Pressure-Retaining Sampler and High-Pressure Systems to Study Deep-Sea Microbes Under in situ Conditions

The pelagic realm of the dark ocean is characterized by high hydrostatic pressure, low temperature, high-inorganic nutrients, and low organic carbon concentrations. Measurements of metabolic activities of bathypelagic bacteria are often underestimated due to the technological limitations in recovering samples and maintaining them under in situ environmental conditions. Moreover, most of the pressure-retaining samplers, developed by a number of different labs, able to maintain seawater samples at in situ pressure during recovery have remained at the prototype stage, and therefore not available to the scientific community. In this paper, we will describe a ready-to-use pressure-retaining sampler, which can be adapted to use on a CTD-carousel sampler. As well as being able to recover samples under in situ high pressure (up to 60 MPa) we propose a sample processing in equi-pressure mode. Using a piloted pressure generator, we present how to perform sub-sampling and transfer of samples in equi-pressure mode to obtain replicates and perform hyperbaric experiments safely and efficiently (with <2% pressure variability). As proof of concept, we describe a field application (prokaryotic activity measurements and incubation experiment) with samples collected at 3,000m-depth in the Mediterranean Sea. Sampling, sub-sampling, transfer, and incubations were performed under in situ high pressure conditions and compared to those performed following decompression and incubation at atmospheric pressure. Three successive incubations were made for each condition using direct dissolved-oxygen concentration measurements to determine the incubation times. Subsamples were collected at the end of each incubation to monitor the prokaryotic diversity, using 16S-rDNA/rRNA high-throughput sequencing. Our results demonstrated that oxygen consumption by prokaryotes is always higher under in situ conditions than after decompression and incubation at atmospheric pressure. In addition, over time, the variations in the prokaryotic community composition and structure are seen to be driven by the different experimental conditions. Finally, within samples maintained under in situ high pressure conditions, the active (16S rRNA) prokaryotic community was dominated by sequences affiliated with rare families containing piezophilic isolates, such as Oceanospirillaceae or Colwelliaceae. These results demonstrate the biological importance of maintaining in situ conditions during and after sampling in deep-sea environments

From In Situ to satellite observations of pelagic Sargassum distribution and aggregation in the Tropical North Atlantic Ocean

International audienceThe present study reports on observations carried out in the Tropical North Atlantic in summer and autumn 2017, documenting Sargassum aggregations using both ship-deck observations and satellite sensor observations at three resolutions (MSI-10 m, OLCI-300 m, VIIRS-750 m and MODIS-1 km). Both datasets reported that in summer, Sargassum aggre-gations were mainly observed off Brazil and near the Caribbean Islands, while they accumulated near the African coast in autumn. Based on in situ observations, we propose a five-class typology allowing standardisation of the description of in situ Sargassum raft shapes and sizes. The most commonly observed Sargassum raft type was windrows, but large rafts composed of a quasi-circular patch hundreds of meters wide were also observed. Satellite imagery showed that these rafts formed larger Sargassum aggregations over a wide range of scales, with smaller aggregations (of tens of m 2 area) nested within larger ones (of hundreds of km 2). Match-ups between different satellite sensors and in situ observations were limited for this dataset, mainly because of high cloud cover during the periods of observation. Nevertheless, comparisons between the two datasets showed that satellite sensors successfully detected Sargassum abundance and aggregation patterns consistent with in situ observations. MODIS and VIIRS sensors were better suited to describing the Sargas-sum aggregation distribution and dynamics at Atlantic scale, while the new sensors, OLCI and MSI, proved their ability to detect Sargassum aggregations and to describe their (sub-) mesoscale nested structure. The high variability in raft shape, size, thickness, depth and biomass density observed in situ means that caution is called for when using satellite maps of Sargassum distribution and biomass estimation. Improvements would require additional in situ and airborne observations or very high-resolution satellite imagery

Formation of pristane from a-tocopherol under simulated anoxic sedimentary conditions : A combination of biotic and abiotic degradative processes

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Salinity shifts in marine sediment: Importance of number of fluctuation rather than their intensities on bacterial denitrifying community

International audienceThe sensitivity of denitrifying community to salinity fluctuations was studied in microcosms filled with marine coastal sediments subjected to different salinity disturbances over time (sediment under frequent salinity changes vs sediment with "stable" salinity pattern). Upon short-term salinity shift, denitrification rate and denitrifiers abundance showed high resistance whatever the sediment origin is. Denitrifying community adapted to frequent salinity changes showed high resistance when salinity increases, with a dynamic nosZ relative expression level. Marine sediment denitrifying community, characterized by more stable pattern, was less resistant when salinity decreases. However, after two successive variations of salinity, it shifted toward the characteristic community of fluctuating conditions, with larger proportion of Pseudomonas-nosZ, exhibiting an increase of nosZ relative expression level. The impact of long-term salinity variation upon bacterial community was confirmed at ribosomal level with a higher percentage of Pseudomonas and lower proportion of nosZII clade genera

Anaerobic bacterial degradation of pristenes and phytenes in marine sediments does not lead to pristane and phytane during early diagenesis

