@phdthesis{AriasAndres2018, author = {Arias Andr{\´e}s, Mar{\´i}a de Jes{\´u}s}, title = {Microbial gene exchange on microplastic particles}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-417241}, school = {Universit{\"a}t Potsdam}, pages = {94}, year = {2018}, abstract = {Plastic pollution is ubiquitous on the planet since several millions of tons of plastic waste enter aquatic ecosystems each year. Furthermore, the amount of plastic produced is expected to increase exponentially shortly. The heterogeneity of materials, additives and physical characteristics of plastics are typical of these emerging contaminants and affect their environmental fate in marine and freshwaters. Consequently, plastics can be found in the water column, sediments or littoral habitats of all aquatic ecosystems. Most of this plastic debris will fragment as a product of physical, chemical and biological forces, producing particles of small size. These particles (< 5mm) are known as "microplastics" (MP). Given their high surface-to-volume ratio, MP stimulate biofouling and the formation of biofilms in aquatic systems. As a result of their unique structure and composition, the microbial communities in MP biofilms are referred to as the "Plastisphere." While there is increasing data regarding the distinctive composition and structure of the microbial communities that form part of the plastisphere, scarce information exists regarding the activity of microorganisms in MP biofilms. This surface-attached lifestyle is often associated with the increase in horizontal gene transfer (HGT) among bacteria. Therefore, this type of microbial activity represents a relevant function worth to be analyzed in MP biofilms. The horizontal exchange of mobile genetic elements (MGEs) is an essential feature of bacteria. It accounts for the rapid evolution of these prokaryotes and their adaptation to a wide variety of environments. The process of HGT is also crucial for spreading antibiotic resistance and for the evolution of pathogens, as many MGEs are known to contain antibiotic resistance genes (ARGs) and genetic determinants of pathogenicity. In general, the research presented in this Ph.D. thesis focuses on the analysis of HGT and heterotrophic activity in MP biofilms in aquatic ecosystems. The primary objective was to analyze the potential of gene exchange between MP bacterial communities vs. that of the surrounding water, including bacteria from natural aggregates. Moreover, the thesis addressed the potential of MP biofilms for the proliferation of biohazardous bacteria and MGEs from wastewater treatment plants (WWTPs) and associated with antibiotic resistance. Finally, it seeks to prove if the physiological profile of MP biofilms under different limnological conditions is divergent from that of the water communities. Accordingly, the thesis is composed of three independent studies published in peer-reviewed journals. The two laboratory studies were performed using both model and environmental microbial communities. In the field experiment, natural communities from freshwater ecosystems were examined. In Chapter I, the inflow of treated wastewater into a temperate lake was simulated with a concentration gradient of MP particles. The effects of MP on the microbial community structure and the occurrence of integrase 1 (int 1) were followed. The int 1 is a marker associated with mobile genetic elements and known as a proxy for anthropogenic effects on the spread of antimicrobial resistance genes. During the experiment, the abundance of int1 increased in the plastisphere with increasing MP particle concentration, but not in the surrounding water. In addition, the microbial community on MP was more similar to the original wastewater community with increasing microplastic concentrations. Our results show that microplastic particles indeed promote persistence of standard indicators of microbial anthropogenic pollution in natural waters. In Chapter II, the experiments aimed to compare the permissiveness of aquatic bacteria towards model antibiotic resistance plasmid pKJK5, between communities that form biofilms on MP vs. those that are free-living. The frequency of plasmid transfer in bacteria associated with MP was higher when compared to bacteria that are free-living or in natural aggregates. Moreover, comparison increased gene exchange occurred in a broad range of phylogenetically-diverse bacteria. The results indicate a different activity of HGT in MP biofilms, which could affect the ecology of aquatic microbial communities on a global scale and the spread of antibiotic resistance. Finally, in Chapter III, physiological measurements were performed to