| 1. |
- Corcoll, Natàlia, 1981, et al.
(författare)
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Comparison of different DNA extraction methods for marine periphyton and its implications for ecotoxico-metagenomics
- 2015
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Ingår i: Poster Presentation at the SETAC Europe 25th Annual Meeting, Barcelona, Spain.
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Konferensbidrag (övrigt vetenskapligt/konstnärligt)abstract
- This is a time with unprecedented capacity of DNA sequencing technologies for metagenomic analysis (identifying new genes and species living in environmental samples) in natural communities. However, sequencing efforts on non-model organisms or communities, such as marine periphyton communities, are still scarce in ecotoxicology. Also, uncertainties in basic aspects, such as choice of appropriate DNA extraction method, are still not resolved. This study aims to compare the effects of different DNA extraction methods for marine periphyton in terms of DNA yield, purity and integrity, as well as eukaryotic and prokaryotic diversity. Cost-effectiveness among the methods was also assessed. Four different methods (the Plant DNAzol reagent (Invitrogen), the Fast DNA Spin kit for soil (MP Biomedicals), the PowerPlant and the PowerBiofilm DNA Isolation Kits (MoBio Laboratories)) commonly used for DNA extraction of eukaryotic and prokaryotic microorganisms from marine periphyton were compared. Results showed that the commercial method Plant DNAzol reagent and the Fast DNA Spin kit for soil appear to be the best to extract high quantities of DNA (> 50 ng/μL). DNA fragments obtained with these methods cover a wide range of sizes, from 250 to 15000 bp. With the PowerPlant and the PowerBiofilm DNA Isolation Kits, lower amounts of DNA were obtained (< 23 ng/μL), but the purity of these extractions, in terms of protein and polysaccharide contamination, was higher. Extractions from these latter kits also produced DNA size distributions with larger sizes, between 3000 and 15000 bp. Obtained DNA products were used for amplicon sequencing of the 18S rRNA and 16S rRNA genes, targeting eukaryotes and prokaryotes, respectively. Such DNA amplicon sequencing will be a powerful tool to compare community composition and biodiversity across samples, and to select DNA extraction method for further metagenomics of periphyton communities exposed to toxicants.
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| 2. |
- Corcoll, Natàlia, 1981, et al.
(författare)
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Comparison of four DNA extraction methods for comprehensive assessment of 16S rRNA bacterial diversity in marine biofilms using high-throughput sequencing
- 2017
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Ingår i: FEMS Microbiology Letters. - : Oxford University Press (OUP). - 1574-6968 .- 0378-1097. ; 364:14
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Tidskriftsartikel (refereegranskat)abstract
- High-throughput DNA sequencing technologies are increasingly used for the metagenomic characterization of microbial biodiversity. However, basic issues, such as the choice of an appropriate DNA extraction method, are still not resolved for non-model microbial communities. This study evaluates four commonly used DNA extraction methods for marine periphyton biofilms in terms of DNA yield, efficiency, purity, integrity and resulting 16S rRNA bacterial diversity. Among the tested methods, the Plant DNAzol® Reagent (PlantDNAzol) and the Fast DNATM SPIN Kit for Soil (FastDNA Soil) methods were best suited to extract high quantities of DNA (77 - 130 μg g wet wt-1). Lower amounts of DNA were obtained (
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| 3. |
- Corcoll, Natàlia, 1981, et al.
