Temporal variability of diazotroph community composition in the upwelling region off NW Iberia.

Knowledge of the ecology of N2-fixing (diazotrophic) plankton is mainly limited to oligotrophic (sub)tropical oceans. However, diazotrophs are widely distributed and active throughout the global ocean. Likewise, relatively little is known about the temporal dynamics of diazotrophs in productive areas. Between February 2014 and December 2015, we carried out 9 one-day samplings in the temperate northwestern Iberian upwelling system to investigate the temporal and vertical variability of the diazotrophic community and its relationship with hydrodynamic forcing. In downwelling conditions, characterized by deeper mixed layers and a homogeneous water column, non-cyanobacterial diazotrophs belonging mainly to nifH clusters 1G (Gammaproteobacteria) and 3 (putative anaerobes) dominated the diazotrophic community. In upwelling and relaxation conditions, affected by enhanced vertical stratification and hydrographic variability, the community was more heterogeneous vertically but less diverse, with prevalence of UCYN-A (unicellular cyanobacteria, subcluster 1B) and non-cyanobacterial diazotrophs from clusters 1G and 3. Oligotyping analysis of UCYN-A phylotype showed that UCYN-A2 sublineage was the most abundant (74%), followed by UCYN-A1 (23%) and UCYN-A4 (2%). UCYN-A1 oligotypes exhibited relatively low frequencies during the three hydrographic conditions, whereas UCYN-A2 showed higher abundances during upwelling and relaxation. Our findings show the presence of a diverse and temporally variable diazotrophic community driven by hydrodynamic forcing in an upwelling system

Development of Microsatellite Markers in the Razor Clam Solen Marginatus (Bivalvia: Solenidae)

[Abstract] Four microsatellite loci in the razor clam Solen marginatus are described. Loci were isolated from the sequences of intersimple sequence repeat (ISSR) markers and an enriched library. Detailed analysis of these sequences led to the design of eight primer pairs. Allelic variation was assessed in 20 individuals from Redondela, Spain. The genetic variation observed in the markers presented here will be useful for future studies on the population structure of Solen marginatus in the wild and for aquaculture of this species.Xunta de Galicia; PGIDT03MA10301PR

Isolation and characterization of polymorphic microsatellite loci in the razor clam "Ensis siliqua"

[Abstract] Five polymorphic microsatellite loci in the razor clam Ensis siliqua are described. A collection consisting of 34 individuals from Finisterre, Spain, was analysed. Loci were isolated from the sequences of intersimple sequence repeat (ISSR) markers. Detailed analysis of 42 ISSR markers led to the design of 16 primer pairs. Five of these yielded consistent and polymorphic products. The number of alleles ranged from five to 23 per locus with the observed heterozygosity ranging from 0.46 to 0.94. Linkage equilibrium was observed in all loci and three of them showed significant deviations from Hardy–Weinberg equilibrium

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

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

Unlocking the in vitroanti- inflammatory and antidiabetic potential of Polygonum maritimum

Context: Several Polygonum species (Polygonaceae) are used in traditional medicine in Asia, Europe and Africa to treat inflammation and diabetes. Objective: Evaluate the in vitro antioxidant, anti-inflammatory and antidiabetic potential of methanol and dichloromethane extracts of leaves and roots of the halophyte Polygonum maritimum L. Material and methods: Antioxidant activity was determined (up to 1mg/mL) as radical-scavenging activity (RSA) of 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), copper (CCA) and iron (ICA) chelating activities and iron reducing power (FRAP). NO production was measured in lipopolysaccharide (LPS)-stimulated macrophages for 24 h at concentrations up to 100 mu g/mL and antidiabetic potential was assessed by alpha-amylase and alpha-glucosidase inhibition (up to 10 g/mL) assays. The phytochemical composition of the extracts was determined by gas chromatography-mass spectrometry (GC-MS). Results: The methanol leaf extract had the highest activity against DPPH center dot (IC50 = 26 mu g/mL) and ABTS1(+)center dot (IC50 = 140 mu g FRAP (IC50 = 48 mu g/mL) and CCA (IC50 = 770 mu g/mL). Only the dichloromethane leaf extract (LDCM) showed anti-inflammatory activity (IC50 = 48 mu g/mL). The methanol root (IC50 = 19 mu g/mL) and leaf (IC50 = 29 mu g/mL) extracts strongly inhibited baker's yeast alpha-glucosidase, but LDCM had higher rat's alpha-glucosidase inhibition (IC50 = 2527 mu g/mL) than acarbose (IC50 = 4638 mu g/mL). GC-MS analysis identified beta-sitosterol, stigmasterol, 1-octacosanol and linolenic acid as possible molecules responsible for the observed bioactivities. Conclusions: Our findings suggest P. maritimum as a source of high-value health promoting commodities for alleviating symptoms associated with oxidative and inflammatory diseases, including diabetes.XtremeBio project - Foundation for Science and Technology (FCT) [PTDC/MAR-EST/4346/2012]; Portuguese National Budget; FCT [CCMAR/Multi/04326/ 2013, IF/00049/2012, SFRH/BPD/86071/2012, UID/Multi/00612/2013

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

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

Identification and characterization of miRNAs in spleens of sheep subjected to repetitive vaccination

Accumulative evidence has shown that short non-coding RNAs such as miRNAs can regulate the innate and adaptive immune responses. Aluminium hydroxide is a commonly used adjuvant in human and veterinary vaccines. Despite its extended use, its mechanism of action is not fully understood and very few in vivo studies have been done to enhance understanding at the molecular level. In this work, we took advantage of a previous long-term experiment in which lambs were exposed to three different treatments by parallel subcutaneous inoculations with aluminium-containing commercial vaccines, an equivalent dose of aluminium or mock injections. Spleen samples were used for miRNA-seq. A total of 46 and 16 miRNAs were found differentially expressed when animals inoculated with commercial vaccines or the adjuvant alone were compared with control animals, respectively. Some miRNAs previously related to macrophage polarization were found dysregulated exclusively by the commercial vaccine treatment but not in the aluminium inoculated animals. The dysregulated miRNAs in vaccine group let-7b-5p, miR-29a-3p, miR-27a and miR-101-3p are candidates for further research, since they may play key roles in the immune response induced by aluminium adjuvants added to vaccines. Finally, protein–protein interaction network analysis points towards leucocyte transendothelial migration as a specific mechanism in animals receiving adjuvant only