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Institute
- Fachbereich 7 (8) (remove)
Studies on the toxicity of chemical mixtures find that components at levels below no-observed-effect concentrations (NOECs) may cause toxicity resulting from the combined effects of mixed chemicals. However, chemical risk assessment frequently focuses on individual chemical substances, although most living organisms are substantially exposed to chemical mixtures rather than single substances. The concepts of additive toxicity, concentration addition (CA), and independent action (IA) models are often applied to predict the mixture toxicity of similarly and dissimilarly acting chemicals, respectively. However, living organisms and the environment may be exposed to both types of chemicals at the same time and location. In addition, experimental acquisition of toxicity data for every conceivable mixture is unfeasible since the number of chemical combinations is extremely large. Therefore, an integrated model to predict mixture toxicity on the basis of single mixture components having various modes of toxic action (MoAs) needs to be developed. The objectives of the present study were to analyze the challenges in predicting mixture toxicity in the environment, and to develop integrated models that overcome the limitations of the existing prediction models for estimating the toxicity of non-interactive mixtures through computational models. For these goals, four sub-topics were generated in this study. Firstly, applicable domains and limitations of existing integrated models were analyzed and grouped into three kinds of categories in this study. There are current approaches used to assess mixture toxicity; however, there is a need for a new research concept to overcome challenges associated with such approaches, which recent studies have addressed. These approaches are discussed with particular emphasis on those studies involved in computational approaches to predict the toxicity of chemical mixtures based on the toxicological data of individual chemicals. Secondly, through a case study and a computational simulation, it was found that the Key Critical Component (KCC) and Composite Reciprocal (CR) methods (as described in the European Union (EU) draft technical guidance notes for calculating the Predicted No Effect Concentration (PNEC) and Derived No Effect Level (DNEL) of mixtures) could derive significantly different results. As the third and fourth sub-topics of this study, the following two integrated addition models were developed and successfully applied to overcome the inherent limitations of the CA and IA models, which could be theoretically used for either similarly or dissimilarly acting chemicals: i) a Partial Least Squares-Based Integrated Addition Model (PLS-IAM), and, ii) a Quantitative Structure-Activity Relationship-Based Two-Stage Prediction (QSAR-TSP) model. In this study, it was shown that the PLS-IAM might be useful to estimate mixture toxicity when the toxicity data of similar mixtures having the same compositions were available. In the case of the QSAR-TSP model, it showed the potential to overcome the critical limitation of the conventional TSP model, which requires knowledge of the MoAs for all chemicals. Therefore, this study presented good potential for the advanced integrated models (e.g., PLS-IAM and QSAR-TSP), while considering various non-interactive constituents that have different MoAs in order to increase the reliance of conventional models and simplify the procedure for risk assessment of mixtures.
Structure of soil organic matter (SOM) is a hot topic of discussion among scientific community for several decades. The mostly discussed models, among many, are polymer model and supramolecular model. While the former considers SOM as macromolecules consisting of amorphous and crystalline domains, the latter explains SOM as a physicochemical entity dominated by weak hydrophobic and H-bond interactions in the secondary level, which holds individual molecules of primary structure together. The weak forces in secondary level impart characteristic mobility of SOM. Very important consequence of this multidimensional formulation is that physicochemical structure plays a crucial role in most biogeochemical functions of SOM, apart from the chemical composition. Recently introduced concept of cation and water molecule mediated bridges between OM molecular segments (CaB and WaMB, respectively) evolved from physicochemical understanding of SOM structure. Even though several indirect evidences were produced for CaB and WaMB during last years, no clear-cut understanding of these processes has been achieved yet. Experimental difficulty due to overlapping effects of equally important CaB-governing parameters such a pH and competing cations raises huge challenge in investigating CaB-related influences. This thesis, therefore, aims to validate an experimental set-up for inducing CaB within OM structures and assessing it from various chemical and physicochemical aspects.
The method involved removal of omnipresent cations and adjustment of pH before cation addition. This helped to separate pH effects and cation effects. Based on results obtained on two different types of organic matter, it can be deduced that multivalent cations can cross-link SOM, given that functional group density of the SOM material is enough for the functional groups to be arranged in sufficient spatial proximity to each other. Physicochemical structural reorganisation during aging causes formation of more and/or stronger CaB and WaMB. As for inducing CaB directly after cation treatment, cationic size and valency were found determinant also for aging effect. A strongly cross-linked system in the beginning is less vulnerable to structural changes and undergoes aging with lower intensity, than an initially weakly cross-linked system. Responsible for the structural changes is, the inherent mobility of SOM within its physicochemical assemblage. Thus the information on structural requirement of CaB and its consequences on OM matrix rigidity will help to obtain insight into the physicochemical SOM structure. Additionally, organic matter quality (assessed by thermal analysis) and pore structure of SOM formed in a set of artificial soils showed that mineral materials are important for the chemical nature of SOM molecules, but not for the physical structure of organo-mineral associations, at least after several months of SOM development.
