gracilis, Monhystera paludicola, Brevitobrilus stefanskii, and Ethmolaimus pratensis related to the latter. Nematode communities in oligotrophic and mesotrophic lakes were characterized by a similar species composition, with pronounced occurrences of Eumonhystera longicaudatula, Semitobrilus cf. pellucidus, Prodesmodora circulata, and Rhabdolaimus terrestris. Overall, the results suggested that lake trophic state is a major factor structuring littoral nematode communities, although intra-lake variations might be of importance as well.

The functional response of a freshwater benthic community to cadmium pollution

Theory predicts that in freshwater communities under chemical stress secondary production will decrease while the rate of biomass turnover (P/B) will increase. However, this concept has never been tested on organisms of smaller size (bacteria, protozoans, small metazoans), although they form the basis of the heterotrophic food web. The present work describes the results of a 7-month microcosm study, in which the effects of low and high toxic stress on an entire sediment community were examined, with cadmium (Cd) as the model pollutant (50 and 400mg Cd kg(-1) dry sediment). While metazoans and protozoans generally followed the expected trend, in bacteria both production and P/B decreased under Cd stress. These observations provide new insights into the functioning of freshwater ecosystems and demonstrate the functional consequences of toxicants on biological systems.

Biomass estimation across the benthic community in polluted freshwater sediment--a promising endpoint in microcosm studies?

With respect to the ecological relevance of endpoints, biomass as an endpoint might be promising regarding ecotoxicological assessments of benthic communities. In a freshwater microcosm study the effect of two cadmium (Cd) concentrations (50 and 400 mg Cd kg(-1) dw) on biomass and abundance of a benthic community were investigated over a period of seven months. Specifically, the sensitivity of both endpoints in distinguishing differential effects was compared. While bacteria were found to be unaffected by Cd, abundance and biomass of protozoans and metazoans decreased. In a short-term comparison, differences between control and Cd treatments were, overall, more pronounced for flagellate biomass and for metazoan abundance with strong differences between the taxonomic groups; furthermore, over the long-term, the differences among organisms and endpoints changed. Based on toxicant sensitivity, the reasonably low variance of the data and the workload involved, biomass can provide a useful additional endpoint in microcosm studies.

A comparative approach using ecotoxicological methods from single‐species bioassays to model ecosystems

Soft sediments are often hotspots of chemical contamination, and a thorough ecotoxicological assessment of this habitat can help to identify the causes of stress and to improve the health of the respective ecosystems. As an important component of the ecologically relevant meiobenthic fauna, nematodes can be used for sediment assessments, with various assay tools ranging from single-species toxicity tests to field studies. In the present study, microcosms containing sediment were used to investigate direct and indirect effects of zinc on natural nematode assemblages, and acute community toxicity tests considering only direct toxicity were conducted. The responses of the various freshwater nematode species in both approaches were compared with those of Caenorhabditis elegans, determined in standardized tests (ISO 10872). At a median lethal concentration (LC50) of 20 mg Zn/L, C. elegans represented the median susceptibility of 15 examined nematode species examined in the acute community toxicity tests. In the microcosms, Zn affected the nematodes dose-dependently, with changes in species composition first detected at 13 mg Zn/kg to 19 mg Zn/kg sediment dry weight. The observed species sensitivities in the microcosms corresponded better to field observations than to the results of the acute community toxicity tests. Environ Toxicol Chem 2016;35:2987-2997.

Toxicity in relation to mode of action for the nematode Caenorhabditis elegans: Acute‐to‐chronic ratios and quantitative structure–activity relationships

