Dark CO2 fixation by chemolithoautotrophic prokaryotes in the deep-water masses of the north-west coast of the Iberian Peninsule

Recent studies suggest that the prokaryotes inhabiting the dark ocean present higher chemolithoautotrophic activity than assumed previously. These chemolithoautotrophic microbes incorporate dissolved inorganic carbon (DIC) as carbon source for biomass production and use reduced inorganic compound as an energy source. We have quantified DIC fixation in the meso- and bathypelagic waters of the northwestern coast of the Iberian Peninsula, ranging from 1.04 to 46.83 mmol C m-2 d-1. Combining microautoradiography and fluorescence in situ hybridization (MICRO-CARD-FISH), we confirmed that both Thaumarchaeota and some bacterial groups such as SAR-11, SAR-202, SAR-406, Alteromonas take up bicarbonate uptake, particularly in the mesopelagic waters. Quantitative PCR analyses clearly showed a higher abundance of thaumarchaeal 16S and low ammonia concentration (LAC)- amoA genes in meso- and lower bathypelagic waters than in surface waters. In contrast, high ammonia concentration (HAC)- amoA genes dominated the subsurface samples. Taken together, both genomic and physiological evidences indicate that some archaeal and bacterial groups may be significant contributors to dark ocean chemoautolithotrophy

Dissolved inorganic carbon fixation of Thaumarchaeota vs. Bacteria in the meso- and upper bathypelagic waters of the world’s oceans differentiated with the use of metabolic inhibitors

Recent studies suggest that the dark ocean prokaryotes fix inorganic carbon at rates substantially higher than assumed. We have studied the contribution of Archaea vs. Bacteria to total prokaryotic fixation of dissolved inorganic carbon (DIC) in the meso- and upper bathypelagic waters of the world’s oceans during the Malaspina circumnavigation expedition carried out between December 2010 and July 2011. We used the metabolic inhibitor Erythromycin, an antibiotic specifically inhibiting growth of Bacteria but not affecting Archaea. Bacteria dominated throughout the water column in the three major ocean basins (54% of the total DAPI counts), decreasing in their relative contribution to total prokaryotic abundance from the surface to the meso- and bathypelagic waters. By contrast, the relative contribution of Thaumarchaeota was generally higher in the meso- and bathypelagic layers than in the surface waters (up to 29% of the total DAPI counts in the Pacific Ocean). Averaged over the entire water column, Thaumarchaeota contributed 8%, 33% and 18% to the total prokaryotic DIC fixation in the Indian, Pacific and Atlantic Ocean, respectively. The contribution of Thaumarchaeota to total prokaryotic DIC fixation increased with depth, particularly in the Atlantic below 1000 m depth and in the lower mesopelagic zone of the Pacific Ocean. Preliminary results from an station in the Atlantic Ocean, combining microautoradiography and fluorescence in situ hybridization (MICRO-CARD-FISH), confirmed that both Thaumarchaeota and some bacterial groups such as SAR 324 take up DIC. Thaumarchaeota and SAR 324 accounted for 7 % and 12% of DIC-positive DAPI-stained cells, respectively, as revealed by MICRO-CARD-FISH. Our results suggest that some phylogenetic groups may be significant contributors to the dark ocean chemoautotrophy

High dark CO2 fixation rates by active chemolithoautotrophic microbes along the water column (100-5000m) off Galicia (NW Iberian margin)

Poster communicationOur results provide evidence for the significant contribution to chemolithotrophy by specific archaeal and bacterial groups in the dark ocean

Bacterial activity and community composition response to the size-reactivity of dissolved organic matter

Heterotrophic bacteria respond dynamically to variations in organic matter availability in the dark ocean. However, our knowledge on how the differences in sized and/or reactivity of dissolved organic matter (DOM) affect the bacterial community dynamics is still scarce. Our study aims to investigate the response of bacterial activity and community composition to the degradability of filtered and of size-fractionated DOM. A natural bacterial community isolated from Mediterrean Water (MW; at 1000 m depth) was inoculated in seawater from the same location subjected to three different treatments: 0.2µm-filtered seawater (control), low molecular weight fraction (LMW) obtained by ultrafiltration, and the combination of low and high molecular weight fractions at the original ratio (H+L). Bacterial abundance and activity was monitored every 24h over 6 days, while bacterial community composition and DOM characterization were assessed at the beginning (day 0), middle (day 4) and at the end of the experiment (day 6). Low (LNA) and high nucleic acid content (HNA) bacterial abundance, as well as leucine incorporation rates, were consistently higher in the H+L incubations than in the LMW treatments, indicating different reactivity of the two organic matter size fractions. Moreover, actively respiring cells, estimated as CTC-positive cells, highly correlated to humic-like substances (FDOM-M; R=0.7, P<0.05, n=9, Spearman Rank Order), particularly in the H+L incubations. Interestingly, LNA cell abundance was highly correlated with the slope ratio (SR) values (R=-0.8, P<0.05, n=9, Spearman Rank Order), indicating that bacteria belonging to the LNA population are tightly linked to the molecular weight or aromaticity of the DOM. Taken together, our results indicate differences in the bio-reactivity of the low and high molecular weight size classes of DOM associated to the phylogenetic composition of the bacterial communities