Furthermore, nanothermal analysis using atomic force microscopy (AFM-nTA) was implemented in soils for the first time to reveal nanoscale thermal properties and their spatial distribution in nano- and micrometer scales. This helped to identify physicochemical processes, such as disruption of WaMB, in low-organic soils, in which bulk methods fail due to their low sensitivity. Further, various types of materials constituting in soils were distinguished with high resolution by advanced application of the method, in combination with other AFM parameters. Attempts were done to identify various materials, with the usage of defined test materials. Above all, the method is potent to reveal microspatial heterogeneity on sample surfaces, which could help understanding process-relevant hotspots, for example.
This thesis thus contributes to the scientific understanding on physicochemical structural dynamics via cross-linking by cations and via nanoscale thermal properties. Direct investigation on CaB demonstrated here will potentially help making a big leap in knowledge about the interaction. The observed aging effects add well to the understanding of supramolecular consideration of SOM. By introducing nanothermal analysis to the field of soil science, it is made possible to face the problem of heterogeneity and spatial distribution of thermal characteristics. Another important achievement of AFM-nTA is that it can be used to detect physicochemical processes, which are of low intensity.
The polysaccharide hydration phenomenon is nowadays the subject of intense research. The interaction of native and modified polysaccharides and polysaccharides-based bioconjugates with water has an important influence on their functional behaviour. Notwithstanding that the hydration phenomenon has been studied for decades, there is still a lack of awareness about the influence of hydration water on the polysaccharide´s structure and consequences for industrial or medicinal applications. The hydration of polysaccharides is often described by the existence of water layers differing in their physical properties depending on the distance from the polysaccharide. Using the differential scanning calorimetry (DSC) such water layers were categorized according their properties upon cooling in hyaluronan (HYA, sodium salt of ß-1,4-linked units of ß-1,3-linked D-glucuronic acid and N-acetyl-D-glucosamine), a model polysaccharide in the present work. The amount of non-freezing water, i.e. water in close proximity of HYA chain which does not freeze et all, was determined around 0.74gH2O/gHYA for HYA with molecular weight from 100 to 740kDa and 0.84gH2O/gHYA for molecular weight of 1390kDa. The amount of freezing-bound water, the water pool which is affected by presence of HYA but freezes, was determined in the range from 0.74 to 2gH2O/gHYA. Above this value only non-freezing and bulk water are present since melting enthalpy measured above this concentration reached the same value as for pure water. Since this approach suffers from several experimental problems, a new approach, based on the evaporation enthalpy determination, was suggested. The analysis of the evaporation enthalpies revealed an additional process associated with apparent energy release taking part below the water content of 0.34gH2O/gHYA. Existence of this phenomenon was observed also for protonated form of HYA. The existence of energy compensating process was confirmed with the Kissinger-Akahira-Sunose method which allowed determination of actual water evaporation/desorption enthalpies in all stages of the evaporation process. In fact, the apparent evaporation enthalpy value increased until water content of 0.34gH2O/gHYA, and then dropped down to lower values which were, still higher than the value of the pure water evaporation enthalpy. By the use of time domain nuclear magnetic resonance (TD-NMR) technique it was revealed that this phenomenon is the plasticisation of HYA.
Further, it was revealed that the non-freezing water determined by the use of DSC consists of two water fractions, i.e. 15% of water structurally integrated, interacting directly with polar sites, and 85% of water structurally restricted, embedded in-between the HYA chains. The occurrence of plasticisation concentration close to equilibrium moisture content provided the possibility to influence the HYA physical structure during the drying. In this way three samples of native HYA, dried under various conditions were prepared and their physical properties were analyzed. The samples differed in kinetics of rehydration, plasticisation concentration, glass transitions, and morphology. The properties of water pool were studied in solutions of 10"25mg HYA/mL as well. The fast filed cycling (FFC) NMR relaxometry showed the existence of three water fractions which correlation times spanned from 10"6 to 10"10 seconds, progressively decreasing in dependency on its distance from HYA chain. The formation of a weak and transient intramolecular water bridge between HYA chains was observed. It was shown that, unlike the inorganic electrolytes, polyelectrolytes hydration is a dynamic process which reflects not only the technique used for the analysis, experimental conditions but also the conformation of the polysaccharide and its "thermal" and "hydration" history.
