押木 守 (オシキ マモル)

工学研究院 環境工学部門 環境工学准教授
Last Updated :2025/01/11

■研究者基本情報

学位

  • 博士(環境学), 東京大学

プロフィール情報

Researchmap個人ページ

研究キーワード

  • 下水処理
  • 活性汚泥
  • 生物膜(バイオフィルム)
  • 生物学的窒素除去
  • 硝化・脱窒
  • 嫌気好気法
  • 部分硝化
  • 嫌気性アンモニウム酸化(anammox)
  • 生物学的リン除去
  • 嫌気性消化
  • メタン発酵
  • 嫌気性原生動物
  • 尿素分解
  • DNA/RNA解析
  • 定量PCR(qPCR)
  • 微少流路デバイス
  • 16S RNA遺伝子アンプリコン解析
  • メタゲノム解析
  • RNASeq解析
  • 遺伝子等温増幅(RPA)
  • 薬剤耐性遺伝子
  • 遺伝子組換え実験
  • 薬剤耐性プラスミド
  • 遺伝子クローニング
  • バイオプラスチック
  • ポリヒドロキシアルカン酸
  • 微生物培養
  • 嫌気性培養
  • 顕微鏡イメージング
  • 安定同位体比測定
  • 安定同位体トレーサー試験
  • 質量分析
  • 嫌気性アンモニウム酸化(anammox)菌
  • アンモニア酸化細菌(AOB)
  • アンモニア酸化古細菌(AOA)
  • 亜酸化窒素(N2O)
  • ヒドロキシルアミン(NH2OH)/ヒドラジン(N2H4)
  • チオシアン分解
  • 鉄酸化硝酸還元

研究分野

  • 社会基盤(土木・建築・防災), 土木環境システム, 生物学的窒素除去、硝化・脱窒、嫌気性アンモニウム酸化(anammox)、薬剤耐性菌、生物電気化学的プロセス
  • 環境・農学, 環境影響評価, 窒素循環、窒素安定同位体、薬剤耐性遺伝子、薬剤耐性菌
  • 環境・農学, 環境負荷低減技術、保全修復技術, 排水処理、生物処理、環境負荷低減
  • ライフサイエンス, 応用微生物学, 窒素代謝微生物、嫌気性アンモニウム酸化(anammox)菌
  • ライフサイエンス, 水圏生命科学, 窒素循環、窒素代謝

■経歴

経歴

  • 2022年04月 - 現在
    JST創発的研究支援事業 創発研究者
  • 2020年04月 - 現在
    北海道大学大学院, 工学研究院環境創成工学部門, 准教授
  • 2016年04月 - 2020年03月
    長岡工業高等専門学校, 環境都市工学科, 准教授
  • 2014年04月 - 2016年03月
    長岡工業高等専門学校, 環境都市工学科, 助教
  • 2011年04月 - 2014年03月
    日本学術振興会特別研究員(PD), 北海道大学大学院工学研究院
  • 2013年01月 - 2014年01月
    Radboud university (オランダ), 理学部, 客員研究員
  • 2010年01月 - 2011年03月
    北海道大学, 大学院・工学研究院, 博士研究員
  • 2009年04月 - 2009年12月
    東京大学大学院新領域創成科学研究科, 東京大学大学院新領域創成科学研究科, 特別研究員
  • 2007年04月 - 2009年03月
    日本学術振興会特別研究員(DC2), 東京大学大学院新領域創成科学研究科

学歴

  • 2006年04月 - 2009年03月, 東京大学, 新領域創成科学研究科, 社会文化環境学専攻 博士課程
  • 2004年04月 - 2006年03月, 東京大学, 新領域創成科学研究科, 社会文化環境学 修士課程
  • 2002年04月 - 2004年03月, 長岡工業高等専門学校, 環境都市工学専攻
  • 1997年04月 - 2002年03月, 長岡工業高等専門学校, 環境都市工学科

委員歴

  • 2018年04月 - 現在
    日本水環境学会, 微生物生態と水環境工学 運営委員, 学協会
  • 2020年05月 - 2026年05月
    ISME Journal Editorial board, 学協会
  • 2023年04月 - 2025年03月
    公益社団法人 土木学会, 環境工学委員会
  • 2021年01月 - 2025年03月
    日本微生物生態学会, 日本微生物生態学会事務局代表幹事
  • 2017年12月 - 2019年08月
    日本水環境学会, Japan-YWP企画委員, 学協会
  • 2016年04月 - 2018年03月
    日本水環境学会, WET部会委員, 学協会

