Identifikation geeigneter Nachweismöglichkeiten von hormonaktiven und reproduktionstoxischen Wirkungen in aquatischen Ökosystemen

Background and objectives There is an urgent need to detect, assess, and reduce effects of hormonally active compounds and endocrine disrupters in aquatic systems, as reflected in national research programs like the Swiss NRP 50 “Endocrine Disruptors” and its consensus platforms. As a medium-term measure, the EU strategy on endocrine disruptors (SEC(2007)1635) uses the Endocrine Disruptor Testing and Assessment (EDTA) Task Force of the Organisation for Economic Co-operation and Development (OECD) along with other research activities. In particular the test methods of the OECD that are currently in validation or already validated may contribute to a better understanding of the extent of endocrine disruption, in particular if they are applied on environmental samples and in the context of risk-assessment strategies, for instance in waste water treatment. This article aims to give an overview and an evaluation on available and validated biological test systems for the detection of endocrine disruptive and reproductive effects in aquatic systems. Based on this a recommendation for a modular ecotoxicological test platform is given. The study focuses on test methods for sex hormone active substances. Material and methods On the basis of an extensive literature search and ongoing international validation efforts by the OECD for methods to detect endocrine disruptive effects, 15 biological test methods (5 in vivo and 10 in vitro) were selected. Comprising, for example, of eight OECD methods and three out of five in-vitro methods mentioned in the Global Water Research Coalition (GWRC) report “Tools to detect estrogenic activity in environmental waters” (Leusch 2008). Experienced users and developers were then asked to rate the test according to given relevant criteria. The resulting criteria profiles were compiled, compared, and evaluated. Results The methods were selected on the basis of validation status, distribution, their suitability for standardisation, and their proven sensitivity for environmental samples. We assumed that specific YES/YAS-procedures and the ER/AR Calux systems achieve the mentioned criteria. In the case that strong cytotoxicity of environmental samples affects the applicability of cellular reporter gene assays, alternatively a molecular receptor binding assay (e.?g. ELRA) could be used. Additional molecular receptor binding assays are currently in validation by the OECD. Modulating effects on steroidgenesis and probably even on aromatase activity can be detected by the OECD validated H295R Steroidgenesis Assay. Its applicability for environmental samples is currently tested. As in-vivo methods different fish assays (e.?g. Fish Screening Assay and the Zebrafish Embryo Test) were evaluated. For the detection of effects on the thyroid-associated hormone system the Amphibian Metamorphosis Assay (XEMA, for Xenopus Metamorphosis Assay) will soon be available as an OECD-Guideline. Because of trends in the international community to avoid tests with vertebrates in the future, only an optional recommendation was given for such tests. Reproduction toxicity, on the other hand, can be sensitively tested by the tests using the gastropod Potamopyrgus antipodarum, which, as invertebrate tests, do not require a permit to perform animal testing procedures. This test is also sensitive for many substances which are able to induce an endocrine disruption in vertebrates. Its validation is currently performed by the OECD. The applicability of the herein proposed test methods of the modular test platform for environmental-sample assessments were previously confirmed by several published scientific studies. Discussion We propose several test methods to be included in an application-oriented modular test platform. These tests should be suitable to efficiently indicate different mechanisms of endocrine disruption and reproduction toxicity, and may be employed in different situations according to their respective advantages. The modular test platform is able to detect impacts on the reproduction relevant effects in invertebrates, amphibians, and fishes. Furthermore the different mode of actions of estrogenic and androgenic receptor binding, steroid genesis and perhaps even the modulation of aromatase are detectable. A research report on the relevance of endocrine substances and pharmaceuticals in aquatic environments (Moltmann et al. 2007) showed that endocrine end-points of in-vivo tests (31 of 71 tested substances) tend to be more sensitive than general ecotoxicological end-points, such as mortality and growth. Consequently, endocrine and general-toxic effects should be detected in an integrative manner by ecotoxicological test platforms. Conclusions By combining literature research with a targeted query for information about the chosen test procedures it was possible to obtain a detailed overview about the current state-of-the-art of science and technology in the detection of hormone-active effects and reproduction toxicity. Ecotoxicological and regulative aspects were considered equally, and the applicability of the test procedures was evaluated. Because of the diversity of endocrine disrupting mechanisms, a modular combination of in-vivo and in-vitro methods in a joint test platform is needed to recognise and differentiate the transitions from hormone-active effects to endocrine disruption. Recommendations and perspectives Our study leads to a proposal of a modular ecotoxicological test platform, which offers an integrative detection of hormone-active and reproduction-relevant effects in the aquatic environment. The modular system presented here allows the switching between test modules according to the continuously developing state-of-the-art of science and technology as well as the incorporation of novel developments. Further standardisation of such methods for regulative applications is recommended.     

