Depressurization and sample processing delays may impact the outcome of shipboard microbial incubations of samples collected from the deep sea. To address this knowledge gap, we developed a remotely operated vehicle (ROV)-powered incubator instrument to carry out and compare results from in situ and shipboard RNA stable isotope probing (RNA-SIP) experiments to identify the key chemolithoautotrophic microbes and metabolisms in diffuse, low-temperature venting fluids from Axial Seamount. All the incubations showed microbial uptake of labeled bicarbonate primarily by thermophilic autotrophic Epsilonbacteraeota that oxidized hydrogen coupled with nitrate reduction. However, the in situ seafloor incubations showed higher abundances of transcripts annotated for aerobic processes, suggesting that oxygen was lost from the hydrothermal fluid samples prior to shipboard analysis. Furthermore, transcripts for thermal stress proteins such as heat shock chaperones and proteases were significantly more abundant in the shipboard incubations, suggesting that depressurization induced thermal stress in the metabolically active microbes in these incubations. Together, the results indicate that while the autotrophic microbial communities in the shipboard and seafloor experiments behaved similarly, there were distinct differences that provide new insight into the activities of natural microbial assemblages under nearly native conditions in the ocean.
The size and biogeochemical impact of the subseafloor biosphere in oceanic crust remain largely unknown due to sampling limitations. We used reactive transport modeling to estimate the size of the subseafloor methanogen population, volume of crust occupied, fluid residence time, and nature of the subsurface mixing zone for two low-temperature hydrothermal vents at Axial Seamount. Monod CH4 production kinetics based on chemostat H2 availability and batch-culture Arrhenius growth kinetics for the hyperthermophile Methanocaldococcus jannaschii and thermophile Methanothermococcus thermolithotrophicus were used to develop and parameterize a reactive transport model, which was constrained by field measurements of H2, CH4, and metagenome methanogen concentration estimates in 20–40 °C hydrothermal fluids. Model results showed that hyperthermophilic methanogens dominate in systems where a narrow flow path geometry is maintained, while thermophilic methanogens dominate in systems where the flow geometry expands. At Axial Seamount, the residence time of fluid below the surface was 29–33 h. Only 1011 methanogenic cells occupying 1.8–18 m3 of ocean crust per m2 of vent seafloor area were needed to produce the observed CH4 anomalies. We show that variations in local geology at diffuse vents can create fluid flow paths that are stable over space and time, harboring persistent and distinct microbial communities.
At deep-sea hydrothermal vents, microbial communities thrive across geochemical gradients above, at, and below the seafloor. In this study, we determined the gene content and transcription patterns of microbial communities and specific populations to understand the taxonomy and metabolism both spatially and temporally across geochemically different diffuse fluid hydrothermal vents. Vent fluids were examined via metagenomic, metatranscriptomic, genomic binning, and geochemical analyses from Axial Seamount, an active submarine volcano on the Juan de Fuca Ridge in the NE Pacific Ocean, from 2013 to 2015 at three different vents: Anemone, Marker 33, and Marker 113. Results showed that individual vent sites maintained microbial communities and specific populations over time, but with spatially distinct taxonomic, metabolic potential, and gene transcription profiles. The geochemistry and physical structure of each vent both played important roles in shaping the dominant organisms and metabolisms present at each site. Genomic binning identified key populations of SUP05, Aquificales and methanogenic archaea carrying out important transformations of carbon, sulfur, hydrogen, and nitrogen, with groups that appear unique to individual sites. This work highlights the connection between microbial metabolic processes, fluid chemistry, and microbial population dynamics at and below the seafloor and increases understanding of the role of hydrothermal vent microbial communities in deep ocean biogeochemical cycles.
Chemical input to the deep sea from hydrothermal systems is a globally distributed phenomenon. Hydrothermal discharge is one of the primary mechanisms by which the Earth’s interior processes manifest themselves at the Earth’s surface, and it provides a source of energy for autotrophic processes by microbes that are too deep to capitalize on sunlight. Much is known about the water-column signature of this discharge from high-temperature mid-ocean Ridge (MOR) environments and their neighboring low-temperature counterparts. Hydrothermal discharge farther away from the ridge, however, has garnered less attention, owing in part to the difficulty in finding this style of venting, which eludes methods of detection that work well for high-temperature ‘black smoker’-type venting. Here we present a case study of the plume from one such ‘invisible’ off-axis environment, The Lost City, with an emphasis on the dissolved volatile content of the hydrothermal plume. Serpentinization and abiotic organic synthesis generate significant concentrations of H2 and CH4 in vent fluid, but these species are unevenly transported to the overlying plume, which itself appears to be a composite of two different sources. A concentrated vent cluster on the talus slope channels fluid through at least eight chimneys, producing a water-column plume with the highest observed concentrations of CH4 in the field. In contrast, a saddle in the topography leading up to a carbonate cap hosts broadly distributed, nearly invisible venting apparent only in its water-column signals of redox potential and dissolved gas content, including the highest observed plume H2. After normalizing H2 and CH4 to the 3He background-corrected anomaly (3HeΔ) to account for mixing and relative amount of mantle input, it appears that, while a minimum of 60% of CH4 is transported out of the system, greater than 90% of the H2 is consumed in the subsurface prior to venting. The exception to this pattern occurs in the plume originating from the area dubbed Chaff Beach, in which somewhat more than 10% of the original H2 remains, indicating that this otherwise unremarkable plume, and others like it, may represent a significant source of H2 to the deep sea.
Dissolved oxygen is often considered the most important single chemical species in the ocean. Despite its central importance to understanding the biogeochemistry of the ocean, the accurate measurement of oxygen in the marine environment remains surprisingly challenging. Commercially available “optode” oxygen instruments are often plagued by data drift issues tied to material choices of the sensing membrane. This is especially true over long instrument deployment periods and in extreme environments. C-DEBI provided funds to build and deploy an optode-like instrument utilizing a UW-developed oxygen crystalline sensing material with superior long-term stability. To successfully meet the projects goals we built the VentO2 instrument, wrote control software for realtime acquisition and data processing, and deployed it on multiple dives using the ROV Jason II at Axial Seamount. We used a spectrometer-based measurement approach for the VentO2, which is a more challenging, but ultimately more powerful approach than the phase measurement used in optodes. The VentO2 instrument was successfully tested for functionality in the laboratory, and successfully recorded oxygen and temperature data on five separate dives to Axial Seamount on the Juan de Fuca Ridge. The resulting data was precise, but not accurate– it tracked the trends in oxygen concentration as measured by the onboard reference instrument, but the magnitude of the data was offset. While we are encouraged by the success to date on a very limited budget, we acknowledge that much additional work is necessary to move the instrument beyond a working proof of concept.
