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Factors triggering the ecotoxicity of metal-based nanoparticles towards aquatic invertebrates
(2015)
Nanoparticles are produced and used in huge amounts increasing their probability to end up in surface waters. There, they are subject to environmentally driven modification processes. Consequently, aquatic life may be exposed to different nanoparticle agglomerate sizes, while after sedimentation benthic organisms are more likely to be affected.
However, most ecotoxicity studies with nanoparticles exclusively investigated implications of their characteristics (e.g. size) on pelagic organisms, ignoring environmentally modified nanoparticles. Therefore, a systematic assessment of factors triggering the fate and toxicity of nanoparticles under environmentally relevant conditions is needed. The present thesis, therefore, investigates the implications of nanoparticle related factors (i.e., inherent material-properties and nanoparticle characteristics) as well as environmental conditions towards the pelagic living organism Daphnia magna and the benthic species Gammarus fossarum. In detail, inert titanium dioxide (nTiO2) and ion-releasing silver nanoparticles (nAg), both of varying particle characteristics (e.g. initial size), were tested for their toxicity under different environmental conditions (e.g. ultraviolet-light (UV-light)).
The results indicate that the toxicity of nTiO2 and nAg is mainly determined by: their adsorption potential onto biota, and their fate in terms of reactive oxygen species or Ag+ ion release. Thus, inherent material-properties, nanoparticle characteristics and environmental conditions promoting or inhibiting these aspects revealed significant implications in the toxicity of nTiO2 and nAg towards daphnids.
Furthermore, the presence of ambient UV-light, for example, adversely affected gammarids at 0.20 mg nTiO2/L, while under darkness no effects occurred even at 5.00 mg nTiO2/L. Hence, the currently associated risk of nanoparticles might be underestimated if disregarding their interaction with environmental parameters
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.
Fate and effects of insecticides in vegetated agricultural drainage ditches and constructed wetlands
(2006)
Studies have shown that runoff and spray-drift are important sources of nonpoint-source pesticide pollution of surface waters. Owing to this, public concern over the presence of pesticides in surface and ground water has resulted in intensive scientific efforts to find economical, yet environmentally sound solutions to the problem. The primary objective of this research was to assess the effectiveness of vegetated aquatic systems in providing buffering between natural aquatic ecosystems and agricultural landscape following insecticide associated runoff and spray-drift events. The first set of studies were implemented using vegetated agricultural ditches, one in Mississippi, USA, using pyrethroids (bifenthrin, lambda-cyhalothrin) under simulated runoff conditions and the other in the Western Cape, South Africa using the organophosphate insecticide, azinphos-methyl (AZP), under natural runoff and spray-drift conditions. The second set of studies were implemented using constructed wetlands, one in the Western Cape using AZP under natural spray-drift conditions and the other in Mississippi, USA using the organophosphate MeP under simulated runoff conditions. Results from the Mississippi-ditch study indicated that ditch lengths of less than 300 m would be sufficient to mitigate bifenthrin and lambda-cyhalothrin. In addition, data from mass balance calculations determined that the ditch plants were the major sink (generally > 90%) and/or sorption site for the rapid dissipation of the above pyrethroids from the water column. Similarly, results from the ditch study in South Africa showed that a 180 m vegetated system was effective in mitigating AZP after natural spray drift and low flow runoff events. Analytical results from the first wetland study show that the vegetated wetland was more effective than the non-vegetated wetland in reducing loadings of MeP. Mass balance calculations indicated approximately 90% of MeP mass was associated with the plant compartment. Ninety-six hours after the contamination, a significant negative acute effect of contamination on abundances was found in 8 out of the 15 macroinvertebrate species in both wetland systems. Even with these toxic effects, the overall reaction of macroinvertebrates clearly demonstrated that the impact of MeP in the vegetated wetland was considerably lower than in the non-vegetated wetland. Results from the constructed wetland study in South Africa revealed that concentrations of AZP at the inlet of the 134 m wetland system were reduced by 90% at the outlet. Overall, results from all of the studies in this thesis indicate that the presence of the plant compartment was essential for the effective mitigation of insecticide contamination introduced after both simulated and natural runoff or spray-drift events. Finally, both the vegetated agricultural drainage ditch and vegetated constructed wetland systems studied would be effective in mitigating pesticide loadings introduced from either runoff or spray-drift, in turn lowering or eliminating potential pesticide associated toxic effects in receiving aquatic ecosystems. Data produced in this research provide important information to reduce insecticide risk in exposure assessment scenarios. It should be noted that incorporating these types of best management practices (BMPs) will decrease the risk of acute toxicity, but chronic exposure may still be an apparent overall risk.
