Fachbereich 7
Refine
Year of publication
Document Type
- Doctoral Thesis (58)
- Article (2)
- Bachelor Thesis (1)
Keywords
- Agriculture (2)
- Bestäubung (2)
- Biodiversität (2)
- Ecotoxicology (2)
- Grundwasserfauna (2)
- Landwirtschaft (2)
- Pesticides (2)
- Pflanzenschutzmittel (2)
- Weinbau (2)
- decomposition (2)
Institute
- Fachbereich 7 (61) (remove)
In the last decades, it became evident that the world is facing an unprecedented, human-induced global biodiversity crisis with amphibians being one of the most threatened species groups. About 41% of the amphibian species are classified as endangered by the IUCN, but even in amphibian species that are listed as "least concern", population declines can be observed on a local level. With land-use change and agrochemicals (i.e. pesticides), two of the main drivers for this amphibian decline are directly linked to intensive agriculture, which is the dominant landscape type in large parts of Europe. Thus, understanding the situation of amphibians in the agricultural landscape is crucial for conservation measures. In the present thesis, I investigated the effects of viticulture on amphibian populations around Landau in der Pfalz (Germany) in terms of habitat use, pesticide exposure, biometric traits as well as genetic and age structure. From the perspective of amphibians, land-use change means usually the destruction of habitats in agricultural landscapes, which often leads to landscape fragmentation. Thus, I followed the question if also vineyards lead to the fragmentation of the landscape and if pesticides that are frequently used in viticulture have to be considered as a factor too, so if there is a chemical landscape fragmentation. Using telemetry, I could show that common toads (Bufo bufo) can be found directly in vineyards, but that they tend to avoid them as habitat. Analysing the genetic structure of common frogs (Rana temporaria) revealed that vineyards have to be considered as a barrier for amphibians. To identify if pesticides contribute to the resulting landscape fragmentation, I conducted an arena choice experiment in the laboratory in which I found evidence for an avoidance of pesticide-contaminated soil. Such an avoidance could be one of the underlying reasons for a potential chemical landscape fragmentation. By combining telemetry data with information about pesticide applications from local wine growers, I could show that a large part of the common toads is likely to come in contact with pesticides. Further, I demonstrated that the agricultural landscape, probably due to the application of pesticides, can have negative effects on the reproduction capacity of common toads. By studying palmate newts (Lissotriton helveticus) I found that adult newts from agricultural ponds are smaller than those from forest ponds. As I did not find differences in the age structure and growth, these differences might be carry-over effects from earlier life stages. While agricultural ponds might be suitable habitats for adult palmate newts, the potential carry-over effect indicates suboptimal conditions for larvae and/or juveniles. I conclude that the best management measure for sustaining amphibians in the agricultural landscape would be a heterogeneous cultural landscape with a mosaic of different habitat patches that work without or at least a reduced amount of pesticides. Green corridors between populations and different habitats would allow migrating individuals to avoid agricultural and thus pesticide-contaminated areas. This would reduce the pesticide exposure risk of amphibians, while preventing the fragmentation of the landscape and thus the isolation of populations.
Ponds in agricultural landscapes are often used by amphibians as breeding habitat. However, the characteristics of agricultural ponds and especially the surrounding area are usually said to be suboptimal for many amphibian species. Using suboptimal habitats might allow a species’ survival and reproduction, but can have negative consequences at the individual and population level. In the present study, we investigated Palmate Newt (Lissotriton helveticus) populations from an intensive wine-growing region in southern Germany and compared them with populations located in a nearby forested area in terms of biometric traits, age and genetic structure. By analyzing over 900 adult newts from 11 ponds, we could show that newts reproducing in forest ponds were larger than newts reproducing in agricultural ponds. We did not find differences in the newt age and growth rate between habitat types. Therefore, differences in the body size of newts might already existed in larvae and/or juveniles, what might be related to a lower habitat quality for larvae and/or juveniles in the agricultural landscape. Body mass, body condition and sexual dimorphic traits (length of the caudal filament and max. height of the tail) correlated with body size, but no additional effect of the habitat type was found. The analysis of microsatellites revealed a higher genetic diversity in forest ponds. However, no clear sign of inbreeding was observed in any agricultural population, suggesting some degree of gene flow between them. We conclude, that agricultural ponds can be suitable habitats for the Palmate Newt and that conservation effort should aim to preserve them. The observed effects on body size indicate the need to increase the quality of the aquatic and terrestrial habitat for early life stages of this newt species in agricultural landscapes.
