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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).
Recent estimates have confirmed that inland waters emit a considerable amount of CH4 and CO2 to the atmosphere at the regional and global scale. But these estimates are based on extrapolated measured data and lack of data from inland waters in arid and semi-arid regions and carbon sources from wastewater treatment plants (WWTPs) as well insufficient resolution of the spatiotemporal variability of these emissions.
Through this study, we analyzed monthly hydrological, meteorological and water quality data from three irrigation and drinking water reservoirs in the lower Jordan River basin and estimated the atmospheric emission rates of CO2. We investigated the effect of WWTPs on surrounding aquatic systems in term of CH4 and CO2 emission by presenting seasonally resolved data for dissolved concentrations of both gases in the effluents and in the receiving streams at nine WWTPs in Germany.
We investigated spatiotemporal variability of CH4 and CO2 emission from aquatic ecosystems by using of simple low-cost tools for measuring CO2 flux and bubble release rate from freshwater systems. Our estimates showed that reservoirs in semi-arid regions are oversaturated with CO2 and acted as net sources to the atmosphere. The magnitude of observed fluxes at the three water reservoirs in Jordan is comparable to those from tropical reservoirs (3.3 g CO2 m-2 d-1). The CO2 emission rate from these reservoirs is linked to changes of water surface area, which is the result of water management practices. WWTPs have been shown to discharge a considerable amount of CH4 (30.9±40.7 kg yr-1) and CO2 (0.06±0.05 Gg yr-1) to their surrounding streams, and emission rates of CH4 and CO2 from these streams are significantly enhanced by effluents of WWTPs up to 1.2 and 8.6 times, respectively.
Our results showed that both diffusive flux and bubble release rate varied in time and space, and both of emission pathways should be included and variability should be resolved adequately in further sampling and measuring strategies. We conclude that future emission measurements and estimates from inland waters may consider water management practices, carbon sources from WWTPs as well spatial and temporal variability of emission.
Das Grundwasser unterliegt zahlreichen Nutzungen, gleichzeitig ist es Lebensraum einer artenreichen, hoch angepaßten Fauna. Verunreinigungen des Grundwassers mit Kontaminationen unterschiedlichsten Ursprungs stellen eine wachsende Problematik dar. Für das Monitoring von Altlastflächen werden bisher überwiegend physiko-chemische Methoden eingesetzt. Als weitere Möglichkeit bietet sich ein grundwasserfaunistisch begründetes Monitoring an. Da Freilanduntersuchungen über das Auftreten und die Verteilung von Grundwasserfauna in Altlastflächen bisher fehlen, widmet sich die vorliegende Dissertation dieser Thematik. Ein grundsätzliches Problem grundwasserfaunistischer Untersuchungen ist das Fehlen standardisierter Sammelmethoden. Daher erwies es sich als notwendig, die für die Untersuchung der Altlastflächen optimale Sammelmethode zu identifizieren. Insoweit liefert ein vorangehender Methodenvergleich die Grundlage für das Altlasten-Projekt. Ziel des Methodenvergleichs war es herauszufinden, ob die in Kluftgrundwasserleitern im Vergleich mit Lockergesteinsleitern festgestellte Artenarmut und die meist niedrigen Abundanzen habitat- oder auch methodenbedingt sind. Unter Verwendung eines phreatobiologischen Netzsammlers, einer pneumatischen Kolbenhubpumpe (System NIEDERREITER) und eines Quellnetzes wurden im Naturraum Pfälzerwald 16 Grundwasser-meßstellen (je 8 in den Kluftaquiferen des Buntsandsteins und den sandig-kiesigen Aquiferen der Talauen) und 8 Quellen, sowie 4 Grundwassermeßstellen in der pfälzischen Rheinebene hydrochemisch und faunistisch beprobt. Die Grundwässer beider Naturräume unterschieden sich sowohl hydrochemisch wie auch in ihrer faunistischen Zusammensetzung signifikant voneinander. Aus methodischer Sicht zeigten alle Sammeltechniken, mit Ausnahme der Quellbeprobung, trotz gewisser Unterschiede, qualitativ vergleichbare Ergebnisse. Ziel des Hauptteils der Arbeit " dem Altlasten-Projekt " war die Gewinnung erster empirischer Daten über das Vorkommen und die Verteilung von Grundwasserfauna in Abhängigkeit von Altlasten. Des weiteren sollten über die Korrelation der Grundwasserfauna mit hydrochemischen Parametern und den Koloniezahlen erste Ansätze für eine bioindikative Eignung gefunden werden. Basierend auf den Ergebnissen des methodischen Teils wurden unter Verwendung des phreatobiologischen Netzsammlers fünf in der pfälzischen Rheinebene liegende Untersuchungsstandorte (sechs Einzelschäden), die Kontaminationen unterschiedlicher Art aufwiesen, untersucht. Dabei handelte es sich um drei militärische und zwei industrielle Altlasten sowie eine Hausmülldeponie. Insgesamt wurden 91 Meßstellen, die sich im Zustrom, direkt im Schaden sowie im Abstrom befanden, zweimalig beprobt. Trotz der ausgeprägten Heterogenität der untersuchten Altlaststandorte sowie geringer Arten- und Taxazahlen und Abundanzen weisen die Verteilungsmuster auf Sensitivitäten der Fauna gegenüber Grundwasserverunreinigungen hin. In bezug auf einzelne Schadstoffe (LCKW, PCE, AKW) deuten sich schadstoffspezifische Verteilungsmuster der Fauna an. Das abundanteste Taxon der Untersuchung - die Nematoda - traten an den LCKW / PCE- kontaminierten Standorten nur in geringsten Abundanzen auf, ebenso wie die Parastenocaridae (Crustacea). Oligochaeta und einzelne Cyclopoida-Arten (Crustacea) scheinen hingegen deutlich höhere Konzentrationen dieses Schadstoffs zu tolerieren. Eine gegensätzliche Faunenverteilung wurde an den AKW- kontaminierten Standorten beobachtet. Hier zeigten sich die Nematoda wesentlich toleranter als alle anderen Taxa, ebenso wie die Parastenocaridae. Hingegen traten die Oligochaeta und Cyclopoida oberhalb geringer Konzentrationen nur noch sporadisch auf. Die abundanteste Cyclopoida-Art der gesamten Untersuchung, Diacyclops languidoides, konnte in Anwesenheit von AKW nicht nachgewiesen werden.
