Biogeosciences www.biogeosciences.net 1726-4170 1726-4189 5 4 2008 10.5194/bg-5-1127-2008 http://www.biogeosciences.net/5/1127/2008/ http://www.biogeosciences.net/5/1127/2008/bg-5-1127-2008.html http://www.biogeosciences.net/5/1127/2008/bg-5-1127-2008.pdf 1127 1144 2008-08-18 Biogeochemical processes and microbial diversity of the Gullfaks and Tommeliten methane seeps (Northern North Sea) G. Wegener gwegener@mpi-bremen.de M. Shovitri K. Knittel H. Niemann M. Hovland A. Boetius Max Planck Institute for Marine Microbiology, Bremen, Germany Alfred Wegener Institute for Polar and Marine Research, Bremerhaven, Germany Statoil, Stavanger, Norway Jacobs University Bremen, Bremen, Germany now at: Institute for Environmental Geosciences, University of Basel, Switzerland Fluid flow related seafloor structures and gas seeps were detected in the North Sea in the 1970s and 1980s by acoustic sub-bottom profiling and oil rig surveys. A variety of features like pockmarks, gas vents and authigenic carbonate cements were found to be associated with sites of oil and gas exploration, indicating a link between these surface structures and the underlying, deep hydrocarbon reservoirs. In this study we performed acoustic surveys and videographic observation at Gullfaks, Holene Trench, Tommeliten, Witch's Hole and the giant pockmarks of the UK Block 15/25, to investigate the occurrence and distribution of cold seep ecosystems in the Northern North Sea. The most active gas seep sites, i.e. Gullfaks and Tommeliten, were investigated in detail. At both sites, gas bubbles escaped continuously from small holes in the seabed to the water column, reaching the upper mixed surface layer. At Gullfaks a gas emitting, flat area of 0.1 km² of sandy seabed covered by filamentous sulfur-oxidizing bacteria was detected. At Tommeliten, we found a patchy distribution of small bacterial mats indicating sites of gas seepage. Below the patches the seafloor consisted of sand from which gas emissions were observed. At both sites, the anaerobic oxidation of methane (AOM) coupled to sulfate reduction (SR) was the major source of sulfide. Molecular analyses targeting specific lipid biomarkers and 16S rRNA gene sequences identified an active microbial community dominated by sulfur-oxidizing and sulfate-reducing bacteria (SRB) as well as methanotrophic bacteria and archaea. Stable carbon isotope values of specific, microbial fatty acids and alcohols from both sites were highly depleted in the heavy isotope ¹³C, indicating that the microbial community incorporates methane or its metabolites. The microbial community composition of both shallow seeps shows high similarities to the deep water seeps associated with gas hydrates such as Hydrate Ridge or the Eel River basin. Amann, R. I., Krumholz, L., and Stahl, D. A.: Fluorescent-oligonucleotide probing of whole cells for determinative, phylogenetic, and environmental studies in microbiology, J. Bacteriol., 172(2), 762–770, 1990. Blumenberg, M., Seifert, R., Reitner, J., Pape, T., and Michaelis, W.: Membrane lipid patterns typify distinct anaerobic methanotrophic consortia, P. Natl. Acad. Sci. USA, 101(30), 11 111–11 116, 2004. Boetius, A., Ravenschlag, K., Schubert, C. J., Rickert, D., Widdel, F., Gieseke, A., Amann, R., Jørgensen, B. B., Witte, U., and