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Organic contaminants, and herbicides in particular, represent a risk for aquatic ecosystems. The primary target of herbicides are producers, the base of food webs, but frequently they end up far from the application point affecting non-target species. Its presence can work as sub-lethal stimulus, which sort the genetic and phenotypic differences within a species. Intraspecific variation allows adaptation to changes in the environment but also to new niches due to variations in species’ sensitivity and biochemical response to a certain chemical. A better understanding of these variations can lead to the development of improved strategies for ecosystem protection. This research aimed to compare a sensitive and a tolerant strain of the freshwater diatom Nitzschia palea to atrazine. Strains were exposed to three concentrations within their tolerance range, during 96 h. The activity of the antioxidant enzymes superoxide dismutase, catalase, glutathione-S-transferases and glutathione peroxidases was determined. In addition, chlorophylls a and c, carotenoids, reduced glutathione, proteins and lipid peroxidation were quantified. Both strains displayed different strategies to deal with atrazine toxicity: while the sensitive strain decreased the oxidative stress, increasing the activity of antioxidant enzymes such as superoxide dismutase, the tolerant strain invested in conjugation pathways and carotenoids’ maintenance. [ABSTRACT FROM AUTHOR]

Benthic diatoms are relevant indicators of the ecological status of the littoral zone of lakes. Their use as bio-indicators is based on their morphological identification at species level using microscopy which is time consuming, requires taxonomic expertise, and is consequently expensive. To overcome these limitations, a molecular approach for diatom identification has been tested with success in rivers. DNA metabarcoding enables species identification from a standardized DNA barcode and high-throughput sequencing (HTS), using DNA reference library. The suitability of the morphological and molecular approaches to assess the diatom community structure and the ecological status of the littoral zone of the largest deep lake in France (Lake Bourget) was compared. 66 sites were sampled in August 2015 along the shoreline, all around the lake. The composition of diatom assemblages was similar with both morphological and molecular approaches, and diatom assemblages were structured by the same environmental factors. However, the ecological status of Lake Bourget differed significantly among approaches since floristic inventories to species level also differed significantly. The main source of this difference was the incompleteness of the DNA reference library. Nevertheless, in a near future, when this constraint will be solved, the use of DNA metabarcoding for biomonitoring purposes seems promising. [ABSTRACT FROM AUTHOR]

Double-stranded DNA viruses infecting eukaryotic algae (e.g., phycodnaviruses) and cyanobacteria (e.g., cyanophages) are now recognized as widespread and ubiquitous in aquatic environments. However, both the diversity and functional roles of these viruses in fresh waters are still poorly understood. We conducted a year-long study in 2011 of the community structure of planktonic virus groups in the upper lit layer of two important freshwater natural ecosystems in France, Lake Annecy (oligotrophic) and Lake Bourget (oligo-mesotrophic). Using PCR-DGGE to target a number of different structural and functional signature genes, i.e., g20, g23, psbA, polB, and mcp, the phytoplankton viruses were shown to display temporal and spatial variability. There were marked seasonal changes in community structure for all viral groups in Lake Bourget, but only for T4-like myoviruses and psbA-containing cyanophages in Lake Annecy. The multivariate statistical analyses revealed that (1) various environmental factors can directly or indirectly explain the community structure observed for each phytoplankton viral group, and (2) temporal patterns of T4-like myovirus community structure were similar between the two lakes. In general, our results (1) suggest that the observed algal virus patterns were associated with significant shifts in phytoplankton biomass and/or structure, which in turn were shaped by the abiotic environment, and (2) support the Bank model proposed by Breitbart and Rohwer (Trends Microbiol 13:278-284, ). This study provides new evidence that freshwater lakes contain a significant diversity of algal viruses, and that the distribution of these viruses strongly mirrors that of their hosts. [ABSTRACT FROM AUTHOR]

