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Institute
Clean water is fundamental to human health and ecosystem integrity. However, water quality deteriorates due to novel anthropogenic pollutants present at microgram per liter concentrations in urban water cycles (termed micropollutants). Wastewater treatment plants (WWTP) have been identified as major point sources for aquatic (micro-)pollutants. Chemical and ecotoxicological analyses have shown that conventional biological WWTPs do not fully remove micropollutants and associated toxicities, which is often because of mobile, polar and/or recalcitrant compounds and transformation products (TPs). To minimize possible environmental risks, advanced wastewater treatment (AWWT) technologies could be a promising mitigation measure. Multiple processes are therefore being developed and evaluated such as ozonation and ozonation followed by granulated activated carbon (GAC) or biological filtration. Assessing the performance of these combined AWWTs was the focus the TransRisk project. Within this project, this thesis accomplished four major goals.
Firstly, the preparation of (waste)water samples was optimised for in vitro bioassays. Acidification, filtration and solid phase extraction (SPE) were tested for their impact on environmentally relevant in vitro endocrine activities, mutagenicity, genotoxicity and cytotoxicity. Significantly different outcomes of these assays were detected comparing neutral and acidified samples. Sample filtration had a lesser impact, but in some cases retention of particle-bound compounds could have caused significant toxicity losses. Out of three SPE sorbents the Telos C18/ENV at sample pH 2.5 extracted highest toxicity, some undetected in aqueous samples. These results indicate that sample preparation needs to be optimised for specific sample matrices and bioassays to avoid false-positive or -negative detects in effect-based analyses.
Secondly, the above listed in vitro toxicities were monitored in a protected region for drinking water production in South-West Germany (2012-2015). Out of 30 sampling sites surface water and groundwater were the least polluted. Nonetheless, a few groundwater samples induced high anti-estrogenic activity that prompted further monitoring. The latter included a waterworks in which no toxicity was detected. Hospital wastewater also had elevated in vitro toxicities and hospitals are, thus, relevant intervention points for source control. The biological WWTPs were effective in removing most of the detected toxicity, and the selected bioassays proved to be pertinent tools for water quality assessment and prioritisation of pollution hotspots.
Thirdly, the in vivo bioassay ISO10872 based on Caenorhabditis elegans (C. elegans) was adapted for this thesis. Using this model, a median effect concentration (EC50) for reproductive toxicity of the polycyclic aromatic hydrocarbon β-naphthoflavone (β- NF) of 114 µg/L was computed which is slightly lower than reported in the scientific literature. β-NF induced cyp-35A3::GFP (a biomarker in transgenic animals) in a time and concentration dependent manner (≤ 21.3–24 fold above controls). β-NF spiked wastewater samples supported earlier hypotheses on particle-bound pollutants. Reproductive toxicity (96 h) and cyp-35A3 induction (24 h) of biologically treated and/or ozonated wastewater extracts and growth promoting effects of GAC/biologically filtered ozonated wastewater extracts were observed. This suggested the presence of residual bioactive/toxic chemicals not included in the targeted chemical analysis. It also highlighted the importance of integrating multiple (apical and molecular) endpoints in wastewater assessments.
Fourthly, five in vitro and the adapted C. elegans bioassay were integrated into a wastewater quality evaluation (developed within TransRisk). Out of the five AWWT options, ozonation (at 1 g O3,applied/g DOC, HRT ~ 18 min) combined with nonaerated GAC filtration was rated most effective for toxicity removal. All five AWWTs largely removed estrogenic and (anti-)androgenic activities, but not anti-estrogenic activity and mutagenicity, which even increased during ozonation. This has been observed in related studies and points towards toxic TPs. These results also emphasized the need for implementing an effective post-treatment for ozonation. The results from a parallel in vivo study with Lumbriculus variegatus and Potamopyrgus antipodarum conducted on site at the WWTP (using flow through systems) were in accordance with the C. elegans results. In this context, it is suggested to further implement C. elegans as sensitive, feasible and ecologically relevant model.
