Relative abundance of prokaryotes in sediments of the Håkon Mosby mud volcano (Z1-Z3, all habitats)

CARD-FISH was performed as previously described in Ruff et al., (2013; doi:10.1371/journal.pone.0072627) with the following modifications. 4-6 µl of 25-fold diluted sediment were used for filtration. Archaeal cell walls were permeabilized with 0.1M HCl for 2 min to detect ANME-3 cells, or Proteinase K solution (15 µg ml-1 (Merck, Darmstadt, Germany) in 0.05 M EDTA (pH 8), 0.1 M Tris-HCl (pH 8), 0.5 M NaCl) for 2-4 min at room temperature for all other archaea. Bacterial cell walls were permeabilized with lysozyme solution (1000kU/ml) for 60 min at 37°. Cells were stained with DAPI (1µg/ml), embedded in mounting medium and counted in 40-60 independent microscopic fields using an Axiophot II epifluorescence microscope (Carl Zeiss, Jena, Germany).

Macrophage migration inhibitory factor (MIF) family in arthropods : cloning and expression analysis of two MIF and one D-dopachrome tautomerase (DDT) homologues in Mud crabs, Scylla paramamosain

5. Acknowledgements This research was supported by grants from the National Natural Science Foundation of China (Nos. 31172438 and U1205123), the Natural Science Foundation of Fujian Province (No. 2012J06008 and 201311180002) and the projects-sponsored by SRF. TW received funding from the MASTS pooling initiative (The Marine Alliance for Science and Technology for Scotland) funded by the Scottish Funding Council (grant reference HR09011) and contributing institutions.

Biogeochemical investigation of sediments at the Hakon Mosby Mud Volcano (Barents Sea)

We have investigated if in a cold seep methane or sulfide is used for chemosynthetic primary production and if significant amounts of the sulfide produced by anaerobic oxidation of methane are oxidized geochemically and hence are not available for chemosynthetic production. Geochemically controlled redox reactions and biological turnover were compared in different habitats of the Håkon Mosby Mud Volcano. The center of the mud volcano is characterized by the highest fluid flow, and most primary production by the microbial community depends on oxidation of methane. The small amount of sulfide produced is oxidized geochemically with oxygen or is precipitated with dissolved iron. In the medium flow peripheral Beggiatoa habitat sulfide is largely oxidized biologically. The oxygen and nitrate supply is high enough that Beggiatoa can oxidize the sulfide completely, and chemical sulfide oxidation or precipitation is not important. An internally stored nitrate reservoir with average concentrations of 110 mmol L-1 enables the Beggiatoa to oxidize sulfide anaerobically. The pH profile indicates sequential sulfide oxidation with elemental sulfur as intermediate. Gray thiotrophic mats associated with perturbed sediments showed a high heterogeneity in sulfate turnover and high sulfide fluxes, balanced by the opposing oxygen and nitrate fluxes so that biological oxidation dominates over geochemical sulfide removal processes. The three habitats indicate substantial small-scale variability in carbon fixation pathways either through direct biological use of methane or through indirect carbon fixation of methane-derived carbon dioxide by chemolithotrophic sulfide oxidation.

Hydrocarbon analyses of submarine mud volcanoes (MV) in the Kumano forearc basin

Twelve submarine mud volcanoes (MV) in the Kumano forearc basin within the Nankai Trough subduction zone were investigated for hydrocarbon origins and fluid dynamics. Gas hydrates diagnostic for methane concentrations exceeding solubilities were recovered from MVs 2, 4, 5, and 10. Molecular ratios (C1/C2<250) and stable carbon isotopic compositions (d13C-CH4 >-40 per mil V-PDB) indicate that hydrate-bound hydrocarbons (HCs) at MVs 2, 4, and 10 are derived from thermal cracking of organic matter. Considering thermal gradients at the nearby IODP Sites C0009 and C0002, the likely formation depth of such HCs ranges between 2300 and 4300 m below seafloor (mbsf). With respect to basin sediment thickness and the minimum distance to the top of the plate boundary thrust we propose that the majority of HCs fueling the MVs is derived from sediments of the Cretaceous to Tertiary Shimanto belt below Pliocene/Pleistocene to recent basin sediments. Considering their sizes and appearances hydrates are suggested to be relicts of higher MV activity in the past, although the sporadic presence of vesicomyid clams at MV 2 showed that fluid migration is sufficient to nourish chemosynthesis-based organisms in places. Distributions of dissolved methane at MVs 3, 4, 5, and 8 pointed at fluid supply through one or few MV conduits and effective methane oxidation in the immediate subsurface. The aged nature of the hydrates suggests that the major portion of methane immediately below the top of the methane-containing sediment interval is fueled by current hydrate dissolution rather than active migration from greater depth.