In der Biologie stellt das Zeichnen eine zentrale Arbeitstechnik dar. Viele Studien konnten auf einen positiven Effekt des Zeichnens für bestimmte Situationen hinweisen. Schülerinnen und Schüler müssen diese Technik jedoch zunächst erlernen. Hierbei können zahlreiche Schwierigkeiten auftreten, die die inhaltliche Auseinandersetzung gefährden. Jedoch wurden sowohl Schwierigkeiten im Umgang mit unterschiedlichen Repräsentationsformen als auch der Zeichenprozess bislang nur lückenhaft untersucht. Die Studie dieser Arbeit hat daher zum Ziel, (I) den Zeichenprozess auf der Ebene der Sichtstruktur zu beschreiben, (II) die manifesten Schwierigkeiten von Lernenden zu erfassen, auf die sie während der Konstruktion biologisch bedeutsamer Repräsentationsformen (Ablaufdiagramme, mikroskopische Zeichnungen) treffen, (III) und auf Grundlage der empirischen Befunde Schülertypen abzuleiten. Vor diesem Hintergrund waren 21 Schülerinnen und Schüler angehalten, jeweils ein Ablaufdiagramm auf Grundlage eines Texts und eine mikroskopische Zeichnung auf Grundlage eines Präparats zu konstruieren und dabei laut zu denken. Fragen zur Vorerfahrung sowie retrospektiv gestellte Fragen zum Vorgehen der Teilnehmenden umrahmten den videografisch dokumentierten Prozess. Die Ergebnisse zeigen, dass der Zeichenprozess mehr als zehn unterschiedliche Tätigkeiten umfassen kann, wobei die Kerntätigkeit des Zeichnens durchschnittlich nur rund ein Drittel des Prozesses ausmacht. Die Prozessstruktur zwischen Fällen variiert erheblich. Weiterhin konnten etwa 30 Schwierigkeiten bzw. Fehler identifiziert werden, die während der Konstruktion beider Repräsentationsformen auftreten. Diese können dabei sowohl einzelne als auch mehrere Tätigkeiten betreffen und zu Tätigkeitsabbrüchen führen. Schwierigkeiten stehen häufig in Verbindung mit Tätigkeiten, die außerhalb der Kerntätigkeit des Zeichnens liegen (z. B. Abgleich mit der Textgrundlage). Bezogen auf Ablaufdiagramme stellt das Verhältnis depiktional bzw. deskriptional dargestellter Textinformationen den Ausgangspunkt der Typisierung dar: Typ I: realistisch abbildend, II: alternierend abbildend und III: schriftorientiert abbildend. Für mikroskopische Zeichnungen war die Häufigkeit des Abgleichs mit dem Objekt grundlegend für die Typisierung: Typ I: oberflächlich abbildend, II: objektorientiert abbildend und III: undifferenziert detailliert abbildend. Die Studie liefert erstmals Kategoriensysteme, die es erlauben, die Prozessstruktur des Zeichnens sichtbar und zwischen Fällen vergleichbar zu machen sowie schwierigkeitsbezogenes Grundlagenwissen zur Konstruktion von Zeichnungen, basierend auf Texten und Beobachtungen. Die Übertragbarkeit der Befunde auf andere Repräsentationsformen ist an vielen Stellen denkbar. Die theoretisch fundierte Systematisierung von Schwierigkeiten kann von weiterführenden Untersuchungsansätze aufgegriffen werden und erlaubt die Verortung situationsangemessener Unterstützungsmaßnahmen.
