Department Environmental Toxicology

Aquatic Biofilms

Biofilms contribute to crucial ecosystem processes in aquatic ecosystems and capture much of the biological complexity occurring in the environment. Importantly, the high microbial diversity, and sensitivities to environmental changes, suggests that the microorganisms composing biofilms are a suitable model to assess contaminant effects on complex ecological systems. 

By using aquatic biofilms, we aim in our group to (i) unravel the underlying mechanisms of toxicity and biological responses to pollutants at the community level, and (ii) develop and optimize standardized risk assessment approaches accounting for the chemical and biological complexity in the environment. In particular, we are interested in understanding adaptation processes in microbial communities, following chronic exposures to pollutants, as these processes can be ultimately used to establish causalities in the environment.

Latest publications

Carles, L., Wullschleger, S., Joss, A., Eggen, R. I. L., Schirmer, K., Schuwirth, N., … Tlili, A. (2022). Wastewater microorganisms impact microbial diversity and important ecological functions of stream periphyton. Water Research, 225, 119119 (13 pp.). doi:10.1016/j.watres.2022.119119, Institutional Repository

Effluents of wastewater treatment plants can impact microbial communities in the receiving streams. However, little is known about the role of microorganisms in wastewater as opposed to other wastewater constituents, such as nutrients and micropollutants. We aimed therefore at determining the impact of wastewater microorganisms on the microbial diversity and function of periphyton, key microbial communities in streams. We used a flow-through channel system to grow periphyton upon exposure to a mixture of stream water and unfiltered or ultra-filtered wastewater. Impacts were assessed on periphyton biomass, activities and tolerance to micropollutants, as well as on microbial diversity. Our results showed that wastewater microorganisms colonized periphyton and modified its community composition, resulting for instance in an increased abundance of Chloroflexi and a decreased abundance of diatoms and green algae. This led to shifts towards heterotrophy, as suggested by the changes in nutrient stoichiometry and the increased mineralization potential of carbon substrates. An increased tolerance towards micropollutants was only found for periphyton exposed to unfiltered wastewater but not to ultra-filtered wastewater, suggesting that wastewater microorganisms were responsible for this increased tolerance. Overall, our results highlight the need to consider the role of wastewater microorganisms when studying potential impacts of wastewater on the receiving water body.
Groh, K., vom Berg, C., Schirmer, K., & Tlili, A. (2022). Anthropogenic chemicals as underestimated drivers of biodiversity loss: scientific and societal implications. Environmental Science and Technology, 56(2), 707-710. doi:10.1021/acs.est.1c08399, Institutional Repository
Guasch, H., Bernal, S., Bruno, D., Almroth, B. C., Cochero, J., Corcoll, N., … Martí, E. (2022). Interactions between microplastics and benthic biofilms in fluvial ecosystems: knowledge gaps and future trends. Freshwater Science, 41(3), 442-458. doi:10.1086/721472, Institutional Repository

Plastics, especially microplastics (<5 mm in length), are anthropogenic polymer particles that have been detected in almost all environments. Microplastics are extremely persistent pollutants and act as long-lasting reactive surfaces for additives, organic matter, and toxic substances. Biofilms are microbial assemblages that act as a sink for particulate matter, including microplastics. They are ubiquitous in freshwater ecosystems and provide key services that promote biodiversity and help sustain ecosystem function. Here, we provide a conceptual framework to describe the transient storage of microplastics in fluvial biofilm and develop hypotheses to help explain how microplastics and biofilms interact in fluvial ecosystems. We identify lines of future research that need to be addressed to better manage microplastics and biofilms, including how the sorption and desorption of environmental contaminants in microplastics affect biofilms and how microbial exchange between microplastics and the biofilm matrix affects biofilm characteristics like antibiotic resistance, speciation, biodiversity, species composition, and function. We also address the uptake mechanisms of microplastics by consumers and their propagation through the food web.
Holzer, M., Mitrano, D. M., Carles, L., Wagner, B., & Tlili, A. (2022). Important ecological processes are affected by the accumulation and trophic transfer of nanoplastics in a freshwater periphyton-grazer food chain. Environmental Science: Nano, 9, 2990-3003. doi:10.1039/D2EN00101B, Institutional Repository

Fundamental knowledge gaps on the bioaccumulation, trophic transfer and effects of nanoplastics in fresh waters limit the estimation of their ecological risks. In the present study, we investigated these unexplored aspects in a simplified stream food chain. Specifically, we used stream periphyton, which drives crucial ecosystem processes and forms the basis of the food web, and an aquatic snail feeding on periphyton as a primary producer and a primary consumer. Importantly, we used metal-doped polystyrene nanoplastics (nominal concentrations of 0.5, 5 and 50 mg plastics L-1), which allowed quantitative assessment of nanoplastics exposure across this complex biological system. Despite the ecological importance of periphyton in fresh waters, we are yet to understand whether microbial diversity and functions in periphyton, as well as grazer populations, are respectively impacted by accumulation of nanoplastics and dietary exposure. We demonstrated that periphyton efficiently accumulates and retains nanoplastics, which are consequently transferred to the grazers through dietary exposure although this did not lead to notable impacts on periphyton communities. Conversely, combined nanoplastics exposure and grazing pressure shifted community composition. Moreover, dietary exposure resulted in inhibited reproduction and reduced growth of the primary consumer, potentially leading to severe consequences for population dynamics due to trophic interactions in streams.
Lamprecht, O., Wagner, B., Derlon, N., & Tlili, A. (2022). Synthetic periphyton as a model system to understand species dynamics in complex microbial freshwater communities. npj Biofilms and Microbiomes, 8, 61 (14 pp.). doi:10.1038/s41522-022-00322-y, Institutional Repository