assess whether microorganisms on MP had a different functional diversity from those in water. General heterotrophic activity such as oxygen consumption was compared in microcosm assays with and without MP, while diversity and richness of heterotrophic activities were calculated by using Biolog® EcoPlates. Three lakes with different nutrient statuses presented differences in MP-associated biomass build up. Functional diversity profiles of MP biofilms in all lakes differed from those of the communities in the surrounding water, but only in the oligo-mesotrophic lake MP biofilms had a higher functional richness compared to the ambient water. The results support that MP surfaces act as new niches for aquatic microorganisms and can affect global carbon dynamics of pelagic environments. Overall, the experimental works presented in Chapters I and II support a scenario where MP pollution affects HGT dynamics among aquatic bacteria. Among the consequences of this alteration is an increase in the mobilization and transfer efficiency of ARGs. Moreover, it supposes that changes in HGT can affect the evolution of bacteria and the processing of organic matter, leading to different catabolic profiles such as demonstrated in Chapter III. The results are discussed in the context of the fate and magnitude of plastic pollution and the importance of HGT for bacterial evolution and the microbial loop, i.e., at the base of aquatic food webs. The thesis supports a relevant role of MP biofilm communities for the changes observed in the aquatic microbiome as a product of intense human intervention.}, language = {en} } @phdthesis{Drago2022, author = {Drago, Claudia}, title = {Microplastics in the environment: Assessing the ingestion and effect of microplastics on freshwater rotifers in an environmental scenario}, doi = {10.25932/publishup-57335}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-573356}, school = {Universit{\"a}t Potsdam}, pages = {xv, 116}, year = {2022}, abstract = {Microplastics in the environments are estimated to increase in the near future due to increasing consumption of plastic product and also due to further fragmentation in small pieces. The fate and effects of MP once released into the freshwater environment are still scarcely studied, compared to the marine environment. In order to understand possible effect and interaction of MPs in freshwater environment, planktonic zooplankton organisms are very useful for their crucial trophic role. In particular freshwater rotifers are one of the most abundant organisms and they are the interface between primary producers and secondary consumers. The aim of my thesis was to investigate the ingestion and the effect of MPs in rotifers from a more natural scenario and to individuate processes such as the aggregation of MPs, the food dilution effect and the increasing concentrations of MPs that could influence the final outcome of MPs in the environment. In fact, in a near natural scenario MPs interaction with bacteria and algae, aggregations together with the size and concentration are considered drivers of ingestion and effect. The aggregation of MPs makes smaller MPs more available for rotifers and larger MPs less ingested. The negative effect caused by the ingestion of MPs was modulated by their size but also by the quantity and the quality of food that cause variable responses. In fact, rotifers in the environment are subjected to food limitation and the presence of MPs could exacerbate this condition and decrease the population and the reproduction input. Finally, in a scenario incorporating an entire zooplanktonic community, MPs were ingested by most individuals taking into account their feeding mode but also the concentration of MPs, which was found to be essential for the availability of MPs. This study highlights the importance to investigate MPs from a more environmental perspective, this in fact could provide an alternative and realistic view of effect of MPs in the ecosystem.