It was demonstrated that some native polysaccharide structures can be easily modified by manipulation of preparation conditions, giving fractions with specific physicochemical properties without necessity of any chemical modification.
Chemical plant protection is an essential element in integrated pest management and hence, in current crop production. The use of Plant Protection Products (PPPs) potentially involves ecological risk. This risk has to be characterised, assessed and managed.
For the coming years, an increasing need for agricultural products is expected. At the same time, preserving our natural resources and biodiversity per se is of equally fundamental importance. The relationship of our economic success and cultural progress to protecting the environment has been made plain in the Ecosystem Service concept. These distinct 'services' provide the foundation for defining ecological protection goals (Specific Protection Goals, SPGs) which can serve in the development of methods for ecological risk characterisation, assessment and management.
Ecological risk management (RM) of PPPs is a comprehensive process that includes different aspects and levels. RM is an implicit part of tiered risk assessment (RA) schemes and scenarios, yet RM also explicitly occurs as risk mitigation measures. At higher decision levels, RM takes further risks, besides ecological risk, into account (e.g., economic). Therefore, ecological risk characterisation can include RM (mitigation measures) and can be part of higher level RM decision-making in a broader Ecosystem Service context.
The aim of this thesis is to contribute to improved quantification of ecological risk as a basis for RA and RM. The initial general objective had been entitled as "… to estimate the spatial and temporal extent of exposure and effects…" and was found to be closely related to forthcoming SPGs with their defined 'Risk Dimension'.
An initial exploration of the regulatory framework of ecological RA and RM of PPPs and their use, carried out in the present thesis, emphasised the value of risk characterisation at landscape-scale. The landscape-scale provides the necessary and sufficient context, including abiotic and biotic processes, their interaction at different scales, as well as human activities. In particular, spatially (and temporally) explicit landscape-scale risk characterisation and RA can provide a direct basis for PPP-specific or generic RM. From the general need for tiered landscape-scale context in risk characterisation, specific requirements relevant to a landscape-scale model were developed in the present thesis, guided by the key objective of improved ecological risk quantification. In principle, for an adverse effect (Impact) to happen requires a sensitive species and life stage to co-occur with a significant exposure extent in space and time. Therefore, the quantification of the Probability of an Impact occurring is the basic requirement of the model. In a landscape-scale context, this means assessing the spatiotemporal distribution of species sensitivity and their potential exposure to the chemical.
The core functionality of the model should reflect the main problem structures in ecological risk characterisation, RA and RM, with particular relationship to SPGs, while being adaptable to specific RA problems. This resulted in the development of a modelling framework (Xplicit-Framework), realised in the present thesis. The Xplicit-Framework provides the core functionality for spatiotemporally explicit and probabilistic risk characterisation, together with interfaces to external models and services which are linked to the framework using specific adaptors (Associated-Models, e.g., exposure, eFate and effect models, or geodata services). From the Xplicit-Framework, and using Associated-Models, specific models are derived, adapted to RA problems (Xplicit-Models).
Xplicit-Models are capable of propagating variability (and uncertainty) of real-world agricultural and environmental conditions to exposure and effects using Monte Carlo methods and, hence, to introduce landscape-scale context to risk characterisation. Scale-dependencies play a key role in landscape-scale processes and were taken into account, e.g., in defining and sampling Probability Density Functions (PDFs). Likewise, evaluation of model outcome for risk characterisation is done at ecologically meaningful scales.
Xplicit-Models can be designed to explicitly address risk dimensions of SPGs. Their definition depends on the RA problem and tier. Thus, the Xplicit approach allows for stepwise introduction of landscape-scale context (factors and processes), e.g., starting at the definitions of current standard RA (lower-tier) levels by centring on a specific PPP use, while introducing real-world landscape factors driving risk. With its generic and modular design, the Xplicit-Framework can also be employed by taking an ecological entity-centric perspective. As the predictive power of landscape-scale risk characterisation increases, it is possible that Xplicit-Models become part of an explicit Ecosystem Services-oriented RM (e.g., cost/benefit level).