Mikrobiologisch-ökotoxikologische Bodenuntersuchungen von zwei im Pflanzenschutz eingesetzten Öl-Präparaten

Goal and Scope

Since many years mineral and plant oils have been applied with relatively high doses to control noxious insects in sevral agricultural and horticultural crops. In addition they have also been used with low doses as adjuvants (additives) together with herbicides to improve their efficacy. Their ecotoxicological effets on soil microorganisms as an important part of the soil fertility, however, have been insufficiently investigated.

Methods

Under laboratory conditions two agricultural soils were treated with a rapeseed oil and a paraffin oil as well as in some cases with biocidal reference compounds (e.g. dinitrophenol herbicides). The applied dosage were related to the concentrations in soil reached after simulated penetration of practical field amounts into upper soil layers. In some cases green manuring was simulated by adding lucerne meal to the soil. During incubation for several weeks biomass-related microbial activities (dehydrogenase activity, substrate-induced short-term respiration) as well as the mineralization of carbon and nitrogen (including nitrification) were measured. In addition some combinations of these activities were investigated.

Results and Conclusion

As compared to ‘normal’ pesticides both oil products differed in their effects on soil microorganisms. Those effects typical for strongly biocidal compounds including plant protection products, namely a dose-related inhibition of microbial biomass and stimulated mineralization (especially of nitrogen) failed with the tested oily compounds. Depending on the high input of organic carbon by these oils and their rapid decomposition a dose-dependent increase of biomass-related parameters and of the carbon mineralization occurred especially with the high doses, whereas the nitrogen mineralization was reduced or even the available nitrate content was nearly eliminated for some time by microbiological immobilization. In comparison with the reference products the two oil preparations caused only little microbiological-ecotoxicological impacts on the soil and even some stimulations of important microbial soil parameters when tested with the extended test system (including the dosages used).

Recommendation and Perspective

As a result of these investigations and of the existing literature we recommend an extension of the test spectrum for those formulated preparations and adjuvants which should be applied at high dosages or if they contain higher amounts of well decomposable organic substances. This is necessary to distinguish dose-related enhanced activities induced by biocidal effects and the subsequent mineralization of this death material (=indirect effects) from ‘real’ stimulations directly produced by the substrate.     

Rückstände ausgewählter Humanarzneimittel in Oberflächenwasserkörpern im Einzugsgebiet Halle (Saale)

A method for the determination of 16 selected pharmaceuticals in surface water using LC-MS/MS has been developed Methods from literature were adapted within the scope of this study. The limit of quantification of 13 out of 16 analytes is below 10 ng/l. The analysis of samples from the Saale river and lakes around Halle demonstrated that Carbamacepin (Median 207 ng/l) and Diclofenac (Median 196 ng/l) are detectable at various sampling stations along the river. The wastewater path was identified as the most relevant source for pharmaceutical residues entering the environment. Besides, the results reveal increased levels of several substances in some smaller anabranches and brooks.     

Wirkungsabschätzungsmethoden und Querverbindungen im Rahmen der Ökobilanz – Unterschiede zum Risk Assessment

This review defines the differences between impact assessment as component of life cycle assessment (LCA) and risk assessment. From the early LCAs, the article describes the development to the LCAs in the current meaning, deals with today’s practice of impact assessment, reviews the philosophy of impact assessment and ends with the “new” impact categories which have not yet found their way into the practice of LCA.     

Biomonitoring von Immissionswirkungen durch verkehrsbedingte Schwermetalle in einem Heideökosystem

In a part of the Lüneburger Heide situated along a motorway, the deposition of several heavy metals caused by traffic were analysed again after eleven years. As a consequence of new techniques in car production, as well as of a change in car emissions, samples of soil, plants and insects collected in 1996 showed clearly higher concentrations of lead and cadmium than those taken in 1985. For the first time, antimony and platinum were investigated. Antimony was detected in all compartments of the heathland ecosystem, while platinum was only measured in mosses, lichens and litter. Mosses were the best indicators to evaluate the impact of the ecosystem by particular emissions from traffic.     

Biologische Effekte von Hexachlorbenzol (HCB) in aquatischen Modellökosystemen

A long-term study war performed on the fate and effects of hexachlorobenzene (HCB) in aquatic mesocosms. Discontinuous dosing of the sparingly soluble test substance up to the limit of its water solubility (5 μg/l at 20°C) did not cause any significant effects on the aquatic communities. From this point of view, a quality objective of 5 μg/l is considered to maintain the structures and functions of aquatic communities on a long-term scale.     

