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Hirota, Akinari*; Kozuka, Mariko*; Fukuda, Akari*; Miyakawa, Kazuya; Sakuma, Keisuke; Ozaki, Yusuke; Ishii, Eiichi; Suzuki, Yohei*
Microbial Ecology, 87, p.132_1 - 132_15, 2024/10
Times Cited Count:0 Percentile:0.00(Ecology)Deep underground galleries are used to access the deep biosphere in addition to mining and other engineering applications such as geological disposal of radioactive wastes. Fracture networks developed in the excavation damaged zone (EDZ) are concerned to accelerate mass transport, where microbial colonization might be possible due to the availability of space and nutrients. In this study, microbial biofilms at EDZ fractures were investigated by drilling from a 350-m deep gallery and subsequent borehole logging at the Horonobe underground research laboratory (URL). By using microscopic and spectroscopic techniques, the dense colonization of microbial cells was demonstrated at the surfaces of the EDZ fractures with high hydraulic conductivities. 16S rRNA gene sequence analysis revealed the dominance of gammaproteobacterial lineages, the cultivated members of which are aerobic methanotrophs. Near-complete Horonobe groundwater genomes affiliated within the methanotrophic lineages were fully equipped with genes involved in aerobic methanotrophy. Although the mediation of aerobic methanotrophy remains to be demonstrated, microbial O production was supported by the presence of genes in the near-complete genomes, such as catalase and superoxide dismutase that produce O
from reactive oxygen species and a nitric oxide reductase gene with the substitutions of amino acids in motifs. It is concluded that the EDZ fractures provide energetically favorable subsurface habitats to microorganisms.
Nishimura, Hiroki*; Kozuka, Mariko*; Fukuda, Akari*; Ishimura, Toyoho*; Amano, Yuki; Beppu, Hikari*; Miyakawa, Kazuya; Suzuki, Yohei*
Environmental Microbiology Reports (Internet), 15(3), p.197 - 205, 2023/06
Times Cited Count:2 Percentile:38.58(Environmental Sciences)The family Methanoperedenaceae archaea mediate anaerobic oxidation of methane (AOM). We newly developed a high-pressure laboratory incubation system and investigated groundwater from 214- and 249-m deep boreholes at Horonobe Underground Research Laboratory, Japan, where the high and low abundances of
Methanoperedenaceae archaea have been revealed, respectively. We incubated the samples amended with or without amorphous Fe(III) and
C-labelled methane at an in-situ pressure of 1.6 MPa. After three to seven-day incubation, AOM activities were not detected from the 249-m sample but from the 214-m sample. The AOM rates were 93.7
40.6 and 27.7
37.5 nM/day with and without Fe(III) amendment. Suspended particulates were not visible in the 249-m sample on the filter, while they were abundant and contained amorphous Fe(III) and Fe(III)-bearing phyllosilicates in the 214-m sample. This supports the in-situ activity of Fe(III)-dependent AOM in the deep subsurface borehole.
Ino, Kohei*; Hernsdorf, A. W.*; Konno, Yuta*; Kozuka, Mariko*; Yanagawa, Katsunori*; Kato, Shingo*; Sunamura, Michinari*; Hirota, Akinari*; Togo, Yoko*; Ito, Kazumasa*; et al.
ISME Journal, 12(1), p.31 - 47, 2018/01
Times Cited Count:53 Percentile:90.56(Ecology)In this study, we found the dominance ofanaerobic methane-oxidizing archaea in groundwater enriched in sulfate and methane from a 300-m deep underground borehole in granitic rock.
Ino, Kohei*; Konno, Yuta*; Kozuka, Mariko*; Hirota, Akinari*; Togo, Yoko*; Fukuda, Akari*; Komatsu, Daisuke*; Tsunogai, Urumu*; Tanabe, Akifumi*; Yamamoto, Satoshi*; et al.
