Shinsuke Kawagucci

Methane, manganese, and helium-3 in newly discovered hydrothermal plumes over the Central Indian Ridge, 18°–20°S

We have investigated newly discovered submarine hydrothermal plumes over the Central Indian Ridge, 18°–20°S. Onshore chemical analyses of methane, its carbon isotope, manganese, and helium-3 in seawater samples obtained from the plumes revealed their detailed geochemical characteristics. One of the newly discovered hydrothermal plumes located over the western wall of the axial valley at Segment 15B (19°33′S), called the Roger Plateau, showed constant CH4/Mn and CH4/3He ratios throughout the plume. The CH4/3He ratio (4 × 106) and δ13C (−17.5‰) are consistent with those of basalt-hosted sediment-free hydrothermal systems, although the CH4/Mn ratio (∼1) is moderately higher. These features are thought to indicate a metal-depleted fluid chemistry. The other hydrothermal field is located within the axial valley at the northern part of the smooth lava plain at Segment 16 (18°20′S), called the Dodo Great Lava Plain, where several plumes were detected. The CH4/Mn and CH4/3He ratios showed large variation through the plumes while δ13C values were almost constant. Geochemical characteristics of venting fluid estimated from those of the plumes were apparently high CH4/Mn (>6) and CH4/3He (>60 × 106) ratios and low δ13C values (<−27.5‰), suggesting possible influences of several methane input processes to the fluid chemistry.

Discovery of New Hydrothermal Activity and Chemosynthetic Fauna on the Central Indian Ridge at 18°–20°S

Indian Ocean hydrothermal vents are believed to represent a novel biogeographic province, and are host to many novel genera and families of animals, potentially indigenous to Indian Ocean hydrothermal systems. In particular, since its discovery in 2001, much attention has been paid to a so-called ‘scaly-foot’ gastropod because of its unique iron-sulfide-coated dermal sclerites and the chemosynthetic symbioses in its various tissues. Despite increasing interest in the faunal assemblages at Indian Ocean hydrothermal vents, only two hydrothermal vent fields have been investigated in the Indian Ocean. Here we report two newly discovered hydrothermal vent fields, the Dodo and Solitaire fields, which are located in the Central Indian Ridge (CIR) segments 16 and 15, respectively. Chemosynthetic faunal communities at the Dodo field are emaciated in size and composition. In contrast, at the Solitaire field, we observed faunal communities that potentially contained almost all genera found at CIR hydrothermal environments to date, and even identified previously unreported taxa. Moreover, a new morphotype of ‘scaly-foot’ gastropod has been found at the Solitaire field. The newly discovered ‘scaly-foot’ gastropod has similar morphological and anatomical features to the previously reported type that inhabits the Kairei field, and both types of ‘scaly-foot’ gastropods genetically belong to the same species according to analyses of their COI gene and nuclear SSU rRNA gene sequences. However, the new morphotype completely lacks an iron-sulfide coating on the sclerites, which had been believed to be a novel feature restricted to ‘scaly-foot’ gastropods. Our new findings at the two newly discovered hydrothermal vent sites provide important insights into the biodiversity and biogeography of vent-endemic ecosystems in the Indian Ocean.

Disturbance of deep-sea environments induced by the M9.0 Tohoku Earthquake

The impacts of the M9.0 Tohoku Earthquake on deep-sea environment were investigated 36 and 98 days after the event. The light transmission anomaly in the deep-sea water after 36 days became atypically greater (∼35%) and more extensive (thickness ∼1500 m) near the trench axis owing to the turbulent diffusion of fresh seafloor sediment, coordinated with potential seafloor displacement. In addition to the chemical influx associated with sediment diffusion, an influx of 13C-enriched methane from the deep sub-seafloor reservoirs was estimated. This isotopically unusual methane influx was possibly triggered by the earthquake and its aftershocks that subsequently induced changes in the sub-seafloor hydrogeologic structures. The whole prokaryotic biomass and the development of specific phylotypes in the deep-sea microbial communities could rise and fall at 36 and 98 days, respectively, after the event. We may capture the snap shots of post-earthquake disturbance in deep-sea chemistry and microbial community responses.

