In the context of ecological risk assessment of chemicals, individual-based population models hold great potential to increase the ecological realism of current regulatory risk assessment procedures. However, developing and parameterizing such models is time-consuming and often ad hoc. Using standardized, tested submodels of individual organisms would make individual-based modelling more efficient and coherent. In this thesis, I explored whether Dynamic Energy Budget (DEB) theory is suitable for being used as a standard submodel in individual-based models, both for ecological risk assessment and theoretical population ecology. First, I developed a generic implementation of DEB theory in an individual-based modeling (IBM) context: DEB-IBM. Using the DEB-IBM framework I tested the ability of the DEB theory to predict population-level dynamics from the properties of individuals. We used Daphnia magna as a model species, where data at the individual level was available to parameterize the model, and population-level predictions were compared against independent data from controlled population experiments. We found that DEB theory successfully predicted population growth rates and peak densities of experimental Daphnia populations in multiple experimental settings, but failed to capture the decline phase, when the available food per Daphnia was low. Further assumptions on food-dependent mortality of juveniles were needed to capture the population dynamics after the initial population peak. The resulting model then predicted, without further calibration, characteristic switches between small- and large-amplitude cycles, which have been observed for Daphnia. We conclude that cross-level tests help detecting gaps in current individual-level theories and ultimately will lead to theory development and the establishment of a generic basis for individual-based models and ecology. In addition to theoretical explorations, we tested the potential of DEB theory combined with IBMs to extrapolate effects of chemical stress from the individual to population level. For this we used information at the individual level on the effect of 3,4-dichloroanailine on Daphnia. The individual data suggested direct effects on reproduction but no significant effects on growth. Assuming such direct effects on reproduction, the model was able to accurately predict the population response to increasing concentrations of 3,4-dichloroaniline. We conclude that DEB theory combined with IBMs holds great potential for standardized ecological risk assessment based on ecological models. / Für die ökologische Risikobewertung von Chemikalien sind individuenbasierte Populationsmodelle ein vielversprechendes Werkzeug um heutige Bewertungen ökologisch realistischer zu gestalten. Allerdings ist die Entwicklung und Parametrisierung derartiger Modelle zeitaufwendig und oft wenig systematisch. Standardisierte, geprüfte Untermodelle, die Einzelorganismen beschreiben, würden die individuenbasierte Modellierung effizienter und kohärenter machen. In meiner Dissertation habe ich daher untersucht, inwieweit sich die Dynamic Energy Budget-Theorie (DEB) als Standardmodell innerhalb individuenbasierter Populationsmodelle eignet, und zwar sowohl für die ökologische Risikobewertung als auch für die theoretische Populationsökologie. Zunächst habe ich eine generische Implementierung der DEB-Theorie im Rahmen individuenbasierter Modellen (IBM) erstellt: DEB-IBM. Dieses Werkzeug nutzend habe ich dann untersucht, ob es mit Hilfe der DEB-Theorie gelingt, ausgehend von den Eigenschaften und Aktivitäten einzelner Individuen, Populationsdynamik vorherzusagen. Wir nutzten dabei Daphnia magna als Modellart, für die Daten auf der Individuenebene verfügbar waren, um das Modell zu parametrisieren, sowie Populationsdaten, mit denen Modellvorhersagen verglichen werden konnten. DEB-Theorie war in der Lage, beobachtete Populationswachstumsraten sowie die maximalen Abundanzen korrekt vorherzusagen, und zwar für verschiedene Umweltbedingungen. Für Phasen des Rückgangs der Population allerdings, wenn die für die Daphnien verfügbare Nahrungsmenge gering war, kam es zu Abweichungen. Es waren deshalb zusätzliche Annahmen über nahrungsabhängige Sterblichkeit von juvenilen Daphnien erforderlich, um die gesamte Populationsdynamik korrekt vorherzusagen. Das resultierende Modell konnte dann, ohne weitere Kalibrierungen, den für Daphnien charakteristischen Wechsel zwischen Populationszyklen mit großen und kleinen Amplituden richtig vorhersagen. Wir folgern daraus, daß Ebenen übergreifende Tests dabei helfen, Lücken in aktuellen Theorien über Einzelorganismen aufzudecken Dies trägt zur Theorieentwicklung bei und liefert Grundlagen für individuenbasierte Modellierung und Ökologie. Über diese Grundlagenfragen hinaus haben wir überprüft, ob DEB-Theorie in Kombination mit IBMs es ermöglicht, den Effekt von chemischem Streß auf Individuen auf die Populationsebene zu extrapolieren. Wir nutzten Daten über die Auswirkungen von 3,4 Dichloroanalin auf einzelne Daphnien, die zeigten daß im Wesentlichen die Reproduktion, nicht aber das Wachstum beeinträchtigt ist. Mit entsprechenden Annahmen konnte unser Modell den Effekt auf Populationsebene, für den unabhängige Daten vorlagen, korrekt vorhersagen. DEB-Theorie in Kombination mit individuenbasierter Modellierung birgt somit großes Potential für einen standardisierten modellbasierten Ansatz in der ökologischen Risikobewertung von Chemikalien.