}, language = {en} } @article{DragoPawlakWeithoff2020, author = {Drago, Claudia and Pawlak, Julia and Weithoff, Guntram}, title = {Biogenic aggregation of small microplastics alters their ingestion by a common freshwater micro-invertebrate}, series = {Frontiers in Environmental Science}, volume = {8}, journal = {Frontiers in Environmental Science}, publisher = {Frontiers Media}, address = {Lausanne}, issn = {2296-665X}, doi = {10.3389/fenvs.2020.574274}, pages = {11}, year = {2020}, abstract = {In recent years, increasing concerns have been raised about the environmental risk of microplastics in freshwater ecosystems. Small microplastics enter the water either directly or accumulate through disintegration of larger plastic particles. These particles might then be ingested by filter-feeding zooplankton, such as rotifers. Particles released into the water may also interact with the biota through the formation of aggregates, which might alter the uptake by zooplankton. In this study, we tested for size-specific aggregation of polystyrene microspheres and their ingestion by a common freshwater rotifer Brachionus calyciflorus. The ingestion of three sizes of polystyrene microspheres (MS) 1-, 3-, and 6-mu m was investigated. Each MS size was tested in combination with three different treatments: MS as the sole food intake, MS in association with food algae and MS aggregated with biogenic matter. After 72 h incubation in pre-filtered natural river water, the majority of the 1-mu m spheres occurred as aggregates. The larger the particles, the higher the relative number of single particles and the larger the aggregates. All particles were ingested by the rotifer following a Type-II functional response. The presence of algae did not influence the ingestion of the MS for all three sizes. The biogenic aggregation of microspheres led to a significant size-dependent alteration in their ingestion. Rotifers ingested more microspheres (MS) when exposed to aggregated 1- and 3-mu m MS as compared to single spheres, whereas fewer aggregated 6-mu m spheres were ingested. This indicates that the small particles when aggregated were in an effective size range for Brachionus, while the aggregated larger spheres became too large to be efficiently ingested. These observations provide the first evidence of a size- and aggregation-dependent feeding interaction between microplastics and rotifers. Microplastics when aggregated with biogenic particles in a natural environment can rapidly change their size-dependent availability. The aggregation properties of microplastics should be taken into account when performing experiments mimicking the natural environment.}, language = {en} } @misc{DragoWeithoff2021, author = {Drago, Claudia and Weithoff, Guntram}, title = {Variable Fitness Response of Two Rotifer Species Exposed to Microplastics Particles}, series = {Zweitver{\"o}ffentlichungen der Universit{\"a}t Potsdam : Mathematisch-Naturwissenschaftliche Reihe}, journal = {Zweitver{\"o}ffentlichungen der Universit{\"a}t Potsdam : Mathematisch-Naturwissenschaftliche Reihe}, number = {1248}, issn = {1866-8372}, doi = {10.25932/publishup-55261}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-552615}, pages = {13}, year = {2021}, abstract = {Plastic pollution is an increasing environmental problem, but a comprehensive understanding of its effect in the environment is still missing. The wide variety of size, shape, and polymer composition of plastics impedes an adequate risk assessment. We investigated the effect of differently sized polystyrene beads (1-, 3-, 6-µm; PS) and polyamide fragments (5-25 µm, PA) and non-plastics items such as silica beads (3-µm, SiO2) on the population growth, reproduction (egg ratio), and survival of two common aquatic micro invertebrates: the rotifer species Brachionus calyciflorus and Brachionus fernandoi. The MPs were combined with food quantity, limiting and saturating food concentration, and with food of different quality. We found variable fitness responses with a significant effect of 3-µm PS on the population growth rate in both rotifer species with respect to food quantity. An interaction between the food quality and the MPs treatments was found in the reproduction of B. calyciflorus. PA and SiO2 beads had no effect on fitness response. This study provides further evidence of the indirect effect of MPs in planktonic rotifers and the importance of testing different environmental conditions that could influence the effect of MPs.}, language = {en} } @article{DragoWeithoff2021, author = {Drago, Claudia and Weithoff, Guntram}, title = {Variable Fitness Response of Two Rotifer Species Exposed to Microplastics Particles}, series = {Toxics}, volume = {9}, journal = {Toxics}, number = {11}, publisher = {MDPI}, address = {Basel}, issn = {2305-6304}, doi = {10.3390/toxics9110305}, pages = {13}, year = {2021}, abstract = {Plastic pollution is an increasing environmental problem, but a comprehensive understanding of its effect in the environment is still missing. The wide variety of size, shape, and polymer composition of plastics impedes an adequate risk assessment. We investigated the effect of differently sized polystyrene beads (1-, 3-, 6-µm; PS) and polyamide fragments (5-25 µm, PA) and non-plastics items such as silica beads (3-µm, SiO2) on the population growth, reproduction (egg ratio), and