■研究活動情報

受賞

  • 2024年03月, 日本水環境学会, ライオン賞               
    活性汚泥に生息する従属栄養硝化-好気性脱窒細菌の特定
  • 2023年03月, ISME Journal, ISME Journal Outstanding Editorial Board Member Award 2022               
  • 2023年03月, 日本水環境学会, ライオン賞               
    石崎翔大;石塚祐介;樋口宏介;押木守;中屋佑紀;佐藤久
  • 2023年03月, 日本水環境学会, ライオン賞               
    小原紀子;樋口宏介;北島正章;岡部聡;押木守
  • 2022年03月, 日本水環境学会, クリタ賞               
    樋口宏介,北島正章,岡部 聡,押木 守
  • 2021年, 長瀬科学技術振興財団, 長瀬研究振興賞               
  • 2019年, 日本微生物生態学会, Microbes & Environments 2018 最優秀論文賞               
    押木 守
  • 2018年, インテリジェント・コスモス, インテリジェント・コスモス奨励賞               
    押木 守
  • 2018年, 第28回日本MRS年次大会, 奨励賞               
    押木 守
  • 2017年, イムラ・ジャパン, 第1回イムラ・ジャパン賞               
    押木 守
  • 2017年, 建設工学研究振興会, 建設工学奨励賞               
    押木 守
  • 2011年, 日本微生物生態学会, 第27回日本微生物生態学会 優秀ポスター賞               
    押木 守
  • 2011年, アナモックスシンポジウム, 第一回国際アナモックスシンポジウム 最優秀講演賞               
    押木 守
  • 2011年, 国際硝化学会, 第2回国際硝化学会・第16回ヨーロッパ窒素循環会議 発表賞               
    押木 守
  • 2008年, 日本水環境学会, 博士研究奨励賞 (オルガノ賞)               
    押木 守
  • 2002年, 全国高専土木学会, 近藤賞               
    押木 守

論文

  • Interspecific competition and adaptation of anammox bacteria at different salinities: Experimental validation of the Monod growth model with salinity inhibition
    Satoshi Okabe, Akimichi Kamizono, Seiya Kawasaki, Kanae Kobayashi, Mamoru Oshiki
    Water Research, 2025年03月
    研究論文(学術雑誌)
  • Thermophilic dark fermentation for hydrogen and volatile fatty acids production from breadcrumbs
    Oshiki, M., Yamaguchi, G., Takahashi, K., Okabe, S., Kawano, S., Nakagawa, J., Fukushima, T.
    Chemical Engineering Journal, 501, 157633, 2024年12月, [査読有り]
    英語, 研究論文(学術雑誌)
  • Salinity Tolerance and Osmoadaptation Strategies in Four Genera of Anammox Bacteria: Brocadia, Jettenia, Kuenenia, and Scalindua
    Satoshi Okabe, Akimichi Kamizono, Lei Zhang, Seiya Kawasaki, Kanae Kobayashi, Mamoru Oshiki
    Environmental Science & Technology, American Chemical Society (ACS), 2024年03月16日, [査読有り], [最終著者]
    研究論文(学術雑誌)
  • Collaborative metabolisms of urea and cyanate degradation in marine anammox bacterial culture
    Mamoru Oshiki, Emi Morimoto, Kanae Kobayashi, Hisashi Satoh, Satoshi Okabe
    ISME Communications, 4, 1, Oxford University Press (OUP), 2024年01月01日, [査読有り], [筆頭著者]
    研究論文(学術雑誌), Abstract

