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Viruses are ubiquitous and cause significant mortality in marine bacterial and archaeal communities. Little is known about the role of viruses in the sub-seafloor biosphere, which hosts a large fraction of all microbes on Earth. We quantified and characterized viruses in sediments from the Baltic Sea. The results show that the Baltic Sea sub-seafloor biosphere harbors highly abundant viruses with densities up to 1.8 × 1010 viruses cm−3. High potential viral production down to 37 meters below seafloor in ca. 6000-years-old sediments and infected prokaryotic cells visible by transmission electron microscopy demonstrate active viral infection. Morphological and molecular data indicate that the highly diverse community of viruses includes both allochthonous input from the overlying seawater and autochthonous production. The detection of cyanophage-like sequences showed that viruses of phototrophic hosts may persist in marine sediments for thousands of years. Our results imply that viruses influence sub-seafloor microbial community dynamics and thereby affect biogeochemical processes in the sub-seafloor biosphere.

Prokaryotic activity and diversity of the sulphate-methane transition zone (SMTZ) of sediments from Aarhus Bay sediments were characterised. This zone had elevated sulphate reduction and anaerobic oxidation of methane, along with some stimulation in the total prokaryotic population, but surprisingly no active methanogenesis (acetoclastic or hydrogenotrophic) was detected. The prokaryotic population was similar to that present in other SMTZs with prokaryotes belonging to the Deltaproteobacteria, Gammaproteobacteria, JS1, Planctomycetes, Chloroflexi, ANME-1, MBG-D/Thermoplasmatales and MCG. Many of these groups were then maintained in a long-term (6 month), heterotrophic (10 mM glucose, acetate), sediment slurry enrichment with repeat low sulphate and acetate additions (∼2 mM). Other prokaryotes were also enriched including methanogens, consistent with methane formation, Firmicutes, Bacteroidetes, Synergistetes and TM6. This slurry was then inoculated into deep-well plates containing a matrix of substrate (glucose, acetate) and sulphate concentrations (85 combinations, 2 media) for further selective enrichment of SMTZ bacteria. These results demonstrate that important SMTZ bacteria can be maintained in long-term cultivation under specific conditions. For example, JS1 always grew in mixed culture with acetate or acetate/glucose plus sulphate. Chloroflexi occurred more widespread, including in the presence of acetate, which had previously not been shown to be a substrate for Chloroflexi subphylum I. However, Chloroflexi was always found in association with other organisms and was more dominant in medium with seawater salt concentrations. These results provide more information about the physiology of a range of SMTZ prokaryotes and shows that many can be maintained under mixed heterotrophic conditions, including those with few or no cultivated representatives.

Submarine mud volcanoes are a significant source of methane to the atmosphere. The Napoli mud volcano, situated in the brine-impacted Olimpi Area of the Eastern Mediterranean Sea, emits mainly biogenic methane particularly at the centre of the mud volcano. Temperature gradients support the suggestion that Napoli is a cold mud volcano with moderate fluid flow rates. Biogeochemical and molecular genetic analyses were carried out to assess the methanogenic activity rates, pathways and diversity in the hypersaline sediments of the centre of the Napoli mud volcano. Methylotrophic methanogenesis was the only significant methanogenic pathway in the shallow sediments (0–40 cm) but was also measured throughout the sediment core, confirming that methylotrophic methanogens could be well adapted to hypersaline environments. Hydrogenotrophic methanogenesis was the dominant pathway below 50 cm; however, low rates of acetoclastic methanogenesis were also present, even in sediment layers with the highest salinity, showing that these methanogens can thrive in this extreme environment. PCR-DGGE and methyl coenzyme M reductase gene libraries detected sequences affiliated with anaerobic methanotrophs (mainly ANME-1) as well as Methanococcoides methanogens. Results show that the hypersaline conditions in the centre of the Napoli mud volcano influence active biogenic methane fluxes and methanogenic/methylotrophic diversity.

The deep subseafloor biosphere supports a diverse population of prokaryotes belonging to the Bacteria and Archaea. Most of the taxonomic groups identified by molecular methods contain mainly uncultured phylotypes. Despite this several cultured strains have been isolated from this habitat, but they probably do not represent the majority of the population. Evidence is starting to suggest that some of the activities measured, such as sulphate reduction and methanogenesis, reflected in geochemical profiles, are carried out by a small subset of the community detected by molecular methods. It is further possible that heterotrophy may be the most important mode of metabolism in subsurface sediments and heterotrophic microorganisms could dominate the uncultured prokaryotic population. Although, heterotrophy is limited by the increasing recalcitrance of organic matter with depth, this may be counteracted by thermal activation of buried organic matter providing additional substrates at depth.