survival of two common aquatic micro invertebrates: the rotifer species Brachionus calyciflorus and Brachionus fernandoi. The MPs were combined with food quantity, limiting and saturating food concentration, and with food of different quality. We found variable fitness responses with a significant effect of 3-µm PS on the population growth rate in both rotifer species with respect to food quantity. An interaction between the food quality and the MPs treatments was found in the reproduction of B. calyciflorus. PA and SiO2 beads had no effect on fitness response. This study provides further evidence of the indirect effect of MPs in planktonic rotifers and the importance of testing different environmental conditions that could influence the effect of MPs.}, language = {en} } @phdthesis{Kettner2018, author = {Kettner, Marie Therese}, title = {Microbial colonization of microplastic particles in aquatic systems}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-418854}, school = {Universit{\"a}t Potsdam}, pages = {139}, year = {2018}, abstract = {The continuously increasing pollution of aquatic environments with microplastics (plastic particles < 5 mm) is a global problem with potential implications for organisms of all trophic levels. For microorganisms, trillions of these floating microplastics particles represent a huge surface area for colonization. Due to the very low biodegradability, microplastics remain years to centuries in the environment and can be transported over thousands of kilometers together with the attached organisms. Since also pathogenic, invasive, or otherwise harmful species could be spread this way, it is essential to study microplastics-associated communities. For this doctoral thesis, eukaryotic communities were analyzed for the first time on microplastics in brackish environments and compared to communities in the surrounding water and on the natural substrate wood. With Illumina MiSeq high-throughput sequencing, more than 500 different eukaryotic taxa were detected on the microplastics samples. Among them were various green algae, dinoflagellates, ciliates, fungi, fungal-like protists and small metazoans such as nematodes and rotifers. The most abundant organisms was a dinoflagellate of the genus Pfiesteria, which could include fish pathogenic and bloom forming toxigenic species. Network analyses revealed that there were numerous interaction possibilities among prokaryotes and eukaryotes in microplastics biofilms. Eukaryotic community compositions on microplastics differed significantly from those on wood and in water, and compositions were additionally distinct among the sampling locations. Furthermore, the biodiversity was clearly lower on microplastics in comparison to the diversity on wood or in the surrounding water. In another experiment, a situation was simulated in which treated wastewater containing microplastics was introduced into a freshwater lake. With increasing microplastics concentrations, the resulting bacterial communities became more similar to those from the treated wastewater. Moreover, the abundance of integrase I increased together with rising concentrations of microplastics. Integrase I is often used as a marker for anthropogenic environmental pollution and is further linked to genes conferring, e.g., antibiotic resistance. This dissertation gives detailed insights into the complexity of prokaryotic and eukaryotic communities on microplastics in brackish and freshwater systems. Even though microplastics provide novel microhabitats for various microbes, they might also transport toxigenic, pathogenic, antibiotic-resistant or parasitic organisms; meaning their colonization can pose potential threats to humans and the environment. Finally, this thesis explains the urgent need for more research as well as for strategies to minimize the global microplastic pollution.}, language = {en} } @article{KimLeifheitMaassetal.2021, author = {Kim, Shin Woong and Leifheit, Eva F. and Maaß, Stefanie and Rillig, Matthias C.