Diatoms (an ecoregional indicator of nutrients, organic mater and micropollutants pollution)

Les diatomées sont des microalgues ubiquistes d'une diversité exceptionnelle. Cela en fait de bons indicateurs de la qualité des écosystèmes aquatiques et sont utilisées depuis plus de 50 ans. Depuis l'année 2000, la Directive Cadre Européenne sur l'Eau impose leur utilisation pour évaluer la qualité écologique des cours d'eau. Un cadre typologique doit être utilisé afin de comparer des rivières comparables entre elles, c'est-à-dire des rivières de mêmes régions bioclimatiques, coulant sur les mêmes substrats géologiques et à des altitudes comparables. Différentes classifications écorégionales ont été définies sur la base de ces paramètres. Nous avons montré qu'à une échelle couvrant 4 pays (Espagne, France, Italie, Suisse) et à une régionale (Nord-est de la France), les écorégions et la géologie sont déterminantes pour expliquer les communautés. Les paramètres caractérisant la pollution sont moins importants. Contrairement à certains auteurs, nous n'avons pas observé d'homogénéisation des communautés lorsque le niveau de pollution augmente. D'autre part nous n'avons pas observé de communautés restreintes géographiquement : cela permettrait de rassembler des écorégions distinctes géographiquement mais présentant les mêmes caractéristiques physiques. Les diatomées présentent une diversité spécifique très importante qui peut être un frein à leur utilisation en routine. Nous avons montré qu'en augmentant la précision de détermination (de la subdivision à l'espèce), les performances d'évaluation de la pollution augmentait mais beaucoup moins que le nombre de taxons. Les performances d'évaluation entre le genre et l'espèce sont d'ailleurs proches, alors qu'il y a dix fois plus d'espèce que de genres. Nous avons montré aussi que des métriques simplificatrices (formes de vie, guildes écologiques) permettaient d'évaluer aussi bien le niveau en nutriment que des indices diatomiques basés sur les espèces. Ces métriques apportent des informations supplémentaires en termes de structure de biofilm qui ne sont accessible aux données en espèce. Enfin, la pollution des rivières par les micropolluants devient une préoccupation sociétale croissante. Nous avons émis l'hypothèse que les diatomées pouvaient être de bons candidats pour évaluer la pression en herbicides. Quatre expérimentations de 2 mois ont été réalisées en mésocosmes lotiques. Nous avons montré que les diatomées vivant entourées de matrices polysaccharidiques épaisses étaient plus résistantes aux pesticides dissous. Au contraire les diatomées présentant une surface cellulaire de contact importante avec l'eau étaient défavorisées. Ce type de métrique pourrait être utilisé in situ à plus large échelle. Nous concluons sur l'intérêt d'intégrer ces métriques à la bioindication par les diatomées. Mais également nous soulignons l'importance de croiser la phylogénie et l'écologie pour mieux comprendre quelles pressions environnementales ont forcées les diatomées à s'adapter. Si ces pressions peuvent être reliées à des pressions anthropiques, la bioindication par les diatomées en sera améliorée.Diatoms are ubiquitous microalgae of an extreme diversity. This made them good indicators of aquatic ecosystems quality and they are used since 50 years for this purpose. Since year 2000, the European Water Framework Directive requires their use to assess the ecological quality of watercourses. A typological framework has to be used in order to compare comparable rivers between each other, that is, rivers of the same bioclimatic regions, flowing on the same geological substrate at similar altitudes. Various ecoregional classifications were defined on the basis of these parameters. We showed at a scale covering 4 countries (Spain, France, Italy and Switzerland) and at a regional scale (north-east France) that ecoregions and geology are determinant to explain communities. Parameters characterizing pollution were less important. Unlike some authors, we did not observe any homogenization of the communities when pollution level was increasing. Moreover, we did not observe geographically restricted communities: this would enable to aggregate ecoregions geographically distinct but presenting the same physical characteristics. Diatoms display a very important specific diversity which can be a problem for their routine use. We showed that when increasing determination precision (from sub-division to species), pollution assessment performances were increasing but much less than the number of taxa. Assessment performances between genus and species are similar anyway, whereas there are ten time more species than genera. We also showed that using simplifying metrics (life-forms, ecological guilds) enable assessing nutrient level as well as diatom indices based on species. These metrics bring additional information about biofilms structure that is not available with species data. At last, micropollutants pollution in rivers is of increasing concern to citizens. We hypothesized that diatoms could be good candidates to assess herbicide pressure. Four experiments lasting 2 months were conducted in lotic mesocosms. We showed that diatoms surrounded by thick exopolysaccharid matrices were more resistant to dissolved pesticides. On the over hand, diatoms presenting an important cell surface contact with water were disadvantaged. This kind of metric could be used in situ at a larger scale. We conclude on the interest to integrate such metrics to diatom bioassessment. But we also strength the importance to cross phylogeny and ecology to better understand which environmental pressure forced diatoms adapt. If these pressures can be related to anthropogenic pressures, diatom bioassessment will be improved.SAVOIE-SCD - Bib.électronique (730659901) / SudocGRENOBLE1/INP-Bib.électronique (384210012) / SudocGRENOBLE2/3-Bib.électronique (384219901) / SudocSudocFranceF