In conclusion, this thesis shows how optimised sample preparation, long-term (in vitro) environmental monitoring, sensitive and ecologically relevant (in vivo) bioassays as well as innovative evaluation concepts, are pivotal in improving the removal of micropollutants and their toxicities with AWWTs. Future research should further develop and evaluate measures at sewer systems, conventional biological, tertiary and other advanced treatment technologies, as well as sociopolitical strategies (e.g., source control or natural conservation) and restoration projects. The effect-based tools optimised in this thesis will support assessing their success.
In the past decades, the use and production of chemicals has been on the rise globally due to increasing industrialization and intensive agriculture; resulting in the occurrence and ecotoxicological risks of chemicals of emerging concern (CECs) in the aquatic compartments. Risks include changes in community structure resulting in the dominance of one species and ecosystem imbalance. When dominant disease-causing organisms are in the environment, the disease transmission is increased. For example, host snails for the schistosomiasis, a human trematode disease, are known to be tolerant to pesticide
exposure compared to the predators. This would therefore result in an increased abundance of snails which consequently increase the disease transmission in the human population.
Kenya, being a low income country faces a lot of challenges with provision of clean water, diseases and sanitation facilities, and increasing population which results in intensive agriculture coupled with pesticide use. Although a lot of research has been carried out on the environmental occurrence and risk of CECs (Chapter 1), most of these studies have been done in developed countries with limited information from Africa. Additionally, research in Africa focused on urban areas with limited number of compounds analyzed and mostly in the water phase, and inadequate information on the effects of CECs on the aquatic organisms. In order to reduce this knowledge gap, this dissertation focused on identification and quantification of CECs present in water, sediment and snails from western Kenya, and the contribution of pesticides to the transmission of schistosomiasis.
Chapter 2 gives a summary of the results and discussion of the dissertation. In Chapter 3, a comprehensive chemical analysis was carried out on 48 water samples to identify compounds, spatial patterns and associated risks for fish, crustacean and algae using toxic unit (TU) approach. A total of 78 compounds were detected with pesticides and biocides being the compounds most frequently detected. Spatial pattern analysis revealed limited compound grouping based on land use. Acute risk for crustaceans and algae were driven by one to three individual compounds. These compounds responsible for toxicity were prioritized as candidate compounds for monitoring and regulation in Kenya.
In Chapter 4, an extension of Chapter 3 was done to cover the CECs present in snails and sediment from the 48 sites. A total of 30 compounds were found in snails and 78 in sediments with 68 additional compounds being found which were not previously detected in water. Higher contaminant concentrations were found in agricultural sites than in areas without anthropogenic activities. The highest acute toxicity (TU 0.99) was determined for crustaceans based on compounds in sediment samples. The risk was driven by diazinon and pirimiphos-methyl. Acute and chronic risks to algae were driven by diuron whereas fish were found to be at low to no acute risk.
In Chapter 5, the effect of pesticide contamination on schistosomiasis transmission was evaluated by applying complimentary laboratory and field studies. In the field studies, the ecological mechanisms through which pesticides and physical chemical parameters affect host snails, predators and competitors were investigated. Pesticide data was obtained from the results in chapter 3. The overall distribution of grazers and predators was not affected by pesticide pollution. However, within the grazers, pesticide pollution increased dominance of host snails. On the contrary, the host-snail competitors were highly sensitive to pesticide exposure. For the laboratory studies, macroinvertebrates including Schistosoma-host snails, competitors and predators were exposed to 6 concentrations levels of imidacloprid and diazinon. Snails showed higher insecticide tolerance compared to competitors and predators. Finally, Chapter 6 summarizes the conclusions of this dissertation, placing it in a broader
context. In this dissertation, a comprehensive chemical characterization and risk assessment of CECs has been carried out in freshwater systems; together with the effects of pesticides on schistosomiasis transmission in rural western Kenya. Results of this dissertation showed that rural areas are contaminated posing a risk to aquatic organisms which contribute to schistosomiasis transmission. This shows the need for regular monitoring and policy formulation to reduce pollutant emissions which contributes negatively to both ecological and human health effects.
