Diversity, community structure, and quantity of eukaryotic phytoplankton revealed using 18S rRNA and plastid 16S rRNA genes and pigment markers: a case study of the Pearl River Estuary.

Understanding consistencies and discrepancies in characterizing diversity and quantity of phytoplankton is essential for better modeling ecosystem change. In this study, eukaryotic phytoplankton in the Pearl River Estuary, South China Sea were investigated using nuclear 18S rRNA and plastid 16S or 23S rRNA genes and pigment analysis. It was found that 18S abundance poorly explained the variations in total chlorophyll a (Chl-a). However, the ratios of log-transformed 18S abundance to Chl-a in the major phytoplankton groups were generally environment dependent, suggesting that the ratio has potential as an indicator of the physiological state of phytoplankton. The richness of 18S-based operational taxonomic units was positively correlated with the richness of 16S-based amplicon sequence variants of the whole phytoplankton community, but insignificant or weak for individual phytoplankton groups. Overall, the 18S based, rather than the 16S based, community structure had a greater similarity to pigment-based estimations. Relative to the pigment data, the proportion of haptophytes in the 18S dataset, and diatoms and cryptophytes in the 16S dataset, were underestimated. This study highlights that 18S metabarcoding tends to reflect biomass-based community organization of eukaryotic phytoplankton. Because there were lower copy numbers of plastid 16S than 18S per genome, metabarcoding of 16S probably approximates cell abundance-based community organization. Changes in biomass organization of the pigment-based community were sensitive to environmental changes. Taken together, multiple methodologies are recommended to be applied to more accurately profile the diversity and community composition of phytoplankton in natural ecosystems. Supplementary Information: The online version contains supplementary material available at 10.1007/s42995-023-00186-x.

Isotopic niches of the Northern shoveler Spatula clypeata in spring migratory stopover sites in the Marais breton, France.

The Northern shoveler, Spatula clypeata, makes several migratory stopovers to reach its breeding site in the best conditions. These stopovers allow the species to replenish their reserves. Therefore, feeding efficiency at such sites is essential. Despite its importance, few studies have been conducted on the spring ecology of the shoveler, particularly on its diet at stopover sites. Therefore, this study focused on the Northern shoveler's feeding habit during its spring migratory stopover in the Marais breton (MB), a wetland situated in Vendée (France, Atlantic coast). The shoveler's plasma and potential food resources were studied using a stable carbon and nitrogen isotope analysis. The study showed that the shoveler feeds primarily on microcrustaceans, especially Cladocera and Copepoda; Chironomidae larvae, Corixidae, Hydrophilidae larvae, and particulate organic matter. This last food source, the POM, had never been highlighted before.

Drivers for primary producers' dynamics: New insights on annual benthos pelagos monitoring in anthropised freshwater marshes (Charente-Maritime, France).

Wetlands, especially marshes, support many services such as carbon catchment control or water purification led by primary producers such as phytoplankton and microphytobenthos (PB). The impact of the sedimentary compartment, as source and sink of essential nutrients for the water column, is often neglected in the study of their dynamics and water purification capacity of the systems. This work compared monthly (between February 2020 and April 2021) the benthic and pelagic primary producers' dynamics in two anthropised freshwater marshes (Marans and Genouillé), with the simultaneous follow-up of physico-chemical parameters of the water column and nutrient fluxes at the sediment-water (SWI) interface. It was suggested a strong contribution of phytoplankton (pumping) and the benthic compartment (denitrification) to the water purification of these two nitrates (NO3-)-rich marshes. Total phytoplankton production fluctuated between ∼5 (winter) and 1500 mg C m-3 d-1 (fall) at Marans and between 40 (winter) and ∼750 mg C m-3 d-1 (spring) at Genouillé. At Marans, soluble reactive phosphorus (SRP) benthic effluxes (-2.101 to -6.102 µmol m-2 d-1 in fall and summer, respectively) coincided with phytoplankton bloom periods. These effluxes were inhibited by NO3- penetration in the sediment (0 to 5.104 µmol m-2 d-1), by inhibiting iron respiration. At Genouillé, inhibition of SRP effluxes depended on denitrification rate and on P stocks in the sediment, where slight SRP effluxes (-101 µmol m-2 d-1) could have co-occurred with slight NO3- influxes (5.102 µmol m-2 d-1) in spring. The presence of PB (between 10-60 and 40-120 mg gsed-1 at Marans and Genouillé, respectively), suggested a strong contribution of the benthic compartment to the total primary production (benthic and pelagic through resuspension processes) in these environments. This work encourages to consider the benthos and the pelagos as a unicum to provide better sustainable management of such systems and limit eutrophication risks in coastal areas.