Worldwide one third to one half of the freshwater crayfish species are threatened with population decline or extinction. Besides habitat deterioration, pollution, and other man-made environmental changes, invasive species and pathogens are major threats to the survival of European crayfish species. Freshwater crayfish are the largest freshwater invertebrates and strongly influence the structure of food webs. The disappearance of crayfish from a water body may change the food web and could have dramatic consequences for an ecosystem.rnOne goal in modern species conservation strategies is the conservation of genetic diversity, since genetic diversity is an advantage for the long-term survival of a species. The main aim of my thesis was to reveal the genetic structure and to identify genetic hotspots of the endangered noble crayfish (Astacus astacus) throughout Europe (part 1 of my thesis). Since the most significant threat to biodiversity of European crayfish species is the crayfish plague pathogen Aphanomyces astaci I studied new aspects in the distribution of A. astaci (part two of my thesis). The results serve as a basis for future conservation programs for freshwater crayfish. In the first part of my thesis I conducted a phylogeographic analysis of noble crayfish using mitochondrial DNA and nuclear microsatellite data. With these methods I aimed to identify its genetic hotspots and to reconstruct the recolonization history of central Europe by this species. I detected high genetic diversities in southestern Europe indicating that noble crayfish outlasted the cold climate phases during the Pleistocene in this region (Appendix 1). Because of the high genetic diversity found there, southeastern Europe is of particular importance for the conservation of noble crayfish. The mitochondrial DNA analysis points to a bifurcated colonization process from the eastern Black Sea basin to a) the North Sea and to b) the Baltic Sea basin (Appendix 2). A second independent refugium that was localized on the Western Balkans did not contribute to the colonization of central Europe. Furthermore, I found that the natural genetic structure is dissolved, probably due to the high human impact on the distribution of noble crayfish (e.g. artificial translocation). In the second part of this thesis using real-time PCR I identified calico crayfish (Orconectes immunis) as the fourth North American crayfish species to be carrier of the agent of the crayfish plague (Appendix 3). Furthermore I detected the crayfish plague pathogen in American spiny-cheek crayfish (Orconectes limosus) and native narrow-clawed crayfish (Astacus leptodactylus) in the lower Danube in Romania (Appendix 4). The distribution of infected spiny-cheek crayfish poses a threat to the native biodiversity in southeastern Europe and shows the high invasion potential of this crayfish species. Moreover, I found that even the native narrow-clawed crayfish in the Danube Delta, about 970 km downstream of the current invasion front of American crayfish, is a carrier of A. astaci (Appendix 5). This finding is of high importance, as the native species do not seem to suffer from the infection. In Appendix 6 I elucidate demonstrate that the absence of the crayfish plague agent is the most likely explanation for the coexistence of populations of European and American crayfish in central Europe. In my thesis I show that the common assumption that all North American crayfish are carrier of A. astaci and that all native crayfish species die when infected with A. astaci does not hold true. The studies presented in my thesis reveal new aspects that are crucial for native crayfish conservation: 1) The genetic diversity of noble crayfish is highest in southeastern Europe where noble crayfish outlasted the last glacial maximum in at least two different refugia. 2) Not all American crayfish populations are carrier of A. astaci and 3) not all Europen crayish populations die shortly after being infected with the crayfish plague pathogen.rnTo conserve native crayfish species and their (genetic) diversity in the long term, further introductions of American crayfish into European waters must be avoided. However, the introduction will only decrease if the commercial trade with non-indigenous crayfish species is prohibited.
Assessment of bat activity in agricultural environments and the evaluation of the risk of pesticides
(2013)
Although agriculture dominates with around 50% area much of Europe- landscape, there is virtually no information on how bats use this farmed environment for foraging. Consequently, little is known about effective conservation measures to compensate potential negative effects of agrarian management practice on the food availability for bats in this habitat. Moreover, there are currently no specific regulatory requirements to include bats in European Union risk assessments for the registration of pesticides since no information about pesticide exposure on this mammal group is available. To evaluate the potential pesticide exposure of bats via ingestion of contaminated insects, information about bat presence and activity in agricultural habitats is required. In order to examine bat activity on a landscape scale it was necessary to establish a suitable survey method. Contrary to capture methods, telemetry, and direct observations, acoustic surveys of bat activity are a logistically feasible and cost-effective way of obtaining bat activity data. However, concerns regarding the methodological designs of many acoustic surveys are expressed in the scientific literature. The reasons are the failing of addressing temporal and spatial variation in bat activity patterns and the limitations of the suitability of the used acoustic detector systems. By comparing different methods and detector systems it was found that the set up of several stationary calibrated detector systems which automatically trigger the ultrasonic recording has the highest potential to produce reliable, unbiased and comparable data sets on the relative activity of bats.
