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Invasive species often have a significant impact on the biodiversity of ecosystems and the species native to it. One of the worst invaders worldwide is Aphanomyces astaci, the causative agent of the crayfish plague, an often fatal disease to crayfish species not native to North America. Aphanomyces astaci originates from North America and was introduced to Europe in the midst of the 19th century. Since then, it spread throughout Europe diminishing the European crayfish populations. The overall aim of this thesis was to evaluate the threat that A. astaci still poses to European crayfish species more than 150 years after its introduction to Europe. In the first part of the thesis, crayfish specimens, which are available in the German pet trade, were tested for infections with A. astaci. Around 13% of the tested crayfish were clearly infected with A. astaci. The study demonstrated the potential danger the pet trade poses for biodiversity through the import of alien species and their potential pathogens, in general. In the second part of the thesis, the A. astaci infection prevalence of crayfish species in wild populations in Europe was tested. While the stone crayfish, Austropotamobius torrentium, showed high susceptibility to different haplogroups of A. astaci, the narrow-clawed crayfish, Astacus leptodactylus, was able to survive infections, even by haplogroup B, which is considered to be highly virulent. In the last part of the thesis, A. astaci was traced back to its original distribution area of North America. While the crayfish plague never had such a devastating effect on crayfish in North America as it had in Europe, the reasons for the success of invasive crayfish within North America are not yet fully understood. It is possible that A. astaci increases the invasion success of some crayfish species. Several populations of the rusty crayfish, Orconectes rusticus, in the Midwest of North America were confirmed to be infected with A. astaci and a new genotype was identified, possibly indicating that each crayfish host is vector of a unique A. astaci genotype, even in North America. Overall, the present thesis provides evidence that A. astaci is still a major threat to the crayfish species indigenous to Europe. Crayfish mass mortalities still occur in susceptible crayfish species like A. torrentium even 150 years after the first introduction of A. astaci. While there are some indications for increased resistances through processes of co-evolution, the continuous introduction of crayfish species to Europe threatens to cause new outbreaks of the crayfish plague through the parallel introduction of new, highly virulent A. astaci strains.
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.
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.
Systemische Neonicotinoide gehören zu den weltweit meist genutzten Insektiziden. Neben ihrer Anwendung in der Landwirtschaft werden sie zunehmend zur Bekämpfung von Baumschädlingen in
der Forstwirtschaft eingesetzt. Die im Herbst von Laubbäumen fallenden Blätter können allerdings
immer noch Neonicotinoide enthalten. Gelangen diese kontaminierten Blätter schließlich in
nahegelegene Bäche werden die wasserlöslichen Neonicotinoide wieder mobilisiert und somit
potenziell aquatische Nicht-Zielorganismen über die Wasserphase exponiert. Obwohl der Standardtestorganismus Daphnia magna (Crustacea; Cladocera) relativ unempfindlich gegenüber
Neonicotinoiden ist, sind viele andere aquatische Invertebraten bereits bei einer Exposition im ng/L- bis niedrigem μg/L-Bereich negativ beeinträchtigt. Besonders laubzersetzende Invertebraten (= Shredder) könnten, zusätzlich zu einer Exposition über die Wasserphase, durch den Eintrag von Neonicotinoid-kontaminiertem Laub in ein Fließgewässer negativ beeinträchtigt werden, da Laub für sie eine essentielle Nahrungsquelle darstellt. Jedoch erhielt dieser Expositionspfad im Zusammenhang mit aquatischen Shreddern und Neonicotinoid-kontaminiertem Pflanzenmaterial bisher kaum Aufmerksamkeit seitens der Forschung und findet keine Berücksichtigung in der aquatischen Umweltrisikobewertung. Das Hauptziel dieser Arbeit war daher (1) Neonicotinoidrückstände in Blättern zu quantifizieren sowie für Shredder relevante Expositionswege zu identifizieren, (2)
ökotoxikologische Effekte einer Exposition über die Wasserphase sowie über die Nahrung für zwei
Modell-Shredder Gammarus fossarum (Amphipoda) und Chaetopteryx villosa (Insecta) zu untersuchen, und schließlich (3) biotische und abiotische Faktoren zu betrachten, welche eine Exposition unter Feldbedingungen potenziell beeinträchtigen könnten.