Phototrophic biofilms, also known as periphyton, are microbial freshwater communities that drive crucial ecological processes in streams and lakes. Gaining a deep mechanistic understanding of the biological processes occurring in natural periphyton remains challenging due to the high complexity and variability of such communities. To address this challenge, we rationally developed a workflow to construct a synthetic community by co-culturing 26 phototrophic species (i.e., diatoms, green algae, and cyanobacteria) that were inoculated in a successional sequence to create a periphytic biofilm on glass slides. We show that this community is diverse, stable, and highly reproducible in terms of microbial composition, function, and 3D spatial structure of the biofilm. We also demonstrate the ability to monitor microbial dynamics at the single species level during periphyton development and how their abundances are impacted by stressors such as increased temperature and a herbicide, singly and in combination. Overall, such a synthetic periphyton, grown under controlled conditions, can be used as a model system for theory testing through targeted manipulation.
Merbt, S. N., Kroll, A., Tamminen, M., Rühs, P. A., Wagner, B., Sgier, L., … Behra, R. (2022). Influence of microplastics on microbial structure, function, and mechanical properties of stream periphyton. Frontiers in Environmental Science, 10, 928247 (17 pp.). doi:10.3389/fenvs.2022.928247, Institutional Repository

Periphyton is a freshwater biofilm composed of prokaryotic and eukaryotic communities that occupy rocks and sediments, forming the base of the food web and playing a key role in nutrient cycling. Given the large surface that periphyton comprises, it may also act as a sink for a diverse range of man-made pollutants, including microplastics (MP). Here we investigated the effect of 1–4 μm and 63–75 µm sized, spherical polyethylene MP with native and ultraviolet (UV)-weathered surface on developing natural stream periphyton communities over 28 days. In order to ensure proper particle exposure, we first tested MP suspension in water or in water containing either Tween 80, extracellular polymeric substances – EPS, fulvic acids, or protein. We found the extract of EPS from natural periphyton to be most suitable to create MP suspensions in preparation of exposure. Upon exposure, all tested types of MP were found to be associated with the periphyton, independent of their size and other properties. While biomass accrual and phenotypic community structure of the photoautotrophs remained unchanged, the prokaryotic and eukaryotic communities experienced a significant change in composition and relative abundances. Moreover, alpha diversity was affected in eukaryotes, but not in prokaryotes. The observed changes were more prominent in periphyton exposed to UV-treated as compared with native surface MP. Mechanical properties, as assessed by compression rheology, showed that MP-exposed periphyton had longer filamentous streamers, higher stiffness, lower force recovery and a higher viscoelasticity than control periphyton. Despite the observed structural and mechanical changes of periphyton, functional parameters (i.e., photosynthetic yield, respiration and nutrient uptake efficiencies) were not altered by MP, indicating the absence of MP toxicity, and suggesting functional redundancy in the communities. Together, our results provide further proof that periphyton is a sink for MP and demonstrate that MP can impact local microbial community composition and mechanical properties of the biofilms. Consequences of these findings might be a change in dislodgement behavior of periphyton, a propagation through the food chains and impacts on nutrient cycling and energy transfer. Hence, taking the omnipresence, high persistence and material and size diversity of MP in the aquatic environment into account, their ecological consequences need further investigation.
Tamminen, M., Spaak, J., Tlili, A., Eggen, R., Stamm, C., & Räsänen, K. (2022). Wastewater constituents impact biofilm microbial community in receiving streams. Science of the Total Environment, 807(3), 151080 (8 pp.). doi:10.1016/j.scitotenv.2021.151080, Institutional Repository

Microbial life in natural biofilms is dominated by prokaryotes and microscopic eukaryotes living in dense association. In stream ecosystems, microbial biofilms influence primary production, elemental cycles, food web interactions as well as water quality. Understanding how biofilm communities respond to anthropogenic impacts, such as wastewater treatment plant (WWTP) effluent, is important given the key role of biofilms in stream ecosystem function. Here, we implemented 16S and 18S rRNA gene sequencing of stream biofilms upstream (US) and downstream (DS) of WWTP effluents in four Swiss streams to test how bacterial and eukaryotic communities respond to wastewater constituents. Stream biofilm composition was strongly affected by geographic location – particularly for bacteria. However, the abundance of certain microbial community members was related to micropollutants in the wastewater – among bacteria, micropollutant-associated members were found e.g. in Alphaproteobacteria, and among eukaryotes e.g. in Bacillariophyta (algal diatoms). This study corroborates several previously characterized responses (e.g. as seen in diatoms), but also reveals previously unknown community responses – such as seen in Alphaproteobacteria. This study advances our understanding of the ecological impact of the current wastewater treatment practices and provides information about potential new marker organisms to assess ecological change in stream biofilms.
Carles, L., Wullschleger, S., Joss, A., Eggen, R. I. L., Schirmer, K., Schuwirth, N., … Tlili, A. (2021). Impact of wastewater on the microbial diversity of periphyton and its tolerance to micropollutants in an engineered flow-through channel system. Water Research, 203, 117486 (14 pp.). doi:10.1016/j.watres.2021.117486, Institutional Repository

Wastewater treatment plants (WWTPs) play an important role in retaining organic matter and nutrients but to a lesser extent micropollutants. Therefore, treated wastewater is recognized as a major source of multiple stressors, including complex mixtures of micropollutants. These can potentially affect microbial communities in the receiving water bodies and the ecological functions they provide. In this study, we evaluated in flow-through channels the consequences of an exposure to a mixture of stream water and different percentages of urban WWTP effluent, ranging from 0% to 80%, on the microbial diversity and function of periphyton communities. Assuming that micropollutants exert a selective pressure for tolerant microorganisms within communities, we further examined the periphyton sensitivity to a micropollutant mixture extracted from passive samplers that were immersed in the wastewater effluent. As well, micropollutants in water and in periphyton were comprehensively quantified. Our results show that micropollutants detected in periphyton differed from those found in water, both in term of concentration and composition. Especially photosystem II inhibitors accumulated in periphyton more than other pesticides. Although effects of other substances cannot be excluded, this accumulation may have contributed to the observed higher tolerance of phototrophic communities to micropollutants upon exposure to 30% and 80% of wastewater. On the contrary, no difference in tolerance was observed for heterotrophic communities. Exposure to the gradient of wastewater led to structural differences in both prokaryotic and eukaryotic communities. For instance, the relative abundance of cyanobacteria was higher with increasing percentage of wastewater effluent, whereas the opposite was observed for diatoms. Such results could indicate that differences in community structure do not necessarily lead to higher tolerance. This highlights the need to consider other wastewater constituents such as nutrients and wastewater-derived microorganisms that can modulate community structure and tolerance. By using engineered flow-through channels that mimic to some extent the required field conditions for the development of tolerance in periphyton, our study constitutes a base to investigate the mechanisms underlying the increased tolerance, such as the potential role of microorganisms originating from wastewater effluents, and different treatment options to reduce the micropollutant load in effluents.
Lyautey, E., Bonnineau, C., Billard, P., Loizeau, J. L., Naffrechoux, E., Tlili, A., … Pesce, S. (2021). Diversity, functions and antibiotic resistance of sediment microbial communities from Lake Geneva are driven by the spatial distribution of anthropogenic contamination. Frontiers in Microbiology, 12, 738629 (15 pp.). doi:10.3389/fmicb.2021.738629, Institutional Repository

