TY - JOUR A1 - Ijiri, Akira A1 - Inagaki, Fumio A1 - Kubo, Yusuke A1 - Adhikari, Rishi Ram A1 - Hattori, Shohei A1 - Hoshino, Tatsuhiko A1 - Imachi, Hiroyuki A1 - Kawagucci, Shinsuke A1 - Morono, Yuki A1 - Ohtomo, Yoko A1 - Ono, Shuhei A1 - Sakai, Sanae A1 - Takai, Ken A1 - Toki, Tomohiro A1 - Wang, David T. A1 - Yoshinaga, Marcos Y. A1 - Arnold, Gail L. A1 - Ashi, Juichiro A1 - Case, David H. A1 - Feseker, Tomas A1 - Hinrichs, Kai-Uwe A1 - Ikegawa, Yojiro A1 - Ikehara, Minoru A1 - Kallmeyer, Jens A1 - Kumagai, Hidenori A1 - Lever, Mark Alexander A1 - Morita, Sumito A1 - Nakamura, Ko-ichi A1 - Nakamura, Yuki A1 - Nishizawa, Manabu A1 - Orphan, Victoria J. A1 - Roy, Hans A1 - Schmidt, Frauke A1 - Tani, Atsushi A1 - Tanikawa, Wataru A1 - Terada, Takeshi A1 - Tomaru, Hitoshi A1 - Tsuji, Takeshi A1 - Tsunogai, Urumu A1 - Yamaguchi, Yasuhiko T. A1 - Yoshida, Naohiro T1 - Deep-biosphere methane production stimulated by geofluids in the Nankai accretionary complex JF - Science Advances Y1 - 2018 U6 - https://doi.org/10.1126/sciadv.aao4631 SN - 2375-2548 VL - 4 IS - 6 PB - American Assoc. for the Advancement of Science CY - Washington ER - TY - JOUR A1 - Adhikari, Rishi Ram A1 - Glombitza, Clemens A1 - Nickel, Julia C. A1 - Anderson, Chloe H. A1 - Dunlea, Ann G. A1 - Spivack, Arthur J. A1 - Murray, Richard W. A1 - Kallmeyer, Jens T1 - Hydrogen Utilization Potential in Subsurface Sediments JF - Frontiers in microbiology N2 - Subsurface microbial communities undertake many terminal electron-accepting processes, often simultaneously. Using a tritium-based assay, we measured the potential hydrogen oxidation catalyzed by hydrogenase enzymes in several subsurface sedimentary environments (Lake Van, Barents Sea, Equatorial Pacific, and Gulf of Mexico) with different predominant electron-acceptors. Hydrogenases constitute a diverse family of enzymes expressed by microorganisms that utilize molecular hydrogen as a metabolic substrate, product, or intermediate. The assay reveals the potential for utilizing molecular hydrogen and allows qualitative detection of microbial activity irrespective of the predominant electron-accepting process. Because the method only requires samples frozen immediately after recovery, the assay can be used for identifying microbial activity in subsurface ecosystems without the need to preserve live material. We measured potential hydrogen oxidation rates in all samples from multiple depths at several sites that collectively span a wide range of environmental conditions and biogeochemical zones. Potential activity normalized to total cell abundance ranges over five orders of magnitude and varies, dependent upon the predominant terminal electron acceptor. Lowest per-cell potential rates characterize the zone of nitrate reduction and highest per-cell potential rates occur in the methanogenic zone. Possible reasons for this relationship to predominant electron acceptor include (i) increasing importance of fermentation in successively deeper biogeochemical zones and (ii) adaptation of H(2)ases to successively higher concentrations of H-2 in successively deeper zones. KW - hydrogenase KW - tritium assay KW - deep biosphere KW - microbial activity KW - Lake Van KW - Barents Sea KW - Equatorial Pacific KW - Gulf of Mexico Y1 - 2016 U6 - https://doi.org/10.3389/fmicb.2016.00008 SN - 1664-302X VL - 7 PB - Frontiers Research Foundation CY - Lausanne ER - TY - GEN A1 - Ijiri, Akira A1 - Inagaki, Fumio A1 - Kubo, Yusuke A1 - Adhikari, Rishi Ram A1 - Hattori, Shohei A1 - Hoshino, Tatsuhiko A1 - Imachi, Hiroyuki A1 - Kawagucci, Shinsuke A1 - Morono, Yuki A1 - Ohtomo, Yoko A1 - Ono, Shuhei A1 - Sakai, Sanae A1 - Takai, Ken A1 - Toki, Tomohiro A1 - Wang, David T. A1 - Yoshinaga, Marcos Y. A1 - Arnold, Gail L. A1 - Ashi, Juichiro A1 - Case, David H. A1 - Feseker, Tomas A1 - Hinrichs, Kai-Uwe A1 - Ikegawa, Yojiro A1 - Ikehara, Minoru A1 - Kallmeyer, Jens