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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.
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.
Terrestrial mud volcanoes (TMVs) represent geochemically diverse habitats with varying sulfur sources and yet sulfur cycling in these environments remains largely unexplored. Here we characterized the sulfur-metabolizing microorganisms and activity in four TMVs in Azerbaijan. A combination of geochemical analyses, biological rate measurements and molecular diversity surveys (targeting metabolic genes aprA and dsrA and SSU ribosomal RNA) supported the presence of active sulfur-oxidizing and sulfate-reducing guilds in all four TMVs across a range of physiochemical conditions, with diversity of these guilds being unique to each TMV. The TMVs varied in potential sulfate reduction rates (SRR) by up to four orders of magnitude with highest SRR observed in sediments where in situ sulfate concentrations were highest. Maximum temperatures at which SRR were measured was 60 degrees C in two TMVs. Corresponding with these trends in SRR, members of the potentially thermophilic, spore-forming, Desulfotomaculum were detected in these TMVs by targeted 16S rRNA analysis. Additional sulfate-reducing bacterial lineages included members of the Desulfobacteraceae and Desulfobulbaceae detected by aprA and dsrA analyses and likely contributing to the mesophilic SRR measured. Phylotypes affiliated with sulfide-oxidizing Gamma- and Betaproteobacteria were abundant in aprA libraries from low sulfate TMVs, while the highest sulfate TMV harboured 16S rRNA phylotypes associated with sulfur-oxidizing Epsilonproteobacteria. Altogether, the biogeochemical and microbiological data indicate these unique terrestrial habitats support diverse active sulfur-cycling microorganisms reflecting the in situ geochemical environment.
Multibeam bathymetry revealed the occurrence of numerous craterlike depressions, so-called pockmarks, on the sea floor of the Hammerfest Basin and the Loppa High, south-western Barents Sea. To investigate whether these pockmarks are related to ongoing gas seepage, microbial processes associated with methane metabolism were analyzed using geochemical, biogeochemical and microbiological techniques. Gravity cores were collected along transects crossing individual pockmarks, allowing a direct comparison between different locations inside (assumed activity center), on the rim, and outside of a pockmark (reference sites). Concentrations of hydrocarbons in the sediment, particularly methane, were measured as headspace (free) gas, and in the occluded and adsorbed gas fraction. Down to a depth of 2.6 m below sea floor (mbsf) sulfate reduction rates were quantified by radiotracer incubations. Concentrations of dissolved sulfate in the porewater were determined as well. Neither the sulfate profiles nor the gas measurements show any evidence of microbial activity or active fluid venting. Methane concentrations and sulfate reduction rates were extremely low or even below the detection limit. The results show that the observed sediment structures are most likely paleo-pockmarks, their formation probably occurred during the last deglaciation.
As part of the International Continental Drilling Program deep lake drilling project Paleo Van, we investigated sulfate reduction (SR) in deep sediment cores of the saline, alkaline (salinity 21.4%0, alkalinity 155 m mEq-1, pH 9.81) Lake Van, Turkey. The cores were retrieved in the Northern Basin (NB) and at Ahlat Ridge (AR) and reached a maximum depth of 220 m. Additionally, 65-75 cm long gravity cores were taken at both sites. SR rates (SRR) were low (<22 nmol cm-3 day-1) compared to lakes with higher salinity and alkalinity, indicating that salinity and alkalinity are not limiting SR in Lake Van. Both sites differ significantly in rates and depth distribution of SR. In NB, SRR are up to 10 times higher than at AR. SR could be detected down to 19 mblf (meters below lake floor) at NB and down to 13 mblf at AR. Although SRR were lower at AR than at NB, organic matter (OM) concentrations were higher. In contrast, dissolved OM in the pore water at AR contained more macromolecular OM and less low molecular weight OM.VVe thus suggest, that OM content alone cannot be used to infer microbial activity at Lake Van but that quality of OM has an important impact as well. These differences suggest that biogeochemical processes in lacustrine sediments are reacting very sensitively to small variations in geological, physical, or chemical parameters over relatively short distances.