Pfannkuche, O.: A marine microbial consortium apparently mediating anaerobic oxidation of methane, Nature, 407, 623–626, 2000. Daims, H., Bruhl, A., Amann, R., Schleifer, K. H., and Wagner, M.: The domain-specific probe EUB338 is insufficient for the detection of all bacteria: Development and evaluation of a more comprehensive probe set, Syst. Appl. Microbiol., 22(3), 434–444, 1999. Dando, P. R., Bussmann, I., Niven, S. J., O'Hara, S. C. M, Schmaljohann, R., and Taylor, L. J.: A methane seep area in the Skagerrak, the habitat of the pogonophore Siboglinum poseidoni and the bivalve mollusc Thyasira sarsi, Mar. Ecol.-Prog. Ser., 107, 157–167, 1994. Eisma, D. and Kalf, J.: Dispersal, concentration and deposition of suspended matter in the North Sea, J. Geol. Soc. London, 144(1), 161–178, 1987. Elvert, M., Boetius, A., Knittel, K., and Jørgensen, B. B.: Characterization of specific membrane fatty acids as chemotaxonomic markers for sulfate-reducing bacteria involved in anaerobic oxidation of methane, Geomicrobiol. J., 20(4), 403–419, 2003. Elvert, M., Hopmans, E. C., Treude, T., Boetius, A., and Suess, E.: Spatial variations of methanotrophic consortia at cold methane seeps: implications from a high-resolution molecular and isotopic approach, Geobiology, 3(3), 195–209, 2005. Elvert, M. and Niemann, H.: Occurrence of unusual steroids and hopanoids derived from aerobic methanotrophs at an active marine mud volcano, Org. Geochem., 39, 167–177, 2008. Fang, J., Shizuka, A., Kato, C., and Schouten, S.: Microbial diversity of cold-seep sediments in Sagami Bay, Japan, as determined by 16S rRNA gene and lipid analyses, FEMS Microbiol. Ecol., 57(3), 429–441, 2005. Hinrichs, K.-U., Hayes, J. M., Silva, S. P., Brewer, P. G. and DeLong, E. F.: Methane-consuming archaebacteria in marine sediments, Nature, 398, 802–805, 1999. Hinrichs, K.-U. and Boetius, A.: The anaerobic oxidation of methane: New insights in microbial ecology and biogeochemistry, in: Ocean margin systems, edited by: Wefer, G., Billett, D., and Hebbeln, D., Springer-Verlag, Berlin and Heidelberg, Germany, 457–477, 2002. Hovland, M.: On the self-sealing nature of marine seeps, Cont. Shelf. Res., 22(16), 2387–2394, 2002. Hovland, M.: Discovery of prolific natural methane seeps at Gullfaks, northern North Sea, Geo-Mar. Lett., 27(2), 197–201, 2007. Hovland, M., Gardner, J. V., and Judd, A. G.: The significance of pockmarks to understanding fluid flow processes and geohazards, Geofluids, 2(2), 127–136, 2002. Hovland, M. and Judd, A. G.: Seabed pockmarks and seepages, Graham and Trotham, London, UK, 293 pp., 1988. Hovland, M., Judd, A. G., and Burke Jr., R. A.: The global flux of methane from shallow submarine sediments, Chemosphere, 26(1-4), 559–578, 1993. Hovland, M. and Risk, M.: Do Norwegian deep-water coral reefs rely on seeping fluids?, Mar. Geol., 198(1-2), 83–96, 2003. Hovland, M. and Sommerville, J. H.: Characteristics of two natural gas seepages in the North Sea, Mar. Petrol. Geol., 2, 319–326, 1985. Inagaki, F., Kuypers, M. M. M., Tsunogai, U., Ishibashi, J., Nakamura, K. Treude, T., Ohkubo, S., Nakaseama, M., Gena, K., Chiba, H., Hirayama, H., Nunoura, T., Takai, K., Jørgensen, B. B., Horikoshi, K., and