Triclosan (TCS) is a personal care product with many fields of application and is of public interest for several years now. Monitoring studies showed that TCS is a ubiquitous chemical in the aquatic environment. Aquatic organisms are exposed to TCS in a broad range of concentrations, from ng L-1 up to lower μg L-1. TCS has a bactericidal effect for various types of gram-positive and gram negative bacteria. TCS targets a specific bacterial fatty acid biosynthetic enzyme, enoyl-[acyl-carrier protein] reductase (Schweizer, 2001). Therefore the terminal reaction in the fatty acid elongation cycle is inhibited (Levy et al., 1999). Although effects on non-target organisms are reported, the Mode of Action (MoA) of TCS is not well examined for those organisms. The aim of this PhD thesis was to investigate effects of TCS on non-target autotrophic organisms at different levels of biological complexity in the aquatic environment. In this thesis microalgae have been found to be very sensitive to TCS. In some cases even higher sensitivities than in bacteria were observed, which is in accordance with published effect data (Harada et al., 2008; Orvos et al., 2002). Similarly to bacteria, high species sensitivity differences were observed for algae (Franz et al., 2008). In bacteria these sensitivity differences can be ascribed to several resistance mechanisms reported in Schweizer (2001). These findings lead to the question about the reasons for species sensitivity differences in algae. A mesocosm study was performed to detect effects of TCS across levels of biological organization and to investigate the impact of sensitivity differences on complex aquatic communities. For that purpose, structural and functional effects parameters were observed.

Um die Effekte von Umweltchemikalien auf Populationen von Fließgewässerorganismen zu untersuchen, sind an der TU Dresden Fließrinnen entwickelt worden. Die Komplexität wurde bewusst reduziert, um die beobachteten Effekte besser kausal analysieren zu können. Nach einer Erweiterung des Nahrungsnetzes um Grazer (Eintagsfliegen-Larven) und der weiteren Optimierung der Methode zur Bestimmung des Aufwuchses (Periphyton) wurden 2 Experimenten mit einem Pflanzenschutzmittel (Fenoxycarb) und einer Industriechemikalie (Bisphenol A) durchgeführt, um die Effekte dieser Chemikalien auf den Aufwuchs, die Eintagsfliegen-Larven Rhithrogena semicolorata und Seratella ignita, sowie Oligochaeten der Art Lumbriculus variegatus zu erfassen. Die Wirkung von Fenoxycarb in 4 Konzentrationen von 0,05 bis 50 µg L-1 wurde über eine Dauer von 101 Tagen untersucht. Auf die physiko-chemischen Parameter, den Aufwuchs und L. variegatus wurden keine Effekte festgestellt. Bei R. semicolorata wurden Effekte auf das Überleben und die Emergenz beobachtet. Für den Endpunkt "Anzahl der toten Larven in der Fließrinne" wurde eine LC50 von 3,3 µg L-1 bestimmt. Die akute Toxizität (48 h LC50) von Fenoxycarb für S. ignita ist größer als 5 mg L-1. Bei Larven von S. ignita wurden 72 Tage nach Applikation Effekte beobachtet werden, die dem Wirkmechanismus von Fenoxycarb entsprechen. In der höchsten Konzentration zeigten ca. 80 % der emergierten Tiere morphologische Abnormalitäten an 4 Segmenten des Abdomens. Die Exposition erfolgte möglicherweise durch an Aufwuchs bzw. Detritus gebundene Substanz oder deren Metabolite, da die Substanz 19 Tage nach Applikation im Wasser nicht mehr nachweisbar war. Im zweiten Experiment wurde Wirkung von Bisphenol A in 3 Konzentrationen von 5 bis 500 µg L-1 (effektiv 0,24 bis 24,1 µg L-1) über eine Dauer von 103 Tagen untersucht. Auf die physiko-chemischen Parameter wurde kein Einfluss festgestellt. Der Aufwuchs in der höchsten Konzentration erreichte nur die Hälfte des Biomasse-Maximums der Kontrolle. Die EC10 beträgt 1,8 und die EC50 21,7 µg L-1 (effektiv). Das Populationswachstum von Lumbriculus variegatus war in den beiden höchsten Konzentrationen im Vergleich zur Kontrolle um ca. 50 % reduziert. Die akute Toxizität (LC50 96 h) für L. variegatus beträgt 5,3 mg L-1. Für das Populationswachstum in den Fließrinnen wurde eine EC10 von 0,11 µg L-1 (effektiv) ermittelt, die um den Faktor 48184 unter der akuten Toxizität liegt. - Anlage: Rohdaten (0,53 MB)- Nutzung: Referat Informationsvermittlung der SLUB"

