We have developed a technique for cultivation of chemolithoautotrophs under high hydrostatic pressures that is successfully applicable to various types of deep-sea chemolithoautotrophs, including methanogens. It is based on a glass-syringe-sealing liquid medium and gas mixture used in conjunction with a butyl rubber piston and a metallic needle stuck into butyl rubber. By using this technique, growth, survival, and methane production of a newly isolated, hyperthermophilic methanogen Methanopyrus kandleri strain 116 are characterized under high temperatures and hydrostatic pressures. Elevated hydrostatic pressures extend the temperature maximum for possible cell proliferation from 116°C at 0.4 MPa to 122°C at 20 MPa, providing the potential for growth even at 122°C under an in situ high pressure. In addition, piezophilic growth significantly affected stable carbon isotope fractionation of methanogenesis from CO 2. Under conventional growth conditions, the isotope fractionation of methanogenesis by M. kandleri strain 116 was similar to values (؊34‰ to؊27‰) previously reported for other hydrogenotrophic methanogens. However, under high hydrostatic pressures, the isotope fractionation effect became much smaller (<؊12‰), and the kinetic isotope effect at 122°C and 40 MPa was ؊9.4‰, which is one of the smallest effects ever reported. This observation will shed light on the sources and production mechanisms of deep-sea methane.carbon isotope fractionation ͉ deep-sea hydrothermal vent ͉ hyperthermophile ͉ methanogenesis ͉ piezophilic M icrobial methanogenesis in the deep sea is a key process in the carbon cycle of Earth. It contributes to the CH 4 pool (free gas and methane hydrate), a potential energy source and alternative to petroleum (1, 2) as well as a strong greenhouse gas with a potential for rapid release (3), in deep-sea and subseafloor sediments. Methanogens are known to have several methanogenic types using different substrates of H 2 , acetate, methanol, CO, and so on. Hyperthermophilic hydrogenotrophic methanogens play a major role in primary production of ecosystems in deep-sea hydrothermal areas in the present Earth (4, 5) and may represent the most ancient type of microorganisms flourishing in the Archean Earth (6-10).Despite the significance of methanogens in the deep-sea and subseafloor ecosystems, the ecophysiological and biogeochemical characteristics of their in situ habitats have been little understood. It has been quite difficult to incorporate high hydrostatic pressures into experiments involving gaseous substrates such as H 2 and CO 2 . If this difficulty can be overcome by any specific apparatus (11,12), the subsequent handling of microbiological experiments under high hydrostatic pressures remains a great technical barrier. Thus, growth characterization of only thermophilic methanogens Methanocaldococcus jannaschii and Methanothermococcus thermolithotrophicus under high pressures has been successfully achieved, and only their piezophilic responses of growth and methane production have been inv...

Subsurface microbial communities supported by geologically and abiologically derived hydrogen and carbon dioxide from the Earth's interior are of great interest, not only with regard to the nature of primitive life on Earth, but as potential analogs for extraterrestrial life. Here, for the first time, we present geochemical and microbiological evidence pointing to the existence of hyperthermophilic subsurface lithoautotrophic microbial ecosystem (HyperSLiME) dominated by hyperthermophilic methanogens beneath an active deep-sea hydrothermal field in the Central Indian Ridge. Geochemical and isotopic analyses of gaseous components in the hydrothermal fluids revealed heterogeneity of both concentration and carbon isotopic compositions of methane between the main hydrothermal vent (0.08 mM and -13.8 per thousand PDB, respectively) and the adjacent divergent vent site (0.2 mM and -18.5 per thousand PDB, respectively), representing potential subsurface microbial methanogenesis, at least in the divergent vent emitting more 13C-depleted methane. Extremely high abundance of magmatic energy sources such as hydrogen (2.5 mM) in the fluids also encourages a hydrogen-based, lithoautotrophic microbial activity. Both cultivation and cultivation-independent molecular analyses suggested the predominance of Methanococcales members in the superheated hydrothermal emissions and chimney interiors along with the other major microbial components of Thermococcales members. These results imply that a HyperSLiME, consisting of methanogens and fermenters, occurs in this tectonically active subsurface zone, strongly supporting the existence of hydrogen-driven subsurface microbial communities.