The (un-)controlled application of olive oil mill wastewater (OMW) has positive and negative effects on soil quality. On the one hand it can be used as fertilizer, on the other hand especially the occurrence of soil water repellency is problematic. Due to this fact the objective of this study was to characterize the effects of OMW application on soil and to investigate the mechanisms that are in combination with changes of soil organic matter quality responsible for soil water repellency depending on the climatic conditions.
At first several locations of uncontrolled OMW disposal were screened for positive and negative im-pacts. Then, laboratory incubation experiments and finally a field experiment in Israel were conducted in order to determine the influence of climatic conditions. Besides standard soil parameters (pH, elec-tric conductivity, total carbon, dissolved organic carbon , specific UV-Absorption) it was focused on the determination of phenolic compounds, the carbon isotope ratio, the water drop penetration time and the contact angle as well as the thermal analysis.
This thesis shows that soil water repellency of OMW-polluted soils depends on the climatic conditions, i.e. the application season. In the laboratory as well as in the field the wettability of the soil was strongly reduced under hot and dry conditions. It was observed, that the stable carbon fraction characterized by a high calorific value is responsible for soil water repellency. In particular, amphiphilic substances, e.g. fatty acids, may interact with soil particles as a consequence of drying. On the contrary, no reduc-tion of the wettability of the soil was determined under moist conditions and degradation of organic matter of the OMW was enhanced. Nevertheless, too strong irrigation or rainfall, e.g. in winter, may leach phenolic ingredients of the OMW into the groundwater.
At the same time the application led to an increase of organic and inorganic nutrients, which should be emphasized as a positive effect. Due to these results, a controlled application of olive oil mill wastewater as alternative, low-cost and sustainable treatment option is recommended. But, instead of the current application season winter, the olive mill wastewater should be stored and not be spread before spring in order to avoid negative impacts on the soil.
By the work presented in this thesis, the CH4 emissions of the River Saar were quantified in space and time continuously and all relevant processes leading to the observed pattern were identified. The direct comparison between reservoir zones and free-flowing intermediate reaches revealed, that the reservoir zones are CH4 emission hot spots and emitted over 90% of the total CH4. On average, the reservoir zones emitted over 80 times more CH4 per square meter than the intermediate reaches between dams (0.23 vs. 19.7 mol CH4 m-2 d-1). The high emission rates measured in the reservoir zones fall into the range of emissions observed in tropical reservoirs. The main reason for this is the accumulation of thick organic rich sediments and we showed that the net sedimentation rate is an excellent proxy for estimating ebullitive emissions. Within the hot spot zones, the ebullitive flux enhanced also the diffusive surface emissions as well as the degassing emissions at dams.
To resolve the high temporal variability, we developed an autonomous instrument for continuous measurements of the ebullition rate over long periods (> 4 weeks). With this instrument we could quantify the variability and identify the relevant trigger mechanisms. At the Saar, ship-lock induces surges and ship waves were responsible for over 85% of all large ebullition events. This dataset was also used to determine the error associated with short sampling periods and we found that with sampling periods of 24 hours as used in other studies, the ebullition rates were systematically underestimated by ~50%. Measuring the temporal variability enabled us to build up a conceptual framework for estimating the temporal pattern of ebullition in other aquatic systems. With respect to the contribution of freshwater systems to the global CH4 emissions, hot spot emission sites in impounded rivers have the potential to increase the current global estimate by up to 7%.
Despite the significant presence of neuroactive substances in the environment, bioassays that allow to detect diverse groups of neuroactive mechanisms of action are not well developed and not properly integrated into environmental monitoring and chemical regulation. Therefore, there is a need to develop testing methods which are amenable for fast and high-throughput neurotoxicity testing. The overall goal of this thesis work is to develop a test method for the toxicological characterization and screening of neuroactive substances and their mixtures which could be used for prospective and diagnostic hazard assessment.
In this thesis, the behavior of zebrafish embryos was explored as a promising tool to distinguish between different neuroactive mechanisms of action. Recently, new behavioral tests have been developed including photomotor response (PMR), locomotor response (LMR) and spontaneous tail coiling (STC) tests. However, the experimental parameters of these tests lack consistency in protocols such as exposure time, imaging time, age of exposure, endpoint parameter etc. To understand how experimental parameters may influence the toxicological interpretation of behavior tests, a systematic review of existing behavioral assays was conducted in Chapter 2. Results show that exposure concentration and exposure duration highly influenced the comparability between different test methods and the spontaneous tail coiling (STC) test was selected for further testing based on its relative higher sensitivity and capacity to detect neuroactive substances (Chapter 2).
STC is the first observable motor activity generated by the developing neural network of the embryo which is assumed to occur as a result of the innervation of the muscle by the primary motor neurons. Therefore, STC could be a useful endpoint to detect effect on the muscle innervation and also the on the whole nervous system. Consequently, important parameters of the STC test were optimized and an automated workflow to evaluate the STC with the open access software KNIME® was developed (Chapter 3).