Acute-to-chronic ratios (ACRs) and quantitative structure-activity relationships (QSARs) are of particular interest in chemical risk assessment. Previous studies focusing on the relationship between the size or variation of ACRs to substance classes and QSAR models were often based on data for standard test organisms, such as daphnids and fish. In the present study, acute and chronic toxicity tests were performed with the nematode Caenorhabditis elegans for a total of 11 chemicals covering 3 substance classes (nonpolar narcotics: 1-propanol, ethanol, methanol, 2-butoxyethanol; metals: copper, cadmium, zinc; and carbamates: methomyl, oxamyl, aldicarb, dioxacarb). The ACRs were variable, especially for the carbamates and metals, although there was a trend toward small and less variable ACRs for nonpolar narcotic substances. The octanol-water partition coefficient was a good predictor for explaining acute and chronic toxicity of nonpolar narcotic substances to C. elegans, but not for carbamates. Metal toxicity could be related to the covalent index χm2r. Overall, the results support earlier results from ACR and QSAR studies with standard freshwater test animals. As such C. elegans as a representative of small soil/sediment invertebrates would probably be protected by risk assessment strategies already in use. To increase the predictive power of ACRs and QSARs, further research should be expanded to other species and compounds and should also consider the target sites and toxicokinetics of chemicals.

Effects of nutrient enrichment on the trophic structure and species composition of freshwater nematodes—a microcosm study

Abstract. We explored changes in trophic structure and species composition of freshwater nematodes in response to nutrient enrichment in microcosms for 16 mo. Succession of functional groups differed significantly among the 5 nutrient treatments, which ranged from oligotrophic to polytrophic conditions, in the 2nd half of the experiment. Differences among nutrient levels were most important for representatives of lower trophic levels, such as bacterial and algal feeders, whereas suction feeders and predators remained relatively unaffected. In contrast to our expectations, density and biomass of bacterial and algal feeders increased in the nutrient-poor treatments because of the presence of a high fraction of edible unicellular green algae and diatoms, whereas large standing stocks of inedible macrophytes developed in the highly nutrient-enriched treatments. Nematode assemblages in microcosms with intermediate or high nutrient levels were characterized by a large proportion of omnivores and predators, respectively, because of relatively low total densities. Our results also suggest a gradual change in species composition along the nutrient gradient because the degree of pairwise assemblage dissimilarity increased with corresponding differences in nutrient enrichment. Our study highlights the potential of bottom-up forces in shaping freshwater nematode assemblages and provides further evidence of pelagic–meiobenthic coupling.

The D3-D5 region of large subunit ribosomal DNA provides good resolution of German limnic and terrestrial nematode communities

Reliable and well-developed DNA barcode databases are indispensable for the identification of microscopic life. However, effectiveness of molecular barcoding in identifying terrestrial specimens, and nematodes in particular, has received little attention. In this study, ca 600 ribosomal large subunit DNA fragments (D3-D5 region) were successfully amplified for 79 limnic and terrestrial nematode species sampled at 147 locations across Germany. Distinctive DNA motifs in the LSU region were identified in 80% of all species examined. For 13 supposedly single morphospecies, 2-7 LSU barcode groups were detected with a wide range of intraspecific variations (0.09-7.9%). This region seems to be more suitable for the assessment of limno-terrestrial nematode diversity than the frequently used mitochondrial gene COI , as amplification success of the latter fragment is low for several nematode species. Our reference database for nematodes may serve as a starting point for applied and fundamental studies for these ubiquitous, ecologically highly relevant, organisms.

Varying Patterns on Varying Scales: A Metacommunity Analysis of Nematodes in European Lakes.

Ecological community patterns are often extremely complex and the factors with the greatest influence on community structure have yet to be identified. In this study we used the elements of metacommunity structure (EMS) framework to characterize the metacommunities of freshwater nematodes in 16 European lakes at four geographical scales (radius ranging from 80 m to 360 km). The site characteristics associated with site scores indicative of the structuring gradient were identified using Spearman rank correlations. The metacommunities of the 174 nematode species included in this analysis mostly had a coherent pattern. The degree of turnover increased with increasing scale. Ordination scores correlated with geographical variables on the larger scales and with the trophic state index on a regional scale. The association of the structuring gradient with spatial variables and the scale-dependent increase in turnover showed that nematode dispersal was limited. The different metacommunity patterns identified at the increasing geographical scales suggested different, scale-related mechanisms of species distribution, with species sorting dominating on smaller and mass effects on larger geographical scales.