In the European Union an environmental risk assessment is required for the registration of new chemicals, biocides, pesticides and pharmaceuticals. In order to avoid the release of potential hazardous substances, various ecotoxicity tests are performed, including acute and chronic fish tests. As a consequence of the new program of the European Union “Registration, Evaluation and Authorisation of Chemicals” (REACH) the number of animal experiments for environmental risk assessment is expected to increase remarkably within the next years. On the other hand there is a strong societal demand for reducing the number of animal tests by using alternative in vitro models. According to EU directives, investigations using non-human vertebrate embryos are considered pain free in vitro methods and are therefore accepted as alternatives to animal experiments. For the acute fish test, the Danio rerio embryo test (DarT) has been established as a replacement method and included in national regulations at least for waste water (German Waste Water Dues Law). However, no alternatives for chronic fish tests are currently available. The overall goal of this thesis was to work towards such a replacement by extending DarT zu Gene-DarT. Toxicants will initially interact at the molecular level with consequences for physiology, fitness and survival. The analysis of gene expression patterns may unravel elements of these molecular events before any phenotypic changes are visible. The hypothesis of this thesis therefore was that chemical-sensitive genes in embryos exposed in a conventional DarT may indicate toxic impact of substances at sub-acute concentrations and thus enhance the sensitivity of the embryo toxicity test. Furthermore, unlike the conventional DarT-endpoints, gene expression analysis will provide insights into mechanistic processes underlying toxicity. The 3,4-dichloroaniline (3,4-DCA), which is used as a reference compound in the DarT, was selected as model chemical in this thesis. In a first step, differentially expressed genes in embryos exposed to 3,4-DCA were identified by microarray technology and RT-PCR techniques. Six dose-dependent significant differentially expressed genes were identified. These genes were involved in biotransformation pathways (cyp1a, ahr2), stress response (nrf2, maft, ho-1) and cell cycle control (fzr1). Differential expression upon 3,4-DCA exposure was detected below the LOEC (lowest observed effect concentration = 6.2 µM) of survival or developmental disorders of the embryo test (0.78 µM and above). For the validation of stage specific sensitivity, genes were also analysed in post-hatched stages. Extension of exposure to post-hatched stages resulted in a differential expression at lower concentrations as for the embryonic stages, indicating an improved sensitivity due to stage-specific sensitivity or exposure time. To confirm the adaptive function of the 3,4-DCA-sensitive genes, embryonic mRNA abundance was experimentally manipulated by knock down and overexpression. By injection of sense (mRNA) or antisense (siRNA) RNA in one-cell-stages of embryos, the transcript levels of genes were transiently enhanced or repressed in embryos exposed to 3,4-DCA. mRNA injection of the genes cyp1a, ho-1 and nrf2 reduced the number of embryos with 3,4-DCA-induced malformations. In contrast, siRNA injections for the same genes led to an increase in the severity and frequency of developmental disorders. The results clearly indicate the adaptive functions of the investigated genes or their corresponding proteins. This study demonstrates that the analysis of chemical-sensitive gene expression shows the potential to increase the sensitivity of conventional toxicity tests. The analysis of gene expression also provides additional mechanistic information for toxic action, e.g. in the presented study, the involvement of Ah-receptor regulated pathways as an adaptive response. Furthermore, the presented data indicate that functional manipulations, using mRNA and siRNA-injection, are suitable to evaluate the role of differentially expressed genes for toxicity.