To appropriately interpret the observed effect of a single chemical and especially mixture effects, requires the understanding of toxicokinetics and biotransformation. Most importantly, the biotransformation capacity of zebrafish embryos might be limited and this could be a challenge for assessment of chemicals such as organophosphates which require a bioactivation step to effectively inhibit the acetylcholinesterase (AChE) enzyme. Therefore, the influence of the potential limited biotransformation on the toxicity pathway of a typical organophosphate, chlorpyrifos, was investigated in Chapter 5. Chlorpyrifos could not inhibit AChE and this was attributed to possible lack of biotransformation in 24 hpf embryos (Chapter 5).
Since neuroactive substances occur in the environment as mixtures, it is therefore more realistic to assess their combined effect rather than individually. Therefore, mixture toxicity was predicted using the concentration addition and independent action models. Result shows that mixtures of neuroactive substances with different mechanisms of action but similar effects can be predicted with concentration addition and independent action (Chapter 4). Apart
from being able to predict the combined effect of neuroactive substances for prospective risk assessment, it is also important to assess in retrospect the combined neurotoxic effect of environmental samples since neuroactive substances are the largest group of chemicals occurring in the environment. In Chapter 6, the STC test was found to be capable of detecting neurotoxic effects of a wastewater effluent sample. Hence, the STC test is proposed as an effect based tool for monitoring environmental acute and neurotoxic effects.
Overall, this thesis shows the utility and versatility of zebrafish embryo behavior testing for screening neuroactive substances and this allows to propose its use for prospective and diagnostic hazard assessment. This will enhance the move away from expensive and demanding animal testing. The information contained in this thesis is of great potential to provide precautionary solutions, not only for the exposure of humans to neuroactive chemicals but for the environment at large.
Larvae of Cx.pipiens coocurred with Cladocera, but the latter established delayed in time. Biotope structure influenced time of species occurrence with ponds at reed-covered wetlands favouring crustacean development, while ponds at grassland biotopes favoured colonization by mosquito larvae. The mechanisms driving the negative effect of crustaceans on mosquito larvae were investigated within an experiment under artificial conditions. Crustacean communities were found to reduce both oviposition and larval development of Cx.pipiens. Crustacean communities of high taxa diversity, including both predatory and competing crustaceans, were more effective compared with crustacean communities dominated by single taxa. Presence of crustacean communities characterised by high taxa diversity increased the sensitivity of Cx.pipiens larvae towards Bti and prolonged the time of recolonization. In a final step the combined approach, using Bti and crustaceans, was evaluated under field conditions. The joint application of Bti and crustaceans was found to reduce mosquito larval populations over the whole observation period, while single application of Bti caused only short-term reduction of mosquito larvae. Single application of crustaceans had no significant effect, because high abundances of prior established mosquito larvae impeded propagation of crustaceans. At combined treatment, mosquito larvae were reduced by Bti application and hence crustaceans were able to proliferate without disturbance by interspecific competition. In conclusion, natural competitors were found to have a strong negative impact on mosquito larval populations. However, a time span of about 2 weeks has to be bridged, before crustacean communities reached a level sufficient for mosquito control. Results of a combined approach, complementing the short-term effect of the biological insecticide Bti with the long-term effect of crustaceans, were promising. Using natural competitors within an integrated control strategy could be an important tool for an effective, environmentally friendly and sustainable mosquito management.
The role of alternative resources for pollinators and aphid predators in agricultural landscapes
(2021)
The world wide decline of insects is often associated with loss of natural and semi-natural habitat caused by intensified land-use. Many insects provide important ecosystem services to agriculture, such as pest control or pollination. To efficiently promote insects on remaining semi-natural habitat we need precise knowledge of their requirements to non-crop habitat. This thesis focuses on identifying
the most important semi-natural habitats (forest edges, grasslands, and semi-open habitats) for pollinators and natural enemies of crop pests with respect to their food resource requirements. Special
attention is given to floral resources and their spatio-temporal distribution in agricultural landscapes.
Floral resource maps might get closer at characterizing landscapes the way they are experienced by insects compared to classical habitat maps. Performance of the two map types was compared on the prediction of wild bees and natural enemies that consume nectar and pollen, identifying habitats of special importance in the process. In wild bees, influences of spatio-temporal floral resource availability were analysed as well as habitat preferences of specific groups of bees. Understanding dietary needs of natural enemies of crop pests requires additional knowledge on prey use. To this end, ladybird gut contents have been analysed by means of high-throughput sequencing for insight into aphid prey-use.
Results showed, that wild bees were predicted better by floral resource maps compared to classical habitat maps. Forest edge area, as well as floral resources in forest edges had positive effects on abundance and diversity of rare bees and important crop pollinators. Similar patterns were retained for grassland diversity. Especially early floral resources seemed to have positive effects on wild bees. Crops and fruit trees produced a resource pulse in April that exceeded floral resource availability in May and June by tenfold. Most floral resources in forest edges appeared early in the season, with the highest floral density per area. Grasslands provided the lowest amount of floral resources but highest diversity, which was evenly distributed over the season.
Despite natural enemies need for floral resources, classical habitat maps performed better at predicting natural enemies of crop pests compared to floral resource maps. Classical habitat maps revealed a positive effect of forest edge habitat on the abundance of pest enemies, which translated into improved aphid control. Results from gut content analysis reveal high portions of pest aphid species and nettle aphids as well as a broader insight into prey spectra retained from ladybirds collected from sticky traps compared to individuals collected by hand. The aphid specific primer designed for this purpose will be helpful for identifying aphid consumption by ladybirds in future studies.
Findings of this thesis show the potential of floral resource maps for understanding interactions of wild bees and the landscape but also indicate that natural enemies are limited by other resources. I would like to highlight the positive effects of forest edges for different groups of bees as well as natural enemies and their performance on pest control.