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.
Based on results of previous investigations of tephra-tuff volcaniclastic deposits and a geophysical survey in the surroundings of the Zelezna hurka Quaternary volcano, West Bohemia, we performed detailed geophysical Surveys using gravimetry, magnetometry and electrical conductivity techniques. Striking anomalies were revealed in a morphological depression near Mytina, West Bohemia, as a strong evidence of the assumed maar-diatreme structure. The sharp isometric gravity low of -2.30 mGal, as well as the corresponding positive magnetic anomaly of 200 nT with a negative rim on its northern side indicate a steeply clipping geological body of low density and containing magnetic rocks/minerals. Magnetic survey also showed pronounced local anomalies outside the depression that can reflect relicts of the tephra rim of the maar. This geophysical evidence was then proven by an exploratory drilling near the centre of the gravity anomaly. Macroscopic on-site evaluation of the core, and more detailed sedimentological, petrochemical, palynological and microbiological laboratory analyses further confirmed the existence of a maar structure filled by 84 m of lake sediments reflecting a Succession of several warm and cold climatic periods. Results Of palynological analyses confirm the presence of a continuous palaeoclimate archive, with at least three successive warmer periods of most probably interstadial character from the upper Quaternary Saalian complex. Therefore. the recovered sediment sequence holds strong potential for in-depth palaeoclimate reconstruction and deep biosphere studies. At the bottom of the Mytina-1 (MY- 1) borehole (84-85.5 M), Country rock debris Was found, containing also volcanic bombs and lapilli. The discovered volcanic Structure is considered to be the first known Quaternary maar-diatreme volcano on the territory of the Bohemian Massif. Because of hidden active magmatic processes in combination with earthquake swarm seismicity ca. 20-30 km north of the Mytina maar, reconstruction of the palaeovolcanological evolution is important for evaluation of hazard potential of the NE and E Part of the Cheb Basin.
We present a method for the rapid and simple extraction of DNA from marine sediments using electroelution. It effectively separates DNA from compounds, including humic substances, that interfere with subsequent DNA quantification and amplification. After extraction of the DNA from the sediment into an aqueous solution, the crude sample is encased in 2% agarose gel and exposed to an electrical current, which draws the DNA out of the gel into a centrifugal filter vial. After electroelution, the sample is centrifuged to remove contaminants <= 100 000 Da. Recovery of DNA using this method is quantitative and does not discriminate on the basis of size, as determined using DNA standards and DNA extracts from environmental samples. Amplification of DNA is considerably improved due to removal of PCR inhibitors. For Archaea, only these purified extracts yielded PCR products. This method allows for the use of relatively large volumes of sediment and is particularly useful for sediments containing low biomass such as deeply buried marine sediments. It works with both organic-rich and -poor sediment, as well as with sediment where calcium carbonate is abundant and sediment where it is limited; consequently, adjustment of protocols is unnecessary for samples with very different organic and mineral contents.
The low-productivity South Pacific Gyre (SPG) is Earth's largest oceanic province. Its sediment accumulates extraordinarily slowly (0.1-1 m per million years). This sediment contains a living community that is characterized by very low biomass and very low metabolic activity. At every depth in cored SPG sediment, mean cell abundances are 3 to 4 orders of magnitude lower than at the same depths in all previously explored subseafloor communities. The net rate of respiration by the subseafloor sedimentary community at each SPG site is 1 to 3 orders of magnitude lower than the rates at previously explored sites. Because of the low respiration rates and the thinness of the sediment, interstitial waters are oxic throughout the sediment column in most of this region. Consequently, the sedimentary community of the SPG is predominantly aerobic, unlike previously explored subseafloor communities. Generation of H-2 by radiolysis of water is a significant electron-donor source for this community. The per-cell respiration rates of this community are about 2 orders of magnitude higher (in oxidation/reduction equivalents) than in previously explored anaerobic subseafloor communities. Respiration rates and cell concentrations in subseafloor sediment throughout almost half of the world ocean may approach those in SPG sediment.