Do microbial planktonic communities reflect the ecological changes of Glorieuses coral reefs (Iles Eparses, Western Indian Ocean)?

Ecological baselines for the structure and functioning of ecosystems in the absence of human activity can provide essential information on their health status. The Glorieuses islands are located in the Western Indian Ocean (WIO) and can be considered as "pristine" ecosystems that have not been subjected to anthropogenic pressure. Their nutrient context and the microbial assemblages were assessed by determining the abundance of heterotrophic prokaryotes (archaea and bacteria), picocyanobacteria, picoeukaryotes, microphytoplankton and protozooplankton communities in five stations, during two contrasted periods (November 2015 and May 2016). Chlorophyll-a concentrations were always under 1 µg/L and associated to very low levels in orthophosphates, nitrate and dissolved organic carbon, revealing an ultra-oligotrophic status for the Glorieuses waters. Picocyanobacteria confirmed the ultra-oligotrophic status with a predominance of Synechococcus. Zeaxanthin associated with the presence of picocyanobacteria represented the major pigment in both surveys. Three indices of diversity (species richness, Shannon and Pielou indexes) from microscopy observations highlighted the difference of diversity in microphytoplankton between the surveys. A focus on a 16S metabarcoding approach showed a high dominance of picocyanobacteria, Alpha- and Gammaproteobacteria, regardless of station or period. Multivariate analyses (co-inertia analyses) revealed a strong variability of ecological conditions between the two periods, with (i) high nutrient concentrations and heterotrophic nanoflagellate abundance in November 2015, and (ii) high heterotrophic prokaryote and picoeukaryote abundance in May 2016. The impact of a category 5 tropical cyclone (Fantala) on the regional zone in April 2016 is also advanced to explain these contrasted situations. Relative importance of top-down factors between bacterial and heterotrophic nanoflagellates was observed in November 2015 with an active microbial food web. All the results indicate that three microbial indexes potentially can be considered to assess the ecological change in Glorieuses marine waters.

Morphological, calorific and nutritive characteristics of 656 freshwater invertebrates taxa.

BACKGROUND: The Freshwater Animal Diversity Assessment (FADA) project estimated that freshwater animal species represent 9.5% of the 1.2 million species described. Knowing that freshwater represents only 0.01% of the earth's surface, these wetlands are suitable habitats for a great part of the world's total biodiversity. However, it has been shown that there is a lack of knowledge on these species, including freshwater invertebrates. Nevertheless, they play a key role in the majority of freshwater ecosystems and in their foodweb networks. Freshwater invertebrates are the food resource of many species, such as fish and birds. The knowledge of their morphological, energetic and nutritive characteristics allows a better understanding of their selection by predators (size, energy intake etc.), but also leads to the improvement of wetland management. Although information about freshwater invertebrates exists in literature, they are generally heterogeneous, dispersed and difficult to collect. To facilitate the accessibility of these data and, thus, optimise and accelerate research projects including freshwater invertebrates, we propose a literature review describing 14 morphological and nutritive characteristics (size, dry weight, gross energy, crude protein etc.) for 656 taxa of freshwater invertebrates. NEW INFORMATION: This dataset is a review from 104 publications from 1935 to 2020, compiling 14 characteristics when available (size, dry weight, gross energy, crude protein etc.) for 656 taxa of freshwater invertebrates.

New insights in bacterial and eukaryotic diversity of microbial mats inhabiting exploited and abandoned salterns at the Ré Island (France).