In the Sonora Margin cold seep ecosystems (Gulf of California), sediments underlying microbial mats harbor high biogenic methane concentrations, fueling various microbial communities, such as abundant lineages of anaerobic methanotrophs (ANME). However, the biodiversity, distribution, and metabolism of the microorganisms producing this methane remain poorly understood. In this study, measurements of methanogenesis using radiolabeled dimethylamine, bicarbonate, and acetate showed that biogenic methane production in these sediments was mainly dominated by methylotrophic methanogenesis, while the proportion of autotrophic methanogenesis increased with depth. Congruently, methane production and methanogenic Archaea were detected in culture enrichments amended with trimethylamine and bicarbonate. Analyses of denaturing gradient gel electrophoresis (DGGE) fingerprinting and reverse-transcribed PCR-amplified 16S rRNA sequences retrieved from these enrichments revealed the presence of active methylotrophic Methanococcoides burtonii relatives and several new autotrophic Methanogenium lineages, confirming the cooccurrence of Methanosarcinales and Methanomicrobiales methanogens with abundant ANME populations in the sediments of the Sonora Margin cold seeps.

The aim of this work was to relate depth profiles of prokaryotic community composition with geochemical processes in the deep subseafloor biosphere at two shallow-water sites on the Peru Margin in the Pacific Ocean (ODP Leg 201, sites 1228 and 1229). Principal component analysis of denaturing gradient gel electrophoresis banding patterns of deep-sediment Bacteria, Archaea, Euryarchaeota and the novel candidate division JS1, followed by multiple regression, showed strong relationships with prokaryotic activity and geochemistry (R2=55–100%). Further correlation analysis, at one site, between the principal components from the community composition profiles for Bacteria and 12 other variables quantitatively confirmed their relationship with activity and geochemistry, which had previously only been implied. Comparison with previously published cell counts enumerated by fluorescent in situ hybridization with rRNA-targeted probes confirmed that these denaturing gradient gel electrophoresis profiles described an active prokaryotic community

The community compositions of Bacteria and Archaea were investigated in deep, sub-seafloor sediments from the highly productive Peru Margin (ODP Leg 201, sites 1228 and 1229, c. 25 km apart) down to nearly 200m below the seafloor using taxonomic (16S rRNA) and functional (mcrA and dsrA) gene markers. Bacterial and archaeal groups identified from clone libraries of 16S rRNA gene sequences at site 1229 agreed well with sequences amplified from bands excised from denaturing gradient gel electrophoresis (DGGE) depth profiles, with the exception of the Miscellaneous Crenarchaeotic Group (MCG). This suggested that the prokaryotic community at site 1228, obtained from DGGE profiling alone, was reliable. Sites were dominated by Bacteria in the Gammaproteobacteria, Chloroflexi (green nonsulphur bacteria) and Archaea in the MCG and South African Gold Mine Euryarchaeotic Group, although community composition changed with depth. The candidate division JS1 was present throughout both sites but was not dominant. The populations identified in the Peru Margin sediments consisted mainly of prokaryotes found in other deep subsurface sediments, and were more similar to communities from the Sea of Okhotsk (pelagic clays) than to those from the low organic carbon Nankai Trough sediments. Despite broad similarities in the prokaryotic community at the two sites, there were some differences, as well as differences in activity and geochemistry. Methanogens (mcrA) within the Methanosarcinales and Methanobacteriales were only found at site 1229 (4 depths analysed), whereas sulphate-reducing prokaryotes (dsrA) were only found at site 1228 (one depth), and these terminal-oxidizing prokaryotes may represent an active community component present at low abundance. This study clearly demonstrates that the deep subsurface sediments of the Peru Margin have a large diverse and metabolically active prokaryotic population.

Diverse microbial communities and numerous energy-yielding activities occur in deeply buried sediments of the eastern Pacific Ocean. Distributions of metabolic activities often deviate from the standard model. Rates of activities, cell concentrations, and populations of cultured bacteria vary consistently from one subseafloor environment to another. Net rates of major activities principally rely on electron acceptors and electron donors from the photosynthetic surface world. At open-ocean sites, nitrate and oxygen are supplied to the deepest sedimentary communities through the underlying basaltic aquifer. In turn, these sedimentary communities may supply dissolved electron donors and nutrients to the underlying crustal biosphere.

