Accumulator-Based Deep-Sea Microbe Gastight Sampling Technique

1.IntroductionPlenty of microbes exist in deep sea(Jiang,2000).They can survive and propagate under theconditions of extraordinary cold or heat and high pressure(Liu,2001).Studies on deep-sea microbesshowthat they have the function of decomposing petroleu…     

Open-ocean deployment of a buoy-mounted aerosol sampler on the Bermuda Testbed Mooring: Aerosol iron and sea salt over the Sargasso Sea

We report results from the first deployment of a buoy-mounted aerosol sampler on the Bermuda Testbed Mooring (BTM) in the Sargasso Sea, in which a time-series of 21 aerosol samples were collected over the period May 5–September 29, 2004. These aerosol samples were analyzed for iron and soluble sodium (as a proxy for sea salt). Also analyzed was a time-series of 22 aerosol samples collected over the same period at the Tudor Hill atmospheric sampling tower on Bermuda. The buoy sampler worked as intended and successfully collected a time-series of aerosol samples, thus demonstrating that moored buoys can be used as oceanic observatories to provide information on the temporal (weekly, monthly and seasonal) variability in the concentration of aerosol iron (and other trace elements) over the surface ocean. The magnitude and time variation of aerosol Fe concentrations calculated from the BTM buoy samples are in close agreement with the corresponding aerosol Fe record from the Tudor Hill tower, which is located approximately 80 km northwest of the mooring site. Both the BTM and Tudor Hill samples record periods of high aerosol iron loadings in late June and late July 2004, reflecting the transport of soil dust from North Africa, with the highest concentration of aerosol iron at the BTM site (0.83 μg m⁻³) measured in late June. Concentrations of sea-salt aerosol calculated from the BTM samples are comparable to values measured over the Sargasso Sea and for samples collected at the Tudor Hill tower. Sea-salt aerosols do not appear to impede the collection of mineral aerosols by the buoy-mounted sampler.     

Large but variable particulate flux in the Antarctic Ocean and its significance for the chemistry of Antarctic water

Settling particles were collected at 1,460 m and 3,760 m depth in the Antarctic Ocean with sediment traps of time series type. The total deployment period of 40 days was divided into four terms of 10 days each. Seawater samples were collected both at deployment and retrieval of the traps at each site. During the 42 days the concentration of silicate in the surface water decreased by 32%, whereas those of nitrate and phosphate decreased by only 4–5%. The total particulate flux in the Antarctic Ocean is the largest among those hitherto observed in the world ocean. The time variation of the particulate flux at 1,460 m depth almost coincided with that at 3,760 m. The settling particles were comprised roughly of 80% biogenic silica, 15% organic matter and 5% other substances including sea salt. The clay fraction was only 0.05% at 1,460 m depth. The settling flux of biogenic silica agrees fairly well with the calculated rate of change in the concentration of silicate in the surface 100 m. Thus it is concluded that preferential propagation of diatoms reduces the concentration of silicate prior to other nutrients in the Antarctic Ocean.     

Target Deployment and Retrieval Using JIAOLONG Manned Submersible in the Depth of 6600 m in Mariana Trench

China''s 7000 m manned submersible JIAOLONG carried out an exploration cruise at the Mariana Trench from June to July 2016. The submersible completed nine manned dives on the north and south area of the Mariana Trench from the depth of 5500 to 6700 m, to investigate the geological, biological and chemical characteristics in the hadal area. During the cruise, JIAOLONG deployed a gas-tight serial sampler to collect the water near the sea bottom regularly. Five days later, the sub located the sampler in another dive and retrieved it successfully from the same location, which is the first time that scientists and engineers finished the high accuracy in-situ deployment and retrieval using a manned submersible with Ultra-Short Base Line (USBL) positioning system at the depth more than 6600 m. In this task, we used not only the USBL system of the manned submersible but also a compound strategy, including five position marks, the sea floor terrain, the depth contour, and the heading of the sub. This paper introduces the compound strategy of the target deployment and retrieval with the practical diving experience of JIAOLONG, and provides a promising technique for other underwater vehicles such as manned submersible or Remote Operated Vehicle (ROV) under similar conditions.     