(författare)
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Copper affects composition and functioning of microbial communities in marine biofilms at environmentally relevant concentrations
- 2019
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Ingår i: Frontiers in Microbiology. - : Frontiers Media SA. - 1664-302X. ; 9
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Tidskriftsartikel (refereegranskat)abstract
- Copper (Cu) pollution in coastal areas is a worldwide threat for aquatic communities. This study aims to demonstrate the usefulness of the DNA metabarcoding analysis in order to describe the ecotoxicological effect of Cu at environmental concentrations on marine periphyton. Additionally, the study investigates if Cu-induced changes in community structure co-occurs with changes in community functioning (i.e. photosynthesis and community tolerance to Cu). Periphyton was exposed for 18 days to five Cu concentrations, between 0.01 and 10 μM, in a semi-static test. Diversity and community structure of prokaryotic and eukaryotic organisms were assessed by 16S and 18S amplicon sequencing, respectively. Community function was studied as impacts on algal biomass and photosynthetic activity. Additionally, we studied Pollution-Induced Community Tolerance (PICT) using photosynthesis as the endpoint. Sequencing results detected an average of 9504 and 1242 OTUs for 16S and 18S, respectively, reflecting the high biodiversity of marine periphytic biofilms. Eukaryotes represent the most Cu-sensitive kingdom, where effects were seen already at concentrations as low as 0.01 µM. The structure of the prokaryotic part of the community was impacted at slightly higher concentrations (0.06 µM), which is still in the range of the Cu concentrations observed in the area (0.08 µM). The current environmental quality standard for Cu of 0.07 µM therefore does not seem to be sufficiently protective for periphyton. Cu exposure resulted in a more Cu-tolerant community, which was accompanied by a reduced total algal biomass, increased relative abundance of diatoms and a reduction of photosynthetic activity. Cu exposure changed the network of associations between taxa in the communities. A total of 23 taxa, including taxa within Proteobacteria, Bacteroidetes, Stramenopiles and Hacrobia, were identified as being particularly sensitive to Cu. DNA metabarcoding is presented as a sensitive tool for community-level ecotoxicological studies that allows to observe impacts simultaneously on a multitude of pro- and eukaryotic taxa, and therefore to identify particularly sensitive, non-cultivable taxa.
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| 4. |
- Corcoll, Natàlia, 1981, et al.
(författare)
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Linking pesticide pollution with periphyton quality in agricultural streams: a fatty-acids approach
- 2018
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Ingår i: SETAC Europe 28th Annual Meeting 13−17 May 2018 | Rome, Italy. Abstract Book.. - 2309-8031.
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Konferensbidrag (refereegranskat)abstract
- Although the environmental risks associated with pesticide pollution in agricultural streams are quite well documented, little is know about its potential effects on periphyton quality. Periphyton provides many of the essential polyunsaturated fatty acids (PUFA) that are needed for organisms at higher trophic levels in river food webs. This study aims to assess the effects of pesticide mixtures on periphyton quality in situ. Three streams (Höje å, Skivarpsån and M42) located in the agriculturally dominated region of Skåne (SE Sweden) were sampled in September and October 2016. The effects of pesticide pollution were assessed by passive field sampling coupled with laboratory ecotoxicity tests, by mixture toxicity modelling to predict which chemical stressors were potentially driving the toxicity, and by examining the fatty acid profiles, pigment content and algal diversity of periphyton communities. Results from water chemical analyses clearly showed higher levels of nutrients and pesticide pollution in Skivarpsån and M42 than in Höje å. Ecotoxicity tests using the passive sampler extracts demonstrated that the pesticide mixtures ocurring at Skivarpsån and M42 were toxic for periphyton communities from Höje å, causing an inhibition of the photosynthetic activity up to 63% and 53%, respectively. Cluster and principal component analyses based on pigments content, algal diversity and fatty acid profiles, clearly separated the periphyton from the three river sites studied. Algal biomass from periphyton of pesticide polluted streams (Skivarpsån and M42) was higher than in Höje å. The nutritive quality of the periphyton differed among streams, and fatty acids considered high-quality such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were also more abundant in pesticide polluted streams (Skivarpsån and M42). Overal, even though results from the lab show that the mixture of pesticides pollution in the studied streams might be toxic for periphyton (i.e. inhibiting the photosynthetic activity), being herbicides the driving chemical stressors. Results from the field, indicate that when the levels of pesticide pollution are low and co-occur with high levels of nutrients pollution, nutrients migh mask pesticides effects on periphyton quantity and quality because compensatory effects from nutrients
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| 5. |
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| 6. |
- Corcoll, Natàlia, 1981, et al.
(författare)
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Risk assessment of chemical pollution in Mediterranean rivers based on classical periphyton bioindicators and new biomarkers
- 2015
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Ingår i: Poster presentation at the SETAC Europe 25th Annual Meeting, Barcelona, Spain.