Lake sediments are natural receptors for a wide range of anthropogenic contaminants including organic matter and toxicants such as trace metals, polycyclic aromatic hydrocarbons, polychlorinated biphenyls that accumulate over time. This contamination can impact benthic communities, including microorganisms which play a crucial role in biogeochemical cycling and food-webs. The present survey aimed at exploring whether anthropogenic contamination, at a large lake scale, can influence the diversity, structure and functions of microbial communities associated to surface sediment, as well as their genetic potential for resistance to metals and antibiotics. Changes in the characteristics of these communities were assessed in surface sediments collected in Lake Geneva from eight sampling sites in October 2017 and May 2018. These sampling sites were characterized by a large concentration range of metal and organic compound contamination. Variation between the two sampling periods were very limited for all sampling sites and measured microbial parameters. In contrast, spatial variations were observed, with two sites being distinct from each other, and from the other six sites. Benthic communities from the most contaminated sampling site (Vidy Bay, near the city of Lausanne) were characterized by the lowest bacterial and archaeal diversity, a distinct community composition, the highest abundance of antibiotic resistance genes and functional (respiration, denitrification, methanogenesis, phosphatase, and beta-glucosidase) activity levels. The second sampling site which is highly influenced by inputs from the Rhône River, exhibited low levels of diversity, a distinct community composition, high abundance of antibiotic resistance genes and the highest bacterial abundance. Overall, our results suggest that local anthropogenic contamination, including organic matter and toxicants, is a major driver of the diversity and functioning of sediment-microbial communities in Lake Geneva. This highlights the need to consider benthic microbial communities and a suite of complementary ecotoxicological endpoints for more effective environmental risk assessments of contaminants in lake sediments.
Tlili, A. (2021). Effects of engineered nanoparticles on plant litter decomposition in streams. In C. M. Swan, L. Boyero, & C. Canhoto (Eds.), The ecology of plant litter decomposition in stream ecosystems (pp. 411-430). doi:10.1007/978-3-030-72854-0_18, Institutional Repository

Engineered nanoparticles (ENPs) are the basis of novel and most promising technologies of the twenty-first century, and their production and use in commercial products is exponentially growing. Consequently, freshwater ecosystems will increasingly be recipients for these nanomaterials, which comes at a potential environmental risk when sensitive organisms assume important functions. This chapter describes the current knowledge on ENP effects on plant litter decomposition, an important process in streams. The main focus is on microbial decomposer communities, with both their structure and functions being affected upon ENP exposure. The consequences on detritus-based food-webs are also discussed in the light of evidence from published literature and a case study, showing that both waterborne and dietary exposures to ENPs are important pathways. Particular attention is given to the multifaceted nature and complexity of the environmental behaviour of ENPs and the significant risks they pose to freshwater ecosystems. Finally, several knowledge gaps are highlighted throughout the chapter and suggestions for future research directions are provided.
Batista, D., Tlili, A., Gessner, M. O., Pascoal, C., & Cássio, F. (2020). Nanosilver impacts on aquatic microbial decomposers and litter decomposition assessed as pollution-induced community tolerance (PICT). Environmental Science: Nano, 7(7), 2130-2139. doi:10.1039/D0EN00375A, Institutional Repository

The growing proliferation of silver nanoparticles (AgNPs) calls for detailed information on ecotoxicological effects, particularly on diverse communities and key ecosystem processes where impacts remain poorly known. This includes the decomposition of plant litter by fungi and bacteria in streams. Impacts are likely to depend on community composition, because species vary in their sensitivities to stressors. Therefore, our goal was to determine if shifts in microbial communities triggered by chronic exposure to low concentrations of nano (<200 μg L-1) and ionic (20 μg L-1) silver increase community tolerance to these contaminants, as described in the pollution-induced community tolerance (PICT) concept. We used stream microbial decomposers associated with leaf litter in microcosms to assess the applicability of this concept by determining tolerance acquisition towards AgNP and ionic Ag in short-term inhibition assays. Endpoints included fungal sporulation, bacterial production, microbial respiration and the potential activity of a protein-degrading enzyme, leucine aminopeptidase. Analyses of microbial communities showed that chronic exposure to the highest AgNP concentrations led to similar communities, and that these were distinct from the control communities. Most important, chronic exposure of fungi and bacteria to both AgNP and ionic Ag also increased tolerance of the microbes, as revealed by notably reduced adverse effects on bacterial production. Overall, our results demonstrate the usefulness of applying the PICT concept to litter decomposers and decomposition as an approach to assess the risks posed by nano and ionic silver to freshwater ecosystems.
Creusot, N., Casado-Martinez, C., Chiaia-Hernandez, A., Kiefer, K., Ferrari, B. J. D., Fu, Q., … Hollender, J. (2020). Retrospective screening of high-resolution mass spectrometry archived digital samples can improve environmental risk assessment of emerging contaminants: a case study on antifungal azoles. Environment International, 139, 105708 (10 pp.). doi:10.1016/j.envint.2020.105708, Institutional Repository