A new gas-tight isobaric sampler for hydrothermal fluids

A new gas-tight isobaric sampler for the collection of hydrothermal fluids venting at the seafloor has been designed, constructed, and tested at a ridge-crest vent site. The new device is constructed of chemically inert titanium, is gas-tight to 450 bar and can be used to sample fluids with temperatures up to 400°C. Compressed gas is used to maintain the sample at seafloor pressure before and during sample withdrawal onboard ship, allowing subsampling without degassing the fluid remaining in the sampler. This feature eliminates the need to collect separate gas-tight and major element samples since a single fluid sample can be analyzed quantitatively for major, trace, semi-volatile, and volatile components. The sampler fill rate is regulated to minimize entrainment of ambient seawater during collection of fluids from environments characterized by low fluid flow such as diffuse hydrothermal vents. In addition to deployment at the ridge-crest, the samplers can be used to collect gas-tight samples from other subseasurface environments such as hydrocarbon seeps, areas of methane-gas hydrate formation, cold seeps associated with serpentinites, regions of groundwater egress to the oceans, and the water column.     

Probability distribution of wave heights attenuated by salt marsh vegetation during tropical cyclone

Zero-crossing wave heights, obtained from the field measurement of random waves propagating through salt marsh vegetation (Spartina alterniflora) during a tropical storm, were analyzed to examine their probability distribution. Wave data (significant wave heights up to 0.4 m in 0.8 m depth) were collected over a two-day period along a 28 m transect using three pressure transducers sampling at 10 Hz. Wave height distribution was observed to deviate from the Rayleigh distribution. The observed probability densities of the larger wave heights were reduced significantly by vegetation, producing wave heights lower than those predicted by the Rayleigh distribution. Assuming Rayleigh distributed wave heights for the incident waves to the vegetation patch, existing vegetation-induced wave attenuation formulations are used to derive a special form of two-parameter Weibull distribution for wave heights in the inundated wetland. The scale parameter of the distribution is theoretically shown to be a function of the shape parameter, which agrees with the measurements, effectively reducing the proposed distribution to a one-parameter type. The derived distribution depends on the local parameters only and fits well to the observed distribution of wave heights attenuated by vegetation. Empirical relationships are developed to estimate the shape parameter from the local wave parameters.     

Linking the Acadian Orogeny with organic-rich black shale deposition: Evidence from the Marcellus Shale

The trace and rare earth elements (REE) analyses were conducted on samples collected from a 30 m core of the Marcellus Shale obtained from Greene County, southwestern Pennsylvania. Our results suggest that organic matter enrichment trends in the Marcellus Shale can be directly linked with the Acadian Orogeny. The Acadian Orogeny has been recognized as a main sediment source for the Marcellus Shale. Synthesis of tectonic history and recent ash bed geochronology, reveals that deposition of the organic carbon-rich (OR) zone (characterized by TOC >4%; located between 2393 m and 2406.5 m core depth) in the studied Marcellus Shale core was coincident with tectonically active and magmatic quiescent period of the Acadian orogeny (ca. 395–380 Ma). This time period also corresponds to the highest rate of mountain building in the Acadian Orogeny. The light rare earth (LREE) and selected trace elemental (e.g., Ta, Cs) composition of the OR zone sediments is similar to that of the bulk continental crust, supporting the lack of magmatic activity in the source area (i.e. Acadian Orogeny). In contrast, subsequent deposition of the organic carbon-poor (OP) sediments (characterized by TOC <4%; located between 2376 m and 2393 m core depth) in the upper Marcellus Shale occurred synchronously with a magmatic active phase (ca. 380–370 Ma) during the Acadian orogeny. The OP zone sediments have LREE and trace elemental composition similar to the average of the upper continental crust, suggesting intrusion of granodiorite rocks during a magmatic active period of Acadian Orogeny. The temporal and geochemical correlation between the Acadian orogenesis and the Marcellus deposition provide evidence for the role of tectonism in the enrichment of organic matter in the Marcellus Shale.     

