Nematode abundance in cold seep sediments of the Darwin mud volcano

During the JC-10 cruise (2007), we sampled the Darwin mud volcano (MV) for meiofaunal community and trophic structure in relation of pore-water geochemistry along a 10 m transect from a seep site on the rim of the crater towards the MV slope. Sediment samples were retrieved by the ROV Isis using push cores. On board and after the pore water extraction, the top 10 cm of the cores were sliced into 1 cm sections and fixed them in 4% formaldehyde for meiofaunal community analysis. In the home laboratory, the formaldehyde-fixed samples were washed over a 32 µm mesh sieve and extracted the meiofauna from the sediment by Ludox centrifugation (Heip et al. 1985). Meiofauna was then sorted, enumerated and identified at coarse taxonomic level. From each slice, ca. 100 nematodes were identified to genus level. Afterwards, abundance of Nematoda were depth integrated over the top 5 cm to gain individual abundances per 10 cm**2. Overall, total nematode biomass in the top 5 cm of the seep sediment core was ~10x higher than that in the core taken 1100 m away. Nematode genus composition varied little among cores and was mainly dominated by Sabatieria.

Microbial community, biomarker concentrations and their isotopic signature in sediment of Gullfaks and Tommeliten methane seeps in the Northern North Sea

Gullfaks is one of the four major Norwegian oil and gas fields, located in the northeastern edge of the North Sea Plateau. Tommeliten lies in the greater Ekofisk area in the central North Sea. During the cruises HE 208 and AL 267 several seep locations of the North Sea were visited. At the Heincke seep at Gullfaks, sediments were sampled in May 2004 (HE 208) using a video-guided multiple corer system (MUC; Octopus, Kiel). The samples were recovered from an area densely covered with bacterial mats where gas ebullition was observed. The coarse sands limited MUC penetration depth to maximal 30 centimeters and the highly permeable sands did not allow for a high-resolution, vertical subsampling because of pore water loss. The gas flare mapping and videographic observation at Tommeliten indicated an area of gas emission with a few small patches of bacterial mats with diameters <50 cm from most of which a single stream of gas bubbles emerged. The patches were spaced apart by 10-100 m. Sampling of sediments covered by bacterial mats was only possible with 3 small push cores (3.8 cm diameter) mounted to ROV Cherokee. These cores were sampled in 3 cm intervals. Lipid biomarker extraction from 10 -17 g wet sediment was carried out as described in detail elsewhere (Elvert et al., 2003; doi:10.1080/01490450303894). Briefly, defined concentrations of cholestane, nonadecanol and nonadecanolic acid with known delta 13C-values were added to the sediments prior to extraction as internal standards for the hydrocarbon, alcohol and fatty acid fraction, respectively. Total lipid extracts were obtained from the sediment by ultrasonification with organic solvents of decreasing polarity. Esterified fatty acids (FAs) were cleaved from the glycerol head group by saponification with methanolic KOH solution. From this mixture, the neutral fraction was extracted with hexane. After subsequent acidification, FAs were extracted with hexane. For analysis, FAs were methylated using BF3 in methanol yielding fatty acid methyl esters (FAMES). The fixation for total cell counts and CARD-FISH were performed on-board directly after sampling. For both methods, sediments were fixed in formaldehyde solution. After two hours, aliquots for CARD-FISH staining were washed with 1* PBS (10mmol/l sodium phosphate solution, 130mmol/l NaCl, adjusted to a pH of 7.2) and finally stored in a 1:1 PBS:ethanol solution at -20°C until further processing. Samples for total cell counts were stored in formalin at 4°C until analysis. For sandy samples, the total cell count/CARD-FISH protocol was optimized to separate sand particles from the cells. Cells were dislodged from sediment grains and brought into solution with the supernatant by sonicating each sample onice for 2 minutes at 50W. This procedure was repeated four times and supernatants were combined. The sediment samples were brought to a final dilution of 1:2000 to 1:4000 and filtered onto 0.2µm GTTP filters (Millipore, Eschbonn, Germany).

Mineralogy of cold seep sediment inhabited by vesicomyid clams in the Japan Deep Sea Trench

Sediment sampling was performed at the center of the clam colony. Immediately after sample recovery onboard, the sediment core was sub-sampled and preserved for later analyses. Pyrite and carbonate content of the sediment was measured by X-ray refraction analysis as previously described (Ertefai et al., 2010).