Risk of herbicide mixtures as a key parameter to explain phytoplankton fluctuation in a great lake: the case of Lake Geneva, Switzerland

Mixture risk assessment predictions have rarely been confronted with biological changes observed in the environment. In this study, long-term monitoring of a European great lake, Lake Geneva, provides the opportunity to assess to what extent the predicted toxicity of herbicide mixtures explains the changes in the composition of the phytoplankton community next to other classical limnology parameters such as nutrients. To reach this goal, the gradient of the mixture toxicity of 14 herbicides regularly detected in the lake was calculated using concentration addition and response addition models. A temporal gradient of toxicity was observed which decreased from 2004 to 2009. Redundancy analysis and partial redundancy analysis showed that this gradient explains a significant portion of the variation in phytoplankton community composition with and without having removed the effect of all other co-variables. Moreover, species that are significantly influenced, positively or negatively, by the decrease of toxicity in the lake over time are highlighted. It can be concluded that the herbicide mixture toxicity is one of the key parameters to explain phytoplankton changes in Lake Genev

Trophic history of French sub-alpine lakes over the last ~150 years: phosphorus reconstruction and assessment of taphonomic biases

Like many lakes worldwide, French sub-alpine lakes (lakes Annecy, Bourget and Geneva) have suffered from eutrophication in the mid-20th century. Although restoration measures have been undertaken and resulted in significant reductions in nutrient inputs and concentrations over the last 30 years in all three lakes, the limnological monitoring does not extend back far enough to establish the reference conditions, as defined by the European Water Framework Directive. The over-arching aim of this work was to reconstruct, using a paleolimnological approach, the pre-eutrophication levels and subsequent temporal changes in the lakes trophic status over the last century. The objectives were three-fold: i) to test whether fossil diatoms archived in deep sediment cores adequately reflect past changes in the planktonic diatom communities for these deep sub-alpine lakes based on data from lake Geneva; ii) to investigate changes in the diatom communities over the last 150 years in the three lakes; and iii) to infer the past total phosphorus (TP) concentrations of the lakes from a diatom based transfer function. Annual paleolimnological and limnological diatom countings for lake Geneva were strongly correlated over the last 30 years. Most notable differences essentially resulted from both taphonomic and depositional biases, as evidenced by the underestimation of thin skeleton species such as Asterionella formosa and Diatoma tenuis in the paleolimnological dataset and the presence of many benthic taxa. The fossil diatom records revealed shifts in the communities in the three lakes over time, most of which were changes typically associated with nutrient enrichment. Indeed, in all three lakes, the proportion of Cyclotella spp. was very high before the 1950s, but these species were then replaced by more eutrophic taxa, such as Stephanodiscus spp, by the mid-20th century. From the 1980s, some but not all diatom species typical of re-oligotrophicated conditions (i.e. Cyclotella sp.) re-appeared in all three lakes. Yet, not all changes that occurred in the diatom communities since then may be attributed to decreasing TP. TP concentrations inferred from weighted averaging with classical deshrinking in lake Annecy and lake Geneva and weighted averaging with inverse deshrinking in Lake Bourget were very close to the monitored values. Comparisons of diatom-inferred TP concentrations to other paleo-proxies for trophic status revealed though that the transfer functions were rather insensitive to changes occurring below 10 ?g L-1, which limits the study ability to set accurate TP reference conditions below this threshold. However, current diatom community compositions arestill rather different from the pre-eutrophication ones, suggesting that reference conditions are still not achieved