Among all nanomaterials, which are intentionally manufactured and applied, nanosilver (nAg) is one of the most frequently applied nanomaterials. It is introduced into wastewater treatment plants (WWTPs) due to its use as antimicrobial resource in household and medical care products and hence concern raised regarding its impact on activated sludge organisms and their purification efficiency. Within this thesis, the effects of nAg on single species, simple food chains and communities related to activated sludge were investigated. Among all tested species in this thesis, the gammaproteobacteria R. planticola was the most sensitive organism regarding the tested nAg material, NM-300K. The environmental risk assessment (ERA), based on an assumed predicted environmental concentration derived from modeled concentrations of other types of nAg, revealed no risk for the activated sludge. This result should be interpreted with care, considering the tolerantly chosen safety factor for calculation of the predicted no effect concentration and the assumptions which were made concerning environmental concentrations. Differences in acute toxic effects of nAg on the ciliate P. tetraurelia were observed depending on the type of medium and the exposure pathway (via medium or via bacterial food). More detailed investigations are required to analyze the distribution, availability and uptake of nAg into ciliates in the respective tests. In chronic experiments concentration response was very steep in the range between the effect concentration determined in acute toxicity testing (resulting in 100% mortality) and a tenfold lower concentration (no effect observed) for both exposure pathways. Community experiments with activated sludge exposed to realistic and high concentrations of nAg revealed acute effects on the protozoan community at high nAg concentration using multivariate statistics for data analysis. In contrast, the sludge biotic index was not meaningful for data interpretation, as no differences were observed between the samples of different treatments. For chronic testing, more preliminary work is required to develop a protocol for artificial wastewater which serves the needs of activated sludge organisms over longer time periods and which retains a typical composition of the activated sludge community.

Um die Effekte von Umweltchemikalien auf Populationen von Fließgewässerorganismen zu untersuchen, sind an der TU Dresden Fließrinnen entwickelt worden. Die Komplexität wurde bewusst reduziert, um die beobachteten Effekte besser kausal analysieren zu können. Nach einer Erweiterung des Nahrungsnetzes um Grazer (Eintagsfliegen-Larven) und der weiteren Optimierung der Methode zur Bestimmung des Aufwuchses (Periphyton) wurden 2 Experimenten mit einem Pflanzenschutzmittel (Fenoxycarb) und einer Industriechemikalie (Bisphenol A) durchgeführt, um die Effekte dieser Chemikalien auf den Aufwuchs, die Eintagsfliegen-Larven Rhithrogena semicolorata und Seratella ignita, sowie Oligochaeten der Art Lumbriculus variegatus zu erfassen. Die Wirkung von Fenoxycarb in 4 Konzentrationen von 0,05 bis 50 µg L-1 wurde über eine Dauer von 101 Tagen untersucht. Auf die physiko-chemischen Parameter, den Aufwuchs und L. variegatus wurden keine Effekte festgestellt. Bei R. semicolorata wurden Effekte auf das Überleben und die Emergenz beobachtet. Für den Endpunkt "Anzahl der toten Larven in der Fließrinne" wurde eine LC50 von 3,3 µg L-1 bestimmt. Die akute Toxizität (48 h LC50) von Fenoxycarb für S. ignita ist größer als 5 mg L-1. Bei Larven von S. ignita wurden 72 Tage nach