Ladderane lipid analysis and pore water and sediment chemistry of sediment cores from the continental shelf and slope off northwest Africa

The presence and abundance of anaerobic ammonium-oxidizing (anammox) bacteria was investigated in continental shelf and slope sediments (300-3000 m water depth) off northwest Africa in a combined approach applying quantitative polymerase chain reaction (q-PCR) analysis of anammox-specific 16S rRNA genes and anammox-specific ladderane biomarker lipids. We used the presence of an intact ladderane monoether lipid with a phosphocholine (PC) headgroup as a direct indicator for living anammox bacteria and compared it with the abundance of ladderane core lipids derived from both living and dead bacterial biomass. All investigated sediments contained ladderane lipids, both intact and core lipids, in agreement with the presence of anammoxspecific 16S rRNA gene copies, indicating that anammox occurs at all sites. Concentrations of ladderane core lipids in core top sediments varied between 0.3 and 97 ng g**-1 sediment, with the highest concentrations detected at the sites located on the shelf at shallower water depths between 300 and 500 m. In contrast, the C20 [3]-ladderane monoether-PC lipid was most abundant in a core top sediment from 1500 m water depth. Both anammox-specific 16S rRNA gene copy numbers and the concentration of the C20 [3]-ladderane monoether-PC lipid increased downcore in sediments located at greater water depths, showing highest concentrations of 1.2 x 10**8 copies g**-1 sediment and 30 pg g**-1 sediment, respectively, at the deepest station of 3000 m water depth. This suggests that the relative abundance of anammox bacteria is higher in sediments at intermediate to deep water depths where carbon mineralization rates are lower but where anammox is probably more important than denitrification.

Lipid analyses of ODP Leg 117 samples

The solvent-extractable organic fractions of sediment samples from six Ocean Drilling Program Leg 117 sites were investigated by gas chromatography and gas chromatography-mass spectrometry. Sediments deposited in the Indus Fan (Site 720) as well as Miocene sediments from the Owen Ridge (Sites 722 and 731) contain almost exclusively organic matter of terrigenous origin. The organic matter in sediments from the Oman Margin (Sites 723, 725, and 728) and in the Pliocene/Pleistocene sections from the Owen Ridge is mainly of a marine origin with variable admixtures of terrigenous material. In these latter samples strong variations of the lipid composition and distribution are noted. However, the interpretation of the relation to potential biological sources is hampered by a lack of information on the possible lipid composition of appropriate organisms.

Geochemistry, microbiology and phospholipid fatty acid content of ODP Site 169-1036 sediments

Phospholipid fatty acids were measured in samples of 60°-130°C sediment taken from three holes at Site 1036 (Ocean Drilling Program Leg 169) to determine microbial community structure and possible community replacement at high temperatures. Five of six samples had similar concentrations of phospholipid fatty acids (2-6 pmol/g dry weight of sediment), and biomass estimates from these measurements compare favorably with direct microscopic counts, lending support to previous microscopic measures of deep sedimentary biomass. Very long-chain phospholipid fatty acids (21 to 30 carbons) were detected in the sediment and were up to half the total phospholipid fatty acid measured; they appear to increase in abundance with temperature, but their significance is not known. Community composition from lipid analysis showed that samples contained standard eubacterial membrane lipids but no detectable archaeal lipids, though archaea would be expected to dominate the samples at high temperatures. Cluster analysis of Middle Valley phospholipid fatty acid compositions shows that lipids in Middle Valley sediment samples are similar to each other at all temperatures, with the exception of very long-chain fatty acids. The data neither support nor deny a shift to a high-temperature microbial community in hot cores, so at the present time we cannot draw conclusions about whether the microbes observed in these hot sediments are active.

Non-isoprenoidal intact polar lipids, isoprenoidal and non-isoprenoidal core lipids, isoprenoidal intact polar lipids of the Black Sea site GeoB15105 sampled during Meteor cruise M84/1 in February 2011

This PhD thesis focused on the analysis and application of microbial membrane lipids as biomarkers in marine sediments. Existing protocols for lipid extraction from marine sediments and biomass were modified. In addition, recent protocols based on high-performance liquid chromatography coupled to mass spectrometry (HPLC-MS) as well as state-of-the-art mass spectrometric analysis by quadrupole time-of-flight (MSqTOF) and the triple quadrupole (MSQQQ) mass spectrometer were used to investigate matrix effects and evaluate the reliability of quantitative analysis in marine environmental samples. The improved lipid extraction and quantification were used to analyze Black Sea water column and sediments samples to a depth of 8 meters from site GeoB 15105 taken during cruise M84/1 (DARCSEAS) with R/V Meteor to apply lipid analysis in benthic bio systems. With this component specific differentiation between planktonic and benthic lipid signature we assessed possible lipid sources. Here, this high detail lipid fingerprinting allowed us to observe changes in the head group and lipid core structures of the intact polar lipids according to the geochemical zonation.