Grassland management has been increasingly intensified throughout centuries since mankind started to control and modify the landscape. Species communities were always shaped alongside management changes leading to huge alterations in species richness and diversity up to the point where land use intensity exceeded the threshold. Since then biodiversity became increasingly lost. Today, global biodiversity and especially grassland biodiversity is pushed beyond its boundaries. Policymakers and conservationists seek for management options which fulfill the requirements of agronomic interests as well as biodiversity conservation alongside with the maintenance of ecosystem processes. However, there is and will always be a trade-off.
Earlier in history, natural circumstances in a landscape mainly determined regionally adapted land use. These regional adaptions shaped islands for many specialist species, and thus diverse species communities, favoring the establishment of a high β-diversity. With the raising food demand, these regional and traditional management regimes became widely unprofitable, and the invention of mineral fertilizers ultimately led to a wide homogenization of grassland management and, as follows, the loss of biotic heterogeneity. In the course of the green revolution, this immediate coherence and the dependency between grassland biodiversity and traditional land use practices becomes increasingly noticed. Indeed, some traditional forms of management such as meadow irrigation have been preserved in a few regions and thus give us the opportunity to directly investigate their long-term relevance for the species communities and ecosystem processes. Traditional meadow irrigation was a common management practice to improve productivity in lowland, but also alpine hay meadows throughout Europe until the 20th century. Nowadays, meadow irrigation is only practiced as a relic in a few remnant areas. In parts of the Queichwiesen meadows flood irrigation goes back to the Middle Ages, which makes them a predestined as a model region to study the long- and short-term effects of lowland meadow irrigation on the biodiversity and ecosystem processes.
Our study pointed out the conservation value of traditional meadow irrigation for the preservation of local species communities as well as the plant diversity at the landscape scale. The structurally more complex irrigated meadows lead to the assumption of a higher arthropod diversity (Orthodoptera, Carabidae, Araneae), which could not be detected. However, irrigated meadows are a significant habitat for moisture dependent arthropod species. In the light of the agronomic potential, flood irrigation could be a way to at least reduce fertilizer costs to a certain degree and possibly prevent overfertilization pulses which are necessarily hazardous to non-target ecosystems. Still, the reestablishment of flood irrigation in formerly irrigated meadows, or even the establishment of new irrigation systems needs ecological and economic evaluation dependent on regional circumstances and specific species communities, at which this study could serve as a reference point.
Aquatic macrophytes can contribute to the retention of organic contaminants in streams, whereas knowledge on the dynamics and the interaction of the determining processes is very limited. The objective of the present study was thus to assess how aquatic macrophytes influence the distribution and the fate of organic contaminants in small vegetated streams. In a first study that was performed in vegetated stream mesocosms, the peak reductions of five compounds were significantly higher in four vegetated stream mesocosms compared to a stream mesocosm without vegetation. Compound specific sorption to macrophytes was determined, the mass retention in the vegetated streams, however, did not explain the relationship between the mitigation of contaminant peaks and macrophyte coverage. A subsequent mesocosm study revealed that the mitigation of peak concentrations in the stream mesocosms was governed by two fundamentally different processes: dispersion and sorption. Again, the reductions of the peak concentrations of three different compounds were in the same order of magnitude in a sparsely and a densely vegetated stream mesocosm, respectively, but higher compared to an unvegetated stream mesocosm. The mitigation of the peak reduction in the sparsely vegetated stream mesocosm was found to be fostered by longitudinal dispersion as a result of the spatial distribution of the macrophytes in the aqueous phase. The peak reduction attributable to longitudinal dispersion was, however, reduced in the densely vegetated stream mesocosm, which was compensated by compound-specific but time-limited and reversible sorption to macrophytes. The observations on the reversibility of sorption processes were subsequently confirmed by laboratory experiments. The experiments revealed that sorption to macrophytes lead to compound specific elimination from the aqueous phase during the presence of transient contaminant peaks in streams. After all, these sorption processes were found to be fully reversible, which results in the release of the primarily adsorbed compounds, once the concentrations in the aqueous phase starts to decrease. Nevertheless, the results of the present thesis demonstrate that the processes governing the mitigation of contaminant loads in streams are fundamentally different to those already described for non-flowing systems. In addition, the present thesis provides knowledge on how the interaction of macrophyte-induced processes in streams contributes to mitigate loads of organic contaminants and the related risk for aquatic environments.
Pelagic oxyclines, the transition zone between oxygen rich surface waters and oxygen depleted deep waters, are a common characteristic of eutrophic lakes during summer stratification. They can have tremendous effects on the biodiversity and the ecosystem functioning of lakes and, to add insult to injury, are expected to become more frequent and more pronounced as climate warming progresses. On these grounds, this thesis endeavors to advance the understanding of formation, persistence, and consequences of pelagic oxyclines: We test, whether the formation of metalimnetic oxygen minima is intrinsically tied to a locally enhanced oxygen consuming process, investigate the relative importance of vertical physical oxygen transport and biochemical oxygen consumption for the persistence of pelagic oxyclines, and finally assess their potential consequences for whole lake cycling. To pursue these objectives, the present thesis nearly exclusively resorts to in situ measurements. Field campaigns were conducted at three lakes in Germany featuring different types of oxyclines and resolved either a short (hours to days) or a long (weeks to months) time scale. Measurements comprised temperature, current velocity, and concentrations of oxygen and reduced substances in high temporal and vertical resolution. Additionally, vertical transport was estimated by applying the eddy correlation technique within the pelagic region for the first time. The thesis revealed, that the formation of metalimnetic oxygen minima does not necessarily depend on locally enhanced oxygen depletion, but can solely result from gradients and curvatures of oxygen concentration and depletion and their relative position to each other. Physical oxygen transport was found to be relevant for oxycline persistence when it considerably postponed anoxia on a long time scale. However, its influence on oxygen dynamics was minor on short time scales, although mixing and transport were highly variable. Biochemical consumption always dominated the fate of oxygen in pelagic oxyclines. It was primarily determined by the oxidative breakdown of organic matter originating from the epilimnion, whereas in meromictic lakes, the oxidation of reduced substances dominated. Beyond that, the results of the thesis emphasize that pelagic oxyclines can be a hotspot of mineralization and, hence, short-circuit carbon and nutrient cycling in the upper part of the water column. Overall, the present thesis highlights the importance of considering physical transport as well as biochemical cycling in future studies.