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.
Diet-related effects of antimicrobials in aquatic decomposer-shredder and periphyton-grazer systems
(2022)
Leaf-associated microbial decomposers as well as periphyton serve as important food sources for detritivorous and herbivorous macroinvertebrates (shredders and grazers) in streams. Shredders and grazers, in turn, provide not only collectors with food but also serve as prey for predators. Therefore, decomposer-shredder and periphyton-grazer systems (here summarized as freshwater biofilm-consumer systems) are highly important for the energy and nutrient supply in heterotrophic and autotrophic stream food webs. However, both systems can be affected by chemical stressors, amongst which antimicrobials (e.g., antibiotics, fungicides and algaecides) are of particular concern. Antimicrobials can impair shredders and grazers not only via waterborne exposure (waterborne effect pathway) but also through dietary exposure and microorganism-mediated alterations in the food quality of their diet (dietary effect pathway). Even though the relevance of the latter pathway received more attention in recent years, little is known about the mechanisms that are responsible for the observed effects in shredders and grazers. Therefore, the first objective of this thesis was to broaden the knowledge of indirect antimicrobial effects in a model shredder and grazer via the dietary pathway. Moreover, although freshwater biofilm-consumer systems are most likely exposed to antimicrobial mixtures comprised of different stressor groups, virtually nothing is known of these mixture effects in both systems. Therefore, the second objective was to assess and predict diet-related antimicrobial mixture effects in a model freshwater biofilm-consumer system. During this thesis, positive diet-related effects of a model antibiotic on the energy processing and physiology of the shredder Gammarus fossarum were observed. They were probably triggered by shifts in the leaf-associated microbial community in favor of aquatic fungi that increased the food quality of leaves for the shredder. Contrary to that, a model fungicide induced negative effects on the energy processing of G. fossarum via the dietary pathway, which can be explained by negative impacts on the microbial decomposition efficiency leading to a reduced food quality of leaf litter for gammarids. For diet-related antimicrobial effects in periphyton-grazer systems, a model algaecide altered the periphyton community composition by increasing nutritious and palatable algae. This resulted in an enhanced consumption and physiological fitness of the grazer Physella acuta. Finally, it was shown that complex horizontal interactions among leaf-associated microorganisms are involved, making diet-related antimicrobial mixture effects in the shredder G. fossarum difficult to predict. Thus, this thesis provides new insights into indirect diet-related effects of antimicrobials on shredders and grazers as well as demonstrates uncertainties of antimicrobial mixture effect predictions for freshwater biofilm-consumer systems. Moreover, the findings in this thesis are not only informative for regulatory authorities, as indirect effects and effects of mixtures across chemical classes are not considered in the environmental risk assessment of chemical substances, but also stimulate future research to close knowledge gaps identified during this work.