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.
Assessment of bat activity in agricultural environments and the evaluation of the risk of pesticides
(2013)
Although agriculture dominates with around 50% area much of Europe- landscape, there is virtually no information on how bats use this farmed environment for foraging. Consequently, little is known about effective conservation measures to compensate potential negative effects of agrarian management practice on the food availability for bats in this habitat. Moreover, there are currently no specific regulatory requirements to include bats in European Union risk assessments for the registration of pesticides since no information about pesticide exposure on this mammal group is available. To evaluate the potential pesticide exposure of bats via ingestion of contaminated insects, information about bat presence and activity in agricultural habitats is required. In order to examine bat activity on a landscape scale it was necessary to establish a suitable survey method. Contrary to capture methods, telemetry, and direct observations, acoustic surveys of bat activity are a logistically feasible and cost-effective way of obtaining bat activity data. However, concerns regarding the methodological designs of many acoustic surveys are expressed in the scientific literature. The reasons are the failing of addressing temporal and spatial variation in bat activity patterns and the limitations of the suitability of the used acoustic detector systems. By comparing different methods and detector systems it was found that the set up of several stationary calibrated detector systems which automatically trigger the ultrasonic recording has the highest potential to produce reliable, unbiased and comparable data sets on the relative activity of bats.
By using the proposed survey method, bat diversity and activity was recorded in different crops and semi-natural habitats in southern Rhineland-Palatinate. Simultaneously, the availability of aerial prey insects was studied by using light and sticky traps. In more than 500 sampling nights about 110,000 call sequences were acoustically recorded and almost 120,000 nocturnal insects were sampled. A total of 14 bat species were recorded, among them the locally rare and critically endangered northern bat (Eptesicus nilssonii) and the barbastelle (Barbastella barbastellum), all of them also occurring over agricultural fields. The agricultural landscape of southern Palatinate is dominated by vineyards, a habitat that was shown to be of low quality for most bat species because of the demonstrated low availability of small aerial insects. By surveying bat activity and food availably in a pair-wise design on several rain water retention ponds and neighbouring vineyards it was demonstrated that aquatic insect emergence in artificial wetlands can provide an important resource subsidy for bats. The creation of artificial wetlands would be a possibility to create important foraging habitats for bats and mitigate negative effects of management practice in the agricultural landscape.
In several other agricultural crops, however, high abundances of suitable prey insects and high bat activity levels, comparable or even higher than in the nearby forests and meadows known to be used as foraging habitats were demonstrated. Especially high bat activity levels were recorded over several fruit orchards and vegetable fields where insects were also present. Both crops are known for high pesticide inputs, and, therefore, a pesticide exposure through ingestion of contaminated insects can not be excluded. To follow the current risk assessment approach for birds and mammals pesticide residues were measured on bat-specific food items in an apple orchard following insecticide applications and bat activity was recorded in parallel. The highest residue values were measured on foliage-dwelling arthropods which may results in a reproductive risk for all bat species that, even to a small extent, include this prey group in their diet. The presence of bats in agricultural landscapes that form a majority of the land area in Europe but also on a global scale leads to exposure of bats by contaminated food and depletion of their food resources by pesticide use. So far conservation efforts for bats focussed on securing hibernation sites and the creation of artificial roost sites since especially the latter were thought to be limiting population growth. However the potential pesticide effects might be also crucial for the population persistence in agricultural landscapes of bats and need to be addressed adequately, especially in risk assessment procedures for the regulation of pesticides.
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.
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.