Microbial community distribution in sediments from the Mount Elbert Gas Hydrate Stratigraphic Test Well, Alaska North Slope

We studied the microbial communities collected from hydrate-bearing sediments on the North Slope of Alaska to determine how abiotic variables (e.g., grain size, hydrate presence, formation fluid gases) may correspond to the type and distribution of microbes in the sediments. The cores were acquired from sub-permafrost, Eocene (46-55 million year old) sediments in the BPXA-DOE-USGS Mount Elbert Gas Hydrate Stratigraphic Test Well within which hydrates are believed to have formed 1.5 mya. Forty samples, eight of which originally contained hydrates, were acquired from depths of ca. 606-666 m below land surface. Five drilling fluid samples acquired from the same depth range were included in the analysis as a control for possible contamination by drilling fluid microbes during the drilling and handling of cores. DNA was extracted from 15 samples (typically <1 ng DNA/g sediment was recovered) and then amplified using polymerase chain reaction with primers specific for bacterial and archaeal 16S rDNA genes, which indicates the likelihood that microbes were present in all analyzed samples. Only bacterial DNA amplicons were detected. Terminal-restriction fragment length polymorphism (T-RFLP) was used to measure bacterial diversity in the respective samples. Non-metric multidimensional scaling (NMS) was used to determine the abiotic variables that may have influenced bacterial diversity. NMS analysis revealed that the microbial taxa present in the sediment were distinct from the taxa present in the drilling fluids suggesting that the sediments were not contaminated by the drilling fluids. Multi-response permutation procedures (MRPP) found no significant difference between three sample groups identified a priori as being from within a hydrate zone, outside of hydrate zone, or on the edge of a hydrate/non-hydrate zone according to downhole nuclear magnetic resonance spectroscopy logs. However, among the several other abiotic parameters that were evaluated mud gas methane concentration and variables related to hydrate presence (e.g., chloride concentration, salinity, and resistivity) appeared to define the arrangement of microbial community signatures in plots resulting from NMS analysis. Communities from hydrate and non-hydrate layers each contained unique taxa as determined by the T-RFLP assay.     

Benthic Disturbance and Impact Experiments in the Central Indian Ocean Basin

As a part of the Environmental Impact Assessment studies for nodule mining, a long-term program has been initiated in the Central Indian Basin. Multidisciplinary studies on geological, biological, physical, and chemical parameters were carried out in an area selected on the basis of baseline data collected in the first phase of the program. A benthic disturbance was simulated with a hydraulic device also used in the previous experiments in the Pacific Ocean. A site of 3,000 ×200 m was repeatedly disturbed by a combination of fluidizing pump and suction pump to dislodge and discharge sediment from the seafloor into the water column 5 m above the seafloor. During 9 days of operation, 26 tows were carried out for 47 h of disturbance, resuspending about 6,000 m 3 of sediment along an 88-km line. Data for postdisturbance impact assessment were collected with sediment traps, deep-towed cameras, seafloor samples, and conductivity-temperature depth sensor (CTD)-rosette observations. Seafloor data, sediment samples, and water column studies were aimed at evaluating the impact of benthic disturbance, on the basis of pre- and postdisturbance data collected during the experiment. Observations show that vertical mixing of sediment as well as its lateral movement and resedimentation because of plume migration alters various parameters and leads to changes in the environment around the area.     