In order to understand the effect of human practices on microbial mats organisation, the study aimed to investigate the biodiversity within microbial mats from exploited and abandoned salterns. Despite several attempts, archaeal 16S rRNA gene fragment sequences were not obtained, indicating that microbial mats were probably dominated by Bacteria with very low abundance of Archaea (< 1%). Thus, the study compared the bacterial and meiofaunal diversity of microbial mats from abandoned and exploited salterns. The higher salinity (101 ± 3.7 psu vs. 51.1 ± 0.7 psu; Welch t-test p < 0.05) of the exploited site maintained lower bacterial diversity in comparison to the abandoned site where the salinity gradient was no longer maintained. However, the microbial mats exhibited similar bacterial class composition while the eukaryotic diversity was significantly higher in the exploited saltern. The abandoned saltern was dominated by sulfate-reducing bacteria and Nematoda, while the exploited saltern was characterized by the presence of halophilic bacteria belonging to Marinobacter, Salinivibrio and Rhodohalobacter genera, and the larger abundance of Hypotrichia (ciliates). Such bacterial and eukaryotic diversity difference might be explained by human actions for salt recovery in exploited salterns such as scraping the surface of microbial mat and increasing salinity renewing the microbial mat each year. Such action decreases the bacterial diversity changing the food web structure that favour the presence of a larger diversity of eukaryotic organisms. Our study provides new insights on microbial mat communities inhabiting salterns, especially the consequences of abandoning saltern exploitation.

Diuron effects on photosynthesis and vertical migration of microphytobenthos: Potential rapid bioassessment of herbicide toxicity in coastal sediments.

The effects of herbicide diuron on photosynthesis and vertical migration of intertidal microphytobenthos (MPB) assemblages were investigated using chlorophyll fluorometry. The results shown diuron ≤ 60 µg L-1 had no obvious effect on MPB vertical migration during 24 h indicated by consistent rhythm. Low concentration of 10 µg L-1 diuron had no significant influence on MPB photosynthesis throughout, however, high concentrations of 40, 50, and 60 µg L-1 had significant impacts exhibited by decreased parameters of maximum relative electron transport rate (rETRmax), maximal PS II quantum yield (Fv/Fm) and non-photochemical quenching (NPQ). For middle concentrations of 20 and 30 µg L-1, above decreased 3 parameters recovered sooner or later after 2 h or 16.5 h. Comparatively, rETRmax, Fv/Fm and NPQ are concentration dependent and more sensitive than other parameters in assessing diuron toxicity. This study revealed the potential of using MPB assemblages and chlorophyll fluorometry for rapid assessing diuron toxicity in coastal sediments.

Coupling between sediment biogeochemistry and phytoplankton development in a temperate freshwater marsh (Charente-Maritime, France): Evidence of temporal pattern.

In freshwater systems, sediment can be an important source for the internal loading of PO4. The limiting character of this element in such system leads to consider this phenomenon in terms of eutrophication risks and water quality stakes. A four-months follow-up (January, March, April and May 2019) was carried out in a strong phosphate (PO4) limited secondary channel from an artificial irrigation system of Charente Maritime (France) to link the mobilization of remineralization products in the upper 6 cm layer of sediment (conventional core slicing/centrifugation and DET probes) and the phytoplankton biomass dynamics in the water column. Results showed congruent patterns between the temporal succession of the organic matter mineralization processes in the sediment and the primary biomass dynamics in the water column. In January and March (considered in winter), PO4 proved to be retained by adsorption onto iron oxides in anoxic sediment since pore water nitrate inhibited for about a month the respiration of metal oxides in the first cm of sediment, thus limiting PO4 availability and the phytoplankton growth. In April and May (early spring), after exhaustion of pore water nitrate, the dissolutive reduction of iron oxides released PO4 into pore water generated a significant diffusive outgoing flux from the sediment to the water column with a maximum in April (-1.10E-04±2.81E-05 nmol cm-2 s-1). This release coincided with the nanophytoplankton bloom (5.50 µg Chla L-1) and a potential increase of PO4 concentration in the water column. This work provides some insight on the importance of benthic-pelagic coupling in anthropogenic systems. This conceptual model has to be deployed on other sites of interest where internal loading of P takes precedence over external inputs and nitrate mitigation drives its benthic recycling and ultimately its bioavailability. This is to be essential to characterize the aquatic environment quality in order to limit eutrophication risks.

Maturation of the European sardine Sardina pilchardus under experimental conditions strengthens bioenergetic estimate.

