Nematode abundance in cold seep sediments of the Nordic Margins (Nyregga, Styregga, Haakon Mosby Mud Volcano)

Cold-seep environments and their associated symbiont-bearing mega faunal communities create islands of primary production for macro-and meiofauna in the otherwise monotonous and nutrient-poor deep-sea environment. To examine the spatial variation and distribution patterns of metazoan meiobenthos in different seepage-related habitats, samples were collected in two regions off Norway: several pockmarks associated with the Storegga Slide including the Nyegga pockmark area, and the active, methane-venting Haakon Mosby Mud Volcano west of the Barents Sea during the Vicking cruise aboard the RV ''PourquoiPas?'' in May-June 2006. Meiofaunal samples at control sites were sampled with a multiple corer, while the other sites were sampled with push cores operated by the ROV Victor6000.The meiofaunal samples were fixed in 4% buffered formaldehyde and washed over a 32 mm-mesh sieve. Metazoan meiofauna were extracted by density gradient centrifugation. All material was fixed with 4% buffered formalin and stained with Rose Bengal. The metazoan meiofauna was sorted out, enumerated and identified down to major taxa under the stereomicroscope. Afterwards, abundances of Nematodes were depth integrated over the top 5 cm to gain individual abundances per 10 cm**2.

Abundance and biomass of phytoplankton in the western part of the Black Sea during Parshin cruise BSRP in September and October 2005

The dataset is composed of 61 samples from 15 stations. The phytoplankton samples were collected by 5l Niskin bottles attached to the CTD system. The sampling depths were selected according to the CTD profile and the in situ fluorometer readings: surface, temperature, salinity and fluorescence gradients and 1 m above the bottom. At some stations phytoplankton net samples (20 µm mesh-size) were collected to assist species biodiversity examination. The samples (1l sea water) were preserved in 4% buffered to pH 8-8.2 with disodiumtetraborate formaldehyde solution and stored in plastic containers. On board at each station few live samples were qualitatively examined under microscope for preliminary analysis of taxonomic composition and dominant species. Taxon-specific phytoplankton abundance were concentrated down to 50 cm**3 by slow decantation after storage for 20 days in a cool and dark place. The species identification was done under light microscope OLIMPUS-BS41 connected to a video-interactive image analysis system at magnification of the ocular 10X and objective - 40X. A Sedgwick-Rafter camera (1ml) was used for counting. 400 specimen were counted for each sample, while rare and large species were checked in the whole sample (Manual of phytoplankton, 2005). Species identification was mainly after Carmelo T. (1997) and Fukuyo, Y. (2000). The cell biovolume of the taxon-specific phytoplankton biomass was determined based on morpho-metric measurement of phytoplankton units and the corresponding geometric shapes as described in detail in (Edier, 1979).

Abundance of mesozooplankton from the S-IT3 cruise in the Sicily Channel in March 2008

The "SESAME_IT3_ZooAbundance_0-50-100m_SZN" dataset contains data of mesozooplankton species composition and abundance (ind. m-3) from samples collected in the Sicily Channel in the early spring of 2008 (17,18 March) during the SESAME-WP2 cruise IT3. Samples were collected by vertical tows with a closing WP2 net (56 cm diameter, 200 µm mesh size) in the following depth layers: 100-200 m, 50-100 m, 0-50 m. Sampling was always performed in light hours with the exception of station S-IT3-03 where zooplankton were collected in dark hours. A flowmeter was applied to the mouth of the net, however, due to its malfunctioning, the volume of filtered seawater was calculated by multiplying the the area by the height of the sampled layer from winch readings. After collection, each sample was split in two halves (1/2) after careful mixing with graduated beakers. Half sample was immediately fixed and preserved in a formaldehyde-seawater solution (4% final concentration) for species composition and abundance. The other half sample was kept fresh for biomass measurements (data already submitted to SESAME database in different files).Here, only the zooplankton abundance of samples in the upper layers 0-50 m and 50-100 m are presented. The abundance data of the samples in the layer 50-100 m will be submitted later in a separate file. The volume of filtered seawater was estimated by multiplying the the area by the height of the sampled layer from winch readings. Identification and counts of specimens were performed on aliquots (1/20-1/5) of the fixed sample or on the total sample (half of the original sample) by using a graduate large-bore pipette. Copepods were identified to the species level and separated into females, males and juveniles (copepodites). All other taxa were identified at the species level when possible, or at higher taxonomic levels. Taxonomic identification was done according to the most relevant and updated taxonomic literature. Total mesozooplankton abundance was computed as sum of all specific abundances determined as explained above.