Generation of Electricity and Illumination by an Environmental Fuel Cell in Deep‐Sea Hydrothermal Vents

Public interest in the generation of power by alternative energy sources in the ocean beyond fossil fuels and nuclear energy has increased in recent years. Power generation in the ocean is also of great interest for the inexpensive and efficient supply of electricity for the survey and exploration of submarine resources. Deep-sea hydrothermal vents are environments that discharge crustal hydrothermal fluids, geologically driven by magmatism and geochemically processed by the high temperatures of rock–seawater interactions and alterations. Hydrothermal fluids enriched with reduced chemicals are mixed with oxidative chemicals in seawater in the vicinity of deep-sea vents. The steep chemical slope between the hydrothermal fluids and ambient seawater has the potential to generate electricity. Therefore, it is likely that electricity is generated in situ between hydrothermal fluids (HF) and seawater (SW), which can be promoted and confirmed by deployment of artificial electrodes and a conductor. Herein, we show the development of an HF–SW fuel cell for deep-sea hydrothermal vents. During the Integrated Ocean Drilling Program (IODP) in 2010, several artificial hydrothermal vents were created by the drilling of wells in the Iheya North hydrothermal field of the Okinawa Trough, Japan. One of the artificial hydrothermal vents (C0014G) had a vigorous discharge with a high fluid temperature (Tmax= 309 8C) at a water depth of 1053 m. In this work, seafloor electrochemical analyses were conducted in the C0014G vent using a remotely operated vehicle (ROV) equipped with a deep-sea potentiostat/galvanostat system (D-Pote), which was covered with an anti-pressure housing and controlled by an onboard computer through data communication. First, the oxidation–reduction potential (ORP) of the ambient seawater and hydrothermal fluids was measured (Figure 1). The average temperature and ORP of the ambient seawater were approximately 4 8C and + 478 mV, respectively, versus the standard hydrogen electrode (SHE), while those of the hydrothermal fluids were approximately 309 8C

Post-drilling changes in seabed landscape and megabenthos in a deep-sea hydrothermal system, the Iheya North field, Okinawa Trough.

There has been an increasing interest in seafloor exploitation such as mineral mining in deep-sea hydrothermal fields, but the environmental impact of anthropogenic disturbance to the seafloor is poorly known. In this study, the effect of such anthropogenic disturbance by scientific drilling operations (IODP Expedition 331) on seabed landscape and megafaunal habitation was surveyed for over 3 years using remotely operated vehicle video observation in a deep-sea hydrothermal field, the Iheya North field, in the Okinawa Trough. We focused on observations from a particular drilling site (Site C0014) where the most dynamic change of landscape and megafaunal habitation was observed among the drilling sites of IODP Exp. 331. No visible hydrothermal fluid discharge had been observed at the sedimentary seafloor at Site C0014, where Calyptogena clam colonies were known for more than 10 years, before the drilling event. After drilling commenced, the original Calyptogena colonies were completely buried by the drilling deposits. Several months after the drilling, diffusing high-temperature hydrothermal fluid began to discharge from the sedimentary subseafloor in the area of over 20 m from the drill holes, ‘artificially’ creating a new hydrothermal vent habitat. Widespread microbial mats developed on the seafloor with the diffusing hydrothermal fluids and the galatheid crab Shinkaia crosnieri endemic to vents dominated the new vent community. The previously soft, sedimentary seafloor was hardened probably due to barite/gypsum mineralization or silicification, becoming rough and undulated with many fissures after the drilling operation. Although the effects of the drilling operation on seabed landscape and megafaunal composition are probably confined to an area of maximally 30 m from the drill holes, the newly established hydrothermal vent ecosystem has already lasted 2 years and is like to continue to exist until the fluid discharge ceases and thus the ecosystem in the area has been altered for long-term.

Cell-Specific Thioautotrophic Productivity of Epsilon-Proteobacterial Epibionts Associated with Shinkaia crosnieri

In this study, we report experimental evidence of the thioautotrophic activity of the epibiotic microbial community associated with the setae of Shinkaia crosnieri, a galatheid crab that is endemic to deep-sea hydrothermal systems in the Okinawa Trough in Japan. Microbial consumption of reduced sulfur compounds under in situ hydrostatic and atmospheric pressure provided evidence of sulfur-oxidizing activity by the epibiotic microbial community; the rate of sulfur oxidation was similar under in situ and decompressed conditions. Results of the microbial consumption of reduced sulfur compounds and tracer experiments using 13C-labeled bicarbonate in the presence and absence of thiosulfate (used as a thioautotrophic substrate) convincingly demonstrated that the epibiotic microbial community on S. crosnieri drove primary production via an energy metabolism that was coupled with the oxidation of reductive sulfur compounds. A combination of tracer experiments, fluorescence in situ hybridization (FISH) and nano-scale secondary ion mass spectrometry (Nano-SIMS) indicated that the filamentous cells of the genus Sulfurovum belonging to the class Epsilonproteobacteria were thioautotrophs in the epibiotic community of S. crosnieri. In conclusion, our results strongly suggest that thioautotrophic production by Sulfurovum members present as the epibiotic microbial community play a predominant role in a probable nutritional ectosymbiosis with S. crosnieri.