Im Rahmen dieser Arbeit konnten Rückstände der Neonicotinoide Imidacloprid, Thiacloprid und
Acetamiprid in Blätter behandelter Schwarzerlen quantifiziert werden. Ein entwickeltes „Worst-Case
Modell“ prognostizierte niedrige Imidaclopridwasserkonzentrationen für einen Bach in welchen Imidacloprid-kontaminierte Blätter eingetragen werden. Jedoch konnte mit Hilfe des Modells die Aufnahme über die Nahrung als ein für aquatische Shredder relevanter Expositionspfad identifiziert werden. Der Konsum von Neonicotinoid-kontaminierten Blättern führte, bei gleichzeitiger Exposition über die Wasserphase (= kombinierte Exposition), in beiden Testorganismen zu stärkeren Effekten als die alleinige Exposition über die Wasserphase. Des Weiteren gelang es in einem weiteren Laborexperiment die beiden Expositionswege mittels einer Durchflussanlage zu separieren. Hierbei führte die separate Exposition von G. fossarum sowohl über die Nahrung (= Konsum von Thiaclopridkontaminierten Blättern) als auch über die Wasserphase zu vergleichbaren Effektgrößen. Zudem ließen sich die unter einer kombinierten Exposition beobachteten Effektgrößen weitestgehend mit dem Referenzmodell der „Unabhängigen Wirkung“ vorhersagen, was eine Wirkung auf unterschiedliche molekulare Zielorte vermuten lässt. Die durch Imidacloprid ausgelöste toxischen Effekte auf G. fossarum konnten schließlich durch eine Behandlung der Blätter mit UV-Strahlung (repräsentativ für Sonnenlicht) sowie durch Leaching in Wasser reduziert werden. Jedoch waren beide Shredder-Spezies nicht dazu in der Lage aktiv eine Aufnahme von Neonicotinoiden über die Nahrung zu vermeiden. Daher geht aus dieser Arbeit die Empfehlung hervor, bereits während der Registrierung von systemischen Pestiziden, auf nahrungsbedingte Effekte zu testen und dadurch aquatische Shredder als auch assoziierte Ökosystemfunktionen (z.B. Laubabbau) zu schützen.
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.
The application of pesticides to agricultural areas can result in transport to adjacent non-target environments. In particular, surface water systems are likely to receive agricultural pesticide input. When pesticides enter aquatic environments, they may pose a substantial threat to the ecological integrity of surface water systems. To minimize the risk to non-target ecosystems the European Union prescribes an ecotoxicological risk assessment within the registration procedure of pesticides, which consists of an effect and an exposure assessment.
This thesis focuses on the evaluation of the exposure assessment and the implications to the complete regulatory risk assessment, and is based on four scientific publications. The main part of the thesis focuses on evaluation of the FOCUS modelling approach, which is used in regulatory risk assessment to predict pesticide surface water concentrations. This was done by comparing measured field concentrations (MFC) of agricultural insecticides (n = 466) and fungicides (n = 417) in surface water to respective predicted environmental concentrations (PEC) calculated with FOCUS step 1 to step 4 at two different levels of field relevance. MFCs were extracted from the scientific literature and were measured in field studies conducted primarily in Europe (publications 1 and 3).
In addition, an alternative fugacity-based multimedia mass-balance model, which needs fewer input parameters and less computing effort, was used to calculate PECs for the same insecticide MFC dataset and compared to the FOCUS predictions (publication 3). Furthermore, FOCUS predictions were also conducted for veterinary pharmaceuticals in runoff from an experimental plot study, to assess the FOCUS predictions for a different class of chemicals with a different relevant entry pathway (publication 2).
In publication 4, the FOCUS step-3 approach was used to determine relevant insecticide exposure patterns. These patterns were analysed for different monitoring strategies and the implications for the environmental risk assessment (publication 4).
The outcome of this thesis showed that the FOCUS modelling approach is neither protective nor appropriate in predicting insecticide and fungicide field concentrations. Up to one third of the MFCs were underpredicted by the model calculations, which means that the actual risk might be underestimated. Furthermore, the results show that a higher degree of realism even reduces the protectiveness of model results and that the model predictions are worse for highly hydrophobic and toxic pyrethroids.
In addition, the absence of any relationship between measured and predicted concentrations questions the general model performance quality (publication 1 and 3). Further analyses revealed that deficiencies in protectiveness and predictiveness of the environmental exposure assessment might even be higher than shown in this thesis, because actual short-term peak concentrations are only detectable with an event-related sampling strategy (publication 4). However, it was shown that the PECs of a much simpler modelling approach are much more appropriate for the prediction of insecticide MFC, especially for calculations with a higher field relevance (publication 3). The FOCUS approach also failed to predict concentrations of veterinary pharmaceuticals in runoff water (publication 2). In conclusion, the findings of this thesis showed that there is an urgent need for the improvement of exposure predictions conducted in the environmental risk assessment of pesticides as a group of highly relevant environmental chemicals, to ensure that the increasing use of those chemicals does not lead to further harmful effects in aquatic ecosystems.