A1 - Kumagai, Hidenori A1 - Lever, Mark Alexander A1 - Morita, Sumito A1 - Nakamura, Ko-ichi A1 - Nakamura, Yuki A1 - Nishizawa, Manabu A1 - Orphan, Victoria J. A1 - Røy, Hans A1 - Schmidt, Frauke A1 - Tani, Atsushi A1 - Tanikawa, Wataru A1 - Terada, Takeshi A1 - Tomaru, Hitoshi A1 - Tsuji, Takeshi A1 - Tsunogai, Urumu A1 - Yamaguchi, Yasuhiko T. A1 - Yoshida, Naohiro T1 - Deep-biosphere methane production stimulated by geofluids in the Nankai accretionary complex T2 - Postprints der Universität Potsdam Mathematisch-Naturwissenschaftliche Reihe N2 - Microbial life inhabiting subseafloor sediments plays an important role in Earth’s carbon cycle. However, the impact of geodynamic processes on the distributions and carbon-cycling activities of subseafloor life remains poorly constrained. We explore a submarine mud volcano of the Nankai accretionary complex by drilling down to 200 m below the summit. Stable isotopic compositions of water and carbon compounds, including clumped methane isotopologues, suggest that ~90% of methane is microbially produced at 16° to 30°C and 300 to 900 m below seafloor, corresponding to the basin bottom, where fluids in the accretionary prism are supplied via megasplay faults. Radiotracer experiments showed that relatively small microbial populations in deep mud volcano sediments (10 2 to 10 3 cells cm −3 ) include highly active hydrogenotrophic methanogens and acetogens. Our findings indicate that subduction-associated fluid migration has stimulated microbial activity in the mud reservoir and that mud volcanoes may contribute more substantially to the methane budget than previously estimated. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 802 KW - multiply-substituted isotopologues KW - marine subsurface sediments KW - carbon isotopic composition KW - submarine mud volcano KW - intact polar lipids KW - fore-arc basin KW - subseafloor sediments KW - microbial lipids KW - Cascadia margin KW - organic-acids Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-427002 SN - 1866-8372 IS - 802 ER - TY - GEN A1 - Adhikari, Rishi Ram A1 - Glombitza, Clemens A1 - Nickel, Julia C. A1 - Anderson, Chloe H. A1 - Dunlea, Ann G. A1 - Spivack, Arthur J. A1 - Murray, Richard W. A1 - D’Hondt, Steven A1 - Kallmeyer, Jens T1 - Hydrogen utilization potential in subsurface sediments T2 - Frontiers in microbiology N2 - Subsurface microbial communities undertake many terminal electron-accepting processes, often simultaneously. Using a tritium-based assay, we measured the potential hydrogen oxidation catalyzed by hydrogenase enzymes in several subsurface sedimentary environments (Lake Van, Barents Sea, Equatorial Pacific, and Gulf of Mexico) with different predominant electron-acceptors. Hydrogenases constitute a diverse family of enzymes expressed by microorganisms that utilize molecular hydrogen as a metabolic substrate, product, or intermediate. The assay reveals the potential for utilizing molecular hydrogen and allows qualitative detection of microbial activity irrespective of the predominant electron-accepting process. Because the method only requires samples frozen immediately after recovery, the assay can be used for identifying microbial activity in subsurface ecosystems without the need to preserve live material. We measured potential hydrogen oxidation rates in all samples from multiple depths at several sites that collectively span a wide range of environmental conditions and biogeochemical zones. Potential activity normalized to total cell abundance ranges over five orders of magnitude and varies, dependent upon the predominant terminal electron acceptor. Lowest per-cell potential rates characterize the zone of nitrate reduction and highest per-cell potential rates occur in the methanogenic zone. Possible reasons for this relationship to predominant electron acceptor include (i) increasing importance of fermentation in successively deeper biogeochemical zones and (ii) adaptation of H(2)ases to successively higher concentrations of H-2 in successively deeper zones. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 447 KW - hydrogenase KW - tritium assay KW - deep biosphere KW - microbial activity KW - Lake Van KW - Barents Sea KW - Equatorial Pacific KW - Gulf of Mexico Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-407678 ER - TY - JOUR A1 - Roy, Hans A1 - Kallmeyer, Jens A1 - Adhikari, Rishi Ram A1 - Pockalny, Robert A1 - Jorgensen, Bo Barker A1 - D'Hondt, Steven T1 - Aerobic microbial respiration in 86-million-year-old deep-sea red clay JF - Science N2 - Microbial communities can subsist at depth in marine sediments without fresh supply of organic matter for millions of years. At threshold sedimentation rates of 1 millimeter per 1000 years, the low rates of microbial community metabolism in the North Pacific Gyre allow sediments to remain oxygenated tens of meters below the sea floor. We found that the oxygen respiration rates dropped from 10 micromoles of O-2 liter(-1) year(-1) near the sediment-water interface to 0.001 micromoles of O-2 liter(-1) year(-1) at 30-meter depth within 86 million-year-old sediment. The cell-specific respiration rate decreased with depth but stabilized at around 10(-3) femtomoles of O-2 cell(-1) day(-1) 10 meters below the seafloor. This result indicated that the community size is controlled by the rate of carbon oxidation and thereby by the low available energy flux. Y1 - 2012 U6 - https://doi.org/10.1126/science.1219424 SN - 0036-8075 VL - 336 IS - 6083 SP - 922 EP - 925 PB - American Assoc. for the Advancement of Science CY - Washington ER - TY - JOUR A1 - Kallmeyer, Jens A1 - Pockalny, Robert A1 - Adhikari, Rishi Ram A1 - Smith, David C. A1 - D'Hondt, Steven T1 - Global distribution of microbial abundance and biomass in subseafloor sediment JF - Proceedings of the National Academy of Sciences of the United States of America N2 - The global geographic distribution of subseafloor sedimentary microbes and the cause(s) of that distribution are largely unexplored. Here, we show that total microbial cell abundance in subseafloor sediment varies between sites by ca. five orders of magnitude. This variation is strongly correlated with mean sedimentation rate and distance from land. Based on these correlations, we estimate global subseafloor sedimentary microbial abundance to be 2.9 center dot 10(29) cells [corresponding to 4.1 petagram (Pg) C and similar to 0.6% of Earth's total living biomass]. This estimate of subseafloor sedimentary microbial abundance is roughly equal to previous estimates of total microbial abundance in seawater and total microbial abundance in soil. It is much lower than previous estimates of subseafloor sedimentary microbial abundance. In consequence, we estimate Earth's total number of microbes and total living biomass to be, respectively, 50-78% and 10-45% lower than previous estimates. KW - deep biosphere KW - cell enumeration KW - global microbial biomass KW - subsurface life Y1 - 2012 U6 - https://doi.org/10.1073/pnas.1203849109 SN - 0027-8424 VL - 109 IS - 40 SP - 16213 EP - 16216 PB - National Acad. of Sciences CY - Washington ER - TY - JOUR A1 - Adhikari, Rishi Ram A1 - Kallmeyer, Jens T1 - Detection and quantification of microbial activity in the subsurface N2 - The subsurface harbors a large fraction of Earth's living biomass, forming complex microbial ecosystems. Without a profound knowledge of the ongoing biologically mediated processes and their reaction to anthropogenic changes it is difficult to assess the long-term stability and feasibility of any type of geotechnical utilization, as these influence subsurface ecosystems. Despite recent advances in many areas of subsurface microbiology, the direct quantification of turnover processes is still in its infancy, mainly due to the extremely low