Environmental risk assessment associated with aquatic and terrestrial contamination is mostly based on predicted or measured environmental concentrations of a limited list of chemicals in a restricted number of environmental compartments. High resolution mass spectrometry (HRMS) can provide a more comprehensive picture of exposure to harmful chemicals, particularly through the retrospective analysis of digitally stored HRMS data. Using this methodology, our study characterized the contamination of various environmental compartments including 154 surface water, 46 urban effluent, 67 sediment, 15 soil, 34 groundwater, 24 biofilm, 41 gammarid and 49 fish samples at 95 sites widely distributed over the Swiss Plateau. As a proof-of-concept, we focused our investigation on antifungal azoles, a class of chemicals of emerging concern due to their endocrine disrupting effects on aquatic organisms and humans. Our results demonstrated the occurrence of antifungal azoles and some of their (bio)transformation products in all the analyzed compartments (0.1-100 ng/L or ng/g d.w.). Comparison of actual and predicted concentrations showed the partial suitability of level 1 fugacity modelling in predicting the exposure to azoles. Risk quotient calculations additionally revealed risk of exposure especially if some of the investigated rivers and streams are used for drinking water production. The case study clearly shows that the retrospective analysis of HRMS/MS data can improve the current knowledge on exposure and the related risks to chemicals of emerging concern and can be effectively employed in the future for such purposes.
Tlili, A., Corcoll, N., Arrhenius, Å., Backhaus, T., Hollender, J., Creusot, N., … Behra, R. (2020). Tolerance patterns in stream biofilms link complex chemical pollution to ecological impacts. Environmental Science and Technology, 54(17), 10745-10753. doi:10.1021/acs.est.0c02975, Institutional Repository

Preventing and remedying fresh waters from chemical pollution is a fundamental societal and scientific challenge. With other nonchemical stressors potentially co-occurring, assessing the ecological consequences of reducing chemical loads in the environment is arduous. In this case study, we comparatively assessed the community structure, functions, and tolerance of stream biofilms to micropollutant mixtures extracted from deployed passive samplers at wastewater treatment plant effluents. These biofilms were growing up- and downstream of one upgraded and two nonupgraded wastewater treatment plants before being sampled for analyses. Our results showed a substantial decrease in micropollutant concentrations by 85%, as the result of upgrading the wastewater treatment plant at one of the sampling sites with activated carbon filtration. This decrease was positively correlated with a loss of community tolerance to micropollutants and the recovery of the community structure downstream of the effluent. On the other hand, downstream biofilms at the nonupgraded sites displayed higher tolerance to the extracts than the upstream biofilms. The observed higher tolerance was positively linked to micropollutant levels both in stream water and in biofilm samples, and to shifts in the community structure. Although more investigations of upgraded sites are needed, our findings point toward the suitability of using community tolerance for the retrospective assessment of the risks posed by micropollutants, to assess community recovery, and to relate effects to causes in complex environmental conditions.
Gil-Allué, C., Tlili, A., Schirmer, K., Gessner, M. O., & Behra, R. (2018). Long-term exposure to silver nanoparticles affects periphyton community structure and function. Environmental Science: Nano, 5(6), 1397-1407. doi:10.1039/C8EN00132D, Institutional Repository

Silver nanoparticles (AgNPs) are widely used as antimicrobial agents. During their life cycle, some of the AgNPs are released into natural environments, where chronic exposure to them can continue to cause harmful effects on microorganisms. However, very little is known about long-term impacts on important ecosystem compartments such as periphyton, a microbial community of algae and bacteria that covers submerged surfaces and contributes importantly to primary production and other ecosystem processes. Thus, the present study focused on assessing the accumulation of citrate-coated AgNPs and dissolved Ag+ in periphyton and on testing chronic effects on periphyton community structure and a range of functional endpoints. Stream periphyton grown in microcosms was exposed to 0.1, 1 and 10 μM AgNPs and 0.1 μM AgNO3 (a source of Ag+ ions) for up to 21 days. By that time, 84 to 98% of the total available silver in the microcosms was strongly associated with the periphyton. The strongest and broadest impacts on functional endpoints were observed at the highest AgNP concentration (10 μM), which caused a decline in algal primary production and microbial respiration but a simultaneous increase in bacterial secondary production and total biomass accrual. The community structure of both phototrophs and heterotrophs was also changed. Overall, our results reveal that periphyton strongly accumulates AgNPs, leading to a shift of community metabolism towards heterotrophy, with possible consequences for trophic transfer in aquatic food webs exposed to Ag contamination.
Jabiol, J., Cornut, J., Tlili, A., & Gessner, M. O. (2018). Interactive effects of dissolved nitrogen, phosphorus and litter chemistry on stream fungal decomposers. FEMS Microbiology Ecology, 94(10), fiy151 (11 pp.). doi:10.1093/femsec/fiy151, Institutional Repository

The enrichment of ecosystems by nutrients such as nitrogen (N) and phosphorus (P) has important ecological consequences. These include effects on plant litter decomposition in forest soils and forested headwater streams, where fungi play a pivotal role. However, our understanding of nutrient relationships on fungal communities associated with decomposing litter remains surprisingly incomplete. We conducted a fully factorial microcosm experiment with known communities of fungal decomposers from streams to assess the importance of dissolved N and P supply, as well as the atomic nutrient ratio (N:P), on fungal community succession, diversity, biomass and reproduction on three leaf-litter species differing in nutrient and lignin concentrations. Fungal biomass accrual and spore production were strongly controlled by external N supply, whereas P supply was much less important. The magnitude of these effects was mediated by litter quality, with stronger effects of dissolved N and P on lignin-poor and high N:P litter. N supply also influenced fungal diversity and species composition, acting as a pacemaker of community succession. Collectively, our data indicate that N was in much greater demand than predicted by standard stoichiometric models. The most parsimonious explanation for this deviation relates to the need of litter fungi to invest large amounts of N into degradative exoenzymes.
Laux, P., Riebeling, C., Booth, A. M., Brain, J. D., Brunner, J., Cerrillo, C., … Luch, A. (2018). Challenges in characterizing the environmental fate and effects of carbon nanotubes and inorganic nanomaterials in aquatic systems. Environmental Science: Nano, 5(1), 48-63. doi:10.1039/c7en00594f, Institutional Repository

