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Institute
Fungicide effects on the structure and functioning of leaf-associated aquatic fungal communities
(2022)
Aquatic hyphomycetes are a polyphyletic group of saprotrophic fungi growing abundantly on submerged leaf litter. In stream ecosystems shaped by allochthonous leaf litter inputs, they play a central functional role as decomposers and food source for other organisms. Fungicides pose a threat to aquatic hyphomycetes and their functions, since these substances are inherently toxic to fungi and contaminate surface waters around the world due to their widespread use in agricultural and urban landscapes. While fungicides’ potential to reduce fungal diversity are discerned, the extent of impacts on biodiversity-ecosystem functioning relationships (B EF) remains unclear. This is partly attributed to methodological constraints in the detection and quantification of single aquatic hyphomycete species within microbial leaf-associated communities. The primary aim of this thesis was, therefore, (1) to assess the ecotoxicological impacts of fungicides on B-EF relationships in aquatic hyphomycete communities. To facilitate this, subordinate aims were to (2) develop DNA-based biomolecular tools (i.e., qPCR assays) to detect and to quantify the biomass of different aquatic hyphomycete species in mixed cultures and (3) to investigate the mechanisms underlying B-EF relationships in the absence of chemical stressors.
In the course of this thesis, qPCR assays were developed for detection and species-specific biomass quantification of ten common aquatic hyphomycete species and successfully validated for application in eco( toxico )logical microcosm experiments. Via a systematic manipulation of fungal diversity, these assays allow the examination of B-EF relationships by assessments of deviations between observed and (monoculture-based) predicted activities in fungal mixed cultures. Taking advantage of these tools in a microcosm experiment, it was uncovered that leaf decomposition results from the additive activity of community members, even though functionally distinct species were present. Colonization dynamics are characterized by complex interactions. Colonization success of aquatic hyphomycetes is higher if co-occurring species are genetically and functionally distinct (i.e., complementary interactions). However, the co-occurrence of aquatic hyphomycete species does not necessarily result in a greater colonization success compared to monocultures, unless bacteria are present. Accordingly, the presence of other microbial groups such as bacteria may induce new fungal diversity-based feedback loops, which ultimately enable coexistence of aquatic hyphomycete species in the environment. Exposure to fungicides revealed substantial differences in sensitivities among aquatic hyphomycetes. The most productive species were able to cope with extremely high fungicide concentrations up to the mg/L-range. In assemblages containing these species, leaf decomposition was maintained under fungicide exposure. Yet, already at environmentally relevant fungicide concentrations, tolerant species displaced more sensitive ones, potentially affecting leaves’ nutritional quality for consumers. This thesis thus indicates that fungicide exposure poses a risk to stream food webs rather than the microbial leaf decomposition process per se.
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.
Environmental processes transforming inorganic nanoparticles: implications on aquatic invertebrates
(2020)
Engineered inorganic nanoparticles (EINPs) are produced and utilized on a large scale and will end up in surface waters. Once in surface waters, EINPs are subjected to transformations induced by environmental processes altering the particles’ fate and inherent toxicity. UV irradiation of photoactive EINPs is defined as one effect-inducing pathway, leading to the formation of reactive oxygen species (ROS), increasing EINP toxicity by exerting oxidative stress in aquatic life. Simultaneously, UV irradiation of photoactive EINP alters the toxicity of co-occurring micropollutants (e.g. pesticides) by affecting their degradation. The presence of natural organic matter (NOM) reduces the agglomeration and sedimentation of EINPs, extending the exposure of pelagic species, while delaying the exposure of benthic species living in and on the sediment, which is suggested as final sink for EINPs. However, the joint impact of NOM and UV irradiation on EINP-induced toxicity, but also EINP-induced degradation of micropollutants, and the resulting risk for aquatic biota, is poorly understood. Although potential effects of EINPs on benthic species are increasingly investigated, the importance of exposure pathways (waterborne or dietary) is unclear, along with the reciprocal pathway of EINPs, i.e. the transport back from aquatic to terrestrial ecosystems. Therefore, this thesis investigates: (i) how the presence of NOM affects the UV-induced toxicity of the model EINP titanium dioxide (nTiO2) on the pelagic organism Daphnia magna, (ii) to which extent UV irradiation of nTiO2 in the presence and absence of NOM modifies the toxicity of six selected pesticides in D. magna, (iii) potential