Effects of hydrostatic pressure on microbial alteration of sinking fecal pellets

We used a new experimental device called PASS (PArticle Sinking Simulator) during MedFlux to simulate changes in in situ hydrostatic pressure that particles experience sinking from mesopelagic to bathypelagic depths. Particles, largely fecal pellets, were collected at 200 m using a settling velocity NetTrap (SV NetTrap) in Ligurian Sea in April 2006 and incubated in high-pressure bottles (HPBs) of the PASS system under both atmospheric and continuously increasing pressure conditions, simulating the pressure change experienced at a sinking rate of 200 m d⁻¹. Chemical changes over time were evaluated by measuring particulate organic carbon (POC), carbohydrates, transparent exopolymer particles (TEP), amino acids, lipids, and chloropigments, as well as dissolved organic carbon (DOC) and dissolved carbohydrates. Microbial changes were evaluated microscopically, using diamidinophenylindole (DAPI) stain for total cell counts and catalyzed reporter deposition-fluorescence in situ hybridization (CARD-FISH) for phylogenetic distinctions. Concentrations (normalized to POC) of particulate chloropigments, carbohydrates and TEP decreased under both sets of incubation conditions, although less under the increasing pressure regime than under atmospheric conditions. By contrast, dissolved carbohydrates (normalized to DOC) were higher after incubation and significantly higher under atmospheric conditions, suggesting they were produced at the expense of the particulate fraction. POC-normalized particulate wax/steryl esters increased only under pressure, suggesting biochemical responses of prokaryotes to the increasing pressure regime. The prokaryotic community initially consisted of 43% Bacteria, 12% Crenarchaea and 11% Euryarchaea. After incubation, Bacteria dominated (90%) the prokaryote community in all cases, with γ-Proteobacteria comprising the greatest fraction, followed by the Cytophaga–Flavobacter cluster and α-Proteobacteria group. Using the PASS system, we obtained chemical and microbial evidence that degradation by prokaryotes associated with fecal pellets sinking through mesopelagic waters is limited by the increasing pressure they experience.     

A suspended-particle rosette multi-sampler for discrete biogeochemical sampling in low-particle-density waters

To enable detailed investigations of early stage hydrothermal plume formation and abiotic and biotic plume processes we developed a new oceanographic tool. The Suspended Particulate Rosette sampling system has been designed to collect geochemical and microbial samples from the rising portion of deep-sea hydrothermal plumes. It can be deployed on a remotely operated vehicle for sampling rising plumes, on a wire-deployed water rosette for spatially discrete sampling of non-buoyant hydrothermal plumes, or on a fixed mooring in a hydrothermal vent field for time series sampling. It has performed successfully during both its first mooring deployment at the East Pacific Rise and its first remotely-operated vehicle deployments along the Mid-Atlantic Ridge. It is currently capable of rapidly filtering 24 discrete large-water-volume samples (30–100 L per sample) for suspended particles during a single deployment (e.g. >90 L per sample at 4–7 L per minute through 1 μm pore diameter polycarbonate filters). The Suspended Particulate Rosette sampler has been designed with a long-term goal of seafloor observatory deployments, where it can be used to collect samples in response to tectonic or other events. It is compatible with in situ optical sensors, such as laser Raman or visible reflectance spectroscopy systems, enabling in situ particle analysis immediately after sample collection and before the particles alter or degrade.     

Seasonal dynamics of inorganic nutrients and phytoplankton biomass in the NW Alboran Sea

The seasonal dynamics of inorganic nutrients and phytoplankton biomass (chlorophyll a), and its relation with hydrological features, was studied in the NW Alboran Sea during four cruises conducted in February, April, July and October 2002. In the upper layers, the seasonal pattern of nutrient concentrations and their molar ratios (N:Si:P) was greatly influenced by hydrological conditions. The higher nutrient concentrations were observed during the spring cruise (2.54 μM NO₃⁻, 0.21 μM PO₄³⁻ and 1.55 μM Si(OH)₄, on average), coinciding with the increase of salinity due to upwelling induced by westerlies. The lowest nutrient concentrations were observed during summer (<0.54 μM NO₃⁻, 0.13 μM PO₄³⁻ and 0.75 μM Si(OH)₄, on average), when the lower salinities were detected. Nutrient molar ratios (N:Si:P) followed the same seasonal pattern as nutrient distribution. During all the cruises, the ratio N:P in the top 20 m was lower than 16:1, indicating a NO₃⁻ deficiency relative to PO₄³⁻. The N:P ratio increased with depth, reaching values higher than 16:1 in the deeper layers (200–300 m). The N:Si ratio in the top 20 m was lower than 1:1, excepting during spring when N:Si ratios higher than 1:1 were observed in some stations due to the upwelling event. The N:Si ratio increased with depth, showing a maximum at 50–100 m (>1.5:1), which indicates a shift towards Si-deficiency in these layers. The Si:P ratio was much lower than 16:1 throughout the water column during the four cruises. In general, the spatial and seasonal variation of phytoplankton biomass showed a strong coupling with hydrological and chemical fields. The higher chlorophyll a concentrations at the depth of the chlorophyll maximum were found in April (2.57 mg m⁻³ on average), while the lowest phytoplankton biomass corresponded to the winter cruise (0.74 mg m⁻³ on average). The low nitrate concentrations together with the low N:P ratios found in the upper layers (top 20 m) during the winter, summer and autumn cruises suggest that N-limitation could occur in these layers during great part of the year. However, N-limitation during the spring cruise was temporally overcome by nutrient enrichment caused by an intense wind-driven upwelling event.     