Viele Gruppen der Lebewesen, insbesondere Insekten breiten sich durch steigende Temperaturen zunehmend in Gebieten aus, in denen sie ursprünglich nicht vorkommen(Novikov und Vaulin 2014; Bebber 2015). Hierbei ist die steigende Temperatur in
verschiedenen Gebieten der Hauptfaktor für Expansionen dieser Arten in Richtung des nördlichen Polarkreises. Einige dieser Arten sind sehr tolerant für verschiedene Variablen und können damit ihr Verbreitungsgebiet deutlich nach Norden hin ausdehnen. Aufgrund steigender Temperaturen werden jedoch andere Arten in ihrem Verbreitungsgebiet eingeschränkt oder ihre Verbreitung verschiebt sich in nördliche Richtung (Ogden und Lindsay 2016; Lawler et al. 2009). Auch für die Verbreitung von Krankheiten spielen Temperaturen, Ausbreitungen oder Verbreitungsverschiebungen eine wichtige Rolle (Mordecai et al. 2019).
So können, durch die Etablierung der passenden Vektoren, bisher nur in wärmeren Gebieten auftretende Krankheiten zukünftig auch in unseren Breitengraden eingeschleppt und
verbreitet werden. Bremsen, invasive Stechmücken aber auch einheimische Mücken tragen alle ein Potential,verschiedenste Krankheitserreger zu verbreiten, auch wenn die Eignung als
Vektor für jede Art unterschiedlich groß ausfällt und manche Arten daher kaum beobachtet und untersucht werden. Mit dem Augenmerk auf sich ändernde Verbreitungsgebiete hinsichtlich zukünftigen klimatischen Veränderungen und sich wandelnden anthropogenen Einflüssen sollten jedoch auch Arten mit bisher geringem Vektorpotential mit in Beobachtungsprogramme aufgenommen werden.
Wir untersuchten in Projekt I auf kontinentaler Skala die Verbreitung von sechs verschiedenen Bremsenarten und konnten sowohl Rückschlüsse auf eine mangelhafte Beobachtung der
Arten ziehen als auch Artpräferenzen hinsichtlich der Landschaftsnutzung, Auswirkungen des Klimas auf die Verbreitung der Art und bisher unbekannte Toleranzen hinsichtlich tiefen Temperaturen und äußerst verkürzten Wärmeperioden aufdecken. Eine Größenordnung niedriger wurde in Projekt II, basierend auf aktuellen und Vergangenen Klimadaten, die zukünftige und aktuelle Verbreitung einer invasiven, sich zukünftig ausbreitenden Stechmückenart innerhalb Deutschlands modelliert. Durch bisherig im Untersuchungsgebiet nur begrenztes Auftreten konnten noch keine Rückschlüsse auf die unterschiedlichen Präferenzen für das Habitat gezogen werden, es können jedoch für zukünftige Berechnungen Habitatpräferenzen aus anderen Gebieten hinzugezogen werden um die Art und ihre fortschreitende Ausbreitung genauer zu beobachten. Auf der kleinsten untersuchten Ebene konnten in Projekt III innerhalb eines Mikrohabitates verschiedenste Rückschlüsse auf limitierende oder förderliche abiotische Faktoren, die teilweise bisherig nicht oder nur geringfügig beobachtet wurden, gezogen werden. Ebenfalls konnten Auswirkungen der umgebenden Landschaft auf die Abundanzen der Tiere beobachtet werden. Mithilfe von verschiedenen Modellen und in Abhängigkeit von Klimakarten, Landbedeckungsdaten und Landnutzung sowie Eigenschaften und Toleranzen der untersuchten Arten lassen sich in verschiedenen Größenordnungen geeignete Habitate von einheimischen sowie invasiven Arten identifizieren und zukünftige Verbreitungen effizient vorhersagen.
