The sapropel record of the eastern Mediterranean Sea

Research on sediments recovered during Ocean Drilling Leg 160 has concentrated on two issues: the first concerned the stratigraphy of sapropel formation, the second was oriented to clarify specific processes that explain sapropel origin. Progress has been made in the construction of stratigraphic composites out of sedimentary sequences from individual holes at each of the palaeoceanographic sites. On the composites, initial work has resulted in the establishment of high-resolution and intermediate-resolution stratigraphies for three sites (963, 964, 967); correlation of sedimentary cycles to astronomical (insolation) cycles extends the stratigraphies to Sites 969 and 966. The sapropel occurrences in the marine and land sequences over the entire Eastern Mediterranean are correlated; with the resolution that can be obtained from isotope studies, groups of sapropels occurred simultaneously over the entire basin. In detail, however, the temporal and facies patterns of sapropel sequences differ between individual sites and depositional basins. The differences may be related to effects of water depth, diagenesis, and post-depositional tectonic attenuation of sequences. Studies on the geochemistry and facies of sapropels agree that anoxic conditions favoured preservation of organic matter in sapropels, caused the enrichment of trace metals associated with sapropels, and helped to preserve primary sedimentary structures. Besides, all evidence is consistent with elevated fluxes of organic matter and associated elements during sapropel events.

Heterotrophic bacteria, Synechococcus, Prochlorococcus and Virus in the eastern Mediterranean Sea in March and April 2008 during SES_GR1

The dataset is based on samples collected in the framework of the project SESAME, in the Ionian, Libyan and Aegean Sea during March- April 2008. The objectives were to measure the standing stocks and calculate the production of the microbial compartment of the food web, describe the vertical distribution pattern and characterize its structure and function through the water column. Heterotrophic bacteria, Synechococcus, Prochlorococcus and Virus abundance: Subsamples for virus, heterotrophic bacteria and cyanobacteria (Synechococcus spp. and Prochlorococcus spp.) counting were analyzed using a FACSCalibur (Becton Dickinson) flow cytometer equipped with a standard laser (488 nm) and filter set and using deionized water as sheath fluid. Fluorescent beads with a diameter of 0.97 µm (Polysciences) were added to each sample as an internal standard, and all parameters were normalized to the beads and expressed as relative units. SYBRGreen I stain (Molecular Probe) was used to stain viral and heterotrophic bacterial DNA. Viruses were counted according to (Brussaard 1984). In order to avoid bulk consentrations of viruses samples we dilluted to Tris-EDTA (pH=8,0) buffer to a final sollution of 1/5 to 1/100. Total abundance and nucleid content classes were calculated using the Paint-A-Gate software (Becton Dickinson).

Bacterial production in the eastern Mediterranean Sea in March and April 2008 during SES_GR1

The dataset is based on samples collected in the framework of the project SESAME, in the Ionian, Libyan and Aegean Sea during March- April 2008. The objectives were to measure the standing stocks and calculate the production of the microbial compartment of the food web, describe the vertical distribution pattern and characterize its structure and function through the water column. Bacterial production was estimated by the 3H-leucine method (Kirchman et al. 1986, Kirchman 1993). At each depth, duplicate samples and a control were incubated with 20 nM L-[4,5 3H]-leucine. Samples were incubated in the dark, at in situ temperature.

Faecal pellet production of copepoda collected in water depth up to 30 meters in the eastern Mediterranean Sea in Oktober 2008 during SES_GR2

Mesozooplankton is collected by vertical tows within the Black sea water body mass layer in the NE Aegean, using a WP-2 200 µm net equipped with a large non-filtering cod-end (10 l). Macrozooplankton organisms are removed using a 2000 µm net. A few unsorted animals (approximately 100) are placed inside several glass beaker of 250 ml filled with GF/F or 0.2 µm Nucleopore filtered seawater and with a 100 µm net placed 1 cm above the beaker bottom. Beakers are then placed in an incubator at natural light and maintaining the in situ temperature. After 1 hour pellets are separated from animals and placed in separated flasks and preserved with formalin. Pellets are counted and measured using an inverted microscope. Animals are scanned and counted using an image analysis system. Carbon- Specific faecal pellet production is calculated from a) faecal pellet production, b) individual carbon: Animals are scanned and their body area is measured using an image analysis system. Body volume is then calculated as an ellipsoid using the major and minor axis of an ellipse of same area as the body. Individual carbon is calculated from a carbon- total body volume of organisms (relationship obtained for the Mediterranean Sea by Alcaraz et al. (2003) divided by the total number of individuals scanned and c) faecal pellet carbon: Faecal pellet length and width is measured using an inverted microscope. Faecal pellet volume is calculated from length and width assuming cylindrical shape. Conversion of faecal pellet volume to carbon is done using values obtained in the Mediterranean from: a) faecal pellet density 1,29 g cm**3 (or pg µm**3) from Komar et al. (1981); b) faecal pellet DW/WW=0,23 from Elder and Fowler (1977) and c) faecal pellet C%DW=25,5 Marty et al. (1994).