International audience; Subsurface sediments of the Sonora Margin (Guaymas Basin), located in proximity of active cold seep sites were explored. The taxonomic and functional diversity of bacterial and archaeal communities were investigated from 1 to 10 meters below the seafloor. Microbial community structure and abundance and distribution of dominant populations were assessed using complementary molecular approaches (Ribosomal Intergenic Spacer Analysis, 16S rRNA libraries and quantitative PCR with an extensive primers set) and correlated to comprehensive geochemical data. Moreover the metabolic potentials and functional traits of the microbial community were also identified using the GeoChip functional gene microarray and metabolic rates. The active microbial community structure in the Sonora Margin sediments was related to deep subsurface ecosystems (Marine Benthic Groups B and D, Miscellaneous Crenarchaeotal Group, Chloroflexi and Candidate divisions) and remained relatively similar throughout the sediment section, despite defined biogeochemical gradients. However, relative abundances of bacterial and archaeal dominant lineages were significantly correlated with organic carbon quantity and origin. Consistently, metabolic pathways for the degradation and assimilation of this organic carbon as well as genetic potentials for the transformation of detrital organic matters, hydrocarbons and recalcitrant substrates were detected, suggesting that chemoorganotrophic microorganisms may dominate the microbial community of the Sonora Margin subsurface sediments.

The microbial community of a deep (to 234 m below the sea floor) sediment gas hydrate deposit (Cascadia Margin Ocean Drilling Program Site 889/890, Leg 146) was analysed for the first time by molecular genetic techniques. Both bacterial and methanogen diversity were determined by phylogenetic analysis of ribosomal DNA sequences. High molecular mass DNA, indicative of active bacteria, was present in all of the samples. Ribosomal RNA genes were amplified from extracted DNA extracted from sediment using bacteria, and methanogen specific PCR primers, the latter designed in this study. Phylogenetic analysis of approximately 400 bacterial clones demonstrated that 96% were members of the Proteobacteria. These clones were affiliated with the alpha, beta and gamma subdivisions, with Caulobacter (Zymomonas group), Ralstonia and Pseudomonas phylotypes predominating. The methanogen clones were of low diversity and clustered in three sub-groups. Two of these sub-groups (contained 96% of the 400 clones) were closely related to Methanosarcina mazeii, while the third sub-group clustered in the Methanobacteriales. This analysis of a deep sediment gas hydrate environment shows a bacteria and methanogen community of limited diversity and confirms that the gas hydrate zone is biogeochemically active. These results are consistent with the presence of bacterial populations capable of methanogenesis throughout the core, and suggest that the methane hydrate at this site is at least partially biogenic in origin.

Deep-sea sediments become apparently more hostile to life with increasing depth as temperature and pressure rise, and organic matter becomes increasingly recalcitrant. Demonstrations of high bacterial populations in deep sediments1,2 may thus appear enigmatic. How, then, can the continued presence of active bacterial populations in deep sediments that are over 10 million years old be explained? Although volatile fatty acids, particularly acetate, are important intermediates in the anaerobic degradation of organic matter3,4, their concentrations are kept very low in sediments (

Life in hypersaline environments is typically limited by bioenergetic constraints. Microbial activity at the thermodynamic edge, such as the anaerobic oxidation of methane (AOM) coupled to sulphate reduction (SR), is thus unlikely to thrive in these environments. In this study, carbon and sulphur cycling was investigated in the extremely hypersaline cold seep sediments of Mercator mud volcano. AOM activity was partially inhibited but still present at salinity levels of 292 g L-1 (c. eightfold sea water concentration) with rates of 2.3 nmol cm-3 day-1 and was even detectable under saturated conditions. Methane and evaporite-derived sulphate comigrated in the ascending geofluids, which, in combination with a partial activity inhibition, resulted in AOM activity being spread over unusually wide depth intervals. Up to 79% of total cells in the AOM zone were identified by fluorescence in situ hybridization (FISH) as anaerobic methanotrophs of the ANME-1. Most ANME-1 cells formed monospecific chains without any attached partner. At all sites, AOM activity co-occurred with SR activity and sometimes significantly exceeded it. Possible causes of these unexpected results are discussed. This study demonstrates that in spite of a very low energy yield of AOM, microorganisms carrying this reaction can thrive in salinity up to halite saturation.