Lower prokaryotic leucine incorporation rates under in situ pressure than under decompressed conditions in the deep north Atlantic

Comunicación oralProkaryotic activity and community composition is highly depth-stratified in the oceanic water column reflecting the increasing recalcitrance of dissolved organic matter and decreasing temperature with depth. The role of increasing hydrostatic pressure in controlling deep ocean microbial activity is less well-studied. To determine the influence in hydrostatic pressure on heterotrophic microbial activity, an in situ incubator was deployed in the North Atlantic Ocean at a depth between 500 to 2000 m. The in situ incubator was programmed to collect and incubate prokaryotes under the water after adding 3H-leucine and to fix a certain volume of the incubated samples at specific time intervals (3 to 10 h depending on the depth). Prokaryotic leucine incorporation obtained under in situ pressure conditions was generally lower than that on decompressed samples incubated on board. Ratios of in situ prokaryotic leucine incorporation to decompressed conditions decreased with increasing depth. Our results suggest that bulk heterotrophic prokaryotic production in the deep sea might be lower than expected

Taurine: an energy "drink" for deep sea microbes

Presentación oralThe wide use of –omics approaches has led to the discovery of novel metabolic pathways in uncultivated marine bacteria. For example, metagenomic and –proteomic studies revealed that taurine might be an important substrate for heterotrophic marine bacteria. Taurine, an organic acid, is widely produced by marine metazoans and some phytoplankton albeit its concentration and turnover in the ocean has not been determined yet. In this study, we determined the role of taurine as carbon and energy source throughout the water column of the open North Atlantic from the epipelagic to the bathypelagic realm. Bulk uptake and respiration of taurine were measured and microautoradiography was combined with catalyzed reporter deposition fluorescence in situ hybridization to evaluate taurine uptake by specific phylogenetic groups. A shift between the dominant use of taurine as a carbon source from the epi- and mesopelagic (about 40% of taurine respired) to the bathypelagic (76% respired) realm was observed. Taken together, our results indicate that taurine is effectively used by marine prokaryotes, especially in the mesopelagic environment where zooplankton, a potential source for taurine, reside during the day

Changes in bacterial activity and community composition in response to water mass mixing

PosterMixing zones and boundaries between different water masses are "hot spots" of marine biodiversity and activity. We aimed to investigate the effects of water mass mixing in the dark-ocean microbial communities by collecting and incubating natural bacterial communities from the Mediterranean Water (MW; at 1000 m depth), the Subpolar Modal Water (SPMW, 500m) and the Labrador Sea Water (LSW, 1800 m), and comparing them with artificially mixed communities. Mixing experiment 1 consisted of incubating at in sity conditions the original LSW and MW communities, plus a mixture of both (MIX1, dilution 1:1), whereas the Mixing experiment 2 included the original prokaryotic communities from SPMW and MW and a mixture of both (MIX2, dilution 1:1). Bacterial abundance and activity was monitored every 24 h over 8 days, while bacterial community composition and DOM characterization were assessed at the beginning (day 0), middle (day 4) and at the end of the experiment (day 8). Live prokaryotic cell abundance was higher in the MIX1 and MIX2 treatments as compared to the original communities. Moreover, MIX bacteria showed slightly higher leucine incorporation rates than MW or LSW. These metabolic responses were accompanied by changes in the optical properties of DOM, suggesting a change in the dynamics of the organic matter. Taken together, our results indicate differences in the bio-reactvity of the organic matter after mixing as compared to the original water masses that could influence the composition and activity of the bacterial community

Microbes mediating the sulfur cycle in the Atlantic Ocean and their link to chemolithoautotrophy