By using the proposed survey method, bat diversity and activity was recorded in different crops and semi-natural habitats in southern Rhineland-Palatinate. Simultaneously, the availability of aerial prey insects was studied by using light and sticky traps. In more than 500 sampling nights about 110,000 call sequences were acoustically recorded and almost 120,000 nocturnal insects were sampled. A total of 14 bat species were recorded, among them the locally rare and critically endangered northern bat (Eptesicus nilssonii) and the barbastelle (Barbastella barbastellum), all of them also occurring over agricultural fields. The agricultural landscape of southern Palatinate is dominated by vineyards, a habitat that was shown to be of low quality for most bat species because of the demonstrated low availability of small aerial insects. By surveying bat activity and food availably in a pair-wise design on several rain water retention ponds and neighbouring vineyards it was demonstrated that aquatic insect emergence in artificial wetlands can provide an important resource subsidy for bats. The creation of artificial wetlands would be a possibility to create important foraging habitats for bats and mitigate negative effects of management practice in the agricultural landscape.
In several other agricultural crops, however, high abundances of suitable prey insects and high bat activity levels, comparable or even higher than in the nearby forests and meadows known to be used as foraging habitats were demonstrated. Especially high bat activity levels were recorded over several fruit orchards and vegetable fields where insects were also present. Both crops are known for high pesticide inputs, and, therefore, a pesticide exposure through ingestion of contaminated insects can not be excluded. To follow the current risk assessment approach for birds and mammals pesticide residues were measured on bat-specific food items in an apple orchard following insecticide applications and bat activity was recorded in parallel. The highest residue values were measured on foliage-dwelling arthropods which may results in a reproductive risk for all bat species that, even to a small extent, include this prey group in their diet. The presence of bats in agricultural landscapes that form a majority of the land area in Europe but also on a global scale leads to exposure of bats by contaminated food and depletion of their food resources by pesticide use. So far conservation efforts for bats focussed on securing hibernation sites and the creation of artificial roost sites since especially the latter were thought to be limiting population growth. However the potential pesticide effects might be also crucial for the population persistence in agricultural landscapes of bats and need to be addressed adequately, especially in risk assessment procedures for the regulation of pesticides.
The conservation of groundwater ecosystems requires an assessment and evaluation scheme that shows the state of the ecosystem. Consequently, faunal and microbial criteria are required for groundwater monitoring, in addition to physico-chemical analyses. To proof the adequacy of the application of groundwater organisms for the biological assessment of groundwater quality, an extensive sampling collection of various groundwater systems was accomplished between 2002 and 2009 in Germany. Key aspects were the examination of the indicator potential of groundwater communities towards surface-groundwater interactions and anthropogenic impacts, as well as the analysis of stygofaunal distribution patterns, as a base and reference for a faunal assessment scheme of groundwater systems. The sampling design considered local, regional, and biogeographic conditions. To test the indicator potential of groundwater organisms on a local and regional scale, groundwater systems in Nordrhein-Westfalen (NRW) and Baden-Württemberg (BW) were examined. The faunal and microbial data from unconsolidated aquifers (Erftgebiet, NRW) show the sensitivity of groundwater organisms against land use effects, e.g. extensive agriculture. Data analyses revealed positive correlations of organic material (estimated amount of detritus, TOC) and nitrate with faunal abundance, species richness, diversity and the proportions of stygoxene species. Moreover, the bacterial abundance and diversity in the groundwater of the Erftgebiet was high compared to oligotrophic groundwater systems, indicating an effect of surface influence due to agricultural land use. The groundwater colonization in the Alb-Donau-Kreis (BW) was analyzed for regional effects (landscape, type of aquifer, hydrogeology) and local effects (comparing single wells). The results show that the fauna reflects the strength of the hydrological exchange on different spatial scales. Furthermore, the fauna reflects the interaction of regional and local conditions. Accordingly, the diversity and abundance of groundwater organisms was influenced by the high connectivity of the karst and unconsolidated alluvial aquifers, the type of land use, covering layers of soil, the age of groundwater, and the sampling depths. In general, faunal and microbial data of the Alb-Donau-Kreis are characteristic for oligotrophic, oligoalimonic groundwaters. The large scale analyses of stygofaunal distribution patterns revealed significant biogeographical differences of the communities. These community patterns of the groundwater fauna do not coincide with existing classification schemes defined for surface landscapes or freshwater systems. The largest differences between faunal surface and subsurface distribution patterns were found between the groundwater of northern and southern Germany and the foothills of the central mountain ranges - all of them regions shaped by the last ice ages.