}, title = {Time-dependent toxicity of tire particles on soil nematodes}, series = {Frontiers in Environmental Science}, volume = {9}, journal = {Frontiers in Environmental Science}, publisher = {Frontiers Media}, address = {Lausanne}, issn = {2296-665X}, doi = {10.3389/fenvs.2021.744668}, pages = {9}, year = {2021}, abstract = {Tire-wear particles (TWPs) are being released into the environment by wearing down during car driving, and are considered an important microplastic pollution source. The chemical additive leaching from these polymer-based materials and its potential effects are likely temporally dynamic, since amounts of potentially toxic compounds can gradually increase with contact time of plastic particles with surrounding media. In the present study, we conducted soil toxicity tests using the soil nematode Caenorhabditis elegans with different soil pre-incubation (30 and 75 days) and exposure (short-term exposure, 2 days; lifetime exposure, 10 days) times. Soil pre-incubation increased toxicity of TWPs, and the effective concentrations after the pre-incubation were much lower than environmentally relevant concentrations. The lifetime of C. elegans was reduced faster in the TWP treatment groups, and the effective concentration for lifetime exposure tests were 100- to 1,000-fold lower than those of short-term exposure tests. Water-extractable metal concentrations (Cr, Cu, Ni, Pb, and Zn) in the TWP-soils showed no correlation with nominal TWP concentrations or pre-incubation times, and the incorporated metals in the TWPs may be not the main reason of toxicity in this study. Our results show that toxic effects of TWPs can be time-dependent, both in terms of the microplastic particles themselves and their interactions in the soil matrix, but also because of susceptibility of target organisms depending on developmental stage. It is vital that future works consider these aspects, since otherwise effects of microplastics and TWPs could be underestimated.}, language = {en} } @phdthesis{Leiser2021, author = {Leiser, Rico}, title = {Biogeochemical processes governing microplastic transport in freshwater reservoirs}, doi = {10.25932/publishup-52024}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-520240}, school = {Universit{\"a}t Potsdam}, pages = {ix, 143}, year = {2021}, abstract = {The presented study investigated the influence of microbial and biogeochemical processes on the physical transport related properties and the fate of microplastics in freshwater reservoirs. The overarching goal was to elucidate the mechanisms leading to sedimentation and deposition of microplastics in such environments. This is of importance, as large amounts of initially buoyant microplastics are found in reservoir sediments worldwide. However, the transport processes which lead to microplastics accumulation in sediments, were up to now understudied. The impact of biofilm formation on the density and subsequent sedimentation of microplastics was investigated in the eutrophic Bautzen reservoirs (Chapter 2). Biofilms are complex microbial communities fixed to submerged surfaces through a slimy organic film. The mineral calcite was detected in the biofilms, which led to the sinking of the overgrown microplastic particles. The calcite was of biogenic origin, most likely precipitated by sessile cyanobacteria within the biofilms. Biofilm formation was also studied in the mesotrophic Malter reservoir. Unlike in Bautzen reservoir, biofilm formation did not govern the sedimentation of different microplastics in Malter reservoir (Chapter 3). Instead autumnal lake mixing led to the formation of sinking aggregates of microplastics and iron colloids. Such colloids form when anoxic, iron-rich water from the hypolimnion mixes with the oxygenated epilimnetic waters. The colloids bind organic material from the lake water, which leads to the formation of large and sinking iron-organo flocs. Hence, iron-organo floc formation and their influence on the buoyancy or burial of microplastics into sediments of Bautzen reservoir was studied in laboratory experiments (Chapter 4). Microplastics of different shapes (fiber, fragment, sphere) and sizes were readily incorporated into sinking iron-organo flocs. By this initially buoyant polyethylene microplastics were transported on top of sediments from Bautzen reservoir. Shortly after deposition, the microplastic bearing flocs started to subside and transported the pollutants into deeper sediment layers. The microplastics were not released from the sediments within two months of laboratory incubation. The stability of floc microplastic deposition was further investigated employing experiments with the iron reducing model organism Shewanella oneidensis (Chapter 5). It was shown, that reduction or re-mineralization of the iron minerals did not affect the integrity of the iron-organo flocs. The organic matrix was stable under iron reducing conditions. Hence, no incorporated microplastics were released from the flocs. As similar processes are likely to take place in natural sediments, this might explain the previous described low microplastic release from the sediments. This thesis introduced different mechanisms leading to the sedimentation of initially buoyant microplastics and to their subsequent deposition in freshwater reservoirs. Novel processes such as the aggregation with iron-organo flocs were identified and the understudied issue of biofilm densification through biogenic mineral formation was further investigated. The findings might have implications for the fate of microplastics within the river-reservoir system and outline the role of freshwater reservoirs as important accumulation zone for microplastics. Microplastics deposited in the sediments of reservoirs might not be transported further by through flowing river. Hence the study might contribute to better risk assessment and transport balances of these anthropogenic contaminants.