International audienceAnaerobic degradation of mixtures of isomeric pristenes and phytenes by sedimentary bacterial communities was investigated. These isoprenoid alkenes were quickly metabolized without formation of the corresponding alkanes. Identification of several bacterial metabolites allowed us to confirm the key role played by hydration in the metabolism of alkenes. Despite the increasing production of 5alpha-stanols during incubation, attesting to the involvement or biohydrogenation, this process did not act significantly on the double bond of pristenes and phytenes. Their resistance is attributed to the lack of binding polar group(s) to anchor the substrate to the enzyme and thus allow the double bond to reach the reductive catalytic site. It therefore seems that microbial hydrogenation of pristenes and phytenes does not account for the presence of pristane and phytane in recent sediments during early diagenesis

Coupling of bacterial nitrification with denitrification and anammox supports N removal in intertidal sediments (Arcachon Bay, France)

International audienceMicrobial activity measurements and molecular approaches were combined to study temporal variation in nitrogen (N) removal mechanisms from muddy and sandy intertidal sediments (Arcachon Bay, France). Stable isotopes ((NH4+)-N-15/(NO3-)-N-15) were used to measure oxidation of NH4+ to NO3-/NO2- and its subsequent reduction to N-2 via denitrification and/or anammox. We found that denitrification mainly fuelled N loss in both sediment types. However in sandy sediments, anammox accounted for relatively higher N-2 production (45%). Nitrification-denitrification (D-n) and nitrification-anammox coupling was observed particularly in muddy sediments. Temporal variations in prokaryotic abundance (Bacteria, Archaea) and of functional groups responsible for nitrification and denitrification were studied by qPCR and for anammoxifiers by cloning. Bacterial density in both sediment types varied between 10(7-8) equivalent cell per g(-1) dry weight sediment and were overall more abundant in muddy sediments. Archaeal equivalent cell varied between 0.5 and 6.5% of the total prokaryotes, with extreme values observed in muddy sediment during October and January respectively. Denitrifiers were 1-2 orders higher than ammonium oxidizers and archeal nitrifiers appeared insignificant compared to betaproteobacterial counterpart. Furthermore, nitrifiers exhibited greater fluctuations in sandy sediments compared to muddy ones. Anammoxifiers were mainly restricted to the "Candidatus Scalindua" group, and were detected in both sediment types and all sampling periods providing an indirect proof on the occurrence of anammox. In muddy sediments, N-2 production presented overall higher rate over sandy sediments and exhibited the same trend as nitrifier abundance. This finding suggests a central role of bacterially-mediated benthic nitrification in supporting dissimilatory processes through generation of NO3-/NO2 and preventing eutrophication in this mesotidal lagoo

Fatty acid composition of bacterial strains associated with living cells of the haptophyte Emiliania huxleyi

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Denitrification prevails over anammox in tropical mangrove sediments (Goa, India)

Denitrification, anammox (Anx) and di-nitrogen fixation were examined in two mangrove ecosystemsthe anthropogenically influenced Divar and the relatively pristine Tuvem. Stratified sampling at 2 cm increments from 0 to 10 cm depth revealed denitrification as the main process of N-2 production in mangrove sediments. At Divar, denitrification was similar to 3 times higher than at Tuvem with maximum activity of 224.51 +/- 6.63 nmol N-2 g(-1) h(-1) at 0-2 cm. Denitrifying genes (nosZ) numbered up to 2 x 10(7) copies g(-1) sediment and belonged to uncultured microorganisms clustering within Proteobacteria. Anammox was more prominent at deeper depths (8-10 cm) mainly in Divar with highest activity of 101.15 +/- 87.73 nmol N-2 g(-1) h(-1) which was 5 times higher than at Tuvem. Di-nitrogen fixation was detected only at Tuvem with a maximum of 12.47 +/- 8.36 nmol N-2 g(-1) h(-1). Thus, in these estuarine habitats prone to high nutrient input, N-2-fixation is minimal and denitrification rather than Anx serves as an important mechanism for counteracting N loading. (C) 2011 Elsevier Ltd. All rights reserved

Spatio-temporal diversity of free-living and particle-attached prokaryotes in the tropical lagoon of Ahe atoll (Tuamotu Archipelago) and its surrounding oceanic waters

International audienceSpatio-temporal variability of prokaryotic water column communities inside and outside a Polynesian tropical lagoon subjected to pearl oysters farming was assessed in terms of abundance by quantitative PCR and diversity by DGGE. Communities and operational taxonomic units (OTUs) were analysed according to dry/rainy seasons and free-living/particle-attached state. Bacterial density was higher in the lagoon compared to ocean and a seasonal trend was observed. No influence of the localisation within lagoon or of the planktonic/attached states was noticed on bacterial abundance and diversity. The OTUs belonged to Cyanobacteria, to heterotrophic groups in Proteobacteria and Flavobacteria. Archaeal abundance showed seasonal tendency and particle-prevalence, but no effect of lagoon or oceanic location was observed. Lagoon and oceanic archaeal diversity were different and Euryarchaeota (MG-II, MBG, and Halobacteria) were detected. During the dry season, planktonic and particle-associated community differed, whereas at rainy season, both communities were similar and included members usually associated with cora