Insgesamt können, basierend auf all diesen Daten, dadurch für alle untersuchten Faktoren Modelle auf andere Gebiete übertragen werden um somit potentielle Verbreitungen dort
vorherzusagen. Auf unseren Daten basierend können so zum Beispiel Modellierungen für die potentielle Ausbreitung der untersuchten Tabaniden innerhalb anderer Kontinente berechnet werden und Monitoringprogramme können die Ergebnisse unserer Studie als Startpunkt aufgreifen, um durch Beprobung an modellierten Standorten die Korrektheit unserer Modelle zu überprüfen und sowohl Landschaftstypen als auch Artzusammensetzung aufzunehmen um das Modell zu bestätigen oder zu verbessern. Die Modellierung der invasiven Art Aedes albopictus bietet die Möglichkeit, diese Art in Zukunft innerhalb der möglichen Ausbreitungskorridore genauer zu beobachten um ihre fortschreitende Verbreitung zu
verifizieren oder eventuelle Änderungen des klimatischen Verlaufes mit einzubinden und das Modell anzupassen. Die Untersuchung des Mikrohabitats von Culex pipiens pipiens und Culex torrentium bietet, auch hinsichtlich anderer Arten in diesem Habitat, eine potente Methode, Vorhersagen für Artvorkommen innerhalb anderer Unterirdischen Objekte zu berechnen. Hier können, bei ausreichend großer Datenlage, eine Vielzahl von Faktoren in die Auswertung mit einfließen.
Die durchgeführten Studien bestätigen die Notwendigkeit für verbesserte Monitoringkonzepte für alle vektorkompetenten Tiergruppen hinsichtlich der sich ändernden klimatischen Bedingungen, des globalen Handels und die sich wandelnde Nutzung der Landschaften durch den Menschen und darin begründete Veränderungen der Artenzusammensetzung eines Habitates, zeigen Möglichkeiten, diese Konzepte mit bisher
ungenutzten Daten aufzubauen und zu verbessern und können gleichzeitig zu deren Verbesserung herangezogen werden.
The Southern Ocean (SO) is one of the most pristine regions of our Planet, characterised by high levels of biodiversity (5% of the global diversity) (David and Saucède 2015) and hosting a unique fauna (up to 90% of SO species are endemic) (De Broyer and Danis 2011; Chown et al. 2015). Yet, the knowledge on SO biodiversity is still far from being completed. In addition, the knowledge on the impact that changing environments have on SO species-richness is very little and for some groups, it is still totally unknown. For instance, most of studies generally focus on one single species such as Antarctic krill (Kawaguchi et al. 2011), Clio pyramidata Linnaeus, 1767 (Orr et al. 2005), Globigerina bulloides d'Orbigny, 1826 (Moy et al. 2009), or only on a high taxonomic level (e.g. phylum, class): Echinodermata, Crustacea, Mollusca, Porifera, Bryozoa, Brachiopoda, Hydrozoa, Ascidiacea, Holoturoidea
(Barnes 1999; Rowden et al. 2015; Post et al. 2017; Gutt et al. 2019; Vause et al. 2019; Pineda-Metz et al. 2020). Ultimately, the influence of sea-ice coverage on benthic species diversity was totally unknown prior to this study. In light of this, the objectives of the thesis are:
1. To expand the knowledge on shelf and deep-sea peracarid assemblage structure and abundance on a small regional (Weddell Sea) and on a large regional (Atlantic sector of the SO and South Atlantic Ocean) geographic scale.
2. To assess the environmental variables driving peracarid assemblage structure and abundance from the above mentioned areas.
3. To investigate SO benthic isopod species diversity from the Atlantic sector of the SO and assess the influence of environmental variables on their species-richness and composition.
4. To describe new possible peracarid species by means of integrative taxonomy, using morphological descriptions and whole genome sequencing analyses to support the species identification.