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Konferensbidrag (övrigt vetenskapligt/konstnärligt)abstract
- Environmental risk of chemical pollution in Mediterranean basins is of special concern due to high emissions faced with low dilution capacities. The aim of this study is to evaluate the risk of chemical pollution in Mediterranean rivers using periphyton responses as a model to assess chemical impact. The study also aims to compare classical periphyton indicators of water quality (e.g. diatoms diversity) with new biomarkers of toxic stress (e.g chl-a fluorescence parameters and antioxidant enzymatic activities). Nineteen sites in four Iberian rivers (Llobregat, Ebro, Júcar and Guadalquivir) were sampled in autumns 2010 and 2011. A total of 116 organic compounds were detected in surface waters (60 pharmaceuticals and hormones compounds, 11 personal care products and industrial compounds, 36 pesticides and 9 perfluorinated compounds), and 6 metals in periphyton matrix. The potential risk of this cocktail of pollutants was estimated based on the “Toxic Units (TUs)” approach for algae. Results of TUs indicate that the Llobregat and Júcar rivers were those with the highest environmental risk for algae. TUs based on metals bioaccumulation in biofilm were two orders of magnitude higher than those based on organic compounds, being Zn and Cu the metals with the highest risk. Results from diatoms community composition allowed to identify sites with high environmental risk. Communities in these sites presented an altered diatoms composition, where sensitive species to pollution were replaced by tolerant ones. Biomarkers, based on antioxidant enzymatic activities (catalase, glutathione reductase and superoxide dismutase) and chl-a fluorescence parameters (photosynthetic activity and non-photochemical quenching), tended to show a less consistent pattern. Overall, results indicate that in all the Mediterranean basins studied there are many sites with an environmental risk from chemical pollution, and that the current use of periphyton biondicators and biomarkers allow to identify sites presenting high levels of pollution, however, new tools are required to evaluate “in situ” situations of moderate and low chemical pollution
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| 7. |
- Guasch, Helena, et al.
(författare)
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The Use of Biofilms to Assess the Effects of Chemicals on Freshwater Ecosystems : Biofilms in ecotoxicology
- 2016
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Ingår i: Aquatic Biofilms: Ecology, Water Quality and Wastewater Treatment .Edited by: Anna M. Romaní, Helena Guasch and M. Dolors Balaguer. - USA : Caister Academic Press. - 9781910190173 ; , s. 126-144
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Bokkapitel (refereegranskat)abstract
- Nowadays, biofilms are one of the principal targets of community ecotoxicology in aquatic ecosystems with a high potential for future use in ecotoxicology. A large set of methods derived from biofilm ecology has successfully been applied in ecotoxicology providing a diverse and comprehensive toolbox. Our ability to quantify the effects of pollution on different biofilm components, allows the direct effects of pollutants on the most sensitive community and their indirect effects on the rest of biofilm components to be evaluated. Biofilms are also a site for biotransfomation and/or transfer of chemicals to other aquatic organisms, supporting a more generalized use of biofilms in environmental chemistry. Investigations aiming to describe processes at biofilm scale, like nutrient dynamics and those including simple food chains, have recently been applied, providing the opportunity of upscaling the effects of pollutants on biofilms to food webs and ecosystems. Finally, biofilm ecotoxicology should now focus on providing the theoretical background for understanding the complex set of responses of natural communities to pollution. This knowledge should also be the basis for guiding the selection of the most appropriate tools and the development of new approaches for a better detection of the impact of pollution on aquatic life.
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| 8. |
- Karlsson, Max, et al.
(författare)
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Phenotypic and genotypic adaptations in a riverine green alga (Chlorophyceae, Selenastraceae) as a response to long-term exposure to chemical stress
- 2019
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Ingår i: SETAC Helsinki - SETAC Europe 29th Annual Meeting, 26 - 30 May 2019, Helsinki, Finland.