In accordance with the faunal data assessed in groundwater, four different stygoregions were defined that are populated by distinct faunal assemblages. These are 1) the "Northern lowlands", 2) the "Central mountain ranges", 3) the "South-western mountain ranges", and 4) the "Southern mountain ranges and northern alps". The study corroborates that stygofaunal and microbial communities are an adequate tool for the qualitative assessment and monitoring of groundwater ecosystems. The best indicators to detect anthropogenic impacts on groundwater ecosystems are the faunal diversity, abundance, the proportions of stygobitic (obligate groundwater species) and stygoxene species (species not obligate for groundwater), and the bacterial abundance. The development of an ecologically based groundwater assessment and management, is crucial for the conservation of our groundwater ecosystems and thus, healthy drinking water. The defintion of "stygoregions" is an important base for the development of an assessment and reference scheme for groundwater ecosystems. The assessment of the ecological state of groundwater systems must be conducted on local scale, because the strength of the local hydrological influence determines the amount of food and oxygen as well as stygoxene animals within the aquifer, and thus shapes the local groundwater communities. Nevertheless, information of the biogeographic and regional conditions is needed as a reference for the type of species and abundances that potentially can occur in certain groundwater systems.
Agricultural pesticides, especially insecticides, are an integral part of modern farming. However, these may often leave their target ecosystems and cause adverse effects in non- target, especially freshwater ecosystems, leading to their deterioration. In this thesis, the focus will be on Insect Growth Regulators (IGRs) that can in many ways cause disruption of the endocrine system of invertebrates. Freshwater invertebrates play important ecological, economic and medical roles, and disruption of their endocrine systems may be crucial, considering the important role hormones play in the developmental and reproductive processes in organisms. Although Endocrine Disruption Chemicals (EDCs) can affect moulting, behaviour, morphology, sexual maturity, time to first brood, egg development time, brood size (fecundity), and sex determination in invertebrates, there is currently no agreement upon how to characterize and assess endocrine disruption (ED). Current traditional ecotoxicity tests for Ecological Risk Assessment (ERA) show limitations on generating data at the population level that may be relevant for the assessment of EDCs, which effects may be sublethal, latent and persist for several generations of species (transgenerational).
It is therefore the primary objective of this thesis to use a test method to investigate adverse effects of EDCs on endpoints concerning development and reproduction in freshwater invertebrates. The full life-cycle test over two generations that includes all sensitive life stages of C. riparius (a sexual reproductive organism) allows an assessment of its reproduction and should be suitable for the investigation of long-term toxicity of EDCs in freshwater invertebrates. C. riparius is appropriate for this purpose because of its short life cycle that enables the assessment of functional endpoints of the organism over several generations. Moreover, the chironomid life cycle consists of a complete metamorphosis controlled by a well-known endocrine mechanism and the endocrine system of insects has been most investigated in great detail among invertebrates. Hence, the full life-cycle test with C. riparius provides an approach to assess functional endpoints (e.g. reproduction, sex ratio) that are population-relevant as a useful amendment to the ERA of EDCs. In the laboratory, C. riparius was exposed to environmentally-relevant concentrations of the selected IGRs in either spiked water or spiked sediment scenario over two subsequent generations.
The results reported in this thesis revealed significant effects of the IGRs on the development and the reproduction of C. riparius with the second (F1) generation showing greater sensitivity. These findings indicated for the first time the suitability of multigenerational testing for various groups of EDCs and strongly suggested considering the full life-cycle of C. riparius as an appropriate test method for a better assessment of EDCs in the freshwater environment. In conclusion, this thesis helps to detect additional information that can be extrapolated at population level and, thus, might contribute to better protection of freshwater ecosystems against the risks of Endocrine Disrupting Chemicals (EDCs.) It may furthermore contribute to changes in the ERA process that are necessary for a real implementation of the new European chemical legislation, REACH (Registration, Evaluation Authorization and Restriction of Chemicals). Finally, significant interactions between temperature, chemical exposure and generation were reported for the first time and, may help predict impacts that may occur in the future, in the field, under predicted climate change scenarios.