Objective outcomes: The present thesis provides new information on the abundance and assemblage structure based on 64766 peracarid crustaceans from different 28 locations within the Atlantic sector of the SO continental shelf and deep sea (Chapters I-II). These locations are characterised by different environmental conditions, for instance different sea-ice concentrations. Results from Chapters I-II confirmed the dominance of peracarid assemblages in the benthos, with amphipods being the most abundant group, followed by isopods. Sea ice was identified as the main driver shaping benthic peracarid assemblage structure (Chapter I). On a larger geographic scale and wider bathymetric range (e.g. including sampling locations from previous studies performed in the South Atlantic Ocean
and at a depth range from 160 to ~6000 m), depth was the main physical variable driving peracarid assemblage structure (Chapter III). In addition, 16157 isopod specimens from the Atlantic sector of the SO were identified to species level at a smaller scale (Chapter IV). In this case, sea ice was identified as the main physical driver affecting isopod diversity and composition among sampling locations (Chapter IV). Reduced concentration of sea ice
causes a decrease in isopod biodiversity, thus climate change was identified as a huge threat for this taxon and for SO benthos in general. During the identification process, two new isopod species were discovered (Chapter V). The two new species (Notopais sp.1 n. sp. and Notopais sp.2 n. sp.) were accurately described and identified by means of integrative taxonomy. This provided the first whole genome sequencing of benthic isopods from the SO and the first complete mitochondrial genome of the genus Notopais (Chapter V). Thanks to the collaboration with the University of Genoa (Dipartimento di Scienze della Terra dell'Ambiente e della Vita, DISTAV, Italy) and the National Antarctic Museum (MNA) in Genoa, two new SO species of the suborder Valvifera G. O. Sars, 1883 were described by means of classical taxonomy. In this case, a molecular approach could not be used because both new species were represented by a single specimen, therefore it was important to preserve the integrity of the holotypes (Chapters VI-VII).
Plastics contain a complex mixture of chemicals including polymers, additives, starting substances and side-products of processing. These plastic chemicals are prone to leach into the packaged goods, in the case of food contact materials (FCMs), or into the natural environment, in the case of plastic debris. Thus, plastics represent an exposure source of chemicals for humans and wildlife alike. While it is widely known that individual plastic chemicals, such as bisphenol A and phthalates, are hazardous, little is known on the overall chemical composition and toxicity of plastics. When fragmented into smaller particles, referred to as microplastics (< 5 mm), the plastic itself can be ingested by many species. It is well established that microplastic ingestion can have negative consequences for a wide range of organisms including invertebrates, but the contribution of plastic chemicals to the toxicity of microplastics is unclear.
Given the above, the present thesis aimed at a comprehensive toxicological, ecotoxicological and chemical characterization of everyday plastics. For a comparative evaluation, 77 plastic products were selected covering 16 material types (e.g., polyethylene) made from petroleum or renewable feedstocks. These products included biodegradable products, FCMs and non-FCMs, as well as raw materials and final products, respectively. In the first two studies, the chemical mixtures contained in the 77 products were extracted with methanol and extracts were analyzed in a set of four in vitro bioassays and by non-target high-resolution gas or liquid chromatography mass spectrometry. Since an exposure only occurs if chemicals actually leach under realistic conditions, in a third study migration experiments with water were conducted for 24 out of the 77 products. The aqueous migrates were assessed in the same way as the methanolic extracts. In addition, the freshwater invertebrate Daphnia magna was exposed chronically to microplastics made of polyvinylchloride (PVC), polyurethane (PUR) and polylactic acid (PLA) to investigate the contribution of chemicals in microplastic toxicity, in a fourth study.
The experimental findings demonstrate that a wide variety of chemicals is present in plastics. A single plastic product can contain up to several thousand chemical features, most of which unique to that product and at the same time unknown. The results also indicate that the majority of these chemical mixtures are toxic in vitro. Accordingly, 65% of the plastic extracts induced baseline toxicity and 42% an oxidative stress response, while 25% had an antiandrogenic and 6% an estrogenic activity. This implies that chemicals causing unspecific toxicity are more prevalent in plastics than such with endocrine effects. These chemicals can also leach from plastics under realistic conditions. Between 17 and 8936 chemical features were detected in a single migrate sample and all 24 tested migrates induced in vitro toxicity. This means that humans and wildlife can actually be exposed to toxic plastic chemicals under realistic conditions. Generally, each product has its individual toxicological and chemical fingerprint. Thus, neither material type, feedstock, biodegradability nor the food contact suitability of a product can serve as a predictor for the toxicity, the chemical composition or complexity of a product. Likewise, this means that bio-based and biodegradable materials are not superior to their petroleum-based counterparts from a toxicological perspective despite being promoted as sustainable alternatives to conventional plastics.
