Peter Heininger

Variability of sediment-contact tests in freshwater sediments with low-level anthropogenic contamination — Determination of toxicity thresholds

Freshwater sediments with low levels of anthropogenic contamination and a broad range of geochemical properties were investigated using various sediment-contact tests in order to study the natural variability and to define toxicity thresholds for the various toxicity endpoints. Tests were performed with bacteria (Arthrobacter globiformis), yeast (Saccharomyces cerevisiae), nematodes (Caenorhabditis elegans), oligochaetes (Lumbriculus variegatus), higher plants (Myriophyllum aquaticum), and the eggs of zebrafish (Danio rerio). The variability in the response of some of the contact tests could be explained by particle size distribution and organic content. Only for two native sediments could a pollution effect not be excluded. Based on the minimal detectable difference (MDD) and the maximal tolerable inhibition (MTI), toxicity thresholds (% inhibition compared to the control) were derived for each toxicity parameter: >20% for plant growth and fish-egg survival, >25% for nematode growth and oligochaete reproduction, >50% for nematode reproduction and >60% for bacterial enzyme activity.

Nematode species at risk--a metric to assess pollution in soft sediments of freshwaters.

Soft sediments are often highly polluted as many of the toxic chemicals introduced into surface waters bind to settling particles. The resulting accumulation of pollutants in the sediments poses a risk for benthic communities. However, pollution induced changes in benthic communities have been difficult to determine when using macro-invertebrates as bioindicators, as these organisms are often absent in soft sediment. The present study therefore examined the ability of meiofaunal organisms, specifically, nematodes, to assess the ecological status of soft sediments. Over a 9-year period, nematode communities present in sediments collected from large rivers and lake Constance in Germany were studied. These sediments showed a large range of physico-chemical properties and anthropogenic contamination. After the degree of metal and organic contamination was translated into ecotoxicologically more relevant toxic units (TUs), multivariate methods were used to classify nematode taxa in species at risk (NemaSPEAR) or not at risk (NemaSPE(not)AR). This approach clearly distinguished the influence of sediment texture from that of the toxic potential of the samples and thus allowed classification of the nematode species according to their sensitivity to or tolerance of toxic stress. Two indices, expressing the proportion of species at risk within a sample (NemaSPEAR[%](metal), NemaSPEAR[%](organic)), were calculated from independent data sets obtained in field and experimental studies and showed good correlations with the toxic potential (field data) or chemical concentrations (microcosm data). NemaSPEAR[%] indices for metal and organic pollution were therefore judged to be suitable for assessing the impact of chemical contamination of freshwater soft sediments.

A new plant-based bioassay for aquatic sediments

Background, Goal and ScopeTo date, standardised bioassays for the assessment of the ecotoxicological potential in sediments and dredged material use test organisms like bacteria, algae and crustaceae. This paper presents the development and application of a novel sediment contact test (whole sediment) withMyriophyllum aquaticum, a representative of rooted aquatic macrophytes. The aim of the present study is to demonstrate the value of a sediment contact test with rooted macrophytes as a supplement to existing test batteries in order to improve the assessment of sediment toxicity.MethodsThe newly developed sediment contact test withMyriophylhim aquaticum was applied to natural whole sediments. For performing the test, whorls ofMyriophyllum aquaticum were directly planted in the native sediment and incubated in the light at 24°C (cf. section results and discussion). The end points of the test were the number of the shoots and the fresh weight of the whole plants. The duckweed growth inhibition test withLemna minor according to ISO/DIS 20079 was performed in pore waters from sediment samples. The results of the sediment contact test withMyriophyllum aquaticum were compared with each other and with those of the aquatic duckweed test.Results and DiscussionA test protocol for the new plant-based sediment contact test using the aquatic plantMyriophyllum aquaticum as an indicator was developed. The best control sediment proved to be the OECD sediment (OECD 207). A test period of 10 days appeared to be sufficient for the test. The increase of biomass and the derived growth rate were found to be the most suitable evaluation parameters. The growth behaviour ofMyriophyllum aquaticum differed depending on the origin of sediments. Therefore, plant-affecting contamination, that is bound in sediments, was indicated.ConclusionsThe novel sediment contact test withMyriophyllum aquaticum can indicate phytotoxic effects in sediments. Therefore, it allows a better assessment of the overall-toxicity in whole sediments.Recommendations and OutlookThe sediment contact test withMyriophyllum aquaticum is a valuable tool for the evaluation of the ecotoxicological risk potential of waters and sediments. It should become a complement to a standardised test battery generally used for the assessment of sediment toxicity.