exposure pathway dependent effects of nTiO2 and silver (nAg) EINPs on the benthic organism Gammarus fossarum, and (iv) the transport of nTiO2 and gold EINPs (nAu) via the merolimnic aquatic insect Chaetopteryx villosa back to terrestrial ecosystems. nTiO2 toxicity in D. magna increased up to 280-fold in the presence of UV light, and was mitigated by NOM up to 12-fold. Depending on the pesticide, UV irradiation of nTiO2 reduced but also enhanced pesticide toxicity, by (i) more efficient pesticide degradation, and presumably (ii) formation of toxic by-products, respectively. Likewise, NOM reduced and increased pesticide toxicity, induced by (i) protection of D. magna against locally acting ROS, and (ii) mitigation of pesticide degradation, respectively. Gammarus’ energy assimilation was significantly affected by both EINPs, however, with distinct variation in direction and pathway dependence between nTiO2 and nAg. EINP presence delayed C. villosa emergence by up to 30 days, and revealed up to 40% reduced lipid reserves, while the organisms carried substantial amounts of nAu (~1.5 ng/mg), and nTiO2 (up to 2.7 ng/mg). This thesis shows, that moving test conditions of EINPs towards a more field-relevant approach, meaningfully modifies the risk of EINPs for aquatic organisms. Thereby, more efforts need to be made to understand the relative importance of EINP exposure pathways, especially since a transferability between different types of EINPs may not be given. When considering typically applied risk assessment factors, adverse effects on aquatic systems might already be expected at currently predicted environmental EINP concentrations in the low ng-µg/L range.
Groundwater is essential for the provision of drinking water in many areas around the world. The ecosystem services provided by groundwater-related organisms are crucial for the quality of groundwater-bearing aquifers. Therefore, if remediation of contaminated groundwater is necessary, the remediation method has to be carefully selected to avoid risk-risk trade-offs that might impact these valuable ecosystems. In the present thesis, the ecotoxicity of the in situ remediation agent Carbo-Iron (a composite of zero valent nano-iron and active carbon) was investigated, an estimation of its environmental risk was performed, and the risk and benefit of a groundwater remediation with Carbo-Iron were comprehensively analysed.
At the beginning of the work on the present thesis, a sound assessment of the environmental risks of nanomaterials was impeded by a lack of guidance documents, resulting in many uncertainties on selection of suitable test methods and a low comparability of test results from different studies with similar nanomaterials. The reasons for the low comparability were based on methodological aspects of the testing procedures before and during the toxicity testing. Therefore, decision trees were developed as a tool to systematically decide on ecotoxicity test procedures for nanomaterials. Potential effects of Carbo-Iron on embryonic, juvenile and adult life stages of zebrafish (Danio rerio) and the amphipod Hyalella azteca were investigated in acute and chronic tests. These tests were based on existing OECD and EPA test guidelines (OECD, 1992a, 2013a, 2013b; US EPA, 2000) to facilitate the use of the obtained effect data in the risk assessment. Additionally, the uptake of particles into the test organisms was investigated using microscopic methods. In zebrafish embryos, effects of Carbo-Iron on gene expression were investigated. The obtained ecotoxicity data were complemented by studies with the waterflea Daphnia magna, the algae Scenedesmus vacuolatus, larvae of the insect species Chironomus riparius and nitrifying soil microorganisms.
In the fish embryo test, no passage of Carbo-Iron particles into the perivitelline space or the embryo was observed. In D. rerio and H. azteca, Carbo-Iron was detected in the gut at the end of exposure, but no passage into the surrounding tissue was detected. Carbo-Iron had no significant effect on soil microorganisms and on survival and growth of fish. However, it had significant effects on the growth, feeding rate and reproduction of H. azteca and on survival and reproduction in D. magna. Additionally, the development rate of C. riparius and the cell volume of S. vacuolatus were negatively influenced.
A predicted no effect concentration of 0.1 mg/L was derived from the ecotoxicity studies based on the no-effect level determined in the reproduction test with D. magna and an assessment factor of 10. It was compared to measured and modelled environmental concentrations for Carbo-Iron after application to an aquifer contaminated with chlorohydrocarbons in a field study. Based on these concentrations, risk quotients were derived. Additionally, the overall environmental risk before and after Carbo-Iron application was assessed to verify whether the chances for a risk-risk trade-off by the remediation of the contaminated site could be minimized. With the data used in the present study, a reduced environmental risk was identified after the application of Carbo-Iron. Thus, the benefit of remediation with Carbo-Iron outweighs potential negative effects on the environment.