The current lack of commonly used protocols for dispersion, characterization, and aquatic toxicity testing of nanomaterials (NMs) has resulted in inconsistent results, which make meaningful comparisons difficult. The need for standardized sample preparation procedures that allow the reproducible generation of relevant test conditions remains a key challenge for studies of the environmental fate and aquatic toxicity of NMs. Together with the further development of optimized and cost-effective analytical techniques for physicochemical characterization that depend on reproducible sample preparation, such methods have the potential to overcome the current uncertainties with regard to NM dispersion properties, effective dose, and particle dissolution. In this review, recent data available on the challenges are summarized, especially those associated with preparing and quantifying NM dispersions, determining NM uptake and accumulation in aquatic organisms, and the transformation of organic and inorganic NM in aquatic species. Additional limitations and challenges that are specific to certain types of NMs are highlighted. The release of highly persistent carbon nanotubes (CNTs) from nanocomposites is determined to be a potential source of environmental contamination. Furthermore, the role of NM dissolution and the contribution of ions versus particles to NM toxicity are discussed. A phenomenon of particular relevance for the environment is photoactivation of NMs. This is elucidated with regard to its consequences in complex aquatic ecosystems. Widespread implementation of standardized protocols alongside the consideration of phenomena associated with different life cycle stages of industrial products is crucial to the future establishment of NM environmental risk assessment.
Sgier, L., Merbt, S. N., Tlili, A., Kroll, A., & Zupanic, A. (2018). Characterization of aquatic biofilms with flow cytometry. Journal of Visualized Experiments (136), e57655 (9 pp.). doi:10.3791/57655, Institutional Repository

Biofilms are dynamic consortia of microorganism that play a key role in freshwater ecosystems. By changing their community structure, biofilms respond quickly to environmental changes and can be thus used as indicators of water quality. Currently, biofilm assessment is mostly based on integrative and functional endpoints, such as photosynthetic or respiratory activity, which do not provide information on the biofilm community structure. Flow cytometry and computational visualization offer an alternative, sensitive, and easy-to-use method for assessment of the community composition, particularly of the photoautotrophic part of freshwater biofilms. It requires only basic sample preparation, after which the entire sample is run through the flow cytometer. The single-cell optical and fluorescent information is used for computational visualization and biological interpretation. Its main advantages over other methods are the speed of analysis and the high-information-content nature. Flow cytometry provides information on several cellular and biofilm traits in a single measurement: particle size, density, pigment content, abiotic content in the biofilm, and coarse taxonomic information. However, it does not provide information on biofilm composition on the species level. We see high potential in the use of the method for environmental monitoring of aquatic ecosystems and as an initial biofilm evaluation step that informs downstream detailed investigations by complementary and more detailed methods.
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      authors => protected'Tlili,&nbsp;A.; Jabiol,&nbsp;J.; Behra,&nbsp;R.; Gil-Allue,&nbsp;C.; Gessner
         ,&nbsp;M.&nbsp;O.' (93 chars)
      title => protected'Chronic exposure effects of silver nanoparticles on stream microbial decompo
         ser communities and ecosystem functions' (115 chars)
      journal => protected'Environmental Science and Technology' (36 chars)
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      description => protected'With the accelerated use of silver nanoparticles (AgNP) in commercial produc
         ts, streams will increasingly serve as recipients of, and repositories for, 
         AgNP. This raises concerns about the potential toxicity of these nanomateria
         ls in the environment. Here we aimed to assess the impacts of chronic AgNP e
         xposure on the metabolic activities and community structure of fungal and ba
         cterial plant litter decomposers as central players in stream ecosystems. Mi
         nimal variation in the size and surface charge of AgNP indicated that nanopa
         rticles were rather stable during the experiment. Five days of exposure to 0
         .05 and 0.5 μM AgNP in microcosms shifted bacterial community structure but
          had no effect on a suite of microbial metabolic activities, despite silver 
         accumulation in the decomposing leaf litter. After 25 days, however, a broad
          range of microbial endpoints, as well as rates of litter decomposition, wer
         e strongly affected. Declines matched with the total silver concentration in
          the leaves and were accompanied by changes in fungal and bacterial communit
         y structure. These results highlight a distinct sensitivity of litter-associ
         ated microbial communities in streams to chronic AgNP exposure, with effects
          on both microbial functions and community structure resulting in notable ec
         osystem consequences through impacts on litter decomposition and further bio
         geochemical processes.' (1390 chars)
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      authors => protected'Tlili,&nbsp;A.; Hollender,&nbsp;J.; Kienle,&nbsp;C.; Behra,&nbsp;R.' (67 chars)
      title => protected'Micropollutant-induced tolerance of <I>in situ</I> periphyton: establishing 
         causality in wastewater-impacted streams' (116 chars)
      journal => protected'Water Research' (14 chars)
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      categories => protected'pollution-induced community tolerance; passive samplers; wastewater treatmen
         t plants; causality; micropollutant mixture; biofilm' (128 chars)
      description => protected'The overarching aim of this field study was to examine causal links between 
         <I>in-situ</I> exposure to complex mixtures of micropollutants from wastewat
         er treatment plants and effects on freshwater microbial communities in the r
         eceiving streams. To reach this goal, we assessed the toxicity of serial dil
         utions of micropollutant mixtures, extracted from deployed passive samplers 
         at the discharge sites of four Swiss wastewater treatment plants, to <I>in s
         itu</I> periphyton from upstream and downstream of the effluents. On the one
          hand, comparison of the sensitivities of upstream and downstream periphyton
          to the micropollutant mixtures indicated that algal and bacterial communiti
         es composing the periphyton displayed higher tolerance towards these micropo
         llutants downstream than upstream. On the other hand, molecular analyses of 
         the algal and bacterial structure showed a clear separation between upstream
          and downstream periphyton across the sites. This finding provides an additi
         onal line of evidence that micropollutants from the wastewater discharges we
         re directly responsible for the change in the community structure at the sam
         pling sites by eliminating the micropollutant-sensitive species and favourin
         g the tolerant ones. What is more, the fold increase of algal and bacterial 
         tolerance from upstream to downstream locations was variable among sampling 
         sites and was strongly correlated to the intensity of contamination by micro
         pollutants at the respective sites. Overall, our study highlights the sensit
         ivity of the proposed approach to disentangle effects of micropollutant mixt
         ures from other environmental factors occurring in the field and, thus, esta
         blishing a causal link between exposure and the observed ecological effects 
         on freshwater microbial communities.' (1784 chars)
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      authors => protected'Stamm,&nbsp;C.; Burdon,&nbsp;F.; Fischer,&nbsp;S.; Kienle,&nbsp;C.; Munz,&nb
         sp;N.; Tlili,&nbsp;A.; Altermatt,&nbsp;F.; Behra,&nbsp;R.; Bürgmann,&nbsp;H
         .; Joss,&nbsp;A.; Räsänen,&nbsp;K.; Eggen,&nbsp;R.' (204 chars)
      title => protected'Einfluss von Mikroverunreinigungen' (34 chars)
      journal => protected'Aqua & Gas' (10 chars)
      year => protected2017 (integer)
      volume => protected97 (integer)
      issue => protected'6' (1 chars)
      startpage => protected'90' (2 chars)
      otherpage => protected'95' (2 chars)
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      description => protected'Im Projekt EcoImpact wurden die ökotoxikologischen und ökologischen Effekt
         e von Mikroverunreinigungen aus Kläranlagen auf aquatische Lebensgemeinscha
         ften untersucht. Chemische und biologische Untersuchungen oberhalb und unter
         halb von Kläranlagen deuten auf Effekte dieser Stoffe hin, die von physiolo
         gischen Antworten der Organismen bis hin zu veränderten Ökosystemfunktione
         n wie z. B. dem Laubabbau reichen. Gezielte Rinnenexperimente mit kontrollie
         rter Wasserqualität unterstützen diese Befunde.' (505 chars)
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      originalId => protected10538 (integer)
      authors => protected'Tlili,&nbsp;A.; Cornut,&nbsp;J.; Behra,&nbsp;R.; Gil-Allué,&nbsp;C.; Gessne
         r,&nbsp;M.&nbsp;O.' (94 chars)
      title => protected'Harmful effects of silver nanoparticles on a complex detrital model system' (74 chars)
      journal => protected'Nanotoxicology' (14 chars)
      year => protected2016 (integer)
      volume => protected10 (integer)
      issue => protected'6' (1 chars)
      startpage => protected'728' (3 chars)
      otherpage => protected'735' (3 chars)
      categories => protected'bacterial and fungal growth; community nanotoxicology; extracellular enzymat
         ic activities; litter decomposition; stream ecosystem' (129 chars)
      description => protected'The rapid proliferation of silver nanoparticles (AgNP) in industry and the e
         nvironment requires realistic toxicity assessments based on approaches that 
         consider the biological complexity of ecosystems. Here we assessed the acute
          toxicity of carbonate-coated AgNP and, for comparison, AgNO<SUB>3</SUB> (Ag
         <SUP>+</SUP>) by using a model system consisting of decomposing plant litter
          and the associated fungal and bacterial decomposers as central players in t
         he functioning of stream ecosystems. Little variation in size and surface ch
         arge during the experiment indicated that the AgNP used were essentially sta
         ble. AgNP disrupted bacterial growth (≤83% reduction in protein biosynthes
         