Only about 10%–30% of the organic matter produced in the epipelagic layers reaches the dark ocean. Under these limiting conditions, reduced inorganic substrates might be used as an energy source to fuel prokaryotic chemoautotrophic and/or mixotrophic activity. The aprA gene encodes the alpha subunit of the adenosine-5′-phosphosulfate (APS) reductase, present in sulfate-reducing (SRP) and sulfur-oxidizing prokaryotes (SOP). The sulfur-oxidizing pathway can be coupled to inorganic carbon fixation via the Calvin–Benson–Bassham cycle. The abundances of aprA and cbbM, encoding RuBisCO form II (the key CO2 fixing enzyme), were determined over the entire water column along a latitudinal transect in the Atlantic from 64°N to 50°S covering six oceanic provinces. The abundance of aprA and cbbM genes significantly increased with depth reaching the highest abundances in meso- and upper bathypelagic layers. The contribution of cells containing these genes also increased from mesotrophic towards oligotrophic provinces, suggesting that under nutrient limiting conditions alternative energy sources are advantageous. However, the aprA/cbbM ratios indicated that only a fraction of the SOP is associated with inorganic carbon fixation. The aprA harbouring prokaryotic community was dominated by Pelagibacterales in surface and mesopelagic waters, while Candidatus Thioglobus, Chromatiales and the Deltaproteobacterium_SCGC dominated the bathypelagic realm. Noticeably, the contribution of the SRP to the prokaryotic community harbouring aprA gene was low, suggesting a major utilization of inorganic sulfur compounds either as an energy source (occasionally coupled with inorganic carbon fixation) or in biosynthesis pathways.En prensa5,84

Dynamic prokaryotic communities in the dark western Mediterranean Sea

Dark ocean microbial dynamics are fundamental to understand ecosystem metabolism and ocean biogeochemical processes. Yet, the ecological response of deep ocean communities to environmental perturbations remains largely unknown. Temporal and spatial dynamics of the meso- and bathypelagic prokaryotic communities were assessed throughout a 2-year seasonal sampling across the western Mediterranean Sea. A common pattern of prokaryotic communities’ depth stratification was observed across the different regions and throughout the seasons. However, sporadic and drastic alterations of the community composition and diversity occurred either at specific water masses or throughout the aphotic zone and at a basin scale. Environmental changes resulted in a major increase in the abundance of rare or low abundant phylotypes and a profound change of the community composition. Our study evidences the temporal dynamism of dark ocean prokaryotic communities, exhibiting long periods of stability but also drastic changes, with implications in community metabolism and carbon fluxes. Taken together, the results highlight the importance of monitoring the temporal patterns of dark ocean prokaryotic communities.Versión del editor2,92

Dynamics of actively dividing prokaryotes in the western Mediterranean Sea

Microbial community metabolism and functionality play a key role modulating global biogeochemical processes. However, the metabolic activities and contribution of actively growing prokaryotes to ecosystem energy fluxes remain underexplored. Here we describe the temporal and spatial dynamics of active prokaryotes in the different water masses of the Mediterranean Sea using a combination of bromodeoxyuridine labelling and 16S rRNA gene Illumina sequencing. Bulk and actively dividing prokaryotic communities were drastically different and depth stratified. Alteromonadales were rare in bulk communities (contributing 0.1% on average) but dominated the actively dividing community throughout the overall water column (28% on average). Moreover, temporal variability of actively dividing Alteromonadales oligotypes was evinced. SAR86, Actinomarinales and Rhodobacterales contributed on average 3–3.4% each to the bulk and 11, 8.4 and 8.5% to the actively dividing communities in the epipelagic zone, respectively. SAR11 and Nitrosopumilales contributed less to the actively dividing than to the bulk communities during all the study period. Noticeably, the large contribution of these two taxa to the total prokaryotic communities (23% SAR11 and 26% Nitrosopumilales), especially in the meso- and bathypelagic zones, results in important contributions to actively dividing communities (11% SAR11 and 12% Nitrosopumilales). The intense temporal and spatial variability of actively dividing communities revealed in this study strengthen the view of a highly dynamic deep ocean. Our results suggest that some rare or low abundant phylotypes from surface layers down to the deep sea can disproportionally contribute to the activity of the prokaryotic communities, exhibiting a more dynamic response to environmental changes than other abundant phylotypes, emphasizing the role they might have in community metabolism and biogeochemical processes.This research has been supported by RADMED-TRES (2015–2019) and ATHAPOC (CTM2014-54374-R) projects, funded by the Spanish Institute of Oceanography and the Spanish Ministry of Economy and Competitiveness, respectively. CM was supported by pre-doctoral FPI fellowship from Conselleria d’Innovació, Recerca i Turisme of the regional Government of the Balearic Islands, co-financed by the European Social Fund as part of the FSE 2014-2020 operational program.Postprin