Details of techniques and approaches for a comprehensive and integrated study of bacteria in deep marine sediment layers are presented. The effectiveness of these techniques is documented by reference to anaerobic sulphate-reducing bacteria and their associated activity in deep sediments from the Japan Sea and Peru Margin. The effect of sample storage and pressure on the results obtained is considered. The characteristics of enrichment cultures of sulphate-reducing bacteria demonstrate that they are metabolically diverse, and that this diversity is maintained with increasing sediment depth. Pure cultures of sulphate-reducing bacteria from 80 and 500 m are barophilic. Maximum growth occurs at their calculated in situ pressure, which demonstrates their deep sea origin. These and other data [1–6] demonstrate the presence of a deep bacterial biosphere (to a depth of at least 500 m) within marine sediments, which is well adapted to this extreme environment. These techniques should be applicable to other subsurface environments where a combination of low bacterial populations and activities provide a major challenge to the study of microbial ecology.

The diversity of bacterial communities in deep marine sediments, up to 503 metres below the sea floor of the Japan Sea, was investigated by sequence analysis of amplified 16S rRNA genes. The use of different sample handling procedures greatly affected the types and diversity of sequences obtained. DNA from sediment samples stored aerobically for up to 24 h before freezing was dominated by sequences belonging to the β- and γ-proteobacteria, many of which appeared to originate from aerobic bacteria. Sub-samples equilibrated anaerobically at 16°C, were then injected with a radiotracer and immediately frozen, to simulate the conditions of a typical control sample from a radiotracer based activity assay, contained mostly α-proteobacterial sequences. Pristine sediment samples taken anaerobically and frozen within 2 h contained the widest diversity of sequences from α-, γ-, δ-proteobacteria and Gram-positive bacteria, which appeared to have originated from predominantly anaerobic or facultative bacteria. It was clear that both samples that were not frozen immediately (within 2 h) showed signs of enrichment of specific bacterial groups. Our results strongly suggest that immediate freezing should always be employed when sediment samples are to be used to assess bacterial diversity by molecular methods.

ALTHOUGH around 70% of the Earth's surface is marine, little is known about the microbiology of underlying sediments, which can be more than a kilometre deep1. Selective degradation of organic matter within sediments over geological time profoundly affects the chemical composition of the ocean and atmosphere2. Microbial processes have a fundamental role in surface sediments3,4, but despite geochemical evidence5, their significance in deeper sediments has not been established6. Here we report the discovery of viable sediment bacterial populations at five Pacific Ocean sites to depths >500m. Bacterial distributions and activities are commensurate with geochemical changes. Bacterial profiles with depth are remarkably consistent, and deviations can be linked to specific environmental factors. The rate of decline in these populations indicates that bacteria are present to even greater depths.

International audience; Marine mud volcanoes are geological structures emitting large amounts of methane from their active centres. The Amsterdam mud volcano (AMV), located in the Anaximander Mountains south of Turkey, is characterized by intense active methane seepage produced in part by methanogens. To date, information about the diversity or the metabolic pathways used by the methanogens in active centres of marine mud volcanoes is limited. (14)C-radiotracer measurements showed that methylamines/methanol, H(2)/CO(2) and acetate were used for methanogenesis in the AMV. Methylotrophic methanogenesis was measured all along the sediment core, Methanosarcinales affiliated sequences were detected using archaeal 16S PCR-DGGE and mcrA gene libraries, and enrichments of methanogens showed the presence of Methanococcoides in the shallow sediment layers. Overall acetoclastic methanogenesis was higher than hydrogenotrophic methanogenesis, which is unusual for cold seep sediments. Interestingly, acetate porewater concentrations were extremely high in the AMV sediments. This might be the result of organic matter cracking in deeper hotter sediment layers. Methane was also produced from hexadecanes. For the most part, the methanogenic community diversity was in accordance with the depth distribution of the H(2)/CO(2) and acetate methanogenesis. These results demonstrate the importance of methanogenic communities in the centres of marine mud volcanoes.