Today’s agriculture heavily relies on pesticides to manage diverse pests and maximise crop yields. Despite elaborate regulation of pesticide use based on a complex environmental risk assessment (ERA) scheme, the widespread use of these biologically active compounds has been shown to be a threat to the environment. For surface waters, pesticide exposure has been observed to exceed safe concentration levels and negatively impact stream ecology leading to the question whether current ERA schemes ensure a sustainable use of pesticides. To answer this, the large-scale “Kleingewässer-Monitoring” (KgM) assessed the occurrence of pesticides and related effects in 124 streams throughout Germany, Central Europe, in 2018 and 2019.
Based on five scientific publications originating from the KgM, this thesis evaluated pesticide exposure in streams, ecological effects and the regulatory implications. More than 1,000 water samples were analysed for over 100 pesticide analytes to characterise occurrence patterns (publication 1). Measured concentrations and effects were used to validate the exposure and effect concentrations predicted in the ERA (publication 2). By jointly analysing real-world pesticide application data and measured pesticide mixtures in streams, the disregard of environmental pesticide mixtures in the ERA was evaluated (publication 3). The toxic potential of mixtures in stream water was additionally investigated using suspect screening for 395 chemicals and a battery of in-vitro bioassays (publication 4). Finally, the results from the KgM stream monitoring were used to assess the capability to identify pesticide risks in governmental monitoring programmes (publication 5).
The results of this thesis reveal the widespread occurrence of pesticides in non-target stream ecosystems. The water samples contained a variety of pesticides occurring in complex mixtures predominantly in short-term peaks after rainfall events (publications 1 & 4). Respective pesticide concentration maxima were linked to declines in vulnerable invertebrate species and exceeded regulatory acceptable concentrations in about 80% of agricultural streams, while these thresholds were still estimated partly insufficient to protect the invertebrate community (publication 2). The co-occurrence of pesticides in streams led to a risk underestimated in the single substance-oriented ERA by a factor of about 3.2 in realistic worst-case scenarios, which is further exacerbated by a high frequency at which non-target organism are exposed to pesticides (publication 3). Stream water samples taken after rainfall caused distinct effects in bioassays which were only explainable to a minor extent by the many analytes, indicating the relevance of unknown chemical or biological mixture components (publication 4). Finally, the regulatory monitoring of surface waters under the Water Framework Directive (WFD) was found to significantly underestimate pesticide risks, as about three quarters of critical pesticides and more than half of streams at risk were overlooked (publication 5).
Essentially, this thesis involves a new level of validation of the ERA of pesticides in aquatic ecosystems by assessing pesticide occurrence and environmental impacts at a scale so far unique. The overall results demonstrate that the current agricultural use of pesticides leads to significant impacts on stream ecology that go beyond the level tolerated under the ERA. This thesis identified the underestimation of pesticide exposure, the potential insufficiency of regulatory thresholds and the general inertia of the authorisation process as the main causes why the ERA fails to meet its objectives. To achieve a sustainable use of pesticides, the thesis proposes substantial refinements of the ERA. Adequate monitoring programmes such as the KgM, which go beyond current government monitoring efforts, will continue to be needed to keep pesticide regulators constantly informed of the validity of their prospective ERA, which will always be subject to uncertainty.
Statistical eco(-toxico)logy
(2017)
Freshwaters are of immense importance for human well-being.
Nevertheless, they are currently facing unprecedented levels of threat from habitat loss and degradation, overexploitation, invasive species and
pollution.
To prevent risks to aquatic ecosystems, chemical substances, like agricultural pesticides, have to pass environmental risk assessment (ERA) before entering the market.
Concurrently, large-scale environmental monitoring is used for surveillance of biological and chemical conditions in freshwaters.
This thesis examines statistical methods currently used in ERA.
Moreover, it presents a national-scale compilation of chemical monitoring data, an analysis of drivers and dynamics of chemical pollution in streams and, provides a large-scale risk assessment by combination with results from ERA.
Additionally, software tools have been developed to integrate different datasets used in ERA.
The thesis starts with a brief introduction to ERA and environmental monitoring and gives an overview of the objectives of the thesis.
Chapter 2 addresses experimental setups and their statistical analyses using simulations.
The results show that current designs exhibit unacceptably low statistical power, that statistical methods chosen to fit the type of data provide higher power and that statistical practices in ERA need to be revised.
In chapter 3 we compiled all available pesticide monitoring data from Germany.
Hereby, we focused on small streams, similar to those considered in ERA and used threshold concentrations derived during ERA for a large-scale assessment of threats to freshwaters from pesticides.
This compilation resulted in the most comprehensive dataset on pesticide exposure currently available for Germany.
Using state-of-the-art statistical techniques, that explicitly take the limits of quantification into account, we demonstrate that 25% of small streams are at threat from pesticides.
In particular neonicotinoid pesticides are responsible for these threats.
These are associated with agricultural intensity and can be detected even at low levels of agricultural use.
Moreover, our results indicated that current monitoring underestimates pesticide risks, because of a sampling decoupled from precipitation events.
Additionally, we provide a first large-scale study of annual pesticide exposure dynamics.
Chapters 4 and 5 describe software solutions to simplify and accelerate the integration of data from ERA, environmental monitoring and ecotoxicology that is indispensable for the development of landscape-level risk assessment.
Overall, this thesis contributes to the emerging discipline of statistical ecotoxicology and shows that pesticides pose a large-scale threat to small streams.
