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To assess microbial assemblages and microbiallydriven biogeochemical processes related to active and past fluid seepage at cold seeps in the Marmara Sea To link biogeochemical processes and microbial assemblages to fluid seepage patterns and dynamics More specifically we aim at : Defining the structure, diversity and functionning of microbial assemblages involved in the transformation of fluid components (e.g., methane oxidation, sulphate reduction, sulphide oxidation...) Understanding how microbial assemblages (and associated processes) respond to varying seepage activity, compostion and chemistry of fluids, flow rates... and how they impact on the precipitation of authigenic minerals and on cold-seep macrofauna Molecular Tools : lipid biomarkers microbial cell membrane lipids as sources information (biomarkers) on the composition and biogeochemistry of microbial assemblages Distinctive ‘taxonomic’ structural features > > community structure Concentrations > > relative abundances of microbial groups > > biomass of extant (active) organisms Stable carbon isotopes > > carbon source utilized, carbon flow OH O OX X’ O O OH X Molecular Tools : lipid biomarkers Lipid biomarkers : analytical methods • Solvent extraction of lipids • Chromatographic isolation of lipid biomarker classes • Gas chromatography (GC) • Gas chromatography-mass spectrometry (GC-MS) • Isotope ratio monitoring-gas chromatography-mass spectrometry (GC-irMS) * • High performance liquid chromatography- mass spectrometry (HPLC-MS) * * in collaboration Undisturbed Sediments Cores Authigenic Crusts Microbial Mats, Benthic Fauna Samples to be taken in contrasted settings : Varying fluid seepage Different benthic activity To be sub-sampled and conditionned on board And to be shared for coupled investigations This contribution to the MARNAUT project is viewed as part of an integrated biogeochemical approach and, as such, it requires strong interdisciplinary coupling More specifically we plan to tightly couple this study with: Microbiology and molecular biology approaches (P. Lopez) Fluid analyses and fluxes, biogeochemical reaction rates, transport/reaction models (V. Aloisi) Authigenic carbonate geochemistry (C. Pierre) Biology of benthic fauna (Ifremer, Brest) Previous and ongoing work at cold seeps in the framework of MEDIFLUX (ESF) BIOZAIRE (IFREMER/TOTAL) HERMES (EU) Biomarkers in Crusts (Olimpi mud volcano area - MEDINAUT) Apolar compounds C25 isoprenoids C20 isoprenoids Cr PMI:0 PMI:1-3 Squ:0 Cr:1 H H C25 isoprenoids PMI:3-5 C30 isoprenoids Squ: 3-6 C35 isoprenoids Bouloubassi et al., (2006) Biomarkers in Crusts (Olimpi mud volcano area-MEDINAUT) Polar compounds OH I X Archaeol O O X’ O X OH O IId II IIa O O X IIc OH Hydroxyarchaeol Ib IIb Ic Bouloubassi et al., (2006); Aloisi et al., 2002; Pancost et al., 2001 d13C ‰ Bouloubassi et al., (2006) B A E B B n-29 Alkane cholesterol BisHomo-ol BisHomo-acid ia15 FAME ia17 FAME Diethers II Diethers I PMI:1-5 PMI DGDT Archaeol OH-Archaeol Biomarkers in Crusts (Olimpi & Anaximander mud volcano area) NAUTINIL 2003 Polar lipid patterns in carbonate crusts NL 20 CC1 Caldera X Archaeol O X’ O Hydroxyarchaeol OH OH O NL 14 CC5 Pockmarks Dialkyl Glycerol Diethers (DGD) O X OH O O X ...and suite of sterols, alcohols, hopanols hopanoïdes crocetane PMI GC/MS : Fraction hydrocarbures n-alcanes (C14 – C34) : essentiellement sources terrigènes Hopanoïdes (hopanes-hopènes, C27 - C32 : sources microbiennes - Bactéries C20 et C25 Isoprénoïdes irréguliers : sources microbiennes - Archées