Distribution and ecology of Didymosphenia geminata (Lyngbye) M. Schmidt (Bacillariophyta) in Trentino watercourses (Northern Italy)

P. 141-160In 2000 the European Parliament and The European Union Council recommended that member countries assess water quality using diatoms, as part of the phytobenthos. In Italy this recommendation has given new ímpetus to the study of diatom communities and the application of biotic indices. During the summer of 2004, a total of 11 rivers of the Trentino province (Northern Italy) were sampled, revealing the presence in low abundances of the diatom Didymosphenia geminata {Lyngbye) M. Schmidt. The sites where this diatom occurred were characterised by a low pollution impact; they were also lake fed or had a regulated flow regime, although with different geologies. Morphological features of D. gemina/a frustules were analysed using LM and SEM and specimens could be referred to the morphotype "geminata". In many countries this species is expanding its distribution and in New Zealand it is considered an invasive alga. This work provides an initial report on the presence of D. geminara in the Trento province, and contributes to increasing the knowledge on its distribution in Ilaly and its ecology in the AlpsS

Recommendations for the preservation of environmental samples in diatom metabarcoding studies

Implementation of DNA metabarcoding for diatoms for environmental monitoring is now moving from a research to an operational phase, requiring rigorous guidelines and standards. In particular, the first steps of the diatom metabarcoding process, which consist of sampling and storage, have been addressed in various ways in scientific and pilot studies and now need to be rationalised. The objective of this study was to compare three currently applied preservation protocols through different storage durations (ranging from one day to one year) for phytobenthos and phytoplankton samples intended for diatom DNA metabarcoding analysis. The experimental design used samples from four freshwater and two marine sites of diverse ecological characteristics. The impact of the sample preservation and storage duration was assessed through diatom metabarcoding endpoints: DNA quality and quantity, diversity and richness, diatom assemblage composition and ecological index values (for freshwater samples). The yield and quality of extracted DNA only decreased for freshwater phytobenthos samples preserved with ethanol. Diatom diversity was not affected and their taxonomic composition predominantly reflected the site origin. Only rare taxa (< 100 reads) differed among preservation methods and storage durations. For biomonitoring purposes, freshwater ecological index values were not affected by the preservation method and storage duration tested (including ethanol preservation), all treatments returning the same ecological status for a site. This study contributes to consolidating diatom metabarcoding. Thus, accompanied by operational standards, the method will be ready to be confidently deployed and prescribed in future regulatory monitoring

Benthic Diatom Communities in an Alpine River Impacted by Waste Water Treatment Effluents as Revealed Using DNA Metabarcoding