Abundance and biomass of phytoplankton in the western part of the Black Sea during the R/V Akademik cruise Ak082001 in August 2001

The dataset is composed of 41 samples from 10 stations. The phytoplankton samples were collected by 5l Niskin bottles attached to the CTD system. The sampling depths were selected according to the CTD profile and the in situ fluorometer readings: surface, temperature, salinity and fluorescence gradients and 1 m above the bottom. At some stations phytoplankton net samples (20 µm mesh-size) were collected to assist species biodiversity examination. The samples (1l sea water) were preserved in 4% buffered to pH 8-8.2 with disodiumtetraborate formaldehyde solution and stored in plastic containers. On board at each station few live samples were qualitatively examined under microscope for preliminary analysis of taxonomic composition and dominant species. The taxon-specific phytoplankton abundance samples were concentrated down to 50 cm**3 by slow decantation after storage for 20 days in a cool and dark place. The species identification was done under light microscope OLIMPUS-BS41 connected to a video-interactive image analysis system at magnification of the ocular 10X and objective - 40X. A Sedgwick-Rafter camera (1ml) was used for counting. 400 specimen were counted for each sample, while rare and large species were checked in the whole sample (Manual of phytoplankton, 2005). Species identification was mainly after Carmelo T. (1997) and Fukuyo, Y. (2000). Total phytoplankton abundance was calculated as sum of taxon-specific abundances. Total phytoplankton biomass was calculated as sum of taxon-specific biomasses. The cell biovolume was determined based on morpho-metric measurement of phytoplankton units and the corresponding geometric shapes as described in detail in (Edier, 1979).

MARECHIARA-phytoplankton long-term time-series (1984-2006) at the fixed coastal station in the Gulf of Naples, Southern Tyrrhenian Sea

The "MARECHIARA-phytoplankton" dataset contains phytoplankton data collected in the ongoing time-series at Stn MC ( 40°48.5' N, 14°15' E) in the Gulf of Naples. This dataset spans over the period 1984-2006 and contains data of phytoplankton species composition and abundance. Phytoplankton sampling was regularly conducted from January 1984 till July 1991 and in 1995-2006. Sampling was interrupted from August 1991 till January 1995. The sampling frequency was fortnightly till 1991 and weekly since 1995. Phytoplankton samples were collected at 0.5 m depth using Niskin bottles and immediately fixed with formaldehyde (0.8-1.6% final concentration) for species identification and counts.

Geochemistry, isotopic ratios and aldehyde concentrations of frost flowers, snow and ice near Barrow, Alaska

Frost flowers, intricate featherlike crystals that grow on refreezing sea ice leads, have been implicated in lower atmospheric chemical reactions. Few studies have presented chemical composition information for frost flowers over time and many of the chemical species commonly associated with Polar tropospheric reactions have never been reported for frost flowers. We undertook this study on the sea ice north of Barrow, Alaska to quantify the major ion, stable oxygen and hydrogen isotope, alkalinity, light absorbance by soluble species, organochlorine, and aldehyde composition of seawater, brine, and frost flowers. For many of these chemical species we present the first measurements from brine or frost flowers. Results show that major ion and alkalinity concentrations, stable isotope values, and major chromophore (NO3- and H2O2) concentrations are controlled by fractionation from seawater and brine. The presence of these chemical species in present and future sea ice scenarios is somewhat predictable. However, aldehydes, organochlorine compounds, light absorbing species, and mercury (part 2 of this research and Sherman et al. (2012, doi:10.1029/2011JD016186)) are deposited to frost flowers through less predictable processes that probably involve the atmosphere as a source. The present and future concentrations of these constituents in frost flowers may not be easily incorporated into future sea ice or lower atmospheric chemistry scenarios. Thinning of Arctic sea ice will likely present more open sea ice leads where young ice, brine, and frost flowers form. How these changing ice conditions will affect the interactions between ice, brine, frost flowers and the lower atmosphere is unknown.

Microbial community composition and bacterioplankton at time series station Helgoland Roads, North Sea

A process of global importance in carbon cycling is the remineralization of algae biomass by heterotrophic bacteria, most notably during massive marine algae blooms. Such blooms can trigger secondary blooms of planktonic bacteria that consist of swift successions of distinct bacterial clades, most prominently members of the Flavobacteriia, Gammaproteobacteria and the alphaproteobacterial Roseobacter clade. This study explores such successions during spring phytoplankton blooms in the southern North Sea (German Bight) for four consecutive years. The surface water samples were taken at Helgoland Island about 40 km offshore in the southeastern North Sea in the German Bight at the station 'Kabeltonne' (54° 11.3' N, 7° 54.0' E) between the main island and the minor island, Düne (German for 'dune') using small research vessels (http://www.awi.de/en/expedition/ships/more-ships.html). Water depths at this site fluctuate from 6 to 10 m over the tidal cycle. Samples were processed as described previously (Teeling et al., 2012; doi:10.7554/eLife.11888.001) in the laboratory of the Biological Station Helgoland within less than two hours after sampling. Assessment of absolute cell numbers and bacterioplankton community composition was carried out as described previously (Thiele et al., 2011; doi:10.1016/B978-0-444-53199-5.00056-7). To obtain total cell numbers, DNA of formaldehyde fixed cells filtered on 0.2 mm pore sized filters was stained with 4',6-diamidino-2-phenylindole (DAPI). Fluorescently labeled cells were subsequently counted on filter sections using an epifluores-cence microscope. Likewise, bacterioplankton community composition was assessed by catalyzedreporter deposition fluorescence in situ hybridization (CARD-FISH) of formaldehyde fixed cells on 0.2 mm pore sized filters.

