Kazuya Miyakawa

Potential for microbial H 2 and metal transformations associated with novel bacteria and archaea in deep terrestrial subsurface sediments

Geological sequestration in deep underground repositories is the prevailing proposed route for radioactive waste disposal. After the disposal of radioactive waste in the subsurface, H2 may be produced by corrosion of steel and, ultimately, radionuclides will be exposed to the surrounding environment. To evaluate the potential for microbial activities to impact disposal systems, we explored the microbial community structure and metabolic functions of a sediment-hosted ecosystem at the Horonobe Underground Research Laboratory, Hokkaido, Japan. Overall, we found that the ecosystem hosted organisms from diverse lineages, including many from the phyla that lack isolated representatives. The majority of organisms can metabolize H2, often via oxidative [NiFe] hydrogenases or electron-bifurcating [FeFe] hydrogenases that enable ferredoxin-based pathways, including the ion motive Rnf complex. Many organisms implicated in H2 metabolism are also predicted to catalyze carbon, nitrogen, iron and sulfur transformations. Notably, iron-based metabolism is predicted in a novel lineage of Actinobacteria and in a putative methane-oxidizing ANME-2d archaeon. We infer an ecological model that links microorganisms to sediment-derived resources and predict potential impacts of microbial activity on H2 consumption and retardation of radionuclide migration.

Characterization and thermodynamic study of humic acid in deep groundwater at Horonobe, Hokkaido, Japan

ABSTRACT In this study, humic substances (humic acid and fulvic acid) were isolated from deep groundwater at −350 m depth of Horonobe, Hokkaido, Japan to compare the characteristic property and reaction mechanism with generic humic acid isolated from surface soils. The size distributions of Horonobe humic substances were analyzed by size exclusion chromatography, flow-field flow fractionation, and total organic carbon measurement with ultrafiltration. All of them indicated small molecular weight and particle size of Horonobe humic acid in comparison with generic humic acids. Additionally, the simple protonation behavior of Horonobe humic substances similar to benzoic acid and/or phenol was revealed by thermodynamic quantities obtained by potentiometry and calorimetry. Consequently, molecular size and the reaction mechanism of Horonobe humic substances are different from generic humic acids, due to the characteristic origin.

A Proposed Method to Estimate In Situ Dissolved Gas Concentrations in Gas-Saturated Groundwater

Gas-saturated groundwater forms bubbles when brought to atmospheric pressure, preventing precise determination of its in situ dissolved gas concentrations. To overcome this problem, a modeling approach called the atmospheric sampling method is suggested here to recover the in situ dissolved gas concentrations of groundwater collected ex situ under atmospheric conditions at the Horonobe Underground Research Laboratory, Japan. The results from this method were compared with results measured at the same locations using two special techniques, the sealed sampler and pre-evacuated vial methods, that have been developed to collect groundwater under its in situ conditions. In gas-saturated groundwater cases, dissolved methane and inorganic carbon concentrations derived using the atmospheric sampling method were mostly within ±4 and ±10%, respectively, of values from the sealed sampler and pre-evacuated vial methods. In gas-unsaturated groundwater, however, the atmospheric sampling method overestimated the in situ dissolved methane concentrations, because the groundwater pressure at which bubbles appear (Pcritical ) was overestimated. The atmospheric sampling method is recommended for use where gas-saturated groundwater can be collected only ex situ under atmospheric conditions.

An Evaluation of the Long-Term Stagnancy of Porewater in the Neogene Sedimentary Rocks in Northern Japan

A groundwater dating for very old porewater using 36Cl and 4He was applied to the Koetoi and Wakkanai formations distributed in the northernmost part in Japan. Measured 36Cl/Cl in the Koetoi Formation was 2.6 ± 2.0 × 10−15 and that in the Wakkanai Formation was 8.1 ± 2.5 × 10−15. These values are similar to 36Cl/Cl in situ secular equilibrium calculated from chemical compositions of core suggesting that Cl− ions and porewater have remained in the formations for much longer than half-life of 36Cl . He concentration in porewater ranged from 1.1 × 10−6 to 2.6 × 10−5 ( ) and it is much higher than water saturated with air indicating that both formations contain very old porewater. However, the possibility of mixing of young water was indicated because He concentration was lower than that calculated by multiplication of in situ He production and time after the uplift. This possibility was also supported by Cl−, δD, and δ18O data. After combining information on 36Cl/Cl, 4He, and δD and δ18O, it was inferred that the porewater in the deep part of the Wakkanai Formation might have been stagnant since the uplift. The porewater in the Koetoi Formation and the shallow part of the Wakkanai Formation were found to be affected by young surface water.

Improvements in Drill-Core Headspace Gas Analysis for Samples from Microbially Active Depths

The IsoJar™ container is widely used in headspace gas analysis for gases adsorbed on cuttings or bore cores from oil and gas fields. However, large variations in the carbon isotopic ratios of CH4 and CO2 are often reported, especially for data obtained from depths of 30‰, whereas samples analyzed within a week of collection showed no such fluctuations. The conventional amount of microbial suppressant (~0.5 ml of 10% benzalkonium chloride (BKC) solution) is insufficient to suppress microbial activity if groundwater is used as filling water. The significant variations in carbon isotopic compositions previously reported were caused by microbial methane oxidation after sampling and contamination by groundwater from different depths. To avoid these problems, we recommend the following: (1) if long-term sample storage is necessary, >10 ml of 10% BKC solution should be added or >0.3% BKC concentration is required to suppress microbial activity; (2) analyses should be performed within one week of sampling; and (3) for CO2 analyses, it is important that samples are not contaminated by groundwater from different depths.