Freshwater ecosystems are continuously affected by anthropogenic pressure. One of the main sources of contamination comes from wastewater treatment plant (WWTP) effluents that contain wide range of micro- and macropollutants. Chemical composition, toxicity levels and impact of treated effluents (TEs) on the recipient aquatic ecosystems may strongly differ depending on the wastewater origin. Compared to urban TEs, hospital ones may contain more active pharmaceutical substances. Benthic diatoms are relevant ecological indicators because of their high species and ecological diversity and rapid response to human pressure. They are routinely used for water quality monitoring. However, there is a knowledge gap on diatom communities’ development and behavior in treated wastewater in relation to prevailing micro- and macropollutants. In this study, we aim to (1) investigate the response of diatom communities to urban and hospital TEs, and (2) evaluate TEs effect on communities in the recipient river. Environmental biofilms were colonized in TEs and the recipient river up- and downstream from the WWTP output to study benthic diatoms using DNA metabarcoding combined with high-throughput sequencing (HTS). In parallel, concentrations of nutrients, pharmaceuticals and seasonal conditions were recorded. Diatom metabarcoding showed that benthic communities differed strongly in their diversity and structure depending on the habitat. TE sites were generally dominated by few genera with polysaprobic preferences belonging to the motile guild, while river sites favored diverse communities from oligotrophic and oligosaprobic groups. Seasonal changes were visible to lower extent. To categorize parameters important for diatom changes we performed redundancy analysis which suggested that communities within TE sites were associated to higher concentrations of beta-blockers and non-steroidal anti-inflammatory drugs in urban effluents vs. antibiotics and orthophosphate in hospital effluents. Furthermore, indicator species analysis showed that 27% of OTUs detected in river downstream communities were indicator for urban or hospital TE sites and were absent in the river upstream. Finally, biological diatom index (BDI) calculated to evaluate the ecological status of the recipient river suggested water quality decrease linked to the release of TEs. Thus, in-depth assessment of diatom community composition using DNA metabarcoding is proposed as a promising technique to highlight the disturbing effect of pollutants in Alpine rivers

Autumn drought drives functional diversity of benthic diatom assemblages of continental intermittent streams

International audienceClimate change is predicted to increase drought occurrence and severity in small continental watercourses. Here, we studied the structure and the functional diversity of benthic diatom assemblages in lowland intermittent and permanent watercourses of the Carpathian Basin. We assumed that the community structure of intermittent and permanent watercourses would be markedly different, and the functional diversity in both would be strongly influenced by autumn drought. We found that intermittent streams were primarily characterized by small-sized generalists and aerophilic taxa, while permanent watercourses were inhabited by large-sized planktic or fast moving groups. The functional richness was significantly lower in intermittent than in permanent streams. This decrease in the functional richness of benthic algal communities may negatively affect the functioning of lotic algal communities. We conclude that diatom assemblages in lowland intermittent watercourses are sensitive indicators of changes in ecosystem properties, and should be considered in appropriate evaluation and management of extreme climatic events on aquatic ecosystems

Metadata standards and practical guidelines for specimen and DNA curation when building barcode reference libraries for aquatic life

DNA barcoding and metabarcoding is increasingly used to effectively and precisely assess and monitor biodiversity in aquatic ecosystems. As these methods rely on data availability and quality of barcode reference libraries, it is important to develop and follow best practices to ensure optimal quality and traceability of the metadata associated with the reference barcodes used for identification. Sufficient metadata, as well as vouchers, corresponding to each reference barcode must be available to ensure reliable barcode library curation and, thereby, provide trustworthy baselines for downstream molecular species identification. This document (1) specifies the data and metadata required to ensure the relevance, the accessibility and traceability of DNA barcodes and (2) specifies the recommendations for DNA harvesting and for the storage of both voucher specimens/samples and barcode data.info:eu-repo/semantics/publishedVersio

DNA barcode reference libraries for the monitoring of aquatic biota in Europe: Gap-analysis and recommendations for future work