Sediment contact tests as a tool for the assessment of sediment quality in German waters

A sediment contact test (SCT) battery consisting of five ecotoxicological test systems was applied to 21 native freshwater sediments characterized by a broad variety of geochemical properties and anthropogenic contamination. Higher plants (Myriophyllum aquaticum), nematodes (Caenorhabditis elegans), oligochaetes (Lumbriculus variegatus), zebrafish embryos (Danio rerio), and bacteria (Arthrobacter globiformis), representing various trophic levels and exposure pathways, were used as test organisms. The test battery detected sediment toxicity caused by anthropogenic pollution, whereas the various tests provided site-specific, nonredundant information to the overall toxicity assessment. Based on the toxicity pattern derived from the test battery, the sediments were classified according to a newly proposed classification system for sediment toxicity assessment. The SCT-derived classification generally agreed well with the application of consensus-based sediment quality guidelines (SQGs), especially with regard to sediments with high toxic potential. For sediments with low to medium toxic potential, the SQGs often underestimated the toxicity that was detected by the SCTs, underpinning the need for toxicity tests in sediment quality assessment.

Assessing effects of the pharmaceutical ivermectin on meiobenthic communities using freshwater microcosms.

Ivermectin is a widely applied veterinary pharmaceutical that is highly toxic to several non-target organisms. So far, little is known about its impact on benthic freshwater species, although its rapid sorption to sediment particles and high persistence in aquatic sediments have raised concerns about the risk for benthic organisms. In the present study, indoor microcosms were used to assess the impact of ivermectin on freshwater meiobenthic communities over a period of 224 days. Microcosm sediments were directly spiked with ivermectin to achieve nominal concentrations of 0.9, 9, and 45 microg kg(-1) dw. Initially measured ivermectin concentrations (day 0) were 0.6, 6.2, and 31 microg kg(-1) dw. In addition to abundance of major meiobenthic organism groups, the nematode community was assessed on the species level, assuming a high risk for free-living nematodes due to their close phylogenetic relationship to the main target organisms of ivermectin, parasitic nematodes. Benthic microcrustaceans (cladocerans, ostracods) and nematodes showed the most sensitive response to ivermectin, while tardigrades profited from the presence of the pharmaceutical. The most pronounced effects on the meiofauna community composition occurred at the highest treatment level (31 microg kg(-1) dw), leading to a no observed effect concentration (NOEC(Community)) of 6.2 microg kg(-1) dw. However, the nematode community was already seriously affected at a concentration of 6.2 microg kg(-1) dw with two bacterivorous genera, Monhystera and Eumonhystera, being the most sensitive, whereas species of omnivorous genera (Tripyla, Tobrilus) increased in abundance after the application of ivermectin. Thus, a NOEC(Community) of 0.6 microg kg(-1) dw was derived for nematodes. Direct and indirect effects of ivermectin on meiobenthic communities could be demonstrated. The pharmaceutical is likely to pose a high risk, because its NOECs are close to predicted environmental concentrations (PECs) in sediments (0.45-2.17 microg kg(-1) dw), resulting in worst case risk quotients (RQs) of 1.05-36.2. This observation lends support to efforts aimed at preventing the repeated entry of ivermectin in aquatic environments and thus its accumulation in sediments. Moreover, this study points out that model ecosystem studies should be part of environmental risk assessments (ERAs) of veterinary medicinal products (VMPs).

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.