Environmental Microbiology Reports (Internet), 8(2), p.285 - 294, 2016/04
Times Cited Count:30 Percentile:69.30(Environmental Sciences)To understand the subsurface biosphere, borehole investigation was conducted for 300-m deep granitic rocks at the Mizunami underground research laboratory, Japan. The initial biomass was the highest with the flourish of aerobic H-oxidizing Hydrogenophaga spp., whereas an uncultivated lineage of the phylum Nitrospirae became predominant after three years with decreasing biomass. The common occurrence of many species of Nitrospirae and Chlorobi phyla at the geographically distinct sites and the exclusive detection of their phylogenetically related environmental sequences from deep groundwaters and terrestrial hot springs, suggest that these bacteria are indigenous and potentially adapted to the deep terrestrial subsurface.
Fukuda, Akari*; Hagiwara, Hiroki; Ishimura, Toyoho*; Kozuka, Mariko*; Ioka, Seiichiro*; Amano, Yuki; Tsunogai, Urumu*; Suzuki, Yohei*; Mizuno, Takashi
Microbial Ecology, 60(1), p.214 - 225, 2010/05
Times Cited Count:30 Percentile:65.10(Ecology)To better understand the geochemical and microbiological relationships, we characterized granitic groundwater collected from a 1,148 to 1,169 -m deep borehole interval at the Mizunami Underground Research Laboratory site, Japan, in 2005 and 2008. Geochemical analyses of the groundwater samples indicated that major electron acceptors, such as NO and SO
, were not abundant, while dissolved organic carbon (not including organic acids), CH
and H
were moderately rich. The most common phylotypes were both related to
spp., the cultivated members of which can utilize minor electron donors, such as aromatic and aliphatic hydrocarbons. Geomicrobiological results suggest that deep granitic groundwater has been stably colonized by
spp. probably owing to the limitation of O
, NO
and organic acids.
Fukuda, Akari; Mizuno, Takashi; Aosai, Daisuke; Hagiwara, Hiroki; Yamamoto, Yuhei; Shingu, Shinya; Ito, Kazumasa*; Suzuki, Yohei*; Kozuka, Mariko*; Konno, Yuta*
no journal, ,
no abstracts in English
Fukuda, Akari; Kozuka, Mariko*; Konno, Yuta*; Aosai, Daisuke; Hagiwara, Hiroki; Mizuno, Takashi; Suzuki, Yohei*
no journal, ,
To quantify microbial influences on the hydrogeochemistry of a 99-1169-m deep granitic aquifer system, we developed methodology for sensitive measurements of a variety of aerobic and anaerobic metabolic activities. Briefly, microbial cells were ca. 30-fold concentrated in groundwater by filtration to incubate with electron acceptors. Aerobic respiration was more than 400 umol/L/year at depths of 99 and 175 m and decreased with increasing depth down to 36 umol/L/year at a depth of 1169 m. Nitrate respiration increased from 99 to 308 m (4.3 to 37 umol/L/year) and decreased with increasing depth down to 0.20 umol/L/year. Sulfate respiration ranging from 1.4 to 3.2 umol/L/year was detected only at depths above 200 m. The depletion of O and NO
and a gradual decrease in sulfate with increasing depth in the granitic aquifer could be attributed to microbial respirations at rates clarified in this study. Our research was partially founded by Nuclear and Industry Safety Agency.
Fukuda, Akari*; Hagiwara, Hiroki; Ishimura, Toyoho*; Kozuka, Mariko*; Ito, Kazumasa*; Tsunogai, Urumu*; Suzuki, Yohei*; Mizuno, Takashi
no journal, ,
JAEA and AIST have been conducting collaborative work on hydrochemical study at MIU construction site. The aim of this collaborative study is to establish the methodology for investigation, analysis and evaluation of colloid/organics/microbes study. This paper reports the biogeochemical study. As the result, it is suggested that groundwater taken from deeper part (1150m depth) is relatively oxidized.