Boetius, A.: Microbial community in a sediment-hosted CO₂ lake of the southern Okinawa Trough hydrothermal system, P. Natl. Acad. Sci. USA, 103(38), 14 164–14 169, 2006. Ishii, K., Mussmann, M., MacGregor, B. J., and Amann, R.: An improved fluorescence in situ hybridization protocol for the identification of bacteria and archaea in marine sediments, FEMS Microbiol. Ecol., 50(3), 203–212, 2004. Jørgensen, B. B. and Fenchel, T.: The sulfur cycle of a marine sediment model system, Mar. Biol., 24(3), 189–201., 1974. Joye, S. B., Boetius, A., Orcutt, B. N., Montoya, J. P., Schulz, H. N., Erickson, M. J., and Lugo, S.: The anaerobic oxidation of methane and sulfate reduction in sediments from Gulf of Mexico cold seeps, Chem. Geol., 205, 219–238, 2004. Judd, A. G., Hovland, M., Dimitrov, L. I., Garcia Gil, S., and Jukes, V.: The geological methane budget at continental margins and its influence on climate change, Geofluids, 2, 109–126, 2002. Judd, A. and Hovland, M.: Seabed fluid flow: the impact on geology, biology and the marine environment, Cambridge University Press, Cambridge, UK, 492 pp., 2007. Kallmeyer, J., Ferdelman, T. G., Weber, A., Fossing, H., and Jørgensen, B. B.: A cold chromium distillation procedure for radiolabeled sulfide applied to sulfate reduction measurements, Limnol. Oceanogr.-Meth., 2, 171–180, 2004. Kane, M. D., Poulsen, L. K., and Stahl, D. A.: Monitoring the enrichment and isolation of sulfate-reducing bacteria by using oligonucleotide hybridization probes designed from environmentally derived 16S rRNA sequences, Appl. Environ. Microb., 59, 682–686, 1993. Knittel, K., Boetius, A., Lemke, A., Eilers, H., Lochte, K., Pfannkuche, O., Linke, P., and Amann, R.: Activity, distribution, and diversity of sulfate reducers and other bacteria in sediments above gas hydrate (Cascadia margin, Oregon), Geomicrobiol. J., 20(4), 269–294, 2003. Knittel, K., Lösekann, T., Boetius, A., Kort, R., and Amann, R.: Diversity and distribution of methanotrophic archaea at cold seeps, Appl. Environ. Microb., 71(1), 467–479, 2005. Krüger, M., Treude, T., Wolters, H., Nauhaus, K., and Boetius, A.: Microbial methane turnover in different marine habitats, Palaeogeogr. Palaeocl., 227, 6–17, 2005. Lane, D. J., Pace, B., Olsen, G. J., Stahl, D. A., Sogin, M. L., and Pace, N. R.: Rapid determination of 16S ribosomal RNA sequences for phylogenetic analyses, P. Natl. Acad. Sci. USA, 82(20), 6955–6959, 1985. Levin, L. A.: Ecology of cold seep sediments: Interactions of fauna with flow, chemistry and microbes, Oceanogr. Mar. Biol. Annu. Rev., 43, 1–46, 2005. Llobet-Brossa, E., Rossello-Mora, R., and Amann, R.: Microbial community composition of Wadden Sea sediments as revealed by fluorescence in situ hybridization, Appl. Environ. Microb., 64(7) 2691–2696, 1998. Lösekann, T., Knittel, K., Nadalig, T., Fuchs, B., Niemann, H., Boetius, A., and Amann, R.: Diversity and abundance of aerobic and anaerobic methane oxidizers at the Haakon Mosby Mud Volcano, Barents Sea, Appl. Environ. Microb., 73(10), 3348–3362, 2007. Ludwig, W., Strunk, O., Westram, R., Richter, L., Meier, H., Yadhukumar, Buchner, A., Lai, T., Steppi, S., Jobb, G., Forster, W., Brettske, I., Gerber, S., Ginhart, A. W., Gross, O., Grumann, S., Hermann, S., Jost, R., Konig, A., Liss, T., Lussmann, R., May, M., Nonhoff, B., Reichel, B., Strehlow, R., Stamatakis, A., Stuckmann, N., Vilbig, A., Lenke, M., Ludwig, T., Bode, A., and Schleifer, K.