Moreover, the present thesis demonstrates that plastic chemicals can be the main driver for microplastic toxicity. Irregular microplastics made of PVC, PUR and PLA adversely affected life-history traits of D. magna in a polymer type- and endpoint-dependent manner at concentrations between 100 and 500 mg L-1 and with a higher efficiency than natural kaolin particles. While the toxicity of PVC was triggered by the chemicals used in the material, the effects of PUR and PLA were induced by the physical properties of the particle.
In addition, in the fifth study, results and observations made during this thesis were integrated inter- and transdisciplinarily with the perspectives of a social scientist and a product manufacturer. This elucidated that knowledge on plastic ingredients is often concealed, is lacking or not applicable in practice. These intransparencies hinder the safety evaluation of plastic products as well as the choice and sale of the least toxic packaging material.
Overall, the present thesis highlights that the chemical safety of plastics and their bio-based and biodegradable alternatives is currently not ensured. Thus, chemicals require more consideration in the toxicity and risk assessment of plastics and microplastics. Product-specific and complex chemical compositions, including unknown compounds, pose a challenge here. Two essential steps towards non-toxic products are to increase transparency along the product life cycle and to reduce the chemical complexity of plastics by communication and regulation. The results of the present thesis indicate that products exist which do not contain toxic chemicals. These can serve to direct the design of safer plastics. Since toxicity and chemical complexity seem to increase with processing, the integration of toxicity testing during the production steps would further support the safe and sustainable production and use of plastic products.
Seit den 1950er Jahren hat sich Plastik als unverzichtbare Ressource im menschlichen Alltag etabliert. Als negative Folge dieses Booms wird seit einigen Jahrzehnten jedoch eine zunehmende Belastung aquatischer Ökosysteme mit Plastikmüll bzw. dessen Degradationsprodukten, sogenanntes „Mikroplastik“ (MP, < 5 mm) bzw. „Nanoplastik“ (NP, < 1 µm), beobachtet. Ziel dieser Arbeit war die Untersuchung des aktuellen Vorkommens von MP in limnischen Gewässern, die Analyse der Interaktion zwischen MP und limnischen Wirbellosenarten und der daraus resultierenden Toxizität sowie eine erste Risikoabschätzung.
Das Vorkommen von Mikroplastik in limnischen Gewässern wurde exemplarisch anhand der Elbe als großes Fließgewässer in Deutschland untersucht. Durch die Auswertung von elf Probestellen entlang des Verlaufs der Mittel- und Unterelbe konnte gezeigt werden, dass die MP-Konzentrationen im Sediment (2,26x10^4 – 2,27x10^7 P m^-3) im Mittel fast 150.000-fach höher sind als in der Wasserphase (0,88–13,24 P m^-3). Sedimente sind somit eine Senke für MP. Die Zusammensetzung der Polymerarten sowie MP-Formen deuten zudem an, dass die Partikel sowohl aus diffusen wie auch aus Punktquellen (z.B. Industrieabwässer) stammen. Im globalen Vergleich können die MP-Konzentrationen in deutschen Fließgewässern z. Z. als durchschnittlich betrachtet werden. Allerdings muss insgesamt davon ausgegangen werden, dass die bisher bestimmten MP-Umweltkonzentrationen die realen Konzentrationen möglicherweise unterschätzen. So zeigte die Elbestudie, dass die Sedimentfeinfraktion < 100 µm einen bedeutenden Polymeranteil enthielt. Die meisten bisher durchgeführten Studien zur Bestimmung von MP-Partikeln in Flüssen haben Partikel < 100 µm jedoch nicht in ihrer Analyse berücksichtigt.