This study aims at (1) experimentally estimating first sexual maturation of the European sardine S. pilchardus, (2) using the results to calibrate existing bioenergetic models. During the 183 days-experiment, fish growth and body condition were assessed by biometry, and gonads were weighed when present. Age, wet weight and total length at first maturity were estimated at 262 days, 10.79 ± 0.75 g, and 11.26 ± 0.21 cm, respectively. Including these traits in biphasic Von Bertalanffy models did not significantly improve simulations for either length or weight data, meaning that energy allocation was not impacted by these traits. The implementation of the results in the Dynamic Energy Budget (DEB) calibration procedure strengthened the parameter set of the existing model, but resulted in significant changes in the energy allocation. Our results are a first step that will allow the design of new experiments to further quantify maturation and reproduction rates in diverse environmental conditions, consolidating DEB model calibration.

Diet variably affects the trophic transfer of trace elements in the oyster Crassostrea gigas.

Although it has been shown that trophic transfer of trace elements in oysters can be influenced by the diet, most of the studies investigating the ability of oysters to bioaccumulate trace elements from their diet are based on experiments using phytoplankton alone. Wild oysters feed also on large bacteria, ciliates or detritic organic matter. The present study aimed at examining the influence of food quality on the assimilation efficiency (AE) of trace elements in the Pacific cupped oyster Crassostrea gigas. Oysters were exposed via their food to the radiotracers of essential (57Co, 54Mn and 65Zn) and non-essential (110mAg, 241Am and 109Cd) trace elements under different diets (protozoan ciliates Uronema marinum and diatoms Thalassiosira pseudonana). Significant differences were found only for Ag and 241Am, with lower AEs measured in oysters fed with ciliates than in individuals fed with diatoms (Ag: 54 ± 3% vs. 67 ± 4% and 241Am: 62 ± 4% vs. 76 ± 4%). Interestingly, no significant difference was found among estimated depuration rates (kel) for all trace elements ingested with the two diets tested. These findings indicate that the differences observed are driven by the digestion process, presumably due to difference of bioavailability of trace elements dependent on the quality of the food ingested.

Ecological succession and resilience of plankton recovering from an acute disturbance in freshwater marshes.

The increase in extreme events such as storms is one of the major threats that coastal ecosystems will have to face in the near future. In such a context, both maturation and ecological successions processes remain at the core of ecology to better anticipate the changes to ecosystem biodiversity and functions facing environmental stressors. However, these concepts are mainly approached through closed experimental studies that oversimplify the mechanisms. A survey was carried out on a 'natural' and open ecosystem subjected to an acute disturbance, i.e. a marine submersion of freshwater drained marshes, occurring after a storm. Plankton biomass, production and taxonomic/functional phytoplankton diversity were followed weekly at four stations over 2 months. Most of the stations were disrupted by this acute disturbance and displayed gradual growth and development, as described in the classical maturation process. The main differences between stations were attributed to the heterogeneity of the communities before the storm, the intensity of the disturbance and the different human actions performed to recover the freshwater environment. The concept of 'ecological resilience' was thus better suited than 'engineering resilience' for such open systems facing constant fluctuations in environmental drivers. With regard to ecological succession, the more impacted stations were marked by a significant change in taxonomic beta-diversity, with numerous stochastic processes, due to taxa dispersion. They first exhibited a convergence in functional traits due to the increase in nutrient availability drained from the catchment basin and then an increase in divergence when nutrients became limited.

Impact of human activities on the spatio-seasonal dynamics of plankton diversity in drained marshes and consequences on eutrophication.