}, language = {en} } @misc{PawlakNoetzelDragoetal.2022, author = {Pawlak, Julia and Noetzel, Dominique Christian and Drago, Claudia and Weithoff, Guntram}, title = {Assessing the toxicity of polystyrene beads and silica particles on the microconsumer Brachionus calyciflorus at different timescales}, series = {Zweitver{\"o}ffentlichungen der Universit{\"a}t Potsdam : Mathematisch-Naturwissenschaftliche Reihe}, journal = {Zweitver{\"o}ffentlichungen der Universit{\"a}t Potsdam : Mathematisch-Naturwissenschaftliche Reihe}, number = {1277}, issn = {1866-8372}, doi = {10.25932/publishup-56996}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-569967}, pages = {1 -- 11}, year = {2022}, abstract = {Environmental pollution by microplastics has become a severe problem in terrestrial and aquatic ecosystems and, according to actual prognoses, problems will further increase in the future. Therefore, assessing and quantifying the risk for the biota is crucial. Standardized short-term toxicological procedures as well as methods quantifying potential toxic effects over the whole life span of an animal are required. We studied the effect of the microplastic polystyrene on the survival and reproduction of a common freshwater invertebrate, the rotifer Brachionus calyciflorus, at different timescales. We used pristine polystyrene spheres of 1, 3, and 6 µm diameter and fed them to the animals together with food algae in different ratios ranging from 0 to 50\% nonfood particles. As a particle control, we used silica to distinguish between a pure particle effect and a plastic effect. After 24 h, no toxic effect was found, neither with polystyrene nor with silica. After 96 h, a toxic effect was detectable for both particle types. The size of the particles played a negligible role. Studying the long-term effect by using life table experiments, we found a reduced reproduction when the animals were fed with 3 µm spheres together with similar-sized food algae. We conclude that the fitness reduction is mainly driven by the dilution of food by the nonfood particles rather than by a direct toxic effect.}, language = {en} } @article{PawlakNoetzelDragoetal.2022, author = {Pawlak, Julia and Noetzel, Dominique Christian and Drago, Claudia and Weithoff, Guntram}, title = {Assessing the toxicity of polystyrene beads and silica particles on the microconsumer Brachionus calyciflorus at different timescales}, series = {Frontiers in Environmental Science}, journal = {Frontiers in Environmental Science}, publisher = {Frontiers}, address = {Lausanne, Schweiz}, issn = {2296-665X}, doi = {10.3389/fenvs.2022.955425}, pages = {1 -- 11}, year = {2022}, abstract = {Environmental pollution by microplastics has become a severe problem in terrestrial and aquatic ecosystems and, according to actual prognoses, problems will further increase in the future. Therefore, assessing and quantifying the risk for the biota is crucial. Standardized short-term toxicological procedures as well as methods quantifying potential toxic effects over the whole life span of an animal are required. We studied the effect of the microplastic polystyrene on the survival and reproduction of a common freshwater invertebrate, the rotifer Brachionus calyciflorus, at different timescales. We used pristine polystyrene spheres of 1, 3, and 6 µm diameter and fed them to the animals together with food algae in different ratios ranging from 0 to 50\% nonfood particles. As a particle control, we used silica to distinguish between a pure particle effect and a plastic effect. After 24 h, no toxic effect was found, neither with polystyrene nor with silica. After 96 h, a toxic effect was detectable for both particle types. The size of the particles played a negligible role. Studying the long-term effect by using life table experiments, we found a reduced reproduction when the animals were fed with 3 µm spheres together with similar-sized food algae. We conclude that the fitness reduction is mainly driven by the dilution of food by the nonfood particles rather than by a direct toxic effect.}, language = {en} }