A Remotely Operated Serial Sampler for Collecting Gas-Tight Fluid Samples

This paper describes the design, construction and preliminary test results for a gas-tight serial sampler intended to be deployed at seafloor for long-term operation to take time-series fluid samples from deep-sea environments such as cold seeps, water column and hydrothermal vents. The serial sampler is a modular system that is based on independent and identical sampling modules, which are designed to collect six 160 ml gas-tight fluid samples maintained at high pressure to a depth of 4000 meters. With two working modes, the sampler can be deployed either with seafloor cabled observatory for remote control or as a stand-alone device for autonomous operation. A prototype of the instrument has been constructed and tested on the MARS cabled observatory for two months. The laboratory and field tests proved the success of the design and construction of the serial sampler, and indicated the potential for future ocean sciences.     

Downward fluxes of settling particles in the deep Cretan Sea (NE Mediterranean)

During the CINCS project (Pelagic–benthic Coupling IN the oligotrophic Cretan Sea—NE Mediterranean), a single mooring with two sediment traps (at 200 and 1515m water depth) and two current meters was deployed in the southern Cretan Sea margin at a depth of 1550 m. A second mooring deployed at the 500 m station was lost, as a result of fishing activities. The duration of the study was 12 months (November 1994 to November 1995) with sampling intervals of 15 or 16 days. The traps were retrieved, serviced and the sedimented material was collected every 6 months. In total, 48 samples were collected (24 from each trap) throughout the study period and fluxes of total particulate mass, opal, organic matter, carbonates, and lithogenic component were measured. Natural radionuclides (²¹⁰Po and ²¹⁰Pb) were determined for all trap samples. Total mass flux and the fluxes of four major constituents increased with depth, the total mass flux reaching values of nearly 550 mg m⁻² d⁻¹ at 1515 m and 187 mg m⁻² d⁻¹ at 200 m depth, following the same patterns observed in other experiments (ECOMARGE, SEEP-I, SEEP-II). The mean annual mass fluxes were 209 and 49.8 mg m⁻² d⁻¹ at the near bottom and near surface trap respectively. This suggests that lateral transport of particulate matter is of importance in the area. Total mass fluxes at the two depths were characterized by different seasonal fluctuations, although a general decreasing trend was observed from the I (winter) to the II (summer) deployment at both depths. This was mainly a result of reductions in aluminosilicate inputs during the summer dry period. At 200 m depth carbonates were more important during winter, because of a large carbonate input consisting mainly of coccoliths of Emiliania huxleyi, while during the summer decreased fluxes of carbonates and aluminosilicates resulted in a reduction of the mass flux. In contrast, at 1515 m depth the lithogenic component was the dominant component during the winter deployment, indicating a terrigenous input. During the summer period the decrease in mass flux was strongly effected by the decrease in aluminosilicates. There was a diminution in the organic carbon content with a concomitant increase in total mass flux, which, together with the almost negligible increase in the annual ²¹⁰Pb activity with depth and the increase of ²¹⁰Po activity with depth could be interpreted as indicating a contribution of resuspended material to the input at 1515 m. The complex mesoscale circulation of the Cretan Sea, consisting of a cyclone (east)–anticyclone (west) system, controls particle transfer in the area. This hydrodynamic system seems to move water masses towards the southern Cretan Sea margin, and consequently carry materials from the open sea to the upper slope and shelf.     