         
          endpoints more sensitive to AgNP than to Ag<SUP>+</SUP>. Fungal reproductio
         n, in contrast, was stimulated by AgNP, but not Ag<SUP>+</SUP>, at concentra
         tions up to 25 μM. Both AgNP and Ag<SUP>+</SUP> also stimulated extracellul
         ar alkaline phosphatase but reduced leucine aminopeptidase, whereas β-gluco
         sidase was stimulated by AgNP and reduced by Ag<SUP>+</SUP>. Importantly, th
         e provision of cysteine, a chelating ligand that complexes free Ag<SUP>+</SU
         P>, failed to alleviate AgNP toxicity to microbial growth, clearly demonstra
         ting particle-mediated toxicity independent of the presence of ionic silver.
          This contrasts with the observed inhibition of leucine aminopeptidase by Ag
         <SUP>+</SUP>, which accounted for 2–6% of the total silver in treatments r
         eceiving AgNP. These results show that although outcomes of AgNP and Ag<SUP>
         +</SUP> exposure assessed by different functional endpoints vary widely, AgN
         P strongly interferes with bacterial growth and a range of other microbial p
         rocesses, resulting in severe consequences for natural microbial communities
          and ecosystem functioning.' (1927 chars)
      serialnumber => protected'1743-5390' (9 chars)
      doi => protected'10.3109/17435390.2015.1117673' (29 chars)
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      originalId => protected8024 (integer)
      authors => protected'Gil-Allué,&nbsp;C.; Schirmer,&nbsp;K.; Tlili,&nbsp;A.; Gessner,&nbsp;M.&nbs
         p;O.; Behra,&nbsp;R.' (96 chars)
      title => protected'Silver nanoparticle effects on stream periphyton during short-term exposures' (76 chars)
      journal => protected'Environmental Science and Technology' (36 chars)
      year => protected2015 (integer)
      volume => protected49 (integer)
      issue => protected'2' (1 chars)
      startpage => protected'1165' (4 chars)
      otherpage => protected'1172' (4 chars)
      categories => protected'' (0 chars)
      description => protected'Silver nanoparticles (AgNP) are increasingly used as antimicrobials in consu
         mer products. Subsequently released into aquatic environments, they are like
         ly to come in contact with microbial communities like periphyton, which play
         s a key role as a primary producer in stream ecosystems. At present, however
         , very little is known about the effects of nanoparticles on processes media
         ted by periphyton communities. We assessed the effects of citrate-coated sil
         ver nanoparticles and silver ions (dosed as AgNO<SUB>3</SUB>) on five functi
         onal end points reflecting community and ecosystem-level processes in periph
         yton: photosynthetic yield, respiration potential, and the activity of three
          extracellular enzymes. After 2 h of exposure in experimental microcosms, Ag
         NP and AgNO<SUB>3</SUB> inhibited respiration and photosynthesis of periphyt
         on and the activities of two of the three extracellular enzymes. Addition of
          a chelating ligand that complexes free silver ions indicated that, in most 
         cases, toxicity of AgNP suspensions was caused by Ag(I) dissolved from the p
         articles. However, these suspensions inhibited one of the extracellular enzy
         mes (leucine aminopeptidase), pointing to a specific nanoparticle effect ind
         ependent of the dissolved Ag(I). Thus, our results show that both silver nan
         oparticles and silver ions have potential to disrupt basic metabolic functio
         ns and enzymatic resource acquisition of stream periphyton.' (1427 chars)
      serialnumber => protected'0013-936X' (9 chars)
      doi => protected'10.1021/es5050166' (17 chars)
      uid => protected8024 (integer)
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      originalId => protected8135 (integer)
      authors => protected'Tlili,&nbsp;A.; Berard,&nbsp;A.; Blanck,&nbsp;H.; Bouchez,&nbsp;A.; Cássio,
         &nbsp;F.; Eriksson,&nbsp;K.&nbsp;M.; Morin,&nbsp;S.; Montuelle,&nbsp;B.; Nav
         arro,&nbsp;E.; Pascoal,&nbsp;C.; Pesce,&nbsp;S.; Schmitt-Jansen,&nbsp;M.; Be
         hra,&nbsp;R.' (240 chars)
      title => protected'Pollution-induced community tolerance (PICT): towards an ecologically releva
         nt risk assessment of chemicals in aquatic systems' (126 chars)
      journal => protected'Freshwater Biology' (18 chars)
      year => protected2016 (integer)
      volume => protected61 (integer)
      issue => protected'12' (2 chars)
      startpage => protected'2141' (4 chars)
      otherpage => protected'2151' (4 chars)
      categories => protected'aquatic ecology; bioindicator; chemical status; ecological status; ecotoxico
         logy' (80 chars)
      description => protected'1. A major challenge in environmental risk assessment of pollutants is estab
         lishing a causal relationship between field exposure and community effects t
         hat integrates both structural and functional complexity within ecosystems.<
         BR/>2. Pollution-induced community tolerance (PICT) is a concept that evalua
         tes whether pollutants have exerted a selection pressure on natural communit
         ies. PICT detects whether a pollutant has eliminated sensitive species from 
         a community and thereby increased its tolerance. PICT has the potential to l
         ink assessments of the ecological and chemical status of ecosystems by provi
         ding causal analysis for effect-based monitoring of impacted field sites.<BR
         />3. Using PICT measurements and microbial community endpoints in environmen
         tal assessment schemes could give more ecological relevance to the tools tha
         t are now used in environmental risk assessment. Here, we propose practical 
         guidance and a list of research issues that should be further considered to 
         apply the PICT concept in the field.' (1024 chars)
      serialnumber => protected'0046-5070' (9 chars)
      doi => protected'10.1111/fwb.12558' (17 chars)
      uid => protected8135 (integer)
      _localizedUid => protected8135 (integer)modified
      _languageUid => protectedNULL
      _versionedUid => protected8135 (integer)modified
      pid => protected124 (integer)