The relationship between depth changes in total organic matter (TOC), biopolymers (“carbohydrates”, “proteins” and lipids) and bacterial populations and their activity, were investigated on two cruises to an upwelling site on the Peru Margin. Samples were obtained from the sediment surface to 8020 cm below sea floor (cbsf). Bacterial concentrations increased from near surface to a maximum at 12.5 cbsf, then rapidly decreased to 20 cbsf and a slower decrease to 8020 cbsf. The bacterial population at 8020 cbsf was 9.5% of the near surface value, but at 3.3 × 108 cells/cm3 it was still very significant. The presence of dividing bacterial cells indicated that a portion of the population was active and this was confirmed by radiotracer measurements. Biopolymers were overestimated in the surface 4 cm (138% of the TOC). By 5.5 cbsf, however, organic matter uncharacterized as biopolymers was present. This increased rapidly to 17% of the TOC at 10 cbsf and then more slowly to 79% at 2229 cbsf. Bacterial populations and activity were significantly correlated with depth changes in both TOC and biopolymers (p < 0.002). Calculation of rates of anaerobic carbon metabolism from the measured rates of sulphate reduction and methanogenesis demonstrated that these processes could account for all (101%) of the decrease in “carbohydrate” and “protein” in the 2.5 to 22.5 cbsf interval, where bacterial activity was intense. These data confirm the importance of anaerobic bacterial processes in these high organic matter sediments. At deeper intervals, 22.5–480 and 480–8020 cbsf, the decrease in “carbohydrate” and “protein” only accounted for 40% and 15% respectively of the anaerobic bacterial metabolism and hence the significant bacterial population present must have been utilizing a portion of the increasing uncharacterized organic matter. There was a negative relationship between the total bacterial population and the percentage of uncharacterized organic matter, suggesting that a portion of the uncharacterized organic matter may have come from dead bacterial cells. Estimates of bacterial necromass production from rates of thymidine incorporation were significantly correlated with increases in uncharacterized organic matter (p < 0.05, 2.5–197.5 cbsf) and could account for 16% of the increase within the top 25 cm. However, as the thymidine incorporation technique underestimates bacterial productivity in anoxic sediments it is likely that a much greater proportion of the increase in uncharacterized organic matter should be attributed to “bacterial necromass” production. It is suggested that bacterial necromass production may be an important mechanism for the production of recalcitrant, and hence preserved, organic matter in sediments in high productivity regions.

Despite the global importance of microbial methane production, knowledge of the methanogens involved and related anaerobic methane-oxidizing Euryarchaea (ANME) is quite limited. This includes the controls on methane production and consumption, which determines the atmospheric concentration of methane, which is an important greenhouse gas involved in climate change. This situation, in part, reflects the difficulty of culturing and studying these fastidious anaerobes. Here, we describe a range of cultivation and cultivation independent approaches that we have used to study methanogens, methanogenesis, and related processes in marine sediments, which contain globally significant methane concentrations.

This biogeochemical, molecular genetic and lipid biomarker study of sediments (4 m cores) from the Skagerrak (Denmark) investigated methane cycling in a sediment with a clear sulfate-methane-transition zone (SMTZ) and where CH4 supply was by diffusion, rather than by advection, as in more commonly studied seep sites. Sulfate reduction removed sulfate by 0.7 m and CH4 accumulated below. 14C-radiotracer measurements demonstrated active H2/CO2 and acetate methanogenesis and anaerobic oxidation of CH4 (AOM). Maximum AOM rates occurred near the SMTZ (3 nmol cm-3 day-1 at 0.75 m) but also continued deeper, overall, at much lower rates. Maximum rates of H2/CO2 and acetate methanogenesis occurred below the SMTZ but H2/CO2 methanogenesis rates were × 10 those of acetate methanogenesis, and this was consistent with initial values of 13C-depleted CH4 (δ13C c.-80‰). Areal AOM and methanogenic rates were similar (1.7 mmol m-2 day-1), hence, CH4 flux is finely balanced. A 16S rRNA gene library from 1.39 m combined with methanogen (T-RFLP), bacterial (16S rRNA DGGE) and lipid biomarker depth profiles showed the presence of populations similar to some seep sites: ANME-2a (dominant), ANME-3, Methanomicrobiales, Methanosaeta Archaea, with abundance changes with depth corresponding to changes in activities and sulfate-reducing bacteria (SRB). Below the SMTZ to 1.7 m CH4 became progressively more 13C depleted (δ13C -82‰) indicating a zone of CH4 recycling which was consistent with the presence of 13C-depleted archaeol (δ13C -55‰). Pore water acetate concentrations decreased in this zone (to 5 μM), suggesting that H2, not acetate, was an important CH4 cycling intermediate. The potential biomarkers for AOM-associated SRB, non-isoprenoidal ether lipids, increased below the SMTZ but this distribution reflected 16S rRNA gene sequences for JS1 and OP8 bacteria rather than those of SRB. At this site peak rates of methane production and consumption are spatially separated and seem to be conducted by different archaeal groups. Also AOM is predominantly coupled to sulfate reduction, unlike recent reports from some seep and gassy sediment sites.