Ökonomische Ansätze zur Ökosystembewertung am Beispiel des Bodens

As it is practically impossible in an industrial society to reduce impacts into ecosystems to a level that would preclude any damages, the need for damage valuation arises. One of the available valuation tools is the economic approach. Subsequent publications present this approach using the example of soils. In the first part, soils as part of ecosystems are considered from an ecological and a conventional economic point of view. In the following so-called ecological-economic perspective, more recent developments in economic valuation research are introduced. It is shown how ecological and economic valuation can complement one another. It is emphasized that economic valuations should be restricted to a critical soil structure which, in economic terms, is determined by the non-substitutable services of the ecological asset.     

Kosovo-Konflikt: Ökologische Auswirkungen

10 years ago, the WWF (World Wide Fund for Nature) began to support the work of environmental protection in the countries of Eastern Europe. A main topic is the Danube-Carpathian Program. With the beginning of the bombardements, connected with great amounts of chemicals released directly and as clouds from the fires and explosions, the question raises: What are the consequences for mankind and nature? Is there a risk for the Danube-Carpathian Program? The WWF feels responsible for reporting about these consequences. In July 1999, a group of two Oil Experts and two scientists (each a chemist and a toxicologist) visited Belgrad, Pan?evo and Novi Sad, took samples of soil and water, and estimated the contents of dangerous substances. The first findings and the first consequences for nature and human health are described.     

Ökologische Aspekte des Phosphor-Kreislaufs

In contrast to the complex N-cycle with its high number of reactive compounds, the global P-cycle is uncomplicated. There is only one reactive compound (PO₄ ^3?), and the atmosphere is practically not involved. Nevertheless, severe ecological imbalances may be induced by anthropogenic perturbations of the P-cycle, especially in aquatic ecosystems. The present knowledge on the global P-cycle (reservoirs, fluxes, residence times) is summarized and the ecological dimension of increased phosphorus inputs into surface waters is discussed in detail. Exemplary for Germany, the role of agriculture is described in this context. The fact that the rock phosphate resources are principally limited and not renewable is of a high ecological and economical relevance. The expected lifetime of the P-reserves is calculated for different estimate-scenarios concerning the magnitude of reserves and consumption. Measures to reduce the P-surplus in agriculture, to increase the efficiency of P-fertilization, and to recycle P more consequently are therefore highly required.     

Ökologische Situation der Region Leipzig-Halle

Air pollution in the region Leipzig-Halle was investigated using two-year-old pine needles (Pinus sylvestris L.) at 39 sites of a biomonitoring net covering an area of ca. 7 500 km². The concentration values of Al, Ca, Cr, Fe, K, Mg, Mn, Pb and V characterize the immission patterns of the period 1990/91. The method is less suited for Cu, Ni and Zn due to the transfer route soil-pine, and for Cd due to the small total content. There are significant spatial and element-specific variations which require distinct evaluation of the pollution situation. The city of Leipzig itself is a significant source for Pb, Cr, Fe and V emission. Mainly as far as the centre of the region is concerned, the Pb concentration values are beyond those of heavily contaminated reference sites. The bioindication system is also applicable for comparable regions in Eastern Europe where the pine is the dominating conifer type. Immission trends can be monitored over ecologically relevant periods of time.     

Ökotoxikologische Charakterisierung von Nonylphenol Isomeren

Background, Aim and Scope

Acute toxicity and estrogenic potency of six nonylphenol isomers, with nearly the same chemical structure and the same physico-chemical properties, were investigated in this study. Thes datasets give some insight into how far structure activity relationships can be used for these modes of action.

Methods

The toxicity of the isomers was investigated by the daphnia immobilisation test as well as the duckweed growth inhibition test. The estrogenic potency of the p-NP isomers was analyzed by MVLN-Reportergen-Assay.

Results

No differences between the isomers were found for baseline toxicity, neither in the daphnia immobilisation test nor in the duckweed growth inhibition test. Estrogenic potency of the isomers in the MVLN reportergen assay was markedly different for the tested isomers. The highest estrogenic potency was found for p353-NP, whereas p262-NP and p22-NP exhibited no estrogenic, but partial antagonistic potency in this assay.

Discussion

The results from this study clearly indicate how far simplification and structure activity relationships can be used in risk analysis. For bioconcentration and baseline toxicity structure activity relationships are possible and will allow simplification of the risk analysis. For special modes of action simple structure activity relationships are per se impossible. This was shown especially for the estrogen receptor interactions, where only small structural differences can lead to different modes of action.

Conclusions

It seems possible to reduce the effort for risk analysis, using combinations of measured data and predictive models. At the moment it seems unlikely that predictive models alone will and can be used for risk analysis.