    Anammox process greatly contributes to nitrogen loss occurring in oceanic oxygen minimum zones (OMZs), where the availability of NH4+ is scarce as compared with NO2−. Remineralization of organic nitrogen compounds including urea and cyanate (OCN−) into NH4+ has been believed as an NH4+ source of the anammox process in oxygen minimum zones. However, urea- or OCN−- dependent anammox has not been well examined due to the lack of marine anammox bacterial culture. In the present study, urea and OCN− degradation in a marine anammox bacterial consortium were investigated based on 15N-tracer experiments and metagenomic analysis. Although a marine anammox bacterium, Candidatus Scalindua sp., itself was incapable of urea and OCN− degradation, urea was anoxically decomposed to NH4+ by the coexisting ureolytic bacteria (Rhizobiaceae, Nitrosomonadaceae, and/or Thalassopiraceae bacteria), whereas OCN− was abiotically degraded to NH4+. The produced NH4+ was subsequently utilized in the anammox process. The activity of the urea degradation increased under microaerobic condition (ca. 32–42 μM dissolved O2, DO), and the contribution of the anammox process to the total nitrogen loss also increased up to 33.3% at 32 μM DO. Urea-dependent anammox activities were further examined in a fluid thioglycolate media with a vertical gradient of O2 concentration, and the active collaborative metabolism of the urea degradation and anammox was detected at the lower oxycline (21 μM DO).
  • Denitrification in low oxic environments increases the accumulation of nitrogen oxide intermediates and modulates the evolutionary potential of microbial populations.
    Kohei Takahashi, Mamoru Oshiki, Chujin Ruan, Kana Morinaga, Masanori Toyofuku, Nobuhiko Nomura, David R Johnson
    Environmental microbiology reports, 16, 1, e13221, 2023年11月30日, [査読有り], [国際誌]
    英語, 研究論文(学術雑誌), Denitrification in oxic environments occurs when a microorganism uses nitrogen oxides as terminal electron acceptors even though oxygen is available. While this phenomenon is well-established, its consequences on ecological and evolutionary processes remain poorly understood. We hypothesize here that denitrification in oxic environments can modify the accumulation profiles of nitrogen oxide intermediates with cascading effects on the evolutionary potentials of denitrifying microorganisms. To test this, we performed laboratory experiments with Paracoccus denitrificans and complemented them with individual-based computational modelling. We found that denitrification in low oxic environments significantly increases the accumulation of nitrite and nitric oxide. We further found that the increased accumulation of these intermediates has a negative effect on growth at low pH. Finally, we found that the increased negative effect at low pH increases the number of individuals that contribute to surface-associated growth. This increases the amount of genetic diversity that is preserved from the initial population, thus increasing the number of genetic targets for natural selection to act upon and resulting in higher evolutionary potentials. Together, our data highlight that denitrification in low oxic environments can affect the ecological processes and evolutionary potentials of denitrifying microorganisms by modifying the accumulation of nitrogen oxide intermediates.
  • 異なる溶存酸素条件が複合微生物系における好気性脱窒細菌群に与える影響               
    上村 光輝, 川上 周司, 押木 守, 青木 仁孝, 土田 勝範, 渡利 高大, 荒木 信夫
    土木学会論文集, 79, 25, 23-25034, 2023年11月, [査読有り]
  • Growth of the Nitrosomonas europaea cells in the biofilm and planktonic growth mode: Responses of extracellular polymeric substances production and transcriptome
    Mamoru Oshiki, Takahiro Saito, Yuki Nakaya, Hisashi Satoh, Satoshi Okabe
    Journal of Bioscience and Bioengineering, Elsevier BV, 2023年11月, [査読有り], [筆頭著者, 責任著者]
    研究論文(学術雑誌)
  • Oxygen tolerance and detoxification mechanisms of highly enriched planktonic anaerobic ammonium-oxidizing (anammox) bacteria
    Satoshi Okabe, Shaoyu Ye, Xi Lan, Keishi Nukada, Haozhe Zhang, Kanae Kobayashi, Mamoru Oshiki
    ISME Communications, 3, 1, Springer Science and Business Media LLC, 2023年05月03日, [査読有り], [最終著者]
    研究論文(学術雑誌), Abstract

    Oxygen is a key regulatory factor of anaerobic ammonium oxidation (anammox). Although the inhibitory effect of oxygen is evident, a wide range of oxygen sensitivities of anammox bacteria have been reported so far, which makes it difficult to model the marine nitrogen loss and design anammox-based technologies. Here, oxygen tolerance and detoxification mechanisms of four genera of anammox bacteria; one marine species (“Ca. Scalindua sp.”) and four freshwater anammox species (“Ca. Brocadia sinica”, “Ca. Brocadia sapporoensis”, “Ca. Jettenia caeni”, and “Ca. Kuenenia stuttgartiensis”) were determined and then related to the activities of anti-oxidative enzymes. Highly enriched planktonic anammox cells were exposed to various levels of oxygen, and oxygen inhibition kinetics (50% inhibitory concentration (IC50) and upper O2 limits (DOmax) of anammox activity) were quantitatively determined. A marine anammox species, “Ca. Scalindua sp.”, exhibited much higher oxygen tolerance capability (IC50 = 18.0 µM and DOmax = 51.6 µM) than freshwater species (IC50 = 2.7–4.2 µM and DOmax = 10.9–26.6 µM). The upper DO limit of “Ca. Scalindua sp.” was much higher than the values reported so far (~20 µM). Furthermore, the oxygen inhibition was reversible even after exposed to ambient air for 12–24 h. The comparative genome analysis confirmed that all anammox species commonly possess the genes considered to function for reduction of O2, superoxide anion (O2•-), and H2O2. However, the superoxide reductase (Sor)-peroxidase dependent detoxification system alone may not be sufficient for cell survival under microaerobic conditions. Despite the fact that anaerobes normally possess no or little superoxide dismutase (Sod) or catalase (Cat), only Scalindua exhibited high Sod activity of 22.6 ± 1.9 U/mg-protein with moderate Cat activity of 1.6 ± 0.7 U/