Lipid, carbon and nitrogen content and fatty acid composition of Temora longicornis females and different food species during the experiment

Temora longicornis, a dominant calanoid copepod species in the North Sea, is characterised by low lipid reserves and high biomass turnover rates. To survive and reproduce successfully, this species needs continuous food supply and thus requires a highly flexible digestive system to exploit various food sources. Information on the capacity of digestive enzymes is scarce and therefore the aim of our study was to investigate the enzymatic capability to respond to quickly changing nutritional conditions. We conducted two feeding experiments with female T. longicornis from the southern North Sea off Helgoland. In the first experiment in 2005, we tested how digestive enzyme activities and enzyme patterns as revealed by substrate SDS-PAGE (sodium dodecylsulfate polyacrylamide gel electrophoresis) responded to changes in food composition. Females were incubated for three days fed ad libitum with either the heterotrophic dinoflagellate Oxyrrhis marina or the diatom Thalassiosira weissflogii. At the beginning and at the end of the experiment, copepods were deep-frozen for analyses. The lipolytic enzyme activity did not change over the course of the experiment but the enzyme patterns did, indicating a distinct diet-induced response. In a second experiment in 2008, we therefore focused on the enzyme patterns, testing how fast changes occur and whether feeding on the same algal species leads to similar patterns. In this experiment, we kept the females for 4 days at surplus food while changing the algal food species daily. At day 1, copepods were offered O. marina. On day 2, females received the cryptophycean Rhodomonas baltica followed by T. weissflogii on day 3. On day 4 copepods were again fed with O. marina. Each day, copepods were frozen for analysis by means of substrate SDS-PAGE. This showed that within 24 h new digestive enzymes appeared on the electrophoresis gels while others disappeared with the introduction of a new food species, and that the patterns were similar on day 1 and 4, when females were fed with O. marina. In addition, we monitored the fatty acid compositions of the copepods, and this indicated that specific algal fatty acids were quickly incorporated. With such short time lags between substrate availability and enzyme response, T. longicornis can successfully exploit short-term food sources and is thus well adapted to changes in food availability, as they often occur in its natural environment due seasonal variations in phyto- and microzooplankton distribution.

Elemental contents, non-carbonate, fatty acids and alcohols analysis from different Holes of IODP Expedition 310

During IODP Expedition 310 (Tahiti Sea Level), drowned Pleistocene-Holocene barrier-reef terraces were drilled on the slope of the volcanic island. The deglacial reef succession typically consists of a coral framework encrusted by coralline algae and later by microbialites; the latter make up < 80% of the rock volume. Lipid biomarkers were analyzed in order to identify organisms involved in reef-microbialite formation at Tahiti, as the genesis of deglacial microbialites and the conditions favoring their formation are not fully understood. Sterols plus saturated and monounsaturated short-chain fatty acids predominantly derived from both marine primary producers (algae) and bacteria comprise 44 wt% of all lipids on average, whereas long-chain fatty acids and long-chain alcohols derived from higher land plants represent an average of only 24 wt%. Bacterially derived mono-O-alkyl glycerol ethers (MAGEs) and branched fatty acids (10-Me-C16:0; iso- and anteiso-C15:0 and -C17:0) are exceptionally abundant in the microbial carbonates (average, 19 wt%) and represent biomarkers of intermediate-to-high specificity for sulfate-reducing bacteria. Both are relatively enriched in 13C compared to eukaryotic lipids. No lipid biomarkers indicative of cyanobacteria were preserved in the microbialites. The abundances of Al, Si, Fe, Mn, Ba, pyroxene, plagioclase, and magnetite reflect strong terrigenous influx with Tahitian basalt as the major source. Chemical weathering of the basalt most likely elevated nutrient levels in the reefs and this fertilization led to an increase in primary production and organic matter formation, boosting heterotrophic sulfate reduction. Based on the observed biomarker patterns, sulfate-reducing bacteria were apparently involved in the formation of microbialites in the coral reefs off Tahiti during the last deglaciation.

Separating ITCZ- and ENSO-related rainfall changes in the Galápagos over the last 3 kyr using D/H ratios of multiple lipid biomarkers