Abstract. Formaldehyde (HCHO), the most abundant carbonyl compound in the atmosphere, is generated as an intermediate product in the oxidation of nonmethane hydrocarbons. Proton transfer reaction mass spectrometry (PTR-MS) has the capability to detect HCHO from ion signals at m/z 31 with high time-resolution. However, the detection sensitivity is low compared to other detectable species, and is considerably affected by humidity, due to back reactions between protonated HCHO and water vapor prior to analysis. We performed a laboratory calibration of PTR-MS for HCHO and examined the detection sensitivity and humidity dependence at various field strengths. Subsequently, we deployed the PTR-MS instrument in a field campaign at Mount Tai

Increasing levels of CO2 in the atmosphere are expected to cause climatic change with negative effects on the earth's ecosystems and human society. Consequently, a variety of CO 2 disposal options are discussed, including injection into the deep ocean. Because the dissolution of CO2 in seawater will decrease ambient pH considerably, negative consequences for deep-water ecosystems have been predicted. Hence, ecosystems associated with natural CO2 reservoirs in the deep sea, and the dynamics of gaseous, liquid, and solid CO 2 in such environments, are of great interest to science and society. We report here a biogeochemical and microbiological characterization of a microbial community inhabiting deep-sea sediments overlying a natural CO 2 lake at the Yonaguni Knoll IV hydrothermal field, southern Okinawa Trough. We found high abundances (>10 9 cm ؊3 ) of microbial cells in sediment pavements above the CO2 lake, decreasing to strikingly low cell numbers (10 7 cm ؊3 ) at the liquid CO2͞CO2-hydrate interface. The key groups in these sediments were as follows: (i) the anaerobic methanotrophic archaea ANME-2c and the Eel-2 group of Deltaproteobacteria and (ii) sulfur-metabolizing chemolithotrophs within the Gamma-and Epsilonproteobacteria. The detection of functional genes related to one-carbon assimilation and the presence of highly 13 C-depleted archaeal and bacterial lipid biomarkers suggest that microorganisms assimilating CO2 and͞or CH4 dominate the liquid CO2 and CO 2-hydrate-bearing sediments. Clearly, the Yonaguni Knoll is an exceptional natural laboratory for the study of consequences of CO 2 disposal as well as of natural CO2 reservoirs as potential microbial habitats on early Earth and other celestial bodies.anaerobic oxidation of methane ͉ chemolithotroph ͉ CO2 disposal ͉ CO 2 hydrate ͉ liquid CO2

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The GEOTRACES Intermediate Data Product 2014 (IDP2014) is the first publicly available data product of the international GEOTRACES programme, and contains data measured and quality controlled before the end of 2013. It consists of two parts: (1) a compilation of digital data for more than 200 trace elements and isotopes (TEls) as well as classical hydrographic parameters, and (2) the eGEOTRACES Electronic Atlas providing a strongly inter-linked on-line atlas including more than 300 section plots and 90 animated 3D scenes. The IDP2014 covers the Atlantic, Arctic, and Indian oceans, exhibiting highest data density in the Atlantic. The TEI data in the IDP2014 are quality controlled by careful assessment of intercalibration results and multi-laboratory data comparisons at cross-over stations. The digital data are provided in several formats, including ASCII spreadsheet, Excel spreadsheet, netCDF, and Ocean Data View collection. In addition to the actual data values the IDP2014 also contains data quality flags and 1-sigma data error values where available. Quality flags and error values are useful for data filtering. Metadata about data originators, analytical methods and original publications related to the data are linked to the data in an easily accessible way. The eGEOTRACES Electronic Atlas is the visual representation of the IDP2014 data providing section plots and a new kind of animated 3D scenes. The basin-wide 3D scenes allow for viewing of data from many cruises at the same time, thereby providing quick overviews of large-scale tracer distributions. In addition, the 3D scenes provide geographical and bathymetric context that is crucial for the interpretation and assessment of observed tracer plumes, as well as for making inferences about controlling processes. (C) 2015 The Authors. Published by Elsevier B.V

[1] We measured the intramolecular distribution of 15 N as well as conventional nitrogen and oxygen isotope ratios in oceanic nitrous oxide (N 2 O) in the western North Pacific for the first time. In contrast to a nearly homogeneous vertical distribution of a bulk nitrogen isotope ratio, a wide variation in site preference for intramolecular 15 N distribution was found, suggesting the subsurface and deep source mixing and the production mechanism in the ocean. The oceanic composition of N 2 O isotopomers quantitatively outlines the ocean as the most significant source after the terrestrial one.

[1] We determined the chemical and isotopic compositions of the liquid CO 2 found on Yonaguni IV knoll hydrothermal site, as well as those in hydrothermal fluid venting from the surrounding chimneys. The d 13 C of both CO 2 and CH 4 in the liquid CO 2 almost coincide with those in the hydrothermal fluid, suggesting that the liquid CO 2 must be derived from the hydrothermal fluid. While showing homogeneous d 13 C, the hydrothermal fluids exhibit wide variation in gas contents. Active phase separation must be taking place within the conduits. Besides, H 2 -depletion in the liquid CO 2 suggests formation of solid CO 2 -hydrate must also precede the venting of liquid CO 2 . In conclusion, liquid CO 2 must be produced through following subseafloor processes: phase separation of hydrothermal fluid due to boiling, formation of solid CO 2 -hydrate due to cooling of vapor phase, and melting of the solid CO 2 -hydrate to liquid CO 2 due to a temperature increase within the sedimentary layer.