Chronic exposure effects of silver nanoparticles on stream microbial decomposer communities and ecosystem functions

With the accelerated use of silver nanoparticles (AgNP) in commercial products, streams will increasingly serve as recipients of, and repositories for, AgNP. This raises concerns about the potential toxicity of these nanomaterials in the environment. Here we aimed to assess the impacts of chronic AgNP exposure on the metabolic activities and community structure of fungal and bacterial plant litter decomposers as central players in stream ecosystems. Minimal variation in the size and surface charge of AgNP indicated that nanoparticles were rather stable during the experiment. Five days of exposure to 0.05 and 0.5 μM AgNP in microcosms shifted bacterial community structure but had no effect on a suite of microbial metabolic activities, despite silver accumulation in the decomposing leaf litter. After 25 days, however, a broad range of microbial endpoints, as well as rates of litter decomposition, were strongly affected. Declines matched with the total silver concentration in the leaves and were accompanied by changes in fungal and bacterial community structure. These results highlight a distinct sensitivity of litter-associated microbial communities in streams to chronic AgNP exposure, with effects on both microbial functions and community structure resulting in notable ecosystem consequences through impacts on litter decomposition and further biogeochemical processes.
Tlili, A.; Jabiol, J.; Behra, R.; Gil-Allue, C.; Gessner, M. O. (2017) Chronic exposure effects of silver nanoparticles on stream microbial decomposer communities and ecosystem functions, Environmental Science and Technology, 51(4), 2447-2455, doi:10.1021/acs.est.6b05508, Institutional Repository

Micropollutant-induced tolerance of in situ periphyton: establishing causality in wastewater-impacted streams

The overarching aim of this field study was to examine causal links between in-situ exposure to complex mixtures of micropollutants from wastewater treatment plants and effects on freshwater microbial communities in the receiving streams. To reach this goal, we assessed the toxicity of serial dilutions of micropollutant mixtures, extracted from deployed passive samplers at the discharge sites of four Swiss wastewater treatment plants, to in situ periphyton from upstream and downstream of the effluents. On the one hand, comparison of the sensitivities of upstream and downstream periphyton to the micropollutant mixtures indicated that algal and bacterial communities composing the periphyton displayed higher tolerance towards these micropollutants downstream than upstream. On the other hand, molecular analyses of the algal and bacterial structure showed a clear separation between upstream and downstream periphyton across the sites. This finding provides an additional line of evidence that micropollutants from the wastewater discharges were directly responsible for the change in the community structure at the sampling sites by eliminating the micropollutant-sensitive species and favouring the tolerant ones. What is more, the fold increase of algal and bacterial tolerance from upstream to downstream locations was variable among sampling sites and was strongly correlated to the intensity of contamination by micropollutants at the respective sites. Overall, our study highlights the sensitivity of the proposed approach to disentangle effects of micropollutant mixtures from other environmental factors occurring in the field and, thus, establishing a causal link between exposure and the observed ecological effects on freshwater microbial communities.
Tlili, A.; Hollender, J.; Kienle, C.; Behra, R. (2017) Micropollutant-induced tolerance of in situ periphyton: establishing causality in wastewater-impacted streams, Water Research, 111, 185-194, doi:10.1016/j.watres.2017.01.016, Institutional Repository