Die Interaktion von MP mit limnischer Biota wurde anhand der Artgruppen der Muscheln (Bivalvia), Schnecken (Gastropoda) sowie Krebstiere (Crustacea) näher untersucht. Die Intensität der Interaktion ist maßgeblich von der Aufnahme von MP durch die verschiedenen Arten abhängig. Anhand von zahlreichen Aufnahmestudien mit verschiedenen limnischen Arten, darunter den Muschelarten Dreissena polymorpha, Sinanodonta woodiana und Anodonta anatina, der Lungenschnecke Lymnaea stagnalis sowie der Amphipodenart Gammarus pulex, wurde nachgewiesen, dass die MP-Aufnahme von den Eigenschaften der exponierten Arten bzw. Individuen, den MP-Charakteristika sowie den Expositionsbedingungen abhängt. Die Experimente mit Muscheln verdeutlichten die rasche Aufnahme, aber auch Exkretion von MP-Partikeln innerhalb weniger Stunden. In allen drei Artgruppen war die Aufnahme konzentrationsabhängig mit zunehmender Aufnahme bei steigenden MP-Konzentrationen. Die Muschelexperimente zeigten jedoch auch, dass eine gleichzeitige Exposition mit anderen Partikeln (z.B. Nahrung) zu einer reduzierten Aufnahme führt. Auch die Größe der Testorganismen beeinflusste die Aufnahme: So nahmen im Fall der Muscheln und Krebse kleinere Individuen (bzw. im Fall der Muscheln auch Arten) relativ pro Körpermasse mehr MP-Partikel auf als größere Individuen bzw. Arten. Für alle untersuchten Arten wurde darüber hinaus gezeigt, dass die MP-Größe relevanten Einfluss auf die Menge an aufgenommenen Partikeln hat.
Ein Vergleich zwischen den Artgruppen zeigte, dass Muscheln als filtrierende Organismen in den Laboruntersuchungen bei gleicher Expositionskonzentration mehr MP aufnahmen als Krebse (Zerkleinerer) und Schnecken (Weidegänger). Im Gegensatz zu Muscheln nutzen Krebstiere und Schnecken allerdings die Grenzschicht zwischen Wasser- und Sedimentphase als Suchraum für ihre Nahrung und sind in der Umwelt (auf Grund des höheren MP-Vorkommens in Sedimenten) somit möglicherweise gegenüber höheren MP-Konzentrationen exponiert als Muscheln. Die Extrapolation der gewonnenen Labordaten sowie der Vergleich mit publizierten Umweltdaten legen allerdings nahe, dass das MP-Vorkommen in Individuen aller drei Artgruppen bisher auf einige wenige MP-Partikel begrenzt ist. Ausgeprägte Unterschiede zwischen den Artgruppen sind bisher nicht erkennbar.
MP-Toxizitätsstudien mit G. pulex, L. stagnalis sowie D. polymorpha konnten trotz der Berücksichtigung einer Vielzahl an Endpunkten (Mortalität, Reproduktion, Nahrungsaufnahme, oxidativer Stress, Energiereserven, Immunzellaktivität) und trotz des Einsatzes zum Teil sehr hoher MP-Konzentrationen weit oberhalb aktueller Umweltkonzentrationen nur sehr wenige MP-induzierte Effekte nachweisen, darunter eine Steigerung der Filtrationsaktivität (D. polymorpha) bzw. Veränderung der Immunfunktion von Hämolymphzellen (L. stagnalis).
Zur weiteren Risikoabschätzung wurden diese Studienergebnisse mit publizierten Daten für marine und limnische Muschel- und Krebsarten in Artenempfindlichkeitsverteilungen (Species Sensitivity Distributions, SSD) zusammengeführt und jeweils eine SSD für Muscheln und Krebstiere erstellt. Die Erstellung einer SSD nur für limnische Arten ist zum jetzigen Zeitpunkt auf Grund der geringen Datenlage noch nicht möglich.