cell abundances. We provide an overview of the currently available techniques for the quantification of microbial turnover processes and discuss their specific strengths and limitations. Most techniques employed so far have focused on specific processes, e.g. sulfate reduction or methanogenesis. Recent studies show that processes that were previously thought to exclude each other can occur simultaneously, albeit at very low rates. Without the identification of the respective processes it is impossible to quantify total microbial activity. Even in cases where all simultaneously occurring processes can be identified, the typically very low rates prevent quantification. In many cases a simple measure of total microbial activity would be a better and more robust measure than assays for several specific processes. Enzyme or molecular assays provide a more general approach as they target key metabolic compounds. Depending on the compound targeted a broader spectrum of microbial processes can be quantified. The two most promising compounds are ATP and hydrogenase, as both are ubiquitous in microbes. Technical constraints limit the applicability of currently available ATP-assays for subsurface samples. A recently developed hydrogenase radiotracer assay has the potential to become a key tool for the quantification of subsurface microbial activity. Y1 - 2010 UR - http://www.sciencedirect.com/science/journal/00092819 U6 - https://doi.org/10.1016/j.chemer.2010.05.003 SN - 0009-2819 ER - TY - THES A1 - Adhikari, Rishi Ram T1 - Quantification of total microbial biomass and metabolic activity in subsurface sediments T1 - Quantification of total microbial biomass and metabolic activity in subsurface sediments N2 - Metabolically active microbial communities are present in a wide range of subsurface environments. Techniques like enumeration of microbial cells, activity measurements with radiotracer assays and the analysis of porewater constituents are currently being used to explore the subsurface biosphere, alongside with molecular biological analyses. However, many of these techniques reach their detection limits due to low microbial activity and abundance. Direct measurements of microbial turnover not just face issues of insufficient sensitivity, they only provide information about a single specific process but in sediments many different process can occur simultaneously. Therefore, the development of a new technique to measure total microbial activity would be a major improvement. A new tritium-based hydrogenase-enzyme assay appeared to be a promising tool to quantify total living biomass, even in low activity subsurface environments. In this PhD project total microbial biomass and microbial activity was quantified in different subsurface sediments using established techniques (cell enumeration and pore water geochemistry) as well as a new tritium-based hydrogenase enzyme assay. By using a large database of our own cell enumeration data from equatorial Pacific and north Pacific sediments and published data it was shown that the global geographic distribution of subseafloor sedimentary microbes varies between sites by 5 to 6 orders of magnitude and correlates with the sedimentation rate and distance from land. Based on these correlations, global subseafloor biomass was estimated to be 4.1 petagram-C and ~0.6 % of Earth's total living biomass, which is significantly lower than previous estimates. Despite the massive reduction in biomass the subseafloor biosphere is still an important player in global biogeochemical cycles. To understand the relationship between microbial activity, abundance and organic matter flux into the sediment an expedition to the equatorial Pacific upwelling area and the north Pacific Gyre was carried out. Oxygen respiration rates in subseafloor sediments from the north Pacific Gyre, which are deposited at sedimentation rates of 1 mm per 