The recent emergence of barcoding approaches coupled to those of next-generation sequencing ( NGS) has raised new perspectives for studying environmental communities. In this framework, we tested the possibility to derive accurate inventories of diatom communities from pyrosequencing outputs with an available DNA reference library. We used three molecular markers targeting the nuclear, chloroplast and mitochondrial genomes ( SSU r DNA, rbc L and cox1) and three samples of a mock community composed of 30 known diatom strains belonging to 21 species. In the goal to detect methodological biases, one sample was constituted directly from pooled cultures, whereas the others consisted of pooled PCR products. The NGS reads obtained by pyrosequencing ( Roche 454) were compared first to a DNA reference library including the sequences of all the species used to constitute the mock community, and second to a complete DNA reference library with a larger taxonomic coverage. A stringent taxonomic assignation gave inventories that were compared to the real one. We detected biases due to DNA extraction and PCR amplification that resulted in false-negative detection. Conversely, pyrosequencing errors appeared to generate false positives, especially in case of closely allied species. The taxonomic coverage of DNA reference libraries appears to be the most crucial factor, together with marker polymorphism which is essential to identify taxa at the species level. Rbc L offers a high resolving power together with a large DNA reference library. Although needing further optimization, pyrosequencing is suitable for identifying diatom assemblages and may find applications in the field of freshwater biomonitoring. [ABSTRACT FROM AUTHOR]

The aim of this study was to determine the influence of environmental variables on the structure of benthic diatom assemblages, and to propose type assemblages of diatoms characterizing unpolluted headwater streams of Luxembourg. A total of 289 diatom samples were collected in the headwater streams of Luxembourg. At each sampling site, physical and chemical variables were also measured. The relationships between environmental variables and the distribution of the taxa were estimated using univariate and multivariate statistical analyses. On the basis of geology and stream water chemistry, Luxembourg could be separated in two regions. The streams in the schistose northern part of Luxembourg presented a low carbonate hardness (median 3.5° F); the streams in the southern part of Luxembourg presented a high carbonate hardness (median 21.8° F) notably connected to the presence of sandstone and limestone substrata. A Twinspan classification carried out on diatom assemblages defined two groups of samples corresponding to these two regions. The carbonate hardness that is related to the nature of the geological substratum appeared to be the major structuring variable for the assemblage composition. Anthropogenic pollution was a secondary structuring variable for diatom assemblages since each group could be subdivided in subgroups presenting statistically different nutrient and organic matter concentrations. The diatom assemblages of these subgroups were characterised by differences of saprobic and trophic preferences. On the basis of these results, two type assemblages of diatoms are proposed for the unpolluted headwater streams in the two regions of Luxembourg. The presence of these different type assemblages in Luxembourg shows the necessity to adapt diatom bioindication to the different regions of the country. [ABSTRACT FROM AUTHOR]

Soil algae are present in all kinds of soils, both arable and virgin, in large amounts and in great diversity. These photosynthetic microorganisms, which are concentrated in the top few centimeters of the soil profile, are organized in a community structure that varies depending on soil type, farming method and pesticide application. We first tested several extraction and conservation methods for soil algae, and a [sup 14]C incorporation procedure to evaluate the photosynthetic activity of these micro-organisms. In a second study, we assessed the sensitivity to atrazine of soil microalgae from two corn fields managed by different agricultural practices (conventional vs. organic). Changes in indigenous diatom communities were monitored, together with photosynthetic tests performed on the whole algal community. Comparison of the data for the treated field with those for the reference, untreated organic cornfield showed that previous atrazine application in the conventional cornfield had changed the species composition of the soil diatom communities. Short-term ecotoxicological tests, using photosynthetic activity as endpoint, also showed that the communities that had developed under pesticide stress were more tolerant to further atrazine application than the control communities. The stress caused by major environmental disturbances, such as ground dressing, was not sufficient to mask the difference between the two crops, suggesting that telluric microalgae could be used as indicators of xenobiotic contamination in soils. These initial findings about using soil microalgal communities as bioindicators are promising. In addition, their photosynthetic activity, which reflects their sensitivity to xenobiotic compounds, seems to be a relevant bioindicator of soil contamination. [ABSTRACT FROM AUTHOR]

We have tested the sensitivity of phytoplankton to the herbicides atrazine and nicosulfuron in experiments conduced in increasingly complex systems, from single strain phytoplankton cultures (microplates) to mesocosms mimicking whole ecosystems. The endpoints used to assess sensitivity to atrazine and nicosulfuron were total biomass increase, photosynthetic efficiency, and community diversity, depending on the system considered. Nicosulfuron appeared to be very much less toxic to phytoplankton than atrazine, in accord with the planned changes in agricultural practices to reduce the effects of surface water contamination on aquatic biota. Nevertheless, nicosulfuron had significant effects in some systems (principally microcosms), whereas the single monocultures were almost insensitive to it. This points out the inaccuracy of standardized toxicity test on phytoplanktonic algae alone for predicting the effects of xenobiotics on natural communities and the need for tests in microcosms and mesocosms to obtain reliable evidence about the toxicity of a given chemical on freshwater aquatic ecosystems. [ABSTRACT FROM AUTHOR]