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

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

Changes in activity and community composition shape bacterial responses to size-fraccionated marine DOM

To study the response of bacteria to different size-fractions of naturally occurring dissolved organic matter (DOM), a natural prokaryotic community from North Atlantic mesopelagic waters (1000 m depth) was isolated and grown in (i) 0.1-µm filtered seawater (CONTROL), (ii) the low-molecular-weight (<1 kDa) DOM fraction (L-DOM), and (iii) the recombination of high- (>1 kDa) and low-molecular-weight DOM fractions (H + L-DOM), to test the potential effect of ultrafiltration on breaking the DOM size continuum. Prokaryotic abundance and leucine incorporation were consistently higher in the H + L-DOM niche than in the L-DOM and CONTROL treatments, suggesting a different interaction with each DOM fraction and the disruption of the structural DOM continuum by ultrafiltration, respectively. Rhodobacterales (Alphaproteobacteria) and Flavobacteriales (Bacteroidetes) were particularly enriched in L-DOM and closely related to the colored DOM (CDOM) fraction, indicating the tight link between these groups and changes in DOM aromaticity. Conversely, some other taxa that were rare or undetectable in the original bacterial community were enriched in the H + L-DOM treatment (e.g., Alteromonadales belonging to Gammaproteobacteria), highlighting the role of the rare biosphere as a seed bank of diversity against ecosystem disturbance. The relationship between the fluorescence of protein-like CDOM and community composition of populations in the H + L-DOM treatment suggested their preference for labile DOM. Conversely, the communities growing on the L-DOM niche were coupled to humic-like CDOM, which may indicate their ability to degrade more reworked DOM and/or the generation of refractory substrates (as by-products of the respiration processes). Most importantly, L- and/or H + L-DOM treatments stimulated the growth of unique bacterial amplicon sequence variants (ASVs), suggesting the potential of environmental selection (i.e., changes in DOM composition and availability), particularly in the light of climate change scenarios. Taken together, our results suggest that different size-fractions of DOM induced niche-specialization and differentiation of mesopelagic bacterial communities.Versión del edito

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

Vertical zonation of bacterial assemblages attributed to physical stratification during the summer relaxation of the coastal upwelling off Galicia (NW Spain)

Research articleWe combined flow cytometry, CARD-FISH, and 16S rRNA gene tag pyrosequencing to investigate bacterioplankton dynamics along a transect in shelf waters off A Coruña (Galicia, NW Spain). Over five days (16-20th July 2012) we sampled during the relaxation of a summer upwelling pulse, providing an opportunity to examine the impact of pulses of cold nutrient-rich water into coastal microbial communities. The hydrographic conditions, characterized by intense density stratification of surface waters and the presence of a deep chlorophyll maximum (DCM) at 20–30 m, were relatively maintained over the sampling period. Indeed, bacterial abundance and composition displayed low day to day variation. Alpha diversity analysis suggested that species richness and diversity increased from coastal to shelf stations and from the surface down to the coastal DCM, which could be caused by the mixing of upwelled bacteria with the coastal surface waters. SAR11, SAR86, and Roseobacter were the most abundant bacteria detected in the samples by using CARD-FISH. The assemblages observed by pyrosequencing displayed a strong vertical zonation along the transect. Rhodobacteraceae (under class Alphaproteobacteria) and Bacteriodetes dominated the surface waters and decreased during the upwelling pulse, while SAR 86 (under class Gammaproteobacteria), Actinobacteria and SAR11 clade increased their relative abundance at the coastal DCM with upwelling relaxation, particularly at the shelf stations. Bacterial assemblages from surface waters were associated with higher temperature and light conditions, while coastal DCM assemblages were rather associated to salinity, inorganic nutrients and a diatom-bloom leading to high chlorophyll-a. Our findings suggest that the vertical variability in environmental conditions induced by the intense density stratification, the exportation of warmer and less saline surface water from the rias to the adjacent shelf, and the fertilizing effect of recently upwelled water at the deeper layer, determined the composition of distinct bacterial assemblages at the subsurface and DCM layers.XUNTA DE GALICIA (O9MMA 027604PR, 10MMA604024PR), IEO (PROYECTO RADIALES) y PLAN NACIONAL (MODUPLAN CTM2011-24008)Versión del editor2,24

Bacterial community composition and optical signature of DOM shape empirical leucine-to-carbon conversion factors in north-eastern Atlantic waters (0-4000 m)

