Oceanic crust comprises the largest hydrogeologic reservoir on Earth, containing fluids in thermodynamic disequilibrium with the basaltic crust. Little is known about microbial ecosystems that inhabit this vast realm and exploit chemically favorable conditions for metabolic activities. Crustal samples recovered from ocean drilling operations are often compromised for microbiological assays, hampering efforts to resolve the extent and functioning of a subsurface biosphere. We report results from the first in situ experimental observatory systems that have been used to study subseafloor life. Experiments deployed for 4 years in young (3.5 Ma) basaltic crust on the eastern flank of the Juan de Fuca Ridge record a dynamic, post-drilling response of crustal microbial ecosystems to changing physical and chemical conditions. Twisted stalks exhibiting a biogenic iron oxyhydroxide signature coated the surface of mineral substrates in the observatories; these are biosignatures indicating colonization by iron oxidizing bacteria during an initial phase of cool, oxic, iron-rich conditions following observatory installation. Following thermal and chemical recovery to warmer, reducing conditions, the in situ microbial structure in the observatory shifted, becoming representative of natural conditions in regional crustal fluids. Firmicutes, metabolic potential of which is unknown but may involve N or S cycling, dominated the post-rebound bacterial community. The archaeal community exhibited an extremely low diversity. Our experiment documented in situ conditions within a natural hydrological system that can pervade over millennia, exemplifying the power of observatory experiments for exploring the subsurface basaltic biosphere, the largest but most poorly understood biotope on Earth.
This study examines the representativeness of low-temperature hydrothermal fluid samples with respect to their chemical and microbiological characteristics. Within this scope, we investigated short-term temporal chemical and microbial variability of the hydrothermal fluids. For this purpose we collected three fluid samples consecutively from the same spot at the Clueless field near 5 degrees S on the southern Mid-Atlantic Ridge over a period of 50 min. During sampling, the temperature was monitored online. We measured fluid chemical parameters, characterized microbial community compositions and used statistical analyses to determine significant differences between the samples. Overall, the three fluid samples are more closely related to each other than to any other tested habitat. Therefore, on a broad scale, the three collected fluid samples can be regarded as habitat representatives. However, small differences are apparent between all samples. One of the Clueless samples even displayed significant differences (P-value < 0.01) to the other two Clueless samples. Our data suggest that the observed variations in fluid chemical and microbial compositions are not reflecting sampling artefacts but are related to short-term fluid variability due to dynamic subseafloor fluid mixing. Recorded temporal changes in fact reflect spatial heterogeneity found in the subsurface as the fluid flows through distinctive pathways. While conservative elements (Cl, Si, Na and K) indicate variable degrees of fluid-seawater mixing, reactive components, including Fe(II), O(2) and H(2)S, show that chemical and microbial reactions within the mixing zone further modify the emanating fluids on short-time scales. Fluids entrain microorganisms, which modify the chemical microenvironment within the subsurface biotopes. This is the first study focusing on short-term microbial variability linked to chemical changes in hydrothermal fluids.
Although it has been more than 30 years since the discovery of deep-sea hydrothermal vents, comprehending the interconnections between hydrothermal venting and microbial life remains a challenge. Here we investigate abiotic-biotic linkages in low-temperature hydrothermal biotopes at Desperate and Lilliput on the southern Mid-Atlantic Ridge. Both sites are basalt-hosted and fluids exhibit the expected chemical signatures. However, contrasting crustal permeabilities have been proposed, supporting pervasive mixing at Desperate but restricting circulation at Lilliput. In Desperate fluids, sulfide and O2 were readily available but H2 hardly detectable. Under incubation conditions (oxic unamended, sulfide-spiked, oxic and anoxic H2 -spiked at 18°C), only sulfide oxidation by Thiomicrospira fuelled biomass synthesis. Microbial phylogenies from Desperate incubation experiments resembled those of the natural samples suggesting that the incubation conditions mimicked the environment. In Lilliput fluids, O2 was limited, whereas sulfide and H2 were enriched. Autotrophy appeared to be stimulated by residual sulfide and by amended H2 . Yet, based on bacterial phylogenies only conditions in anoxic H2 -spiked Lilliput incubations appeared similar to parts of the Lilliput habitat. In anoxic H2 -spiked Lilliput enrichments Campylobacteraceae likely supported biomass production through H2 oxidation. We argue that the diverging circulation patterns arising from different subseafloor permeabilities act as major driving forces shaping these biotope structures.
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