Fukuda, Akari; Kozuka, Mariko*; Konno, Yuta*; Aosai, Daisuke; Shingu, Shinya; Hagiwara, Hiroki; Mizuno, Takashi; Suzuki, Yohei*
no journal, ,
no abstracts in English
Suzuki, Yohei*; Fukuda, Akari; Konno, Yuta*; Kozuka, Mariko*; Hagiwara, Hiroki; Aosai, Daisuke; Takeno, Naoto*; Mizuno, Takashi
no journal, ,
Nishimura, Hiroki*; Kozuka, Mariko*; Fukuda, Akari*; Ishimura, Toyoho*; Amano, Yuki; Beppu, Hikari*; Miyakawa, Kazuya; Suzuki, Yohei*
no journal, ,
The family Methanoperedenaceae archaea mediate anaerobic oxidation of methane (AOM). We newly developed a high-pressure laboratory incubation system and investigated groundwater from 214- and 249-m deep boreholes at Horonobe Underground Research Laboratory, Japan, where the high and low abundances of Methanoperedenaceae archaea have been revealed, respectively. We incubated the samples amended with or without amorphous Fe(III) and C-labelled methane at an in-situ pressure of 1.6 MPa. After three to seven-day incubation, AOM activities were not detected from the 249-m sample but from the 214-m sample. The AOM rates were 93.7
40.6 and 27.7
37.5 nM/day with and without Fe(III) amendment. Suspended particulates were not visible in the 249-m sample on the filter, while they were abundant and contained amorphous Fe(III) and Fe(III)-bearing phyllosilicates in the 214-m sample. This supports the in-situ activity of Fe(III)-dependent AOM in the deep subsurface borehole.
Nishimura, Hiroki*; Kozuka, Mariko*; Fukuda, Akari*; Ishimura, Toyoho*; Amano, Yuki; Beppu, Hikari*; Miyakawa, Kazuya; Suzuki, Yohei*
no journal, ,
no abstracts in English
Fukuda, Akari; Mizuno, Takashi; Aosai, Daisuke; Hagiwara, Hiroki; Yamamoto, Yuhei; Shingu, Shinya; Takeno, Naoto*; Suzuki, Yohei*; Konno, Yuta*; Kozuka, Mariko*
no journal, ,
no abstracts in English
Suzuki, Yohei*; Fukuda, Akari*; Kozuka, Mariko*; Ishimura, Toyoho*; Tsunogai, Urumu*; Hagiwara, Hiroki; Mizuno, Takashi
no journal, ,
no abstracts in English
Ito, Kazumasa*; Suzuki, Yohei*; Fukuda, Akari*; Kozuka, Mariko*; Mizuno, Takashi; Hagiwara, Hiroki
no journal, ,
no abstracts in English
Fukuda, Akari; Kozuka, Mariko*; Aosai, Daisuke; Hagiwara, Hiroki; Mizuno, Takashi; Suzuki, Yohei*
no journal, ,
no abstracts in English
Konno, Yuta*; Fukuda, Akari; Kozuka, Mariko*; Komatsu, Daisuke*; Tsunogai, Urumu*; Aosai, Daisuke; Mizuno, Takashi; Suzuki, Yohei*
no journal, ,
Our knowledge of biogeochemical processes mediated in those associated with freshwater should be integrated. We collected the groundwater samples from 200 m to 1150 m depths below ground level at Mizunami Underground Research Laboratory located in central Japan. We measured the concentration and both hydrogen and carbon isotopic compositions of dissolved methane, total inorganic carbon (TIC) and molecular hydrogen. The origin of methane could be speculated to be not biogenic CO reduction or acetate fermentation but thermogenic or abiogenic. In contrast, C1/C2+C3 ratios supported biogenic methane production. The 300 m deep samples were enriched in hydrogen and acetate and depleted in sulfate, which is characteristic of acetogenesis. From these results, it is suggested that the production of acetate rather than methane could be dominant biogeochemical processes in the reducing portions of freshwater aquifers in granitic rocks.