Abundance and biomass of mesozooplankton in the eastern Mediterranean Sea in March and April 2008 during SES_GR1

The dataset is based on samples taken during March-April 2008 in Libyan Sea, in Southern Aegean Sea and in Northern Aegean Sea. Sampling volume was estimated by the net mouth surface and the towing distance for WP-2. Taxon-specific mesozooplankton abundance and total abundance: The sample was split on board in two halves by using the beaker approach. The first sub-sample was immediately fixed and preserved in a seawater formalin solution containing about 4% buffered formaldehyde to allow the determination of species composition abundance. Pipette for the subsamples used in the taxonomic analysis of zooplankton under binocular microscope.

Ingestion rate and egg production of copepods collected in the upper 20m in the eastern Mediterranean Sea in April 2008 during SES_GR2

The ingestion on ciliates and phytoplankton dataset is based on samples taken during October 2008 in Northern Aegean Sea, the area influenced by the Black Sea water outflow. A Lagrangian experiment was established and copepod ingestion was estimated from experiments performed at stations according to the different positions of drifters during the cruise. Copepods for the experiments were obtained with slow non-quantitative tows from the upper 20 m layer of the water column using 200 µm mesh size nets fitted with a large non-filtering cod end. For the grazing experiments we used the following copepod species: Clausocalanus furcatus, and Temoraa stylifera according to the relevant reference (Bamstedt et al. 2000). Copepod clearance rates on ciliates were calculated according to Frost equations (Frost 1972). Ingestion rates were calculated by multiplying clearance rates by the initial standing stocks (Bamstedt et al. 2000). The egg production dataset is based on samples taken during October 2008 in Northern Aegean Sea, the area influenced by the Black Sea water outflow. A Lagrangian experiment was established and copepod egg production was estimated from experiments performed at stations according to the different positions of drifters during the cruise. Egg production rates of the dominant calanoid copepods were determined by incubation of fertilised females (eggs female/day) collected in the 0-20m layer. Copepod egg production was measured for the copepods Clausocalanus furcatus, Temora stylifera. On board experiments for the estimation of copepod egg production were taken place. For the estimation of copepod production (mgC/m**2/day), lengths (copepods and eggs) were converted to body carbon (Hopcroft et al., 1998) and production was estimated from biomass and weight-specific egg production rates, by assuming that those rates are representative for juvenile specific growth rates (Berggreen et al., 1988).

(Table 1) Position and astronomical ages of sedimentary cycles in the eastern Mediterranean Sea

An astronomically calibrated timescale has recently been established [Hilgen, 1991, doi:10.1016/0012-821X(91)90082-S; doi:10.1016/0012-821X(91)90206-W] for the Pliocene and earliest Pleistocene based on the correlation of dominantly precession controlled sedimentary cycles (sapropels and carbonate cycles) in Mediterranean marine sequences to the precession time series of the astronomical solution of Berger and Loutre [1991, doi:10.1016/0277-3791(91)90033-Q ] (hereinafter referred to as Ber90). Here we evaluate the accuracy of this timescale by (1) comparing the sedimentary cycle patterns with 65°N summer insolation time series of different astronomical solutions and (2) a cross-spectral comparison between the obliquity-related components in the 65°N summer insolation curves and high-resolution paleoclimatic records derived from the same sections used to construct the timescale. Our results show that the carbonate cycles older than 3.5 m.y. should be calibrated to one precession cycle older than previously proposed. Application of the astronomical solution of Laskar [1990, doi:10.1016/0019-1035(90)90084-M], (hereinafter referred to as La90) with present-day values for the dynamical ellipticity of the Earth and tidal dissipation by the Sun and Moon results in the best fit with the geological record, indicating that this solution is the most accurate from a geological point of view. Application of Ber90, or La90 solutions with dynamical ellipticity values smaller or larger than the present-day value, results in a less obvious fit with the geological record. This implies that the change in the planetary shape of the Earth associated with ice loading and unloading near the poles during the last 5.3 million years was too small to drive the precession into resonance with the perturbation term, s6-g6+g5, of Jupiter and Saturn. Our new timescale results in a slight but significant modification of all ages of the sedimentary cycles, bioevents, reversal boundaries, chronostratigraphic boundaries, and glacial cycles. Moreover, a comparison of this timescale with the astronomical timescales of ODP site 846 [Shackleton et al., 1995, doi:10.2973/odp.proc.sr.138.106.1995; doi:10.2973/odp.proc.sr.138.117.1995] and ODP site 659 [Tiedemann et al., 1994, doi:10.1029/94PA00208] indicates that all obliquity-related glacial cycles prior to ~4.7 Ma in ODP sites 659 and 846 should be correlated with one obliquity cycle older than previously proposed.