Einfluss von Mikroverunreinigungen

Im Projekt EcoImpact wurden die ökotoxikologischen und ökologischen Effekte von Mikroverunreinigungen aus Kläranlagen auf aquatische Lebensgemeinschaften untersucht. Chemische und biologische Untersuchungen oberhalb und unterhalb von Kläranlagen deuten auf Effekte dieser Stoffe hin, die von physiologischen Antworten der Organismen bis hin zu veränderten Ökosystemfunktionen wie z. B. dem Laubabbau reichen. Gezielte Rinnenexperimente mit kontrollierter Wasserqualität unterstützen diese Befunde.
Stamm, C.; Burdon, F.; Fischer, S.; Kienle, C.; Munz, N.; Tlili, A.; Altermatt, F.; Behra, R.; Bürgmann, H.; Joss, A.; Räsänen, K.; Eggen, R. (2017) Einfluss von Mikroverunreinigungen, Aqua & Gas, 97(6), 90-95, Institutional Repository

Harmful effects of silver nanoparticles on a complex detrital model system

The rapid proliferation of silver nanoparticles (AgNP) in industry and the environment requires realistic toxicity assessments based on approaches that consider the biological complexity of ecosystems. Here we assessed the acute toxicity of carbonate-coated AgNP and, for comparison, AgNO3 (Ag+) by using a model system consisting of decomposing plant litter and the associated fungal and bacterial decomposers as central players in the functioning of stream ecosystems. Little variation in size and surface charge during the experiment indicated that the AgNP used were essentially stable. AgNP disrupted bacterial growth (≤83% reduction in protein biosynthesis, EC50 = 0.3 μM), clearly affected fungal growth (≤61% reduction in ergosterol synthesis, EC50 = 47 μM) with both endpoints more sensitive to AgNP than to Ag+. Fungal reproduction, in contrast, was stimulated by AgNP, but not Ag+, at concentrations up to 25 μM. Both AgNP and Ag+ also stimulated extracellular alkaline phosphatase but reduced leucine aminopeptidase, whereas β-glucosidase was stimulated by AgNP and reduced by Ag+. Importantly, the provision of cysteine, a chelating ligand that complexes free Ag+, failed to alleviate AgNP toxicity to microbial growth, clearly demonstrating particle-mediated toxicity independent of the presence of ionic silver. This contrasts with the observed inhibition of leucine aminopeptidase by Ag+, which accounted for 2–6% of the total silver in treatments receiving AgNP. These results show that although outcomes of AgNP and Ag+ exposure assessed by different functional endpoints vary widely, AgNP strongly interferes with bacterial growth and a range of other microbial processes, resulting in severe consequences for natural microbial communities and ecosystem functioning.
Tlili, A.; Cornut, J.; Behra, R.; Gil-Allué, C.; Gessner, M. O. (2016) Harmful effects of silver nanoparticles on a complex detrital model system, Nanotoxicology, 10(6), 728-735, doi:10.3109/17435390.2015.1117673, Institutional Repository

Silver nanoparticle effects on stream periphyton during short-term exposures

Silver nanoparticles (AgNP) are increasingly used as antimicrobials in consumer products. Subsequently released into aquatic environments, they are likely to come in contact with microbial communities like periphyton, which plays a key role as a primary producer in stream ecosystems. At present, however, very little is known about the effects of nanoparticles on processes mediated by periphyton communities. We assessed the effects of citrate-coated silver nanoparticles and silver ions (dosed as AgNO3) on five functional end points reflecting community and ecosystem-level processes in periphyton: photosynthetic yield, respiration potential, and the activity of three extracellular enzymes. After 2 h of exposure in experimental microcosms, AgNP and AgNO3 inhibited respiration and photosynthesis of periphyton and the activities of two of the three extracellular enzymes. Addition of a chelating ligand that complexes free silver ions indicated that, in most cases, toxicity of AgNP suspensions was caused by Ag(I) dissolved from the particles. However, these suspensions inhibited one of the extracellular enzymes (leucine aminopeptidase), pointing to a specific nanoparticle effect independent of the dissolved Ag(I). Thus, our results show that both silver nanoparticles and silver ions have potential to disrupt basic metabolic functions and enzymatic resource acquisition of stream periphyton.
Gil-Allué, C.; Schirmer, K.; Tlili, A.; Gessner, M. O.; Behra, R. (2015) Silver nanoparticle effects on stream periphyton during short-term exposures, Environmental Science and Technology, 49(2), 1165-1172, doi:10.1021/es5050166, Institutional Repository

Pollution-induced community tolerance (PICT): towards an ecologically relevant risk assessment of chemicals in aquatic systems

1. A major challenge in environmental risk assessment of pollutants is establishing a causal relationship between field exposure and community effects that integrates both structural and functional complexity within ecosystems.
2. Pollution-induced community tolerance (PICT) is a concept that evaluates whether pollutants have exerted a selection pressure on natural communities. PICT detects whether a pollutant has eliminated sensitive species from a community and thereby increased its tolerance. PICT has the potential to link assessments of the ecological and chemical status of ecosystems by providing causal analysis for effect-based monitoring of impacted field sites.
3. Using PICT measurements and microbial community endpoints in environmental assessment schemes could give more ecological relevance to the tools that are now used in environmental risk assessment. Here, we propose practical guidance and a list of research issues that should be further considered to apply the PICT concept in the field.
Tlili, A.; Berard, A.; Blanck, H.; Bouchez, A.; Cássio, F.; Eriksson, K. M.; Morin, S.; Montuelle, B.; Navarro, E.; Pascoal, C.; Pesce, S.; Schmitt-Jansen, M.; Behra, R. (2016) Pollution-induced community tolerance (PICT): towards an ecologically relevant risk assessment of chemicals in aquatic systems, Freshwater Biology, 61(12), 2141-2151, doi:10.1111/fwb.12558, Institutional Repository

Contact

Prof. Dr. Kristin Schirmer Group leader and deputy head of department Tel. +41 58 765 5266 Send Mail

Team members

Sara Isabel Bastos Goncalves Postdoctoral researcher Tel. +41 58 765 5657 Send Mail
Sarah Descloux P.hD. Student Tel. +41 58 765 5450 Send Mail

Introduction video

"Characterization of Aquatic Biofilms with Flow Cytometry"