With 47% land coverage in 2016, agricultural land was one of the largest terrestrial biomes in Germany. About 70% of the agricultural land was cropped area with associated pesticide applications. Agricultural land also represents an essential habitat for amphibians. Therefore, exposure of amphibians to agrochemicals, such as fertilizers and pesticides, seems likely. Pesticides can be highly toxic for amphibians, even a fraction of the original application rate may result in high amphibian mortality.
To evaluate the potential risk of pesticide exposure for amphibians, the temporal coincidence of amphibian presence on agricultural land and pesticide applications (N = 331) was analyzed for the fire-bellied toad (Bombina bombina), moor frog (Rana arvalis), spadefoot toad (Pelobates fuscus) and crested newt (Triturus cristatus) during spring migration. In 2007 and 2008, up to 80% of the migrating amphibians temporally coincided with pesticide applications in the study area of Müncheberg, about 50 km east of Berlin. Pesticide interception by plants ranged between 50 to 90% in winter cereals and 80 to 90% in winter rape. The highest coincidence was observed for the spadefoot toad, where 86.6% of the reproducing population was affected by a single pesticide in winter rape during stem elongation with 80% pesticide interception by plants. Late migrating species, such as the fire-bellied toad and the spadefoot toad, overlapped more with pesticide applications than early migrating species, such as the moor frog, did. Under favorable circumstances, the majority of early migrants may not coincide with the pesticide applications of arable fields during spring migration.
To evaluate the potential effect of pesticide applications on populations of the common frog (Rana temporaria), a landscape genetic study was conducted in the vinicultural area of Southern Palatinate. Due to small sample sizes at breeding sites within viniculture, several DNA sampling methods were tested. Furthermore, the novel repeated randomized selection of genotypes approach was developed to utilize genetic data from siblings for more reliable estimates of genetic parameters. Genetic analyses highlighted three of the breeding site populations located in viniculture as isolated from the meta-population. Genetic differentiation among breeding site populations in the viniculture (median pairwise FST=0.0215 at 2.34 km to 0.0987 at 2.39 km distance) was higher compared to genetic differentiation among breeding site populations in the Palatinate Forest (median pairwise FST=0.0041 at 5.39 km to 0.0159 at 9.40 km distance).
The presented studies add valuable information about the risk of pesticide exposure for amphibians in the terrestrial life stage and possible effects of agricultural land on amphibian meta-populations. To conserve endemic amphibian species and their (genetic) diversity in the long run, the risk assessment of pesticides and applied agricultural management measures need to be adjusted to protect amphibians adequately. In addition, other conservation measures such as the creation of new suitable breeding site should be considered to improve connectivity between breeding site populations and ensure the persistence of amphibians in the agricultural land.
Systemic neonicotinoids are one of the most widely used insecticide classes worldwide. In addition to their use in agriculture, they are increasingly applied on forest trees as a protective measure against insect pests. However, senescent leaves containing neonicotinoids might, inter alia during autumn leaf fall, enter nearby streams. There, the hydrophilic neonicotinoids may be remobilized from leaves to water resulting in waterborne exposure of aquatic non-target organisms. Despite the insensitivity of the standard test species Daphnia magna (Crustacea, Cladocera) toward neonicotinoids, a potential risk for aquatic organisms is evident as many other aquatic invertebrates (in particular insects and amphipods) display adverse effects when exposed to neonicotinoids in the ng/L- to low µg/L-range. In addition to waterborne exposure, in particular leaf-shredding invertebrates (= shredders) might be adversely affected by the introduction of neonicotinoid-contaminated leaves into the aquatic environment since they heavily rely on leaf litter as food source. However, dietary neonicotinoid exposure of aquatic shredders has hardly received any attention from researchers and is not considered during aquatic environmental risk assessment. The primary aim of this thesis is, therefore, (1) to characterize foliar neonicotinoid residues and exposure pathways relevant for aquatic shredders, (2) to investigate ecotoxicological effects of waterborne and dietary exposure on two model shredders, namely Gammarus fossarum (Crustacea, Amphipoda) and Chaetopteryx villosa (Insecta, Trichoptera), and (3) to identify biotic and abiotic factors potentially modulating exposure under field conditions.