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Konferensbidrag (övrigt vetenskapligt/konstnärligt)abstract
- Herbicide pollution is a common problem in agricultural streams due to their extensive use and their unwanted effects on non-target species inhabiting these systems. Selanastraceae is a family of green algae that are particularly dominant in freshwater, making them a key component in the base of the food chain and an important part of the oxygen production. Therefore, it is crucial from a conservation point of view to accurately monitor the ecotoxicological impact of herbicides on this group of algae, not only on a short-term scale (e.g. days or weeks), but also in a long-term perspective (e.g. months and years). In this work, phenotypic and genotypic differences are studied between different strains of a population of Kirchneriella (Chlorophyceae, Selenastraceae) isolated from a stream polluted by herbicides over a 20-year period (Skivarpsån, SE Sweden). According to mixture toxicity modelling performed on data obtained from the Swedish pesticide monitoring program, the main drivers of algal toxicity in this river are the herbicides diflufenican (carotenoid synthesis inhibitor) and isoproturon (PSII inhibitor). Three strains of the isolated algae are examined in this study: two strains from the field that have been exposed to different levels of herbicides and one laboratory strain that has never been exposed to herbicide pollution. The taxonomy of this algae is investigated by light microscopy and DNA barcoding analyses (chloroplast 23S rRNA gene). Phenotypic differences between the strains are examined as impacts on growth rate, cell size, photosynthetic activity (chl fluorescence measurements), pigment content and sensitivity to the specific herbicides mentioned earlier (EC50s). Genotypic differences are studied as single nucleotide polymorphisms thought to be induced through exposure to diflufenican and isoproturon on pds and psbA genes via PCR amplification, cloning and sequencing. The results obtained from this project will contribute to a better understanding of genetic and phenotypic adaptations in green algae as a response to herbicide pollution (evolutionary toxicology) and their implications for ecological functions.
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| 9. |
- Morin, Soizic, et al.
(författare)
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Cumulative Stressors Trigger Increased Vulnerability of Diatom Communities to Additional Disturbances
- 2015
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Ingår i: Microbial Ecology. - : Springer Science and Business Media LLC. - 0095-3628 .- 1432-184X. ; 70:3, s. 585-595
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Tidskriftsartikel (refereegranskat)abstract
- Chronic, non-lethal stressors occurring gradually (in space or time) can result in cumulative impacts that are more dramatic than higher intensities or occasional critical levels of any single one of these stressors. The negative effects of the chronic stressors trigger lasting impacts that may grow in intensity and become problematic over time and/or to higher trophic levels. In rivers, aquatic organisms experience this type of cumulative stress along the up- to downstream gradient in natural and anthropogenic contaminants generally observed in inhabited watersheds. Diatoms are a major component of the periphyton in rivers; their richness and diversity in natural communities are directly related to their varied ecological preferences and sensitivity to disturbance. In this study, we monitored from 2003 to 2008 the changes in the diversity of taxonomic and non-taxonomic features along a small river (Riou-Mort, South West France), at three sites: one site upstream considered as a reference for this watershed, one intermediate site with high nutrient load, and one downstream site exposed to both nutrient and metal pollution. The cumulative impacts of nutrients plus metals led to a gradual decrease in species richness and diversity, and in a potential capacity to cope with additional stresses, e.g., climate change-related ones. This is reflected by a decrease in species richness downstream, more dramatic in the hot summer of 2003 than in cooler summers. With the increasingly protective environmental regulations (e.g., Water Framework Directive in Europe), accumulation of stresses on aquatic resources are recommended to receive increasing attention, in particular considering the expected changes in climate.
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| 10. |
- Ponsati, L, et al.
(författare)
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Multiple-stressor effects on river biofilms under different hydrological conditions
- 2016
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Ingår i: Freshwater Biology. - : Wiley. - 0046-5070. ; 61:12, s. 2102-2115
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Tidskriftsartikel (refereegranskat)abstract
- .We studied the relative importance of environmental factors (river flow, nutrients, dissolved organic matter) and organic micro-pollutants [herbicides, insecticides, industrial organic compounds, personal care products, antibiotics and pharmaceuticals, (PhCs)] to the structure and function of epilithic biofilms under various hydrological conditions and land uses. .Biofilms from four river basins in the Iberian Peninsula (Llobregat, Ebro, Júcar and Guadalquivir) were analysed in summer–autumn during a wet period and base-flow conditions. The sites experienced different levels of pollution. .We used variance partitioning (Redundancy Detrended Analysis, RDA) to link environmental stressors to the structure and function of biofilms, which were assessed as diatom community composition, algal biomass, photosynthetic activity, bacterial density and extracellular phosphatase activity. .RDAs showed that industrial organic compounds, herbicides and PhC products were the pollutants most strongly associated with measures of biofilm structure and function, whereas dissolved inorganic nitrogen, dissolved organic carbon and hydrological variability were the environmental factors most strongly associated with biofilm responses. .Thicker biofilms developing during base flow were most affected by pollutants and nutrients. In contrast, thinner biofilms developing during periods of high river flow were less affected by chemicals.
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