Helium anomalies suggest a fluid pathway from mantle to trench during the 2011 Tohoku-Oki earthquake

Geophysical evidence suggests that fluids along fault planes have an important role in generating earthquakes; however, the nature of these fluids has not been well defined. The 2011 magnitude 9.0 Tohoku-Oki earthquake ruptured the interface between the subducting Pacific plate and the overlying Okhotsk plate. Here we report a sharp increase in mantle-derived helium in bottom seawater near the rupture zone 1 month after the earthquake. The timing and location indicate that fluids were released from the mantle on the seafloor along the plate interface. The movement of the fluids was rapid, with a velocity of ~4 km per day and an uncertainty factor of four. This rate is much faster than what would be expected from pressure-gradient propagation, suggesting that over-pressurized fluid is discharged along the plate interface.

Heterogeneous composition of key metabolic gene clusters in a vent mussel symbiont population.

Chemosynthetic symbiosis is one of the successful systems for adapting to a wide range of habitats including extreme environments, and the metabolic capabilities of symbionts enable host organisms to expand their habitat ranges. However, our understanding of the adaptive strategies that enable symbiotic organisms to expand their habitats is still fragmentary. Here, we report that a single-ribotype endosymbiont population in an individual of the host vent mussel, Bathymodiolus septemdierum has heterogeneous genomes with regard to the composition of key metabolic gene clusters for hydrogen oxidation and nitrate reduction. The host individual harbours heterogeneous symbiont subpopulations that either possess or lack the gene clusters encoding hydrogenase or nitrate reductase. The proportions of the different symbiont subpopulations in a host appeared to vary with the environment or with the host’s development. Furthermore, the symbiont subpopulations were distributed in patches to form a mosaic pattern in the gill. Genomic heterogeneity in an endosymbiont population may enable differential utilization of diverse substrates and confer metabolic flexibility. Our findings open a new chapter in our understanding of how symbiotic organisms alter their metabolic capabilities and expand their range of habitats.

Diversity and methane oxidation of active epibiotic methanotrophs on live Shinkaia crosnieri

Shinkaia crosnieri is a galatheid crab that predominantly dwells in deep-sea hydrothermal systems in the Okinawa Trough, Japan. In this study, the phylogenetic diversity of active methanotrophs in the epibiotic microbial community on the setae of S. crosnieri was characterized by reverse transcription-polymerase chain reaction (RT-PCR) of a functional gene (pmoA) encoding a subunit of particulate methane monooxygenase. Phylogenetic analysis of pmoA transcript sequences revealed that the active epibiotic methanotrophs on S. crosnieri setae consisted of gammaproteobacterial type Ia and Ib methanotrophs. The effect of different RNA stabilization procedures on the abundance of pmoA and 16S rRNA transcripts in the epibiotic community was estimated by quantitative RT-PCR. Our novel RNA fixation method performed immediately after sampling effectively preserved cellular RNA assemblages, particularly labile mRNA populations, including pmoA mRNA. Methane consumption in live S. crosnieri was also estimated by continuous-flow incubation under atmospheric and in situ hydrostatic pressures, and provided a clear evidence of methane oxidation activity of the epibiotic microbial community, which was not significantly affected by hydrostatic pressure. Our study revealed the significant ecological function and nutritional contribution of epibiotic methanotrophs to the predominant S. crosnieri populations in the Okinawa Trough deep-sea hydrothermal systems. In conclusion, our study gave clear facts about diversity and methane oxidation of active methanotrophs in the epibiotic community associated with invertebrates.

Molecular evidence of digestion and absorption of epibiotic bacterial community by deep-sea crab Shinkaia crosnieri.

The hydrothermal vent crab Shinkaia crosnieri is considered to obtain nutrition from the epibiotic bacteria found on the setae, but previous studies have not shown how nutrients can be transferred from the epibionts to the host. In this study, microscopic observations of S. crosnieri intestinal components detected autofluorescent setae fragments and pigmentation derived from the digestion of epibionts in a dye-stained epibiont tracer experiment. An in vitro digestion experiment with epibiotic populations using an intestinal extract demonstrated the degradation of epibiotic cells by digestive enzymes. A phylogenetic analysis showed that many of the bacterial 16S ribosomal RNA gene sequences obtained from the intestine were closely related to the sequences of the epibionts, thus they were probably derived from the epibionts. A stable isotope tracer experiment also indicated that 13C assimilated by the epibionts provided a carbon (nutrition) source for the host. Both activity measurements and isotope studies showed that chemosynthetic metabolism by the gut microbial components were inactive. Together with the feeding behaviour of living S. crosnieri, these results indicate that S. crosnieri ingests the epibionts using maxillipeds and assimilates them via its digestive organs as a nutrient source. The results of this study elucidate the mechanism of nutritional transfer in ectosymbiosis between chemosynthetic bacteria and deep-sea invertebrates.