Effective identification of species using short DNA fragments (DNA barcoding and DNA metabarcoding) requires reliable sequence reference libraries of known taxa. Both taxonomically comprehensive coverage and content quality are important for sufficient accuracy. For aquatic ecosystems in Europe, reliable barcode reference libraries are particularly important if molecular identification tools are to be implemented in biomonitoring and reports in the context of the EU Water Framework Directive (WFD) and the Marine Strategy Framework Directive (MSFD). We analysed gaps in the two most important reference databases, Barcode of Life Data Systems (BOLD) and NCBI GenBank, with a focus on the taxa most frequently used in WFD and MSFD. Our analyses show that coverage varies strongly among taxonomic groups, and among geographic regions. In general, groups that were actively targeted in barcode projects (e.g. fish, true bugs, caddisflies and vascular plants) are well represented in the barcode libraries, while others have fewer records (e.g. marine molluscs, ascidians, and freshwater diatoms). We also found that species monitored in several countries often are represented by barcodes in reference libraries, while species monitored in a single country frequently lack sequence records. A large proportion of species (up to 50%) in several taxonomic groups are only represented by private data in BOLD. Our results have implications for the future strategy to fill existing gaps in barcode libraries, especially if DNA metabarcoding is to be used in the monitoring of European aquatic biota under the WFD and MSFD. For example, missing species relevant to monitoring in multiple countries should be prioritized for future collaborative programs. We also discuss why a strategy for quality control and quality assurance of barcode reference libraries is needed and recommend future steps to ensure full utilisation of metabarcoding in aquatic biomonitoring.This paper is a deliverable of the European Cooperation in Science and Technology (COST) Action DNAqua-Net (CA15219) Working Group 1, led by Torbjørn Ekrem and Fedor Čiampor. Thanks to the University of Minho and University of Pécs for hosting workshops and working group meetings. We also thank staff at National Environment Agencies and others that provided national checklists of taxa used in biomonitoring, and otherwise assisted with checklist proof-reading: Jarmila Makovinská and Emília Mišíková Elexová (Slovakia); Steinar Sandøy and Dag Rosland (Norway); Mišel Jelič (Croatia); Marlen Vasquez (Cyprus); Adam Petrusek (Czech Republic); Kristel Panksep (Estonia); Panagiotis Kaspiditis (Greece); Matteo Montagna (Italy); Marija Katarzyte (Lithuania); Ana Rotter (Slovenia); Rosa Trabajo (Spain); Florian Altermatt (Switzerland); Kristian Meissner (Finland), Rigers Bakiu (Albania), Valentina Stamenkovic and Jelena Hinic (Macedonia); Patricia Mergen (Belgium); Gael Denys & the French Biodiversity Agency (France); Mary Kelly-Quinn (Ireland); Piotr Panek and Andrzej Zawal (Poland); Cesare Mario Puzzi (Italy); Carole Fitzpatrick (United Kingdom); Simon Vitecek (Austria); Ana Filipa Filipe (Portugal); Peter Anton Stæhr & Anne Winding (Denmark); Michael Monaghan (Germany); Alain Dohet, Lionel L'Hoste, Nora Welschbillig & Luc Ector (Luxembourg), Lujza Keresztes, (Romania). The authors also want to thank Dirk Steinke for providing the original European ERMS list for marine taxa and Florian Malard for comments on the manuscript. The preparation of the AMBI checklist was carried out in the scope of a Short-term Scientific Mission (ECOST-STSM-CA15219-150217- 082111) granted to SD visiting AZTI, Spain. ZC was supported by grants EFOP-3.6.1.-16-2016-00004 and 20765-3/2018/FEKUTSTRAT. TE was supported by the NorBOL-grant (226134/F50) from the Research Coun cil of Norway. BR, FL and MFG contributed through support from the GBOL project, which is generously funded by the German Federal Min istry of Education and Research (FKZ 01LI1101 and 01LI1501). MG contributed through support of the Polish National Science Centre, grants N N303 5794 39 and 2014/15/B/NZ8/00266. SF was funded by the project PORBIOTA - Portuguese E-Infrastructure for Information and Research on Biodiversity (POCI-01-0145-FEDER-022127), supported by Operational Thematic Program for Competitiveness and Internationalization (POCI), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (FEDER)