-H.: ARB: a software environment for sequence data, Nucleic Acids Res., 32(4), 1363–1371, 2004. Luff, R. and Wallmann, K.: Fluid flow, methane fluxes, carbonate precipitation and biogeochemical turnover in gas hydrate-bearing sediments at Hydrate Ridge, Cascadia Margin: Numerical modeling and mass balances, Geochim. Cosmochim. Ac., 67(18), 3403–3421, 2003. Manz, W., Eisenbrecher, M., Neu, T. R., and Szewzyk, U.: Abundance and spatial organization of gram-negative sulfate-reducing bacteria in activated sludge investigated by in situ probing with specific 16S rRNA targeted oligonucleotides, FEMS Microbiol. Ecol., 25, 43–61, 1998. Massana, R., Murray, A., Preston, C., and DeLong, E.: Vertical distribution and phylogenetic characterization of marine planktonic Archaea in the Santa Barbara Channel, Appl. Environ. Microb., 63(1), 50–56, 1997. Meyer-Reil, L.-A.: Benthic response to sedimentation events during autumn to spring at a shallow water station in the Western Kiel Bight, Mar. Biol., 77(3), 247–256, 1983. Michaelis, W., Seifert, R., Nauhaus, K., Treude, T., Thiel, V., Blumenberg, M., Knittel, K., Gieseke, A., Peterknecht, K., Pape, T., Boetius, A., Amann, R., Jørgensen, B. B., Widdel, F., Peckmann, J., Pimenov, N. V., and Gulin, M. B.: Microbial reefs in the Black Sea fueled by anaerobic oxidation of methane, Science, 297, 1013–1015, 2002. Mills, H. J., Hodges, C., Wilson, K., MacDonald, I. R., and Sobecky, P. A.: Microbial diversity in sediments associated with surface-breaching gas hydrate mounds in the Gulf of Mexico, FEMS Microbiol. Ecol., 46(1), 39–52, 2003. Moss, C. W. and Lambert-Fair, M. A.: Location of double bonds in monounsaturated fatty acids of Campylobacter cryaerophila with dimethyl disulfide derivatives and combined gas chromatography-mass spectrometry, J. Clin. Microbiol., 27(7), 1467–1470, 1989. Muyzer, G., Teske, A., Wirsen, C. O., and Jannasch, H. W.: Phylogenetic relationships of \textitThiomicrospira species and their identification in deep-sea hydrothermal vent samples by denaturing gradient gel electrophoresis of 16S rDNA fragments, Arch. Microbiol., 164(3), 165–172, 1995. Nauhaus, K., Boetius, A., Krüger, M., and Widdel, F.: In vitro demonstration of anaerobic oxidation of methane coupled to sulphate reduction in sediments from a marine gas hydrate area, Environ. Microbiol., 4(5), 296–305, 2002. Nauhaus, K., Albrecht, M., Elvert, M., Boetius, A., and Widdel, F.: In vitro cell growth of marine archaeal-bacterial consortia during anaerobic oxidation of methane with sulfate, Environ. Microbiol., 9(1), 187–196, 2007. Nelson, D. C. and Jannasch, H. W.: Chemoautotrophic growth of a marine \textitBeggiatoa in sulfide-gradient cultures, Arch. Microbiol., 136(4), 262–269, 1983. Nelson, D. C., Jørgensen, B. B., and Revsbech, N. P.:. Growth pattern and yield of a chemoautotrophic \textitBeggiatoa sp. in oxygen-sulfide microgradients, Appl. Environ. Microb., 52(2), 225–233, 1986. Nelson, D. C., Wirsen, C. O., and Jannasch, H. W.: Characterization of large, autotrophic \textitBeggiatoa