Faecal pellet production of copepoda collected in water depth up to 100 meters in the eastern Mediterranean Sea in March and April 2008 during SES_GR1

The SES_UNLUATA_GR1-Mesozooplankton faecal pellet production rates dataset is based on samples taken during March and April 2008 in the Northern Libyan Sea, Southern Aegean Sea and in the North-Eastern Aegean Sea. Mesozooplankton is collected by vertical tows within the 0-100 m layer or within the Black sea water body mass layer in the case of the NE Aegean, using a WP-2 200 µm net equipped with a large non-filtering cod-end (10 l). Macrozooplankton organisms are removed using a 2000 µm net. A few unsorted animals (approximately 100) are placed inside several glass beaker of 250 ml filled with GF/F or 0.2 µm Nucleopore filtered seawater and with a 100 µm net placed 1 cm above the beaker bottom. Beakers are then placed in an incubator at natural light and maintaining the in situ temperature. After 1 hour pellets are separated from animals and placed in separated flasks and preserved with formalin. Pellets and are counted and measured using an inverted microscope. Animals are scanned and counted using an image analysis system. Carbon- Specific faecal pellet production is calculated from a) faecal pellet production, b) individual carbon: Animals are scanned and their body area is measured using an image analysis system. Body volume is then calculated as an ellipsoid using the major and minor axis of an ellipse of same area as the body. Individual carbon is calculated from a carbon- total body volume of organisms (relationship obtained for the Mediterranean Sea by Alcaraz et al. (2003) divided by the total number of individuals scanned and c) faecal pellet carbon: Faecal pellet length and width is measured using an inverted microscope. Faecal pellet volume is calculated from length and width assuming cylindrical shape. Conversion of faecal pellet volume to carbon is done using values obtained in the Mediterranean from: a) faecal pellet density 1,29 g cm**3 (or pg µm**3) from Komar et al. (1981); b) faecal pellet DW/WW=0,23 from Elder and Fowler (1977) and c) faecal pellet C%DW=25,5 Marty et al. (1994).

Faecal pellet production of copepoda collected at different water depth in the eastern Mediterranean Sea during SES_GR2

The SES_GR2-Mesozooplankton faecal pellet production rates dataset is based on samples taken during August and September 2008 in the Northern Libyan Sea, Southern Aegean Sea and the North-Eastern Aegean Sea. Mesozooplankton is collected by vertical tows within the 0-100 m layer or within the Black sea water body mass layer in the case of the NE Aegean, using a WP-2 200 µm net equipped with a large non-filtering cod-end (10 l). Macrozooplankton organisms are removed using a 2000 µm net. A few unsorted animals (approximately 100) are placed inside several glass beaker of 250 ml filled with GF/F or 0.2 µm Nucleopore filtered seawater and with a 100 µm net placed 1 cm above the beaker bottom. Beakers are then placed in an incubator at natural light and maintaining the in situ temperature. After 1 hour pellets are separated from animals and placed in separated flasks and preserved with formalin. Pellets are counted and measured using an inverted microscope. Animals are scanned and counted using an image analysis system. Carbon- Specific faecal pellet production is calculated from a) faecal pellet production, b) individual carbon: Animals are scanned and their body area is measured using an image analysis system. Body volume is then calculated as an ellipsoid using the major and minor axis of an ellipse of same area as the body. Individual carbon is calculated from a carbon- total body volume of organisms (relationship obtained for the Mediterranean Sea by Alcaraz et al. (2003) divided by the total number of individuals scanned and c) faecal pellet carbon: Faecal pellet length and width is measured using an inverted microscope. Faecal pellet volume is calculated from length and width assuming cylindrical shape. Conversion of faecal pellet volume to carbon is done using values obtained in the Mediterranean from: a) faecal pellet density 1,29 g cm**3 (or pg µm**3) from Komar et al. (1981); b) faecal pellet DW/WW=0,23 from Elder and Fowler (1977) and c) faecal pellet C%DW=25,5 Marty et al. (1994).