The functional diversity of two planktonic functional compartments, the nano-microphytoplankton and the mesozooplankton was used in order to better understand i) the drained marshes functioning and their related ecological functions, ii) the impacts of human control (replenishment) and human activities on the catchment basin (urbanization and catchment basin size). It was based on a monthly seasonal survey on 7 freshwater drained marshes. Both nano-microphyto- and mesozooplankton displayed high seasonal variations linked to the environmental fluctuations and human control on sea lock gates. Winter presented the lower biomasses of both compartments. Winter that is characterized by low water temperature, low light availability and high flood is actually related to the dominance of tychopelagic phytoplankton and K-strategists zooplankton. Spring and summer were characterized by i) the succession of pelagic large cells, small cells and then taxa with alternatives food strategies due to nitrogen limitation and phosphorous desorption from the sediment leading to eutrophication processes and ii) the dominance of r-strategists for mesozooplankton. The artificial summer replenishment acts positively on water quality by decreasing the eutrophication processes since the nitrogen inputs limit the proliferation of phytoplankton mixotrophs and diazotrophs and increase the ecological efficiency during the warm period. Both small and large catchment basins may lead to summer eutrophication processes in drained marshes since the largest ones imply higher hydrodynamic features at the root of large inputs of nitrogen nutrient favoring the phytoplankton development while the smallest ones exhibit hypoxia problems due to high proliferation of macrophytes. Urbanized marshes are less subjected to eutrophication during summer than non urbanized marshes due to more recurrent nutrient inputs from urban waste. However they exhibited a lower ecological efficiency. The results suggest that a better management of the hydrodynamics of such anthropogenic systems can avoid eutrophication risks on coastal areas.

Influence of food (ciliate and phytoplankton) on the trophic transfer of inorganic and methyl-mercury in the Pacific cupped oyster Crassostrea gigas.

Diet is an important route of mercury (Hg) uptake in marine organisms. Trophic transfer of Hg throughout the food webs may be influenced by various factors, including diet and Hg speciation. Bivalves such as oysters are widely used as bioindicators of trace element pollution such as Hg. Nevertheless, our current knowledge regarding their ability to accumulate Hg from their diet is mainly based on experiments performed using phytoplankton. In their natural environment, oysters feed on a variety of food items including ciliates, detritus, in addition to phytoplankton. The present study aimed at examining the influence of diet composition on the trophic transfer of inorganic Hg (iHg) and methylmercury (MeHg) in the Pacific cupped oyster Crassostrea gigas. The pulse-chase feeding method was used with two radiolabeled food items: a heterotrophic protist (Uronema marinum) and a phytoplanktonic diatom (Thalassiosira pseudonana). Depuration of dietary Hg in the oysters was followed for 50 d. Kinetic parameters including assimilation efficiency (AE) and efflux rate constant (ke) were calculated. Our results showed that oysters fed on ciliates assimilated 96 ±â€¯1% and 31 ±â€¯2% of the ingested MeHg and iHg, respectively whereas these elements were similarly assimilated in the oysters fed on phytoplankton (78 ±â€¯3% and 86 ±â€¯4% for MeHg and iHg, respectively). Mercury assimilation in oyster is thus diet dependent (significant differences in AE, p < 0.05), metal species-dependent and likely resulting from variations in Hg bioavailability in the two food items tested and a gut passage time-dependent of the ingested matrix.

First evidence of the presence and activity of archaeal C3 group members in an Atlantic intertidal mudflat.

The phylogenetic assignment of archaeal communities is constantly evolving, and the recent discovery of new phyla that grouped into superphyla has provided novel insights into archaeal ecology and evolution in ecosystems. In intertidal sediments, archaea are known to be involved in key functional processes such as organic matter turnover, but the ecological relevance of the rarest archaeal groups is poorly investigated, due partly to the lack of cultivated members. The high resolution of microbial diversity provided by high-throughput sequencing technologies now allows the rare biosphere to be described. In this work, we focused on the archaeal C3 group, showing that this phylum is not only present (at the DNA level) independently of sediment depth but also active (at the RNA level) in specific sediment niches depending on vertical physicochemical gradients. Moreover, we highlight the ambiguous phylogenetic affiliation of this group, indicating the need of further research to get new insights into the role of the C3 group.

Disturbance Increases Microbial Community Diversity and Production in Marine Sediments.