Environmental monitoring can provide a post-authorisation feedback to ERA.
However, to protect freshwater ecosystems ERA and environmental monitoring need to be further refined and we provide software solutions to utilise existing data for this purpose.
Wild bees are essential for the pollination of wild and cultivated plants. However, within the
last decades, the increasing intensification of modern agriculture has led to both a reduction and fragmentation as well as a degradation of the habitats wild bees need. The resulting loss of pollinators and their pollination poses an immense challenge to global food production. To support wild bees, the availability of flowering resources is essential. However, the flowering period of each resource is temporally limited and has different effects on pollinators and their pollination, depending on the time of their flowering.
Therefore, to efficiently promote and manage wild bee pollinators in agricultural landscapes, we identified species-specific key floral resources of three selected wild bee species and their spatial and temporal availability (CHAPTERS 2, 3 & 4). We examined, which habitat types predominantly provide these resources (CHAPTERS 3 & 4). We also investigated whether floral resource maps based on the use of these key resources and their spatial and temporal availability explain the abundance and development of the selected wild bees (CHAPTERS 3 & 4) and pollination (CHAPTER 5) better than habitat maps, that only indirectly account for the availability of floral resources.
For each of the species studied, we were able to identify different key pollen sources, predominantly woody plants in the early season (April/May) and increasingly herbaceous plants in the later season (June/July; CHAPTERS 2, 3 & 4). The open woody semi-natural habitats of our agricultural landscapes provided about 75% of the floral resources for the buff-tailed bumblebees, 60% for the red mason bees, and 55% for the horned mason bees studied, although they accounted for only 3% of the area (CHAPTERS 3 & 4). In addition, fruit orchards provided about 35% of the floral resources for the horned mason bees on 4% of the landscape area (CHAPTER 3). We showed that both mason bee species benefited from the resource availability in the surrounding landscapes (CHAPTER 3). Yet this was not the case for the bumblebees (CHAPTER 4). Instead, the weight gain of their colonies, the number of developed queen cells and their colony survival were higher with increasing proximity to forests. The proximity to forests also had a positive effect on the mason bees studied (CHAPTER 3). In addition, the red mason bees benefited from herbaceous semi-natural habitats. The proportion of built-up areas had a negative effect on the horned mason bees, and the proportion of arable land on the red mason bees. The habitat maps explained horned mason bee abundances equally well as the floral resource maps, but red mason bee abundances were distinctly better explained by key floral resources. The pollination of field bean increased with higher proportions of early floral resources, whereas synchronous floral resources showed no measurable reduction in their pollination (CHAPTER 5). Habitat maps also explained field bean pollination better than floral resource maps. Here, pollination increased with increasing proportions of built-up areas in the landscapes and decreased with increasing proportions of arable land.
Our results highlight the importance of the spatio-temporal availability of certain key species as resource plants of wild bees in agricultural landscapes. They show that habitat maps are ahead of, or at least equal to, spatio-temporally resolved floral resource maps in predicting wild bee development and pollination. Nevertheless, floral resource maps allow us to draw more accurate conclusions between key floral resources and the organisms studied. The proximity to forest edges had a positive effect on each of the three wild bee species studied. However, besides pure food availability, other factors seem to co-determine the occurrence of wild bees in agricultural landscapes.
Soil organic matter (SOM) is a key component responsible for sequestration of organic molecules in soil and regulation of their mobility in the environment. The basic structure of SOM is a supramolecular assembly responding dynamically to the environmental factors and the presence of interacting molecules. Despite of the advances in the understanding of sorption processes, the relation between sorbate molecules, SOM supramolecular structure and its dynamics is limited. An example of a dynamic nature of SOM is a physicochemical matrix aging that is responsible for SOM structural arrangement. The underlying process of the physicochemical aging is the formation of water molecule bridges (WaMB) between functional groups of molecular segments. Since WaMB influence the stiffness of SOM structure, it was hypothesized that formation of WaMB contributes to the sequestration of organic molecules. However, this hypothesis has not been tested experimentally until now. Furthermore, the knowledge about the influence of organic molecules on WAMB is based solely on computer modeling studies. In addition, the influence of organic molecules on some physical phases forming SOM is not well understood. Especially, the interactions between organic molecules and crystalline phases represented by aliphatic crystallites, are only presumed. Thus, the investigation of those interactions in unfractioned SOM is of high importance.
In order to evaluate the involvement of WaMB in the sequestration of organic molecules and to increase our understanding about interactions of organic chemicals with WaMB or aliphatic crystallites, the following hypotheses were tested experimentally. 1) Similarly to crystalline phases in synthetic polymers, aliphatic crystallites, as a part of SOM, cannot be penetrated by organic molecules. 2) The stability of WaMB is determined by the ability of surrounding molecules to interact with water forming WaMB. 3) WaMB prevent organic molecules to leave the SOM matrix and contribute thus to their immobilization. In order to test the hypotheses 1 and 2, a set of experiments including treatment of soils with chosen chemicals was prepared. Interaction abilities of these chemicals were characterized using interaction parameters from the Linear Solvation Energy Relationship theory. WaMB characteristics were monitored using Differential Scanning Calorimetry (DSC) allowing to measure the WaMB thermal stability and the rigidity of SOM matrix; which in turn was determined by the heat capacity change. In addition, DSC and 13C NMR spectroscopy assessed thermal properties and the structure of aliphatic crystallites. The spiking of samples with a model compound, phenol, and measurements of its desorption allowed to link parameters of the desorption kinetics with WaMB characteristics.