Bioaccumulation in aquatic systems: methodological approaches, monitoring and assessment

Bioaccumulation, the accumulation of a chemical in an organism relative to its level in the ambient medium, is of major environmental concern. Thus, monitoring chemical concentrations in biota are widely and increasingly used for assessing the chemical status of aquatic ecosystems. In this paper, various scientific and regulatory aspects of bioaccumulation in aquatic systems and the relevant critical issues are discussed. Monitoring chemical concentrations in biota can be used for compliance checking with regulatory directives, for identification of chemical sources or event-related environmental risk assessment. Assessing bioaccumulation in the field is challenging since many factors have to be considered that can affect the accumulation of a chemical in an organism. Passive sampling can complement biota monitoring since samplers with standardised partition properties can be used over a wide temporal and geographical range. Bioaccumulation is also assessed for regulation of chemicals of environmental concern whereby mainly data from laboratory studies on fish bioaccumulation are used. Field data can, however, provide additional important information for regulators. Strategies for bioaccumulation assessment still need to be harmonised for different regulations and groups of chemicals. To create awareness for critical issues and to mutually benefit from technical expertise and scientific findings, communication between risk assessment and monitoring communities needs to be improved. Scientists can support the establishment of new monitoring programs for bioaccumulation, e.g. in the frame of the amended European Environmental Quality Standard Directive.

A semi-fluid gellan gum medium improves nematode toxicity testing

This study examined an alternative test medium for nematodes that use gellan gum as the gelling agent instead of agar. The semi-fluid consistency of the gel-like component nematode growth gellan gum (CNGG) supports three-dimensional distribution of the nematodes and food bacteria, but still allows free movement of the former. Moreover, flexible preparation of the medium and easy recovery of the test organisms are possible. Here, the effects of the nematicides ivermectin (pharmaceutical) and aldicarb (pesticide) and of the metal cadmium on the growth and reproduction of the free-living nematodes Caenorhabditis elegans and Panagrolaimus cf. thienemanni were studied in CNGG media. Results were compared to those obtained with the standard liquid test media in order to evaluate the applicability of CNGG for nematode toxicity testing. The sensitivity of P. cf. thienemanni to all three substances was found to be higher than that of C. elegans, but both nematodes showed the highest sensitivity to ivermectin exposure. This raises concerns about the risk posed by the pharmaceutical to non-target nematodes. In contrast to ivermectin bioassays carried out in CNGG medium, those conducted in liquid medium resulted in wide-ranging variability between and within replicates. Thus, CNGG seems to be particularly valuable for testing hydrophobic substances with a high sorption affinity as it favors their sorption to food bacteria and minimizes contact with the surfaces of the test vessels. However, the medium was less suitable for deriving toxicity thresholds for cadmium and may likewise not be an appropriate choice for testing other metals. The medium introduced herein was shown to be appropriate for sublethal nematode toxicity testing and likely provides a convenient environment for testing other nematode species. Besides improved testing of hydrophobic substances, CNGG also offers advantages for long-term studies, such as full life-cycle experiments, in which fresh medium is regularly needed. Moreover it may be beneficial for testing other poorly soluble or insoluble substances, such as nanoparticles.