spp. abundant at hydrothermal vents of the Guaymas Basin, Appl. Environ. Microb., 55(11), 2909–2917, 1989. Niemann, H. and Elvert, M.: Diagnostic lipid biomarker and stable carbon isotope signatures of microbial communities mediating the anaerobic oxidation of methane with sulphate, Chem. Geol., in press, 2008. Niemann, H., Elvert, M., Hovland, M., Orcutt, B., Judd, A., Suck, I., Gutt, J., Joye, S., Damm, E., Finster, K., and Boetius, A.: Methane emission and consumption at a North Sea gas seep (Tommeliten area), Biogeosciences, 2, 335–351, 2005. Niemann, H., Lösekann, T., de Beer, D., Elvert, M., Nadalig, T., Knittel, K., Amann, R., Sauter, E. J., Schlüter, M., Klages, M., Foucher, J. P., and Boetius, A.: Novel microbial communities of the Haakon Mosby mud volcano and their role as a methane sink, Nature, 443, 854–858, 2006. Omoregie, E. O., Mastalerz ,V., de Lange, G., Straub, K. L., Kappler, A., Røy, H., Stadnitskaia, A., Foucher, J. P., and Boetius, A.: Biogeochemistry and community composition of iron- and sulfur-precipitating microbial mats at the Chefren Mud Volcano (Nile Deep Sea Fan, Eastern Mediterranean), Appl. Environ. Microb., 74(10), 3198–3215, 2008 Orphan, V. J., Hinrichs, K.-U., Ussler, W., Paull, C. K., Taylor, L. T., Sylva, S. P., Hayes, J. M., and Delong, E. F.: Comparative analysis of methane-oxidizing archaea and sulfate-reducing bacteria in anoxic marine sediments, Appl. Environ. Microb., 67(4), 1922–1934, 2001a. Orphan, V. J., House, C. H., Hinrichs, K.-U., McKeegan, K. D., and DeLong, E. F.: Methane-consuming archaea revealed by directly coupled isotopic and phylogenetic analysis, Science, 293, 484–487, 2001b. Orphan, V. J., House, C. H., Hinrichs, K.-U., McKeegan, K. D., and DeLong, E. F.: Multiple archaeal groups mediate methane oxidation in anoxic cold seep sediments, P. Natl. Acad. Sci. USA, 99(11), 7663–7668, 2002. Ourisson, G., Albrecht, P., and Rohmer, M.: The hopanoids, Paleochemistry and biochemistry of a group of natural products, Pure Appl. Chem., 51, 709–729, 1979. Pernthaler, A., Pernthaler, J., and Amann, R.: Fluorescence in situ hybridization and catalyzed reporter deposition for the identification of marine bacteria, Appl. Environ. Microb., 68(6), 3094–3101, 2002. Ravenschlag, K., Sahm, K., Pernthaler, J., and Amann, R.: High bacterial diversity in permanently cold marine sediments, Appl. Environ. Microb., 65(9), 3982–3989, 1999. Reeburgh, W.: Oceanic methane biogeochemistry, Chem. Rev., 107(2), 486–513, 2007. Rehder, G., Keir, R. S., Suess, E., and Pohlmann, T.: The multiple sources and patterns of methane in North Sea waters, Aquat. Geochem., 4(3), 403–427, 1998. Rise, L., Saettem, J., Fanavoll, S., Thorsnes, T., Ottesen, D., and Boe, R.: Sea-bed pockmarks related to fluid migration from Mesozoic bedrock strata in the Skagerrak offshore Norway, Mar. Petrol. Geol., 16(7), 619–631, 1999. Ritger, S., Carson, B., and Suess, E.: Methane-derived authigenic carbonates formed by subduction-induced pore-water expulsion along the Oregon/Washington margin, Geol. Soc. Am. Bull., 98(2), 147–156, 1987. Rohmer, M., Bouvier-Nave, P., and Ourisson, G.: Distribution of hopanoid triterpenes in prokaryotes, J. Gen. Microbiol., 130, 1137–1150, 1984. Rusch, A., Forster, S., and