The use of agricultural plastic covers has become common practice for its agronomic benefits such as improving yields and crop quality, managing harvest times better, and increasing pesticide and water use efficiency. However, plastic covers are suspected of partially breaking down into smaller debris and thereby contributing to soil pollution with microplastics. A better understanding of the sources and fate of plastic debris in terrestrial systems has so far been hindered by the lack of adequate analytical techniques for the mass-based and polymer-selective quantification of plastic debris in soil. The aim of this dissertation was thus to assess, develop, and validate thermoanalytical methods for the mass-based quantification of relevant polymers in and around agricultural fields previously covered with fleeces, perforated foils, and plastic mulches. Thermogravimetry/mass spectrometry (TGA/MS) enabled direct plastic analyses of 50 mg of soil without any sample preparation. With polyethylene terephthalate (PET) as a preliminary model, the method limit of detection (LOD) was 0.7 g kg−1. But the missing chromatographic separation complicated the quantification of polymer mixtures. Therefore, a pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS) method was developed that additionally exploited the selective solubility of polymers in specific solvents prior to analysis. By dissolving polyethylene (PE), polypropylene (PP), and polystyrene (PS) in a mixture of 1,2,4-trichlorobenzene and p-xylene after density separation, up to 50 g soil became amenable to routine plastic analysis. Method LODs were 0.7–3.3 mg kg−1, and the recovery of 20 mg kg−1 PE, PP, and PS from a reference loamy sand was 86–105%. In the reference silty clay, however, poor PS recoveries, potentially induced by the additional separation step, suggested a qualitative evaluation of PS. Yet, the new solvent-based Py-GC/MS method enabled a first exploratory screening of plastic-covered soil. It revealed PE, PP, and PS contents above LOD in six of eight fields (6% of all samples). In three fields, PE levels of 3–35 mg kg−1 were associated with the use of 40 μm thin perforated foils. By contrast, 50 μm PE films were not shown to induce plastic levels above LOD. PP and PS contents of 5–19 mg kg−1 were restricted to single observations in four fields and potentially originated from littering. The results suggest that the short-term use of thicker and more durable plastic covers should be preferred to limit plastic emissions and accumulation in soil. By providing mass-based information on the distribution of the three most common plastics in agricultural soil, this work may facilitate comparisons with modeling and effect data and thus contribute to a better risk assessment and regulation of plastics. However, the fate of plastic debris in the terrestrial environment remains incompletely understood and needs to be scrutinized in future, more systematic research. This should include the study of aging processes, the interaction of plastics with other organic and inorganic compounds, and the environmental impact of biodegradable plastics and nanoplastics.
Agriculture requires a sustainable intensification to feed the growing world population without exacer-bating soil degradation and threatening soil quality. Globally, plastic mulching (PM) is increasingly used to improve crop growth and yields and consequently agronomic productivity. However, recent literature reported also critical aspects of PM for soil quality and showed contradictory outcomes. This might result from the numerous applications of PM in different climates across various crops, soils and agri-cultural techniques. Thus, a closer look is necessary on how PM influences soil processes under certain climate and cultivation conditions to obtain a comprehensive understanding of its effects, which is im-portant to evaluate PM in terms of a sustainable agriculture.
The aim of this PhD thesis was to understand how multiannual PM influences soil properties and pro-cesses under the temperate, humid Central European cultivation conditions and to evaluate the resulting consequences for soil quality. I designed a three-year field study to investigate the influence of PM (black polyethylene, 50 μm) on microclimate, structural stability, soil organic matter (SOM) and the concentrations of selected fungicides and mycotoxins in three soil layers (0–10, 10–30 and 30–60 cm) compared to straw mulching (SM). Both mulching types were applied in a drip-irrigated ridge-furrow system in strawberry cultivation.
PM shifted the soil microclimate to higher soil temperatures and lower soil moistures. The higher soil temperature seems thus to be the key factor for the increased crop growth and yields under the present humid climate. The reduced soil moisture under PM indicated that under PM the impeded rainfall infil-tration had a stronger effect on the water balance than the reduced evaporation. This indicate an ineffi-cient rainwater use in contrast to arid climates. PM changed the water cycling in the ridges from down-ward directed water flows to lateral water flows from furrows to ridges. This reduced nitrogen leaching in the topsoil (0–10 cm) in the strawberry establishment period. The plastic mulches avoided aggregate breakdown due to rapid soil wetting and excess water during rainfalls and thus maintained a loose and stable soil structure in the surface soil, which prevents soil compaction and made soil less prone to erosion. PM changed carbon fluxes and transformation so that a larger total and more stable SOM was observed. Thus, the higher belowground biomass productivity under PM compensated the impeded aboveground biomass input and the temperature-induced SOM decomposition. However, SM increased the labile and total SOM in the topsoil after the first experiment year and promoted microbial growth due to the aboveground biomass incorporation. PM reduced fungicide entry into soil compared to SM and reduced consequently the fungal biomass reduction and the biosynthesis of the mycotoxin deoxyni-valenol. The modified microclimate under PM did not increase mycotoxin occurrence. In this context, PM poses no risk for an increased soil contamination, impairing soil quality. This PhD thesis demon-strated that the PM effects on soil can vary depending on time, season and soil depth, which emphasizes the importance to include soil depth and time in future studies.