1000 years, showed that microbial communities could survive for millions of years without fresh supply of organic carbon. Contrary to the north Pacific Gyre oxygen was completely depleted within the upper few millimeters to centimeters in sediments of the equatorial upwelling region due to a higher supply of organic matter and higher metabolic activity. So occurrence and variability of electron acceptors over depth and sites make the subsurface a complex environment for the quantification of total microbial activity. Recent studies showed that electron acceptor processes, which were previously thought to thermodynamically exclude each other can occur simultaneously. So in many cases a simple measure of the total microbial activity would be a better and more robust solution than assays for several specific processes, for example sulfate reduction rates or methanogenesis. Enzyme or molecular assays provide a more general approach as they target key metabolic compounds. Since hydrogenase enzymes are ubiquitous in microbes, the recently developed tritium-based hydrogenase radiotracer assay is applied to quantify hydrogenase enzyme activity as a parameter of total living cell activity. Hydrogenase enzyme activity was measured in sediments from different locations (Lake Van, Barents Sea, Equatorial Pacific and Gulf of Mexico). In sediment samples that contained nitrate, we found the lowest cell specific enzyme activity around 10^(-5) nmol H_(2) cell^(-1) d^(-1). With decreasing energy yield of the electron acceptor used, cell-specific hydrogenase activity increased and maximum values of up to 1 nmol H_(2) cell^(-1) d^(-1) were found in samples with methane concentrations of >10 ppm. Although hydrogenase activity cannot be converted directly into a turnover rate of a specific process, cell-specific activity factors can be used to identify specific metabolism and to quantify the metabolically active microbial population. In another study on sediments from the Nankai Trough microbial abundance and hydrogenase activity data show that both the habitat and the activity of subseafloor sedimentary microbial communities have been impacted by seismic activities. An increase in hydrogenase activity near the fault zone revealed that the microbial community was supplied with hydrogen as an energy source and that the microbes were specialized to hydrogen metabolism. N2 - Mikrobielle Gesellschaften und ihre aktiven Stoffwechselprozesse treten in einer Vielzahl von Sedimenten unterschiedlichster Herkunft auf. In der Erforschung dieser tiefen Biosphäre werden derzeit Techniken wie Zellzählungen, Aktivitätsmessungen mit Radiotracer-Versuchen und Analysen der Porenwasserzusammensetzung angewendet, darüber hinaus auch molekularbiologische Analysen. Viele dieser Methoden stoßen an ihre Nachweisgrenze, wenn Sedimente mit geringer Zelldichte und mikrobieller Aktivität untersucht werden. Bei der Untersuchung von Stoffwechselprozessen mit herkömmlichen Techniken kommt dazu, dass von mehreren Prozessen, die zeitgleich ablaufen können, jeweils nur einer erfasst wird. Deswegen wäre die Entwicklung einer neuartigen Messtechnik für die gesamte mikrobielle Aktivität ein wesentlicher Fortschritt für die Erforschung der tiefen Biosphäre. Ein vielversprechender Ansatz, um die gesamte lebende Biomasse auch in Proben mit geringer Aktivität zu bestimmen, ist eine Hydrogenase-Enzym-Versuchsanordnung mit Tritium als quantifizierbarer Messgröße. In dieser Doktorarbeit wurde die gesamte mikrobielle Biomasse und Aktivität von unterschiedlichen Sedimentproben einerseits mit herkömmlichen Methoden (Zellzählungen, Analyse der Porenwasserzusammensetzung) als auch mit einer neu entwickelten Hydrogenase-Enzym-Versuchsanordnung quantifiziert. Mit einer großen Anzahl eigener Zellzählungsdaten von Sedimenten aus dem Äquatorialpazifik und dem Nordpazifik und ergänzenden publizierten Daten konnte gezeigt