The investigation showed that the WaMB stability is significantly reduced by the presence of molecules with H-donor/acceptor interaction abilities. The matrix rigidity associated with WaMB was mainly influenced by the McGowan’s volume of surrounding molecules, suggesting the importance of dispersion forces. The desorption kinetics of phenol followed a first order model with two time constants. Both of them showed a relation with WaMB stability, which supports the hypothesis that WaMB contribute to the physical immobilization of organic molecules. The experiments targeted to the crystallites revealed their structural change from the ordered to the disordered state, when in contact with organic chemicals. This manifested in their melting point depression and the decrease of overall crystallinity. Described structural changes were caused by molecules interacting with specific as well as non-specific forces, which suggests that aliphatic crystallites can be penetrated and modified by molecules with a broad range of interaction abilities.
This work shows that chosen organic molecules interact with constituents of SOM as exemplified on WaMB and aliphatic crystallites, and cause measurable changes of their structure and properties. These findings show that the relevance of aliphatic crystallites for sorption in soil may have been underestimated. The results support the hypothesis that physicochemical matrix aging significantly contributes to the immobilization of organic chemicals in SOM.
The estimation of the potential risk of pesticide entries into streams - and therefore the potential risk for the ecosystems - is an important requirement for the planning of risk mitigation strategies. Especially on the landscape level the required event triggered sampling methods are conjuncted with considerable efforts with regard to input data, time and personnel. To circumvent these problems simulation models form a reasonable alternative. The aims of this work were (A) the development of a simulation tool for the estimation of pesticide entries into surface waters on the landscape level, and (B) the application of the simulator for an exposure- and risk-assessment as well as the assessment of negative effects of pesticides on aquatic communities. Section 1 - Exposure-, Risk- and Effects In sections 1.1 and 1.2 the simulation model was applied to a multitude of small and medium sized streams in an agricultural impacted study area around the city of Braunschweig, Germany. Section 1.3 gives an overview of the simulators field of application and the general system structure. Section 1.1 - Scenario based simulation of runoff-related pesticide entries into small streams on a landscape level (English publication, p. 27): In this paper we present a simulation tool for the simulation of pesticide entry from arable land into adjacent streams. We used the ratio of exposure to toxicity (REXTOX) model proposed by the OECD which was extended to calculate pesticide concentrations in adjacent streams. We simulated the pesticide entry on the landscape level at 737 sites in small streams situated in the central lowland of Germany. The most significant model parameters were the width of the no-application-zone and the degree of plant-interception. The simulation was carried out using eight different environmental scenarios, covering variation of the width of the no-application-zone, climate and seasonal scenarios. The highest in-stream concentrations were predicted at a scenario using no (0 m) buffer zone in conjunction with increased precipitation. According to the predicted concentrations, the risk for the aquatic communities was estimated based on standard toxicity tests and the application of a safety factor. Section 1.2 - Linking land use variables and invertebrate taxon richness in small and medium-sized agricultural streams on a landscape level (English publication, p. 50): In this study the average numbers of invertebrate species across an arable landscape in central Germany (surveys from 15 years in 90 streams at 202 sites) were assessed for their correlation with environmental factors such as stream width, land use (arable land, forest, pasture, settlement), soil type and agricultural derived stressors. The stress originating from arable land was estimated by the factor "risk of runoff", which was derived from a runoff-model (rainfall induced surface runoff). Multivariate analysis explained 39.9% of the variance in species number, revealing stream width as the most important factor (25.3%) followed by risk of runoff (9.7%). Section 1.3 - Informationssystem zur ökotoxikologischen Bewertung der Gewässergüte in Bezug auf Pflanzenschutzmitteleinträge aus der Landwirtschaft - Systemaufbau und Anwendungsmöglichkeiten (German publication, p. 61): Section 1.3 contains a short overview of the simulation tool, the field of application and some examples of use, covering the effects of the width of the buffer zone as well as the creation of risk maps on the landscape level. Section 2 - The simulation tool An important aspect for the employment of a simulation model in the context of risk assessment is the applicability in practice: the accessibility of the needed input data, the conversion of the mathematical model into a software application that can be run on any current personnel computer and also an appropriate end-user documentation of the system. Section 1.4 - Informationssystem zur ökotoxikologischen Bewertung der Gewässergüte in Bezug auf Pflanzenschutzmitteleinträge aus der Landwirtschaft - Simulationsmodell und Systemaufbau (German report, p. 67): In this section a general overview of the simulation model as well as the schematic system structure given. Section 1.5 - Benutzerhandbuch (German report, p. 71): The user manual contains details concerning the installation of the system, generation of the required input data and the general use of the system. Moreover it presents some application examples (what-if analyses). Section 1.6 - Technical documentation (German report, p. 104): The technical documentation describes internal structures and processes of the simulation system. Section 1.6 provides information regarding the required structure of input/output tables.
Inland waters play an active role in the global carbon cycle. They collect carbon from upstream landmasses and transport it downstream until it finally reaches the ocean. Along this path, manifold processing steps are evident, resulting in (permanent) retention of carbon by sediment burial as well as loss by evasion to the atmosphere. Constraining these carbon fluxes and their anthropogenic perturbation is an urgent need. In this context, attention needs to be set on a widespread feature of inland waters: their partial desiccation. This results in the emergence of formerly inundated sediments to the atmosphere, referred to as dry inland waters. One observed feature of dry inland waters are disproportional high carbon dioxide (CO2) emissions. However, this observation was so far based on local case studies and knowledge on the global prevalence and fundamental mechanisms of these emissions is lacking. Against this background, this thesis aims to provide a better understanding of the magnitude and mechanisms of carbon emissions from dry inland waters on the global and local scale and to assess the impact of dry inland waters on the global carbon cycle. The specific research questions of this thesis were: (1) How do gaseous carbon emissions from dry inland waters integrate into the global carbon cycle and into global greenhouse gas (GHG) budgets? (2) What effect do seasonal and long term drying have on the carbon cycling of inland waters? The thesis revealed that dry inland waters emit disproportional large amounts of CO 2 on a global scale and that these emissions share common drivers across ecosystems. Quantifying global reservoir drawdown and upscaling carbon fluxes to the global scale suggests that reservoirs emit more carbon than they bury, challenging the current understanding of reservoirs as net carbon sinks. On the local scale, this thesis revealed that both, heterogeneous emission pattern between different habitats and seasonal variability of carbon emissions from the drawdown area, needs to be considered. Further, this thesis showed that re-mobilization of buried carbon upon permanent desiccation of water bodies can explain the observed emission rates, supporting the hypothesis of a positive feedback-loop between climate change and desiccation of inland waters. Overall, the present thesis highlights the importance of adding emissions from dry inland waters as a pathway to the global carbon cycle of inland waters.