Applikation Effekte beobachtet werden, die dem Wirkmechanismus von Fenoxycarb entsprechen. In der höchsten Konzentration zeigten ca. 80 % der emergierten Tiere morphologische Abnormalitäten an 4 Segmenten des Abdomens. Die Exposition erfolgte möglicherweise durch an Aufwuchs bzw. Detritus gebundene Substanz oder deren Metabolite, da die Substanz 19 Tage nach Applikation im Wasser nicht mehr nachweisbar war. Im zweiten Experiment wurde Wirkung von Bisphenol A in 3 Konzentrationen von 5 bis 500 µg L-1 (effektiv 0,24 bis 24,1 µg L-1) über eine Dauer von 103 Tagen untersucht. Auf die physiko-chemischen Parameter wurde kein Einfluss festgestellt. Der Aufwuchs in der höchsten Konzentration erreichte nur die Hälfte des Biomasse-Maximums der Kontrolle. Die EC10 beträgt 1,8 und die EC50 21,7 µg L-1 (effektiv). Das Populationswachstum von Lumbriculus variegatus war in den beiden höchsten Konzentrationen im Vergleich zur Kontrolle um ca. 50 % reduziert. Die akute Toxizität (LC50 96 h) für L. variegatus beträgt 5,3 mg L-1. Für das Populationswachstum in den Fließrinnen wurde eine EC10 von 0,11 µg L-1 (effektiv) ermittelt, die um den Faktor 48184 unter der akuten Toxizität liegt. - Anlage: Rohdaten (0,53 MB)- Nutzung: Referat Informationsvermittlung der SLUB"

Among all nanomaterials, which are intentionally manufactured and applied, nanosilver (nAg) is one of the most frequently applied nanomaterials. It is introduced into wastewater treatment plants (WWTPs) due to its use as antimicrobial resource in household and medical care products and hence concern raised regarding its impact on activated sludge organisms and their purification efficiency. Within this thesis, the effects of nAg on single species, simple food chains and communities related to activated sludge were investigated. Among all tested species in this thesis, the gammaproteobacteria R. planticola was the most sensitive organism regarding the tested nAg material, NM-300K. The environmental risk assessment (ERA), based on an assumed predicted environmental concentration derived from modeled concentrations of other types of nAg, revealed no risk for the activated sludge. This result should be interpreted with care, considering the tolerantly chosen safety factor for calculation of the predicted no effect concentration and the assumptions which were made concerning environmental concentrations. Differences in acute toxic effects of nAg on the ciliate P. tetraurelia were observed depending on the type of medium and the exposure pathway (via medium or via bacterial food). More detailed investigations are required to analyze the distribution, availability and uptake of nAg into ciliates in the respective tests. In chronic experiments concentration response was very steep in the range between the effect concentration determined in acute toxicity testing (resulting in 100% mortality) and a tenfold lower concentration (no effect observed) for both exposure pathways. Community experiments with activated sludge exposed to realistic and high concentrations of nAg revealed acute effects on the protozoan community at high nAg concentration using multivariate statistics for data analysis. In contrast, the sludge biotic index was not meaningful for data interpretation, as no differences were observed between the samples of different treatments. For chronic testing, more preliminary work is required to develop a protocol for artificial wastewater which serves the needs of activated sludge organisms over longer time periods and which retains a typical composition of the activated sludge community.