We present a 3000-yr rainfall reconstruction from the Galápagos Islands that is based on paired biomarker records from the sediment of El Junco Lake. Located in the eastern equatorial Pacific, the climate of the Galápagos Islands is governed by movements of the Intertropical Convergence Zone (ITCZ) and the El Niño-Southern Oscillation (ENSO). We use a novel method for reconstructing past ENSO- and ITCZ-related rainfall changes through analysis of molecular and isotopic biomarker records representing several types of plants and algae that grow under differing climatic conditions. We propose that ?D values of dinosterol, a sterol produced by dinoflagellates, record changes in mean rainfall in El Junco Lake, while dD values of C34 botryococcene, a hydrocarbon unique to the green alga Botryococcus braunii, record changes in rainfall associated with moderate-to-strong El Niño events. We use these proxies to infer changes in mean rainfall and El Niño-related rainfall over the past 3000 yr. During periods in which the inferred change in El Niño-related rainfall opposed the change in mean rainfall, we infer changes in the amount of ITCZ-related rainfall. Simulations with an idealized isotope hydrology model of El Junco Lake help illustrate the interpretation of these proxy reconstructions. Opposing changes in El Niño- and ITCZ-related rainfall appear to account for several of the largest inferred hydrologic changes in El Junco Lake. We propose that these reconstructions can be used to infer changes in frequency and/or intensity of El Niño events and changes in the position of the ITCZ in the eastern equatorial Pacific over the past 3000 yr. Comparison with El Junco Lake sediment grain size records indicates general agreement of inferred rainfall changes over the late Holocene.

Stable isotope analysis and biomarkers of carbonates from the Columbia River in southwestern Washington, USA

Exotic limestone masses with silicified fossils, enclosed within deep-water marine siliciclastic sediments of the Early to Middle Miocene Astoria Formation, are exposed along the north shore of the Columbia River in southwestern Washington, USA. Samples from four localities were studied to clarify the origin and diagenesis of these limestone deposits. The bioturbated and reworked limestones contain a faunal assemblage resembling that of modern and Cenozoic deep-water methane-seeps. Five phases make up the paragenetic sequence: (1) micrite and microspar; (2) fibrous, banded and botryoidal aragonite cement, partially replaced by silica or recrystallized to calcite; (3) yellow calcite; (4) quartz replacing carbonate phases and quartz cement; and (5) equant calcite spar and pseudospar. Layers of pyrite frequently separate different carbonate phases and generations, indicating periods of corrosion. Negative d13Ccarbonate values as low as -37.6 per mill V-PDB reveal an uptake of methane-derived carbon. In other cases, d13Ccarbonate values as high as 7.1 per mill point to a residual, 13C-enriched carbon pool affected by methanogenesis. Lipid biomarkers include 13C-depleted, archaeal 2,6,10,15,19-pentamethylicosane (PMI; d13C: -128 per mill), crocetane and phytane, as well as various iso- and anteiso-carbon chains, most likely derived from sulphate-reducing bacteria. The biomarker inventory proves that the majority of the carbonates formed as a consequence of sulphate-dependent anaerobic oxidation of methane. Silicification of fossils and early diagenetic carbonate cements as well as the precipitation of quartz cement - also observed in other methane-seep limestones enclosed in sediments with abundant diatoms or radiolarians - is a consequence of a preceding increase of alkalinity due to anaerobic oxidation of methane, inducing the dissolution of silica skeletons. Once anaerobic oxidation of methane has ceased, the pH drops again and silica phases can precipitate.

Lipid composition of particulate matter measured on water bottle samples during POLARSTERN cruise ARK-XI/1

The lipid composition of particulate matter in oceanic environments can provide informations on the nature and origin of the organic matter as well as on their transformation processes. Molecular characteristics for lipids in the Arctic environment have been used as indicators of the sources and transformation of organic particulate matter (Smith et al., 1997; Fahl and Stein, 1997, 1999). However, the features of the lipid composition of particulate matter in the Arctic with its high seasonality of ice Cover and primary productivity has been studied insufficiently. Lipids are one of the most important compounds of organic matter. On the one hand, the composition of lipids is a result of the variability of biological sources (phyto- and zooplankton, higher plants, bacteria etc.). On the other hand, the lipid composition of particulate matter is undergone significant alteration during vertical transport. The organic matter balance in the Arctic marginal seas, such as the Kara and Laptev seas, is characterized by the significant supply of dissolved and particulate material by the major Eurasian rivers - Ob, Yenisei and Lena (Cauwet and Sidorov, 1996; Gordeev et al., 1996, Martin et al., 1993). In relation to the world's ocean the primary productivity values are lower in the Arctic seas due to the ice-cover. However local increased values of primary productivity can be connected with the melting processes inducing increased phytoplankton growth near ice-edge (Nelson et al., 1989; Fahl and Stein, 1997) and enhanced river supply of nutrients, These features can influence the proportion of allochtonous and autochtonous components of the organic matter in the Arctic marginal seas (Fahl and Stein, 1997; Stein and Fahl, 1999). Furthermore, increased lipid contents in aquatic environments were found near density discontinuities (Parish et al., 1988). Although being less informative than lipid studies on the molecular level the character of lipid composition analysis on the group could also be used for studying of particulate organic matter and its transformation in sedimentation processes in the Arctic. In this paper the investigation of the characteristics of lipid composition performed by Alexandrova and Shevchenko (1997) in Arctic seas was continued.