During the course of this thesis, ecotoxicologically relevant foliar residues of the neonicotinoids imidacloprid, thiacloprid and acetamiprid were quantified in black alder trees treated at field relevant levels. A worst-case model – developed to simulate imidacloprid water concentrations resulting from an input of contaminated leaves into a stream – predicted only low aqueous imidacloprid concentrations (i.e., ng/L-range). However, the model identified dietary uptake as an additional exposure pathway relevant for shredders up to a few days after the leaves’ introduction into the stream. When test organisms were simultaneously exposed (= combined exposure) to neonicotinoids leaching from leaves into the water and via the consumption of contaminated leaves, adverse effects exceeded those observed under waterborne exposure alone. When exposure pathways were separated using a flow-through system, dietary exposure towards thiacloprid-contaminated leaves caused similar sublethal adverse effects in G. fossarum as observed under waterborne exposure. Moreover, the effect sizes observed under combined exposure were largely predictable using the reference model “independent action”, which assumes different molecular target sites to be affected. Dietary toxicity for shredders might, however, be reduced under field conditions since UV-induced photodegradation and leaching decreased imidacloprid residues in leaves and thereby the toxicity for G. fossarum. In contrast, both shredders were found unable to actively avoid dietary exposure. This thesis thus recommends considering dietary exposure towards systemic insecticides, such as neonicotinoids, already during their registration to safeguard aquatic shredders, associated ecosystem functions (e.g., leaf litter breakdown) and ultimately ecosystem integrity.
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.
The European weatherfish (Misgurnus fossilis) is a benthic freshwater fish species belonging to the family Cobitidae, that is subjected to a considerable decline in many regions across its original distribution range. Due to its cryptic behavior and low economic value, the causes of threat to weatherfish remained partly unknown and the species is rarely at the center of conservation efforts. In order to address these concerns, the overall aim of the present thesis was to provide a comprehensive approach for weatherfish conservation, including the development of stocking measures, investigations on the species autecology and the evaluation of potential threats. The first objective was to devise and implement a regional reintroduction and stock enhancement program with hatchery-reared weatherfish in Germany. Within this program (2014-2016), a total number of 168,500 juvenile weatherfish were stocked to seven water systems. Recaptures of 45 individuals at two reintroduction sites supported the conclusion that the developed stocking strategy was appropriate. In order to broaden the knowledge about weatherfish autecology and thereby refining the rearing conditions and the selection of appropriate stocking waters, the second objective was to investigate the thermal requirements of weatherfish larvae. Here, the obtained results revealed that temperatures higher than previously suggested were tolerated by larvae, whereas low temperatures within the range of likely habitat conditions increased mortality rates. As weatherfish can be frequently found in agriculturally impacted waters (e.g. ditch systems), they are assumed to have an increased probability to be exposed to chemical stress. Since the resulting risk has not yet been investigated with a focus on weatherfish, the third objective was to provide a methodical foundation for toxicity testing that additionally complies with the requirements of alternative test methods. For this purpose, the acute fish embryo toxicity test was successfully transferred to weatherfish and first results exhibited that sensitivity of weatherfish towards a tested reference substance (3,4-dichloroaniline) was highest compared to other species. On the basis of these findings, the fourth objective was to apply weatherfish embryos for multiple sediment bioassays in order to investigate teratogenic effects derived from sediment-associated contaminants. In this context, weatherfish revealed particular sensitivity to water extractable substances, indicating that sediment contamination might pose a considerable risk. Moreover, as an endangered benthic fish species with high ecological relevance for European waters that are specifically exposed to hazardous contaminants, the weatherfish might be a prospective species for an ecological risk assessment of sediment toxicity. Overall, the present thesis contributed to the conservation of weatherfish by considering a variety of aspects that interact and reinforce one another in order to achieve improvements for the species situation.
Agricultural land-use may lead to brief pulse exposures of pesticides in edge-of-field streams, potentially resulting in adverse effects on aquatic macrophytes, invertebrates and ecosystem functions. The higher tier risk assessment is mainly based on pond mesocosms which are not designed to mimic stream-typical conditions. Relatively little is known on exposure and effect assessment using stream mesocosms.