Oral communicationMicrobial heterotrophic activity is a major process regulating the flux of dissolved organic matter (DOM) in the ocean. DOM quantity and quality strongly influence its microbial utilization and fate in the ocean. In order to broaden the vertical resolution of leucine-to-carbon conversion factors (CFs), needed for converting substrate incorporation into biomass production by heterotrophic bacteria, nine dilution experiments were performed in the north Atlantic. We found a very consistent depth-stratification in empirical CFs values from epipelagic to bathypelagic waters (3.95 &#177; 0.05 to 0.90 &#177; 0.51 kg C mol Leu-1). Our results demonstrated that the customarily used CF of 1.55 kg C mol Leu-1 in oceanic waters leads to an underestimation of prokaryotic heterotrophic production in epi- and mesopelagic waters, while it causes a severe overestimation in bathypelagic waters. Pearson correlations showed that CFs were related not only to hydrographic variables but also to specific phylogenetic groups and DOM quality and quantity indicators. Furthermore, a multiple linear regression model predicting CFs from relatively simple hydrographic and optical spectroscopic measurements is provided. Taken together, our results suggest that differences in CFs throughout the water column might be mostly associated to the quality of DOM affecting the response of particular phylogenetic groups.ASL

Short term variability of bacterial communities in the shelf waters off Galicia (NW Iberian Peninsula): Patterns and drivers shaping the diversity

PósterEcological and biogeochemical processesEcological and biogeochemical processes in the ocean are dependent on a diverse assemblage of microbes, including members of Bacteria (Glöckner et al, 2012). These bacterial assemblages occurs in vast numbers and represent a huge genetic diversity, fulfilling a wide of ecological roles in the marine system such as carbon geochemical cycle and energy transfer into higher trophic levels.We combined flow cytometry, CARD-FISH and 16S rRNA gene tag pyrosequencing to investigate the short-term variability of the bacterial communities in samples collected along a transect in the shelf off NW Iberian Peninsula over one week

Diversity and abundance of planktonic communities in the deep waters off the galician coast (NW Spain)

Comunicación oralPlanktonic communities play pivotal roles within marine ecosystems, affecting their structure, functioning and services. Although they have been extensively studied in the epipelagic ocean, the knowledge about these communities in the dark ocean is rather short. In this study, we explored patterns of abundance and biomass of a wide variety of taxonomic groups from the prokaryotes to mesozooplankton in the epi-, meso- and bathypelagic waters off the Galician coast. As expected, ciliate and zooplankton abundances are depleted in the bathypelagic waters relative to abundances of prokaryotes and nanoflagellates. The rate of decrease of zooplankton biomass with depth is twice as that of prokaryotes and nanoflagellates, indicating that relative contribution of mesozooplancton to the total plankton biomass decreases with depth. Overall, the diversity of prokaryotes in the dark ocean is almost as high as in the epipelagic layer, although the phylotypes are different. The major fraction of epipelagic ciliates belongs to alloricate genera, whereas tintinnids dominate the deep ciliate populations. Small copepods were dominant in the epi- and meso-pelagic zone. By contrast, foraminiferans, big copepods and myctophic fishes were more abundant in the deep ocean

Optical properties of dissolved organic matter relate to different dept-specific patterns of archaeal and bacterial community structure in the North Atlantic Ocean

ArticleProkaryotic abundance, activity and community composition were studied in the euphotic, intermediate and deep waters off the Galician coast (NW Iberian margin) in relation to the optical characterization of dissolved organic matter (DOM). Microbial (archaeal and bacterial) community structure was vertically stratified. Among the Archaea, Euryarchaeota, especially Thermoplasmata, was dominant in the intermediate waters and decreased with depth, whereas marine Thaumarchaeota, especially Marine Group I, was the most abundant archaeal phylum in the deeper layers. The bacterial community was dominated by Proteobacteria through the whole water column. However, Cyanobacteria and Bacteroidetes ocurrence was considerable in the upper layer and SAR202 was dominant in deep waters. Microbial composition and abundande were not shaped by the quantity of dissolved organic carbon, but instead they revealed a strong connection with the DOM quality. Archaeal communities were mainly related to the fluorescence of DOM (which indicates respiration of labile DOM and generation of refractory subproducts), while bacterial communities were mainly linked to the aromaticity/age of the DOM produced along the water column. Taken together, our results indicate that the microbial community composition is associated with the DOM composition of the water masses, suggesting that distinct microbial taxa have the potential to use and/or produce specific DOM compounds.Versión del edito