Microbial community composition in sediments of the Hydrate Ridge (Cascadian Margin) collected during RV SONNE cruises SO143 (1999) and SO148-1 (2000)

Cold seep environments such as sediments above outcropping hydrate at Hydrate Ridge (Cascadia margin off Oregon) are characterized by methane venting, high sulfide fluxes caused by the anaerobic oxidation of methane, and the presence of chemosynthetic communities. This investigation deals with the diversity and distribution of sulfate-reducing bacteria, some of which are directly involved in the anaerobic oxidation of methane as syntrophic partners of the methanotrophic archaea. The composition and activity of the microbial communities at methane vented and nonvented sediments are compared by quantitative methods including total cell counts, fluorescence in situ hybridization (FISH). Bacteria involved in the degradation of particulate organic carbon (POC) are as active and diverse as at other productive margin sites of similar water depths. The availability of methane supports a two orders of magnitude higher microbial biomass (up to 9.6×10**10cells/cm**3). Sediment samples were obtained during RV SONNE cruises SO143-2 and SO148-1 at the crest of southern Hydrate Ridge at the Cascadia convergent margin off the coast of Oregon. Sediment cores of 20 - 40 cm length were obtained using a video-guided multiple corer from gas hydrate bearing sediments and from reference sites not enriched in methane in the surface sediments. Samples for total cell counts were obtained from 1 cm core slices, fixed with 2% formaldehyde and stored cold (4°C) and the quantification of aggregates was done via epifluorescence microscopy after staining the sediments with Acridine Orange Direct Counts (AODC) according to the method of Meyer- Reil (1983, doi:10.1007/BF00395813). Total cell counts were defined as the sum of single cells plus the aggregated cells in the syntrophic consortia. DAPI staining was used to measure ANME2/DSS aggregate sizes via epifluorescence microscopy of FISH-treated samples. For FISH, subsamples of sediment cores were sliced into 1 cm intervals and fixed for 2-3 h with 3% formaldehyde (final concentration), washed twice with 1×PBS (10 mM sodium phosphate; 130 mM NaCl), and finally stored in 1×PBS/EtOH (1:1) at -20°C.

Microbial community composition of sandy intertidal sediments of Sylt-Rømø Basin, Wadden Sea

Molecular biological methods were used to investigate the microbial diversity and community structure in intertidal sandy sediments near the island of Sylt (Wadden Sea) at a site which was characterized for transport and mineralization rates in de Beer et al., (2005, hdl:10013/epic.21375). The sampling was performed during low tide in the middle of the flat, approximately 40 m in the offshore direction from the high water line on October 6, 1999, March 7, 2000, and July 5, 2000. Two parallel cores were collected from each season for molecular analyses. Within 2 h after sampling the sediment cores were sub-sampled and fixed in formaldehyde for FISH analysis. The cells were hybridized, stained with 4',6'-diamidino-2-phenylindole (DAPI) and microscopically counted as described previously [55]. Details of probes and formamide concentrations which were used are shown in further details. Counts are reported as means calculated from 10-15 randomly chosen microscopic fields corresponding to 700-1000 DAPI-stained cells. Values were corrected for the signals counted with the probe NON338. Fluorescence in situ hybridization (FISH)with group-specific rRNA-targeted oligonucleotide probes were used to characterize the microbial community structure over depth (0-12 cm) and seasons (March, July, October). We found high abundances of bacteria with total cell numbers up to 3×109 cells ml-1 and a clear seasonal variation, with higher values in July and October versus March. The microbial community was dominated by members of the Planctomycetes, the Cytophaga/Flavobacterium group, Gammaproteobacteria, and bacteria of the Desulfosarcina/Desulfococcus group. The high abundance (1.5×10**7 - 1.8×10**8 cells/ml accounting for 3-19% of all cells) of presumably aerobic heterotrophic polymer-degrading planctomycetes is in line with the high permeability, deep oxygen penetration, and the high rates of aerobic mineralization of algal biomass measured in the sandy sediments by de Beer et al., (2005, hdl:10013/epic.21375). The high and stable abundance of members of the Desulfosarcina/Desulfococcus group, both over depth and season, suggests that these bacteria may play a more important role than previously assumed based on low sulfate reduction rates in parallel cores de Beer et al., (2005).