Disturbance strongly impacts patterns of community diversity, yet the shape of the diversity-disturbance relationship remains a matter of debate. The topic has been of interest in theoretical ecology for decades as it has practical implications for the understanding of ecosystem services in nature. One of these processes is the remineralization of organic matter by microorganisms in coastal marine sediments, which are periodically impacted by disturbances across the sediment-water interface. Here we set up an experiment to test the hypothesis that disturbance impacts microbial diversity and function during the anaerobic degradation of organic matter in coastal sediments. We show that during the first 3 weeks of the experiment, disturbance increased both microbial production, derived from the increase in microbial abundance, and diversity of the active fraction of the community. Both community diversity and phylogenetic diversity increased, which suggests that disturbance promoted the cohabitation of ecologically different microorganisms. Metagenome analysis also showed that disturbance increased the relative abundance of genes diagnostic of metabolism associated with the sequential anaerobic degradation of organic matter. However, community composition was not impacted in a systematic way and changed over time. In nature, we can hypothesize that moderate storm disturbances, which impact coastal sediments, promote diverse, and productive communities. These events, rather than altering the decomposition of organic matter, may increase the substrate turnover and, ultimately, remineralization rates.

Trophic relationships between metazooplankton communities and their plankton food sources in the Iles Eparses (Western Indian Ocean).

Coral reef and atoll lagoons are among the most diversified marine ecosystems but also the most affected by the combined effects of climate change and human activities. The Iles Eparses (Scattered Islands) in the Western Indian Ocean have been little affected by human pressure and can be considered to be "pristine" ecosystems. Metazooplankton plays a major role in the functioning and productivity of aquatic ecosystems, and this study was undertaken: (i) to determine the spatial abundance, distribution and species composition of metazooplankton, (ii) to assess the effect of metazooplankton grazing on pico- and nanophytoplankton and (iii) to analyze the trophic positions of metazooplankton by using the stable isotope signatures of a wide variety of taxa and particulate organic matter from the Iles Eparses and Mayotte. Tromelin Island (which is not located in the Mozambique Channel) had the lowest metazooplankton abundance with no cyanobacteria Trichodesmium spp. or mollusks (pteropods) presence, and with δ(15)N signatures of organisms that were higher than for the islands in the Mozambique Channel. Trichodesmium spp. was found in the Mozambique Channel and the plankton food web was probably based preferentially on these cyanobacteria with lower δ(15)N signatures indicating direct or indirect trophic transfer of diazotrophic nitrogen to metazooplankton. Three of the islands were distinct: Europa had the highest proportion of copepods, with oithonids being dominant, which is typical of rich mangrove systems, while Juan de Nova and Mayotte seemed to be the sites most affected by human activity with a high abundance of appendicularians and distinct particulate organic matter ∂(13)C signatures. Grazing experiments showed that food could be a limiting factor for metazooplankton in the Iles Eparses. However, the effect of metazooplankton grazing on phytoplankton appeared to be very low (0.01-2.32% of the total phytoplankton per day).

Growth form defines physiological photoprotective capacity in intertidal benthic diatoms.

In intertidal marine sediments, characterized by rapidly fluctuating and often extreme light conditions, primary production is frequently dominated by diatoms. We performed a comparative analysis of photophysiological traits in 15 marine benthic diatom species belonging to the four major morphological growth forms (epipelon (EPL), motile epipsammon (EPM-M) and non-motile epipsammon (EPM-NM) and tychoplankton (TYCHO)) found in these sediments. Our analyses revealed a clear relationship between growth form and photoprotective capacity, and identified fast regulatory physiological photoprotective traits (that is, non-photochemical quenching (NPQ) and the xanthophyll cycle (XC)) as key traits defining the functional light response of these diatoms. EPM-NM and motile EPL showed the highest and lowest NPQ, respectively, with EPM-M showing intermediate values. Like EPL, TYCHO had low NPQ, irrespective of whether they were grown in benthic or planktonic conditions, reflecting an adaptation to a low light environment. Our results thus provide the first experimental evidence for the existence of a trade-off between behavioural (motility) and physiological photoprotective mechanisms (NPQ and the XC) in the four major intertidal benthic diatoms growth forms using unialgal cultures. Remarkably, although motility is restricted to the raphid pennate diatom clade, raphid pennate species, which have adopted a non-motile epipsammic or a tychoplanktonic life style, display the physiological photoprotective response typical of these growth forms. This observation underscores the importance of growth form and not phylogenetic relatedness as the prime determinant shaping the physiological photoprotective capacity of benthic diatoms.

An efficient and rapid method for the enumeration of heterotrophic prokaryotes in coastal sediments by flow cytometry.