Huettel, M.: Bacteria, diatoms and detritus in an intertidal sandflat subject to advective transport across the water-sediment interface, Biogeochemistry, 55(1), 1–27, 2001. Rusch, A., Huettel, M., Reimers, C. E., Taghon, G. L., and Fuller, C. M.: Activity and distribution of bacterial populations in Middle Atlantic Bight shelf sands, FEMS Microbiol. Ecol., 44(1), 89–100, 2003. Stahl, D. A. and Amann, R.: Development and application of nucleic acid probes, in: Nucleic acid techniques in bacterial systematics, edited by: Stackebrandt, E. and Goodfellow, M., John Wiley and Sons Ltd., Chichester, UK, 205–248, 1991. Teske, A., Hinrichs, K. U., Edgcomb, V., de Vera Gomez, A., Kysela, D., Sylva, S. P., Sogin, M. L., and Jannasch, H. W.: Microbial diversity of hydrothermal sediments in the Guaymas Basin: evidence for anaerobic methanotrophic communities, Appl. Environ. Microb., 68(4), 1994–2007, 2002. Trasher, J., Fleet, A. J., Hay, S. J., Hovland, M., and Düpenbecker, S.: Understanding geology as the key to using seepage in exploration: spectrum of seepage styles, in: Schumacher, D. and Abrams, M. A., Hydrocarbon migration and its near-surface expression, AAPG Memoir, 223–241, 1996. Treude, T., Boetius, A., Knittel, K., Nauhaus, K., Elvert, M., Krüger, M., Lösekann, T., Wallmann, K., Jørgensen, B. B., Widdel, F., and Amman, R.: Anaerobic oxidation of methane at Hydrate Ridge (OR), Geochim. Cosmochim. Ac., 67(18), 491–491, 2003. Treude, T., Knittel, K., Blumenberg, M., Seifert, R., and Boetius, A.: Subsurface microbial methanotrophic mats in the Black Sea, Appl. Environ. Microb., 71(10), 6375–6378, 2005a. Treude, T., Krüger, M., Boetius, A., and Jørgensen, B. B.: Environmental control on anaerobic oxidation of methane in the gassy sediments of Eckernfoerde Bay (German Baltic), Limnol. Oceanogr., 50(6), 1771–1786, 2005b. Treude, T., Orphan, V., Knittel, K., Gieseke, A., House, C., and Boetius, A.: Consumption of methane and CO₂ by methanotrophic microbial mats from gas seeps of the anoxic Black Sea, Appl. Environ. Microbiol., 73, 2271–2283, 2007. Valentine, D. L. and Reeburgh, W. S.: New perspectives on anaerobic methane oxidation, Environ. Microbiol., 2(5), 477–487, 2000. Widdel, F. and Bak, F.: Gram-negative mesophilic sulfate-reducing bacteria, in: The prokaryotes, edited by: Balows, A., Dworkin, M., Harder, W., Schleifer, K.-H., Springer, Berlin and Heidelberg, Germany, and New York, USA, 3352–3378, 1992. Widdel, F., Musat, F., Knittel, K., and Galushko, A.: Anaerobic degradation of hydrocarbons with sulphate as electron acceptor, in: Sulphate-reducing bacteria: environmental and engineered systems, edited by: Barton, L. L. and Hamilton, W. A., Cambridge University Press, Cambridge, UK, 265–304, 2007. Wieringa, E. B. A., Overmann, J., and Cypionka, H.: Detection of abundant sulphate-reducing bacteria in marine oxic sediment layers by a combined cultivation and molecular approach, Environ. Microbiol., 2(4), 417–427, 2000. Zhang, C. L., Huang, Z., Cantu, J., Pancost, R. D., Brigmon, R. L., Lyons, T. W., and Sassen, R.: Lipid biomarkers and carbon isotope signatures of a microbial (\textitBeggiatoa) mat associated with gas hydrates in the Gulf of Mexico, Appl. Environ. Microb., 71(4), 2106–2112, 2005.