werden, dass Zellzahlen sich in ihrer globalen geographischen Verteilung je nach Bohrlokation um 5 bis 6 Größenordnungen unterscheiden. Dabei bestehen Korrelationen zur Sedimentationsrate und zur Entfernung zum Land, mit deren Hilfe sich die Gesamtbiomasse in Tiefseesedimenten zu 4,1 Petagramm-C abschätzen lässt. Das entspricht ~0,6 % der Gesamtbiomasse der Erde und ist damit erheblich weniger als in früheren Schätzungen angegeben. Trotz der Korrektur auf diesen Wert spielt die Biomasse der tiefen Biosphäre weiterhin eine erhebliche Rolle in biogeochemischen Kreisläufen. Um die Zusammenhänge zwischen Aktivität der Mikroben, der Häufigkeit ihres Auftretens und Zustrom von organischem Material zu verstehen, wurde eine Expedition ins Auftriebsgebiet des Äquatorialpazifiks und zum nordpazifischen Wirbel durchgeführt. Daten der Sauerstoffaufnahme in Sedimenten des nordpazifischen Wirbels, die mit Sedimentationsraten von 1 mm pro 1000 Jahren abgelagert werden, zeigen, dass mikrobielle Gesellschaften über Millionen von Jahren ohne Zufuhr von frischem organischen Kohlenstoff überleben konnten. Im Gegensatz zum nordpazifischen Wirbel wird in Sedimenten des äquatorialpazifischen Auftriebsgebiets Sauerstoff bei höherer mikrobieller Aktivität und Verfügbarkeit organischer Verbindungen oberflächennah in den ersten Milli- bis Zentimetern komplett umgesetzt. Auftreten und Variabilität von Elektronenakzeptoren nach Tiefe und Bohrlokation machen die tiefe Biosphäre zu einer komplexen Umgebung für die Quantifizierung der gesamten mikrobiellen Aktivität. Aktuelle Studien zeigen das verschiedene Elektronenakzeptorprozesse gleichzeitig ablaufen können, obwohl man bisher davon ausgegangen war, dass diese sich thermodynamisch ausschließen. In vielen Fällen wäre also eine einfache Methode zur Messung der gesamten mikrobiellen Aktivität eine bessere und verlässlichere Lösung aktueller Analyseaufgaben als Messungen mehrerer Einzelprozesse wie beispielsweise Sulfatreduktion und Methanogenese. Enzym-oder Molekular-Versuchsanordnungen sind ein prozessumfassender Ansatz, weil hier Schlüsselkomponenten der Stoffwechselprozesse untersucht werden. Das Hydrogenase-Enzym ist eine solche Schlüsselkomponente und in Mikroben allgegenwärtig. Deshalb kann die Quantifizierung seiner Aktivität mit der neu entwickelten Hydrogenase-Enzym-Versuchsanordnung als Parameter für die gesamte mikrobielle Aktivität der lebenden Zellen verwendet werden. Hydrogenase-Aktivitäten wurden in Sedimenten unterschiedlicher Lokationen (Vansee, Barentssee, Äquatorialpazifik, und Golf von Mexico) gemessen. In Sedimentproben, die Nitrat enthielten, haben wir mit ca. 10^(-5) nmol H_(2) cell^(-1) d^(-1) die geringste zellspezifische Hydrogenase-Aktivität gefunden. Mit geringerem Energiegewinn des genutzten Elektronenakzeptors steigt die zellspezifische Hydrogenase-Aktivität. Maximalwerte von bis zu 1 nmol H_(2) cell^(-1) d^(-1) wurden in Sedimentproben mit >10 ppm Methankonzentration gefunden. Auch wenn die Hydrogenase-Aktivität nicht direkt in die Umsatzrate eines spezifischen Prozesses konvertierbar ist, können zellspezifische Aktivitätsfaktoren verwendet werden, um die metabolisch aktive Mikrobenpopulation zu quantifizieren. In einer weiteren Studie mit Sedimenten des Nankai-Grabens zeigen Daten der Zelldichte und der Hydrogenase-Aktivität einen Einfluss von seismischen Ereignissen auf Lebensraum und Aktivität der mikrobiellen Gesellschaften. Ein Anstieg der Hydrogenase-Aktivität nahe der Verwerfungszone machte deutlich, dass die mikrobiellen Gesellschaften mit Wasserstoff als Energiequelle versorgt wurden und dass die Mikroben auf einen Wasserstoff-Stoffwechsel spezialisiert waren. KW - Hydrogenase KW - Tritium Versuchsanordnung KW - Untergrunduntersuchung der Biosphäre KW - Hydrogenase KW - Tritium Assay KW - Subsurface Biosphere Y1 - 2013 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-67773 ER -