X-ray fluorescent (XRF) ratios and diatom groups of sediment core GeoB13813-4

The continental shelf adjacent to the Río de la Plata (RdlP) exhibits extremely complex hydrographic and ecological characteristics which are of great socioeconomic importance. Since the long-term environmental variations related to the atmospheric (wind fields), hydrologic (freshwater plume), and oceanographic (currents and fronts) regimes are little known, the aim of this study is to reconstruct the changes in the terrigenous input into the inner continental shelf during the late Holocene period (associated with the RdlP sediment discharge) and to unravel the climatic forcing mechanisms behind them. To achieve this, we retrieved a 10 m long sediment core from the RdlP mud depocenter at 57 m water depth (GeoB 13813-4). The radiocarbon age control indicated an extremely high sedimentation rate of 0.8 cm per year, encompassing the past 1200 years (AD 750-2000). We used element ratios (Ti / Ca, Fe / Ca, Ti / Al, Fe / K) as regional proxies for the fluvial input signal and the variations in relative abundance of salinity-indicative diatom groups (freshwater versus marine-brackish) to assess the variability in terrigenous freshwater and sediment discharges. Ti / Ca, Fe / Ca, Ti / Al, Fe / K and the freshwater diatom group showed the lowest values between AD 850 and 1300, while the highest values occurred between AD 1300 and 1850. The variations in the sedimentary record can be attributed to the Medieval Climatic Anomaly (MCA) and the Little Ice Age (LIA), both of which had a significant impact on rainfall and wind patterns over the region. During the MCA, a weakening of the South American summer monsoon system (SAMS) and the South Atlantic Convergence Zone (SACZ), could explain the lowest element ratios (indicative of a lower terrigenous input) and a marine-dominated diatom record, both indicative of a reduced RdlP freshwater plume. In contrast, during the LIA, a strengthening of SAMS and SACZ may have led to an expansion of the RdlP river plume to the far north, as indicated by higher element ratios and a marked freshwater diatom signal. Furthermore, a possible multidecadal oscillation probably associated with Atlantic Multidecadal Oscillation (AMO) since AD 1300 reflects the variability in both the SAMS and SACZ systems.

(Table 2) Age determination of brock-red sandy mud beds and detrital carbonate beds in the Scotian margin

Piston cores from the continental margin off Nova Scotia show up to four discrete intervals of "brick-red sandy mud", which are up to 20 cm thick. The ages of these intervals are bracketed by several radiocarbon dates, and three fall in the range 12.5-14.1 ka (radiocarbon years with -0.4 kyr reservoir correction). The youngest dates from ~10.4 ka, placing it within the Younger Dryas. The distribution of the beds and their petrographic character indicate a source in the Gulf of Saint Lawrence. The grain size of these beds suggests that they comprise a coarse component transported by ice rafting that diminishes distally and a fine component that represents suspension fallout from a surface plume and resulting nepheloid layers. Graded brick-red beds in some cores were probably redeposited from turbidity currents. The lowermost bed on the Laurentian Fan and East Scotian Rise is immediately overlain by a carbonate-rich interval that can be identified all around the margin of the Grand Banks. This interval is correlated with detrital carbonate bed DC-1 in the Labrador Sea and Heinrich event H1 in the North Atlantic. The sequential occurrence of the two beds suggests that they may be a response to the same trigger, probably sea level rise, but that the Gulf of Saint Lawrence source was more easily destabilized.

Temperature measurements, gas chemistry, and gas hydrates of the Dvurechenskii mud volcano

The sediment temperature distribution at mud volcanoes provides insights into their activity and into the occurrence of gas hydrates. If ambient pressure and temperature conditions are close to the limits of the gas hydrate stability field, the sediment temperature distribution not only limits the occurrence of gas hydrates, but is itself influenced by heat production and consumption related to the formation and dissociation of gas hydrates. Located in the Sorokin Trough in the northern Black Sea, the Dvurechenskii mud volcano (DMV) was in the focus of detailed investigations during the M72/2 and M73/3a cruises of the German R/V Meteor and the ROV Quest 4000 m in February and March 2007. A large number of in-situ sediment temperature measurements were conducted from the ROV and with a sensor-equipped gravity corer. Gas hydrates were sampled in pressurized cores using a dynamic autoclave piston corer (DAPC). The thermal structure of the DMV suggests a regime of fluid flow at rates decreasing from the summit towards the edges of the mud volcano, accompanied by intermittent mud expulsion at the summit. Modeled gas hydrate dissociation temperatures reveal that the gas hydrates at the DMV are very close to the stability limits. Changes in heat flow due to variable seepage rates probably do not result in changes in sediment temperature but are compensated by gas hydrate dissociation and formation.

Pore water measurements of sediment cores from the Helgoland mud area, North Sea

Iron reduction in subseafloor sulfate-depleted and methane-rich marine sediments is currently a subject of interest in subsurface geomicrobiology. While iron reduction and microorganisms involved have been well studied in marine surface sediments, little is known about microorganisms responsible for iron reduction in deep methanic sediments. Here, we used quantitative PCR (Q-PCR)-based 16S rRNA gene copy numbers and pyrosequencing-based relative abundances of bacteria and archaea to investigate covariance between distinct microbial populations and specific geochemical profiles in the top 5 m of sediment cores from the Helgoland mud area, North Sea. We found that gene copy numbers of bacteria and archaea were specifically higher around the peak of dissolved iron in the methanic zone (250-350 cm. The higher copy numbers at these depths were also reflected by the relative sequence abundances of members of the candidate division JS1, methanogenic and Methanohalobium/ANME-3 related archaea. The distribution of these populations was strongly correlated to the profile of pore-water Fe2+ while that of Desulfobacteraceae corresponded to the pore-water sulfate profile. Furthermore, specific JS1 populations also strongly co-varied with the distribution of Methanosaetaceae in the methanic zone. Our data suggest that the interplay among JS1 bacteria, methanogenic archaea and Methanohalobium/ANME-3-related archaea may be important for iron reduction and methane cycling in deep methanic sediments of the Helgoland mud area and perhaps in other methane-rich depositional environments. .