Thus the present thesis evaluates the appliacability of the stream mesocosms to mimic stream-typical pulse exposures, to assess resulting effects on flora and fauna and to evaluate aquatic-terrestrial food web coupling. The first objective was to mimic stream-typical pulse exposure scenarios with different durations (≤ 1 to ≥ 24 hours). These exposure scenarios established using a fluorescence tracer were the methodological basis for the effect assessment of an herbicide and an insecticide. In order to evaluate the applicability of stream mesocosms for regulatory purposes, the second objective was to assess effects on two aquatic macrophytes following a 24-h pulse exposure with the herbicide iofensulfuron-sodium (1, 3, 10 and 30 µg/L; n = 3). Growth inhibition of up to 66 and 45% was observed for the total shoot length of Myriophyllum spicatum and Elodea canadensis, respectively. Recovery of this endpoint could be demonstrated within 42 days for both macrophytes. The third objective was to assess effects on structural and functional endpoints following a 6-h pulse exposure of the pyrethroid ether etofenprox (0.05, 0.5 and 5 µg/L; n = 4). The most sensitive structural (abundance of Cloeon simile) and functional (feeding rates of Asellus aquaticus) endpoint revealed significant effects at 0.05 µg/L etofenprox. This concentration was below field-measured etofenprox concentrations and thus suggests that pulse exposures adversely affect invertebrate populations and ecosystem functions in streams. Such pollutions of streams may also result in decreased emergence of aquatic insects and potentially lead to an insect-mediated transfer of pollutants to adjacent food webs. Test systems capable to assess aquatic-terrestrial effects are not yet integrated in mesocosm approaches but might be of interest for substances with bioaccumulation potential. Here, the fourth part provides an aquatic-terrestrial model ecosystem capable to assess cross-ecosystem effects. Information on the riparian food web such as the contribution of aquatic (up to 71%) and terrestrial (up to 29%) insect prey to the diet of the riparian spider Tetragnatha extensa was assessed via stable isotope ratios (δ13C and δ15N). Thus, the present thesis provides the methodological basis to assess aquatic-terrestrial pollutant transfer and effects on the riparian food web.
Overall the results of this thesis indicate, that stream mesocosms can be used to mimic stream-typical pulse exposures of pesticides, to assess resulting effects on macrophytes and invertebrates within prospective environmental risk assessment (ERA) and to evaluate changes in riparian food webs.
Leaf litter breakdown is a fundamental process in aquatic ecosystems, being mainly mediated by decomposer-detritivore systems that are composed of microbial decomposers and leaf-shredding, detritivorous invertebrates. The ecological integrity of these systems can, however, be disturbed, amongst others, by chemical stressors. Fungicides might pose a particular risk as they can have negative effects on the involved microbial decomposers but may also affect shredders via both waterborne toxicity and their diet; the latter by toxic effects due to dietary exposure as a result of fungicides’ accumulation on leaf material and by negatively affecting fungal leaf decomposers, on which shredders’ nutrition heavily relies. The primary aim of this thesis was therefore to provide an in-depth assessment of the ecotoxicological implications of fungicides in a model decomposer-detritivore system using a tiered experimental approach to investigate (1) waterborne toxicity in a model shredder, i.e., Gammarus fossarum, (2) structural and functional implications in leaf-associated microbial communities, and (3) the relative importance of waterborne and diet-related effects for the model shredder.
Additionally, knowledge gaps were tackled that were related to potential differences in the ecotoxicological impact of inorganic (also authorized for organic farming in large parts of the world) and organic fungicides, the mixture toxicity of these substances, the field-relevance of their effects, and the appropriateness of current environmental risk assessment (ERA).
In the course of this thesis, major differences in the effects of inorganic and organic fungicides on the model decomposer-detritivore system were uncovered; e.g., the palatability of leaves for G. fossarum was increased by inorganic fungicides but deteriorated by organic substances. Furthermore, non-additive action of fungicides was observed, rendering mixture effects of these substances hardly predictable. While the relative importance of the waterborne and diet-related effect pathway for the model shredder seems to depend on the fungicide group and the exposure concentration, it was demonstrated that neither path must be ignored due to additive action. Finally, it was shown that effects can be expected at field-relevant fungicide levels and that current ERA may provide insufficient protection for decomposer-detritivore systems. To safeguard aquatic ecosystem functioning, this thesis thus recommends including leaf-associated microbial communities and long-term feeding studies using detritus feeders in ERA testing schemes, and identifies several knowledge gaps whose filling seems mandatory to develop further reasonable refinements for fungicide ERA.