Flow cytometry offers an easy and powerful way to assess multi-parametric data in different domains, notably in the environmental sciences. Because evaluating heterotrophic prokaryotic abundance is crucial to understand an ecosystem's functioning, we propose a quick and efficient protocol for (1) cell's detachment in muddy coastal sediments followed by (2) enumeration of prokaryotes by flow cytometry compared to epifluorescence microscopy and (3) a type of storage adapted for benthic samples. First, sample preparation by incubation in a detergent mix containing sodium pyrophosphate (0.01M final concentration) and Tween 80 (0.1% final concentration) drastically increased cell detachment from sediment particles (+130.40%) compared to extraction with sodium pyrophosphate only. Cell sorting allowed to control the efficiency of the extraction as few cells were found attached to sediment particles in epifluorescence microscopy after sorting. Flow cytometry gave consistent results with strong reliability by counting 1.81 times more cells compared to epifluorescence microscopy. Thirdly, results revealed that sediment samples fixed with formaldehyde and then liquid-N2 frozen and directly stored at -80°C can be analyzed within 3months. In routine, our method of extraction and counting allowed to evaluate 83.67% of the real abundance in a sediment sample. Finally, this optimized technique was applied on sandy and muddy coastal and freshwater sediments and allowed us to prove the high efficiency of this new method. Flow cytometry is a fast, replicable and low-cost method for counting heterotrophic prokaryotes, even for sediment samples. The two-step method that we developed enables high frequency analyses (30 samples in less than 4h).

Key features of intertidal food webs that support migratory shorebirds.

The migratory shorebirds of the East Atlantic flyway land in huge numbers during a migratory stopover or wintering on the French Atlantic coast. The Brouage bare mudflat (Marennes-Oléron Bay, NE Atlantic) is one of the major stopover sites in France. The particular structure and function of a food web affects the efficiency of carbon transfer. The structure and functioning of the Brouage food web is crucial for the conservation of species landing within this area because it provides sufficient food, which allows shorebirds to reach the north of Europe where they nest. The aim of this study was to describe and understand which food web characteristics support nutritional needs of birds. Two food-web models were constructed, based on in situ measurements that were made in February 2008 (the presence of birds) and July 2008 (absence of birds). To complete the models, allometric relationships and additional data from the literature were used. The missing flow values of the food web models were estimated by Monte Carlo Markov Chain--Linear Inverse Modelling. The flow solutions obtained were used to calculate the ecological network analysis indices, which estimate the emergent properties of the functioning of a food-web. The total activities of the Brouage ecosystem in February and July are significantly different. The specialisation of the trophic links within the ecosystem does not appear to differ between the two models. In spite of a large export of carbon from the primary producer and detritus in winter, the higher recycling leads to a similar retention of carbon for the two seasons. It can be concluded that in February, the higher activity of the ecosystem coupled with a higher cycling and a mean internal organization, ensure the sufficient feeding of the migratory shorebirds.

To what extent do food preferences explain the trophic position of heterotrophic and mixotrophic microbial consumers in a Sphagnum peatland?

Although microorganisms are the primary drivers of biogeochemical cycles, the structure and functioning of microbial food webs are poorly studied. This is the case in Sphagnum peatlands, where microbial communities play a key role in the global carbon cycle. Here, we explored the structure of the microbial food web from a Sphagnum peatland by analyzing (1) the density and biomass of different microbial functional groups, (2) the natural stable isotope (δ(13)C and δ(15)N) signatures of key microbial consumers (testate amoebae), and (3) the digestive vacuole contents of Hyalosphenia papilio, the dominant testate amoeba species in our system. Our results showed that the feeding type of testate amoeba species (bacterivory, algivory, or both) translates into their trophic position as assessed by isotopic signatures. Our study further demonstrates, for H. papilio, the energetic benefits of mixotrophy when the density of its preferential prey is low. Overall, our results show that testate amoebae occupy different trophic levels within the microbial food web, depending on their feeding behavior, the density of their food resources, and their metabolism (i.e., mixotrophy vs. heterotrophy). Combined analyses of predation, community structure, and stable isotopes now allow the structure of microbial food webs to be more completely described, which should lead to improved models of microbial community function.