Faecal pellet production of copepoda collected in water depth up to 20 meters in the eastern Mediterranean Sea in March and April 2008 during SES_GR1

The SES_GR1-Mesozooplankton faecal pellet production rates dataset is based on samples taken during April 2008 in the North-Eastern Aegean Sea. Mesozooplankton is collected by vertical tows within the Black sea water body mass layer in the NE Aegean, using a WP-2 200 µm net equipped with a large non-filtering cod-end (10 l). Macrozooplankton organisms are removed using a 2000 µm net. A few unsorted animals (approximately 100) are placed inside several glass beaker of 250 ml filled with GF/F or 0.2 µm Nucleopore filtered seawater and with a 100 µm net placed 1 cm above the beaker bottom. Beakers are then placed in an incubator at natural light and maintaining the in situ temperature. After 1 hour pellets are separated from animals and placed in separated flasks and preserved with formalin. Pellets are counted and measured using an inverted microscope. Animals are scanned and counted using an image analysis system. Carbon- Specific faecal pellet production is calculated from a) faecal pellet production, b) individual carbon: Animals are scanned and their body area is measured using an image analysis system. Body volume is then calculated as an ellipsoid using the major and minor axis of an ellipse of same area as the body. Individual carbon is calculated from a carbon- total body volume of organisms (relationship obtained for the Mediterranean Sea by Alcaraz et al. (2003) divided by the total number of individuals scanned and c) faecal pellet carbon: Faecal pellet length and width is measured using an inverted microscope. Faecal pellet volume is calculated from length and width assuming cylindrical shape. Conversion of faecal pellet volume to carbon is done using values obtained in the Mediterranean from: a) faecal pellet density 1,29 g cm**3 (or pg µm**3) from Komar et al. (1981); b) faecal pellet DW/WW=0,23 from Elder and Fowler (1977) and c) faecal pellet C%DW=25,5 Marty et al. (1994).

Niche differentiation among mat-forming, sulfide-oxidizing bacteria in chemosynthetic ecosystems of the Nile Deep Sea Fan (Eastern Mediterranean Sea)

Sulfidic muds of cold seeps on the Nile Deep Sea Fan are populated by different types of mat-forming sulfide-oxidizing bacteria. The predominant sulfide oxidizers of three different mats were identified by microscopic and phylogenetic analyses as (i) Arcobacter species producing cotton-ball-like sulfur precipitates, (ii) large filamentous sulfur bacteria including Beggiatoa species, or (iii) single, spherical cells resembling Thiomargarita species. High resolution in situ microprofiles revealed different geochemical settings selecting for different mat types. Arcobacter mats occurred where oxygen and sulfide overlapped at the bottom water interface. Filamentous sulfide oxidizers were associated with non-overlapping, steep gradients of oxygen and sulfide. A dense population of Thiomargarita was favored by temporarily changing supplies of oxygen and sulfide. These results indicate that the decisive factors in selecting for different mat-forming bacteria within one deep-sea province are spatial or temporal variations in energy supply. Furthermore, the occurrence of Arcobacter spp.-related 16S rRNA genes in the sediments below all three types of mats, as well as on top of brine lakes of the Nile Deep Sea Fan, indicates that this group of sulfide oxidizers can switch between different life modes depending on the geobiochemical habitat setting.

Abundance and biomass of ciliates in the eastern Mediterranean Sea in April 2008

The dataset was obtained on samples taken from 6 stations in the Dardanelles Straits, Marmara Sea and Bosporus Straits. These experiments were set up according to DoW of SESAME project. Ciliate abundance: Borax-buffered formalin (final concentration 2% formaldehyde). Samples for ciliate counting were stored at 4°C in the dark until observation. For ciliate identification and enumeration, 100 ml samples were left for 24 h in sedimentation cylinders and then observed under an inverted epifluorescence microscope. Ciliate biomass: Ciliate cell sizes were measured and converted into cell volumes using appropriate geometric formulae (Peuto-Moreau 1991). For biomass estimation, the conversion factor 140 fgC µm**3 was used (Putt and Stoecker (1989), doi:10.4319/lo.1989.34.6.1097)).

Biogeochemical investigation of cold seep sediments in the Eastern Mediterranean Sea

Cold seep ecosystems are highly productive, fragmented ecosystems of the deep-sea floor. They form worldwide where methane reaches the surface seafloor, and are characterized by rich chemosynthetic communities fueled by the microbial utilization of hydrocarbons. Here we investigated with in situ (benthic chamber, microprofiler) and ex situ (pore water constituents, turnover rates of sulfate and methane, prokaryote abundance) techniques reduced sites from three different seep ecosystems in the Eastern Mediterranean deep-sea. At all three cold seep systems, the Amon Mud Volcano, Amsterdam Mud Volcano and the Nile Deep Sea Fan Pockmark area, we observed and sampled patches of highly reduced, methane-seeping sulfidic sediments which were separated by tens to hundreds of (kilo)meters with non-reduced oxygenated seafloor areas. All investigated seep sites were characterized by gassy, sulfidic sediments of blackish color, of which some were overgrown with thiotrophic bacterial mats. Fluxes of methane and oxygen, as well as sulfate reduction rates varied between the different sites.