Compared to semiarid and arid regions, the PM effects found in this PhD thesis were small, absent or in another way. I attributed this to the fact that PM under humid climate reduced instead of increased soil moisture and that SM had due to straw und strawberry canopy a similar ‘covering effect’ as PM. Thus, generalizing the PM effects on soil across different climates seems hardly possible as they differ in type and extent depending on climate. A differentiated consideration is hence necessary to evaluate the PM effects on soil quality. I conclude that PM under temperate, humid climate might contribute to reduce soil degradation (e.g., SOM depletion, erosion, nutrient leaching, soil compaction and soil contamina-tion), which sustains soil quality and helps to enable a sustainable agricultural intensification. However, further research is necessary (1) to support my findings on a larger scale, longer time periods and across various soil and crop types, (2) to address remaining open questions and (3) to develop optimization to overcome the critical aspects of PM (e.g. macro- and microplastic waste in soil, mulch disposal).
Invasive species play increasing roles worldwide. Invasions are considered successful when species establish and spread in their exotic range. Subsequently, dispersal is a major determinant of species’ range dynamics. Mermessus trilobatus, native to North America, has rapidly spread in Europe via aerial dispersal. Here we investigated the interplay of ecological and evolutionary processes behind its colonisation success.
First, we examined two possible ecological mechanisms. Similar to other invasive invertebrates, the colonisation success of Mermessus trilobatus might be related to human-induced habitat disturbance. Opposite to this expectation, our results showed that densities of Mermessus trilobatus decreased with soil disturbance in grasslands suggesting that its invasion success was not connected to a ruderal strategy. Further, invasive species often escape the ecological pressures from novel enemies in their exotic ranges. Unexpectedly, invasive Mermessus trilobatus was more sensitive to a native predator than native Erigone dentipalpis during our predator susceptibility trials. This indicates that the relation between the invasive spider and its native predator is dominated by prey naïveté rather than enemy release.
The remaining three chapters of the thesis investigated the dispersal behaviour of this invasive species. Hitherto, studies of passive aerial dispersal used wind as the primary dispersal-initiating factor despite a recent demonstration of the effects of the atmospheric electric fields on spiders’ pre-dispersal behaviour. During our experiments, only the wind facilitated the flight, although electric fields induced pre-dispersal behaviour in spiders. Consequently, studies around passive aerial dispersal should control electric fields but use wind as a stimulating factor.
Rapidly expanding species might be disproportionately distributed in their exotic range, with an accumulation of dispersive genotypes at the leading edge of their range. Such imbalanced spatial segregation is possible when the dispersal behaviour of expanding species is heritable. Our results showed that the dispersal traits of Mermessus trilobatus were heritable through both parents and for both sexes with recessive inheritance of high dispersal ability in this species.
Following the heritability experiments, we documented an accelerated spread of Mermessus trilobatus in Europe and tested whether dispersal, reproduction or competing ability was at the source of this pattern. Our results showed that the accumulation of more mobile but not reproductive or competitive genotypes at the expansion front of this invasive species gave rise to an accelerated range expansion by more than 1350 km in under 45 years.
Invasive Mermessus trilobatus is inferior to native sympatric species with respect to competing ability (Eichenberger et al., 2009), disturbance tolerance and predation pressure. Nevertheless, the species successfully established in its exotic range and spread by accelerating its expansion rate. Rapid reproduction that balances the high ecological pressures might be the other potential mechanism behind its colonisation success in Europe and deserves further investigation.
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.
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.