Although most plastic pollution originates on land, current research largely remains focused on aquatic ecosystems. Studies pioneering terrestrial microplastic research have adapted analytical methods from aquatic research without acknowledging the complex nature of soil. Meanwhile, novel methods have been developed and further refined. However, methodical inconsistencies still challenge a comprehensive understanding of microplastic occurrence and fate in and on soil. This review aims to disentangle the variety of state-of-the-art sample preparation techniques for heterogeneous solid matrices to identify and discuss best-practice methods for soil-focused microplastic analyses. We show that soil sampling, homogenization, and aggregate dispersion are often neglected or incompletely documented. Microplastic preconcentration is typically performed by separating inorganic soil constituents with high-density salt solutions. Not yet standardized but currently most used separation setups involve overflowing beakers to retrieve supernatant plastics, although closed-design separation funnels probably reduce the risk of contamination. Fenton reagent may be particularly useful to digest soil organic matter if suspected to interfere with subsequent microplastic quantification. A promising new approach is extraction of target polymers with organic solvents. However, insufficiently characterized soils still impede an informed decision on optimal sample preparation. Further research and method development thus requires thorough validation and quality control with well-characterized matrices to enable robust routine analyses for terrestrial microplastics.
The increasing, anthropogenic demand for chemicals has created large environmental problems with repercussions for the health of the environment, especially aquatic ecosystems. As a result, the awareness of the public and decision makers on the risks from chemical pollution has increased over the past half-century, prompting a large number of studies in the field of ecological toxicology (ecotoxicology). However, the majority of ecotoxicological studies are laboratory based, and the few studies extrapolating toxicological effects in the field are limited to local and regional levels. Chemical risk assessment on large spatial scales remains largely unexplored, and therefore, the potential large-scale effects of chemicals may be overlooked.
To answer ecotoxicological questions, multidisciplinary approaches that transcend classical chemical and toxicological concepts are required. For instance, the current models for toxicity predictions - which are mainly based on the prediction of toxicity for a single compound and species - can be expanded to simultaneously predict the toxicity for different species and compounds. This can be done by integrating chemical concepts such as the physicochemical properties of the compounds with evolutionary concepts such as the similarity of species. This thesis introduces new, multidisciplinary tools for chemical risk assessments, and presents for the first time a chemical risk assessment on the continental scale.
After a brief introduction of the main concepts and objectives of the studies, this thesis starts by presenting a new method for assessing the physiological sensitivity of macroinvertebrate species to heavy metals (Chapter 2). To compare the sensitivity of species to different heavy metals, toxicity data were standardized to account for the different laboratory conditions. These rankings were not significantly different for different heavy metals, allowing the aggregation of physiological sensitivity into a single ranking.
Furthermore, the toxicological data for macroinvertebrates were used as input data to develop and validate prediction models for heavy metal toxicity, which are currently lacking for a wide array of species (Chapter 3). Apart from the toxicity data, the phylogenetic information of species (evolutionary relationships among species) and the physicochemical parameters for heavy metals were used. The constructed models had a good explanatory power for the acute sensitivity of species to heavy metals with the majority of the explained variance attributed to phylogeny. Therefore, the integration of evolutionary concepts (relatedness and similarity of species) with the chemical parameters used in ecotoxicology improved prediction models for species lacking experimental toxicity data. The ultimate goal of the prediction models developed in this thesis is to provide accurate predictions of toxicity for a wide range of species and chemicals, which is a crucial prerequisite for conducting chemical risk assessment.
The latter was conducted for the first time on the continental scale (Chapter 4), by making use of a dataset of 4,000 sites distributed throughout 27 European countries and 91 respective river basins. Organic chemicals were likely to exert acute risks for one in seven sites analyzed, while chronic risk was prominent for almost half of the sites. The calculated risks are potentially underestimated by the limited number of chemicals that are routinely analyzed in monitoring programmes, and a series of other uncertainties related with the limit of quantification, the presence of mixtures, or the potential for sublethal effects not covered by direct toxicity.
Furthermore, chemical risk was related to agricultural and urban areas in the upstream catchments. The analysis of ecological data indicated chemical impacts on the ecological status of the river systems; however, it is difficult to discriminate the effects of chemical pollution from other stressors that river systems are exposed to. To test the hypothesis of multiple stressors, and investigate the relative importance of organic toxicants, a dataset for German streams is used in chapter 5. In that study, the risk from abiotic (habitat degradation, organic chemicals, and nutrients enrichment) and biotic stressors (invasive species) was investigated. The results indicated that more than one stressor influenced almost all sites. Stream size and ecoregions influenced the distribution of risks, e.g., the risks for habitat degradation, organic chemicals and invasive species increased with the stream size; whereas organic chemicals and nutrients were more likely to influence lowland streams. In order to successfully mitigate the effects of pollutants in river systems, co-occurrence of stressors has to be considered. Overall, to successfully apply integrated water management strategies, a framework involving multiple environmental stressors on large spatial scales is necessary. Furthermore, to properly address the current research needs in ecotoxicology, a multidisciplinary approach is necessary which integrates fields such as, toxicology, ecology, chemistry and evolutionary biology.
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.