Satellite remote sensing of the geographical distribution of suspended particle size in an energetic shelf sea

Natural and created Spartina brackish marsh habitats in the Guadalupe Estuary, adjacent to the Aransas National Wildlife Refuge, Texas, USA were surveyed during spring, summer, and fall 2004 to evaluate the equivalence of nekton assemblages in an old (>30 years) created marsh. During each season, six replicate samples were collected in each marsh type using a 1-m² drop sampler. Multivariate analysis revealed significant differences in nekton assemblage structure among marsh type, both within and across seasons. Species richness was significantly higher in the natural marsh in spring and summer but not in fall. Several species that were dominant in the natural marsh but rare or absent in the created marsh had strong correlations with the presence of oyster substrate that was only encountered in natural marsh samples. Although cumulative richness was greater in the natural marsh, eight species were collected only from the created marsh. Shrimp and fish biomass was significantly higher in natural marsh. Analysis of the density, biomass and size structure of three commercially important crustaceans indicated that the created marsh supported similar biomass of some species (white shrimp, blue crab); however, the size structure of some populations was variable among marshes (blue crab, brown shrimp). We conclude that lower substrate complexity (specifically oyster) and soil organic content in the created marsh reduced measures of nekton similarity and recommend that these features be addressed in future restoration efforts.     

Diel microbial variations at a coastal Northern Adriatic station affected by Po River outflows

Diel sampling was performed during an early spring survey in the Northern Adriatic Sea at a coastal station off the Po River delta. Samples were taken every 6 h at spring tide maxima and minima in the sub-superficial layer, at the maximum fluorescence depth (∼3 m). Variations in microbial community structure and its processes were assessed by considering heterotrophic bacteria, picocyanobacteria, viruses, exoenzymatic activities, microphytoplankton, nanoplankton and bacterial/cyanobacterial Denaturing Gradient Gel Electrophoresis (DGGE) profiles. A considerable diatom bloom, mostly supported by Skeletonema marinoi was detected. All microbial parameters except viruses, showed a sinusoidal trend with a 12 h period; only picocyanobacteria expressed relative maxima during high tide, showing a phase in opposition to the other parameters. No substantial changes in DGGE band patterns were detected. Even though the results showed bacterial activities to be influenced by the phytoplankton bloom, all microbial parameters' diel trends (except viruses) preferentially followed the tidal fluctuation rather than the light:dark cycle.     

Sediment transport in the Yellow Sea and East China Sea

Eight survey cruises in different seasons have been conducted in the Yellow Sea (YS) and East China Sea (ECS) during the period from 2000 to 2008. Suspended sediment concentration (SSC) and hydrological data were collected during each cruise. Data analysis showed that total suspended sediment mass was approximately 0.18 × 10⁹ tons in the surveyed area during spring and autumn seasons. Highly turbid waters were found in the shallow waters between the Subei coast, the Changjiang estuary and the Zhejiang coast with seasonal variations.     

Authigenic carbonates from methane seeps of the northern Congo fan: Microbial formation mechanism

Authigenic carbonates were collected from methane seeps at Hydrate Hole at 3113 m water depth and Diapir Field at 2417 m water depth on the northern Congo deep-sea fan during RV Meteor cruise M56. The carbonate samples analyzed here are nodules, mainly composed of aragonite and high-Mg calcite. Abundant putative microbial carbonate rods and associated pyrite framboids were recognized within the carbonate matrix. The δ¹³C values of the Hydrate Hole carbonates range from ?62.5‰ to ?46.3‰ PDB, while the δ¹³C values of the Diapir Field carbonate are somewhat higher, ranging from ?40.7‰ to ?30.7‰ PDB, indicating that methane is the predominant carbon source at both locations. Relative enrichment of ¹⁸O (δ¹⁸O values as high as 5.2‰ PDB) are probably related to localized destabilization of gas hydrate. The total content of rare earth elements (REE) of 5% HNO₃-treated solutions derived from carbonate samples varies from 1.6 ppm to 42.5 ppm. The shale-normalized REE patterns all display positive Ce anomalies (Ce/Ce* > 1.3), revealing that the carbonates precipitated under anoxic conditions. A sample from Hydrate Hole shows a concentric lamination, corresponding to fluctuations in δ¹³C values as well as trace elements contents. These fluctuations are presumed to reflect changes of seepage flux.