Anwendung differenzierter Untersuchungsmethoden zur Sedimentbewertung

ZusammenfassungDem Gewässersediment kommt als Habitat einer artenreichen Biozönose und als Reaktionsort vielfältiger Stoffumsetzungen eine besondere Bedeutung zu. Aufgrund des hohen Akkumulationspotenzials für Schadstoffe beeinträchtigen anthropogene Einflüsse in erhöhtem Maße die Sedimentbeschaffenheit und führen zur Störung des natürlichen Sedimentzustands. Zur Sicherung des Schutzgutes Aquatische Lebensgemeinschaft sind diese Veränderungen zu erfassen und zu bewerten und die stofflichen Ursachen zu hinterfragen. Seit Ende der 80er Jahre besteht verstärkt der Anspruch, Sedimente möglichst umfassend zu beurteilen. Nur über einen integrierten Bewertungsansatz, in dem Toxizitätsnachweise unter standardisierten Laborbedingungen, Schadstoffdaten und biologischein situ-Befunde zu einer Gesamtaussage verbunden werden, kann sich diesem Problem genähert werden. Entscheidend für die Aussagekraft der Untersuchungsergebnisse ist die Auswahl von Testverfahren, die die Schadstoffwirkungen differenziert anzeigen können. Vorgestellt werden Ergebnisse aus einem umfangreichen Untersuchungsansatz. Es wurden sowohl DIN-Verfahren mit Porenwässern und Eluaten als auch Tests mit dem Gesamtsediment in die Untersuchungen integriert. Um eine größere Vielzahl an Schadstoffen im Sediment zu erfassen, wurden auch Lösungsmittelextrakte und deren Fraktionen in verschiedenen Biotests eingesetzt. Chemische Untersuchungen erfolgten zur Erfassung der Strukturparameter und prioritärer Schadstoffe. Im non-target screening konnte eine Vielzahl weiterer Stoffe und Stoffklassen ermittelt werden. Die Modellorganismen der Biotests reagierten sehr differenziert und teilweise sehr stark auf die Kontaminationen in den einzelnen Untersuchungsmedien. Speziell in den Sedimentextrakten erwiesen sich einige Fraktionen und damit Stoffe und Stoffgruppen als besonders toxisch. Im Ergebnis konnte gezeigt werden, dass die ausgewählten chemischen, ökotoxikologischen und biologischen Untersuchungen zu einer bewertenden Gesamtaussage beitragen. Zur Optimierung der sehr umfangreichen Untersuchungspalette sollten weitere, möglichst sensitive Biotests und benthosbiologische Parameter, geprüft werden.AbstractThe aquatic sediments have special significance as habitats of species-rich biocoenoses and as the place where manifold transformation processes occur. Because of their high potential for accumulation of contaminants, sediments are particularly sensitive to anthropogenic impacts, which may disturb the natural state of sediments. In order to protect the aquatic life community, such impacts must be identified, assessed, and the chemical causes of toxic effects must be found. Since the end of the 1980s, the interest for a possibly comprehensive assessment of sediments has increased. This problem can be tackled only by an integrated approach, which combines the detection of toxicity under standardized laboratory conditions with chemical data and biologicalin situ studies into a holistic view. The information value of the results depends essentially on the selection of a diversity of test methods, which are able to indicate contaminant effects differentially. This paper presents results from a comprehensive test approach, which integrates standardized methods (DIN) with pore waters and eluates, as well as tests using the whole sediment. To cover a wider variety of contaminants in the sediment, solvent extracts and fractions thereof were also examined by different bioassays. Chemical analyses examined the structural parameters and identified priority contaminants. Non-target screening could detect a variety of further substances and substance classes. The model organisms of the bioassays responded very differentially and sometimes very intensively to the contaminants in the various investigation media. Especially in sediment extracts, some fractions, and consequently also substances and groups of substances, proved to be particularly toxic. It could be shown that the selected chemical, ecotoxicological, and biological study methods contribute to a holistic assessment. Further, possibly very sensitive bioassays and benthos-biological parameters should be examined aiming to optimize the very wide battery of tests.

Estrogenic effects along the river saale

Sediments along the river Saale, one of the main tributaries of the river Elbe, were characterized with the yeast estrogen screen to elucidate possible sources of endocrine-disrupting compounds that might contribute to the downstream contamination of the river Elbe. At two sampling sites, elevated levels of estrogenic activity up to 55,000 ng ethinylestradiol equivalents per kilogram sediment dry weight were detected in the respective sediment extracts. Aliquots of the sediment extracts were analyzed for 4-nonylphenols and natural steroidal estrogens as possible candidates with an estrogenic potential. The maximal concentrations of 4-iso-nonylphenol and estrone were 115 mg/kg dry weight and 20 µg/kg dry weight at the sampling site Luppe, which showed in accordance the highest biological activity. Under consideration of compound concentration and compound specific estrogenic activity the 4-iso-nonylphenols contributed most to the observed estrogenic effect. A strong correlation between the measured estrogenic activity and the concentration of the sediment-associated 4-iso-nonylphenol underlines the relevance of this compound class as a xenoestrogen in the catchment area of the river Saale.