Chemical analyses of the pore waters from hundreds of deep ocean sediment cores have over decades provided evidence for ongoing processes that require biological catalysis by prokaryotes. This sub-seafloor activity of microorganisms may influence the surface Earth by changing the chemistry of the ocean and by triggering the emission of methane, with consequences for the marine carbon cycle and even the global climate. Despite the fact that only about 1% of the total marine primary production of organic carbon is available for deep-sea microorganisms, sub-seafloor sediments harbour over half of all prokaryotic cells on Earth. This estimation has been calculated from numerous microscopic cell counts in sediment cores of the Ocean Drilling Program. Because these counts cannot differentiate between dead and alive cells, the population size of living microorganisms is unknown. Here, using ribosomal RNA as a target for the technique known as catalysed reporter deposition-fluorescence in situ hybridization (CARD-FISH), we provide direct quantification of live cells as defined by the presence of ribosomes. We show that a large fraction of the sub-seafloor prokaryotes is alive, even in very old (16 million yr) and deep (> 400 m) sediments. All detectable living cells belong to the Bacteria and have turnover times of 0.25-22 yr, comparable to surface sediments.

Diversity of Bacteria and Archaea was studied in deep marine sediments by PCR amplification and sequence analysis of 16S rRNA and methyl co-enzyme M reductase (mcrA) genes. Samples analysed were from Ocean Drilling Program (ODP) Leg 190 deep subsurface sediments at three sites spanning the Nankai Trough in the Pacific Ocean off Shikoku Island, Japan. DNA was amplified, from three depths at site 1173 (4.15, 98.29 and 193.29 mbsf; metres below the sea floor), and phylogenetic analysis of clone libraries showed a wide variety of uncultured Bacteria and Archaea. Sequences of Bacteria were dominated by an uncultured and deeply branching 'deep sediment group' (53% of sequences). Archaeal 16S rRNA gene sequences were mainly within the uncultured clades of the Crenarchaeota. There was good agreement between sequences obtained independently by cloning and by denaturing gradient gel electrophoresis. These sequences were similar to others retrieved from marine sediment and other anoxic habitats, and so probably represent important indigenous bacteria. The mcrA gene analysis suggested limited methanogen diversity with only three gene clusters identified within the Methanosarcinales and Methanobacteriales. The cultivated members of the Methanobacteriales and some of the Methanosarcinales can use CO2 and H2 for methanogenesis. These substrates also gave the highest rates in 14C-radiotracer estimates of methanogenic activity, with rates comparable to those from other deep marine sediments. Thus, this research demonstrates the importance of the 'deep sediment group' of uncultured Bacteria and links limited diversity of methanogens to the dominance of CO2/H2 based methanogenesis in deep sub-seafloor sediments.

Subsurface bacteria also occur in hydrothermal sediments with large temperature gradients (up to 12 °C/m) and with population numbers similar to non-hydrothermal sites at temperatures from psychrophilic to mesophilic. At greater depths and temperatures, populations decline rapidly, but they are still significant up to hyperthermophilic temperatures and are even stimulated by subsurface seawater flow. These results suggest that temperature alone does not limit bacteria in non-hydrothermal sediments until about 4 km, and evidence exists that bacterial processes may even be sustained by interaction with thermogenic processes as temperatures increase during deep burial. Experiments demonstrate that in the presence of readily degradable organic substrates, actively growing bacteria can move faster than sediment deposition; hence, these bacteria are not necessarily trapped and buried. However, bacterial growth decreases with depth to such an extent that subsurface bacteria would not be able to keep up with sedimentation rate and hence would be buried. In some circumstances, such as in sapropel layers with high organic matter in the Mediterranean, bacteria may be buried within a specific deposition horizon. Subsurface bacteria can utilize old and recalcitrant organic matter, but only very slowly, and they seem to have a strategy of high biomass and low growth rate, commensurate with their geological habitat of generally low energy flux.