...
Tissue translocation, multigenerational and population effects of microplastics in Daphnia magna
(2021)
The last century saw the widespread adoption of plastic materials throughout nearly every aspect of our lives. Plastics are synthetic polymers that are made up of monomer chains. The properties of the monomer in conjunction with chemical additives allow plastics to have a sheer endless variety of features and use cases. They are cheap, lightweight, and extremely durable. Plastic materials are often engineered for single-use and in conjunction with high production volumes and insufficient waste management and recycling across the globe, this leads to a large number of plastics entering the environment. Marine ecosystems are considered sinks. However, freshwater ecosystems as entry pathways are highly affected by plastic waste as well. Throughout the past decade, the impact of plastic waste on human and environmental health has received a lot of attention from the ecotoxicological community as well as the public. Small plastic fragments (< 1 mm called microplastics) are a large part of this emerging field of research. Within this, the water flea Daphnia magna is probably the most common organism that is used to assess microplastics toxicity. As a filter-feeding organism, it indiscriminately ingests particles from the water column and is thus highly susceptible to microplastics. For this thesis, we identified some gaps in the available data on the ecotoxicity of microplastics to daphnids. To illuminate some of those gaps the present thesis was aimed at five main aspects:
(1) Tissue translocation of spherical microplastics in Daphnia magna
(2) Investigation of the toxicity of irregularly shaped microplastics
(3) Multigenerational and population effects of microplastics
(4) Comparison of the toxicity of microplastics and natural particles
(5) Effects of particle-aging on microplastics toxicity
The thesis is comprised of three peer-reviewed articles and one so-far unpublished study as “additional results”. The first study was aimed at understanding tissue translocation of spherical microplastics to lipid storage droplets of daphnids. The crossing of biological membranes is discussed as a prerequisite to eliciting tissue damage and an inflammatory response. Previously, researchers reported the translocation of fluorescently labeled spherical microplastics to lipid storage droplets of daphnids, even though no plausible biological mechanism to explain this occurrence. Therefore, in order to learn more about this process and potentially illuminate the mechanism we replicated the study. We were able to observe a fluorescence signal inside the lipid droplets only after increasing the exposure concentrations. Nonetheless, it appeared to be independent of particles. This led to the hypothesis, that the lipophilic fluorescent dye uncoupled from the particles and subsequently accumulated in lipid storage droplets. The hypothesis was further confirmed through an additional experiment with a silicone-based passive sampling device showing that the fluorescence occurred both independent of particles and digestive processes. Accordingly, we concluded that the reported findings were a microscopic artifact caused by the uncoupling of the dye from the particles. Therefore, a fluorescence signal alone is not a sufficient proxy to assume that particles have translocated. It needs to be coupled with additional methods to ensure that the observation is indeed caused by the translocation of particles.
It is still unclear whether the toxicity profile of microplastics is different from that of naturally occurring particles or if they are “just another particle”, as there are innumerable amounts in the natural environment surrounding an organism. The goal of the second study was to compare the toxicity of irregularly shaped polystyrene microplastics to that of the natural particle kaolin. The environment is full of natural non-food particles that daphnids ingest more or less indiscriminately and therefore are well adapted to deal with. Daphnids have a short generation time and usually experience food limitation in nature. Therefore, short-term studies only looking at acute toxicity with ad libitum food availability are not representative of the exposure scenario in nature. For a more realistic scenario, we, therefore, used a four-generation multigenerational design under food limitation to investigate how effects translate from one generation to the next. We observed concentration-dependent effects of microplastics but not of natural particles on mortality, reproduction, and growth. Some of the effects increased from generation to generation, leading to the extinction of two treatment groups. Here, microplastics were more toxic than natural particles. At least part of this difference can be explained by physical properties leading to the quick sedimentation of the kaolin, while microplastics remained in the water column. Nonetheless, buoyancy and sedimentation would also affect exposure in the environment and are likely different for most microplastics than for most naturally occurring particle types.
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