Triclosan (TCS) is a personal care product with many fields of application and is of public interest for several years now. Monitoring studies showed that TCS is a ubiquitous chemical in the aquatic environment. Aquatic organisms are exposed to TCS in a broad range of concentrations, from ng L-1 up to lower μg L-1. TCS has a bactericidal effect for various types of gram-positive and gram negative bacteria. TCS targets a specific bacterial fatty acid biosynthetic enzyme, enoyl-[acyl-carrier protein] reductase (Schweizer, 2001). Therefore the terminal reaction in the fatty acid elongation cycle is inhibited (Levy et al., 1999). Although effects on non-target organisms are reported, the Mode of Action (MoA) of TCS is not well examined for those organisms. The aim of this PhD thesis was to investigate effects of TCS on non-target autotrophic organisms at different levels of biological complexity in the aquatic environment. In this thesis microalgae have been found to be very sensitive to TCS. In some cases even higher sensitivities than in bacteria were observed, which is in accordance with published effect data (Harada et al., 2008; Orvos et al., 2002). Similarly to bacteria, high species sensitivity differences were observed for algae (Franz et al., 2008). In bacteria these sensitivity differences can be ascribed to several resistance mechanisms reported in Schweizer (2001). These findings lead to the question about the reasons for species sensitivity differences in algae. A mesocosm study was performed to detect effects of TCS across levels of biological organization and to investigate the impact of sensitivity differences on complex aquatic communities. For that purpose, structural and functional effects parameters were observed.

In the European Union an environmental risk assessment is required for the registration of new chemicals, biocides, pesticides and pharmaceuticals. In order to avoid the release of potential hazardous substances, various ecotoxicity tests are performed, including acute and chronic fish tests. As a consequence of the new program of the European Union “Registration, Evaluation and Authorisation of Chemicals” (REACH) the number of animal experiments for environmental risk assessment is expected to increase remarkably within the next years. On the other hand there is a strong societal demand for reducing the number of animal tests by using alternative in vitro models. According to EU directives, investigations using non-human vertebrate embryos are considered pain free in vitro methods and are therefore accepted as alternatives to animal experiments. For the acute fish test, the Danio rerio embryo test (DarT) has been established as a replacement method and included in national regulations at least for waste water (German Waste Water Dues Law). However, no alternatives for chronic fish tests are currently available. The overall goal of this thesis was to work towards such a replacement by extending DarT zu Gene-DarT. Toxicants will initially interact at the molecular level with consequences for physiology, fitness and survival. The analysis of gene expression patterns may unravel elements of these molecular events before any phenotypic changes are visible. The hypothesis of this thesis therefore was that chemical-sensitive genes in embryos exposed in a conventional DarT may indicate toxic impact of substances at sub-acute concentrations and thus enhance the sensitivity of the embryo toxicity test. Furthermore, unlike the conventional DarT-endpoints, gene expression analysis will provide insights into mechanistic processes underlying toxicity. The 3,4-dichloroaniline (3,4-DCA), which is used as a reference compound in the DarT, was selected as model chemical in this thesis. In a first step, differentially expressed genes in embryos exposed to 3,4-DCA were identified by microarray technology and RT-PCR techniques. Six dose-dependent significant differentially expressed genes were identified. These genes were involved in biotransformation pathways (cyp1a, ahr2), stress response (nrf2, maft, ho-1) and cell cycle control (fzr1). Differential expression upon 3,4-DCA exposure was detected below the LOEC (lowest observed effect concentration = 6.2 µM) of survival or developmental disorders of the embryo test (0.78 µM and above). For the validation of stage specific sensitivity, genes were also analysed in post-hatched stages. Extension of exposure to post-hatched stages resulted in a differential expression at lower concentrations as for the embryonic stages, indicating an improved sensitivity due to stage-specific sensitivity or exposure time. To confirm the adaptive function of the 3,4-DCA-sensitive genes, embryonic mRNA abundance was experimentally manipulated by knock down and overexpression. By injection of sense (mRNA) or antisense (siRNA) RNA in one-cell-stages of embryos, the transcript levels of genes were transiently enhanced or repressed in embryos exposed to 3,4-DCA. mRNA injection of the genes cyp1a, ho-1 and nrf2 reduced the number of embryos with 3,4-DCA-induced malformations. In contrast, siRNA injections for the same genes led to an increase in the severity and frequency of developmental disorders. The results clearly indicate the adaptive functions of the investigated genes or their corresponding proteins. This study demonstrates that the analysis of chemical-sensitive gene expression shows the potential to increase the sensitivity of conventional toxicity tests. The analysis of gene expression also provides additional mechanistic information for toxic action, e.g. in the presented study, the involvement of Ah-receptor regulated pathways as an adaptive response. Furthermore, the presented data indicate that functional manipulations, using mRNA and siRNA-injection, are suitable to evaluate the role of differentially expressed genes for toxicity.

Gewässer sind Lebensgrundlage, jedoch gleichzeitig Schadstoffsenken für eine Vielzahl von Kontaminanten. Biologische Wirkungstests und das Biomonitoring aquatischer Proben sind daher besonders wichtig, um Umwelt-Gefahrenpotenziale erkennen zu können. Der "Comet Assay" (Einzelzell-Gelelektrophorese) ist ein Indikator von DNA-Strangbrüchen und wurde hier als Test auf gentoxische Wirkungen erprobt und angewandt. Mit bekannten, gentoxischen Substanzen wurden Nachweisgrenzen und Dosis-Wirkungs-Beziehungen für die Zelllinien RTG-2 und RTL-W1 (aus der Regenbogenforelle, Oncorhynchus mykiss) in vitro ermittelt und methodische Parameter an die Zellen angepasst. Der Test reagierte sehr sensitiv auf 4-Nitrochinolin-1-oxid. Die Substanz war daher geeignet, um in weiteren Versuchen als Positivkontrolle zu dienen. Zur Bewertung der Messdaten wurde ein geeignetes statistisches Verfahren gefunden, das auch historische Kontrollen mit einbezog. Der zeitliche Verlauf der DNA-Schädigung des Testsystems mit RTG-2-Zellen wurde ermittelt, und durch Inhibition der DNA-Reparatur mit Aphidicolin wurden Zusammenhänge zwischen der Entstehung von DNA-Strangbrüchen, der DNA-Reparaturkapazität sowie der Metabolisierungskapazität untersucht. In einer zweiten Phase wurden unbehandelte Wasserproben aus Rhein, Elbe sowie weitere Oberflächenwasserproben mit dem Comet Assay an RTG-2-Zellen getestet. Bei 15 von 49 Proben zeigten sich gentoxische Effekte. In einer dritten Phase wurden Erythrozyten von freilebenden Döbeln, Leuciscus cephalus, aus der Mosel mit dem Comet Assay untersucht. Die Fische von drei Messstellen zeigten erhöhte Werte von DNA-Schädigungen, gegenüber einer vierten, stromabwärts gelegenen Messstation. Korrelationen mit den Ergebnissen zusätzlicher Biomarker ergaben sich nur teilweise. Chemische Analysen von Wasser- oder Gewebeproben ließen keine Rückschlüsse auf verursachende Kontaminanten zu - gerade dies unterstreicht jedoch die Wichtigkeit biologischer Tests bei komplexen Proben. / Bodies of Water are both vital resources and pollutant sinks for a multitude of contaminants. Therefore, biological effect tests and biomonitoring of aquatic samples are of particular importance to detect potential environmental hazards. The "comet assay" (single cell gel electrophoresis) is an indicator for DNA strand breaks and was explored and applied as a genotoxicity test in the present study. Known genotoxic substances were used to determine the detection limits and dose-response relationships for the cell lines RTG-2 and RTL-W1 (from rainbow trout, Oncorhynchus mykiss) in vitro, and to adapt methodological parameters to the cells. The test was very sensitive to 4-Nitroquinoline-1-oxide. This substance was therefore well-suited to serve as positive control in further experiments. In order to evaluate the measurement data, an appropriate statistical procedure was developed, which also took "historical" controls into account. The time course of DNA damage in the test system using RTG-2 cells was determined, and relationships between the origin of DNA strand breaks, DNA repair capacity and the metabolizing capacity of the cells was investigated by means of inhibition of DNA repair with Aphidicoline. In the second stage, native water samples from the rivers Rhine and Elbe and further surface waters were tested with the comet assay, using RTG-2 cells. 15 out of 49 samples showed genotoxic effects. In a third stage, erythrocytes of feral chub, Leuciscus cephalus, from the Moselle river were examined with the comet assay. The fish from three measuring stations showed elevated values of DNA damage compared to fish sampled from a downstream station. There were only partly correlations with the results from additional biomarkers. Chemical analyses of water and tissue samples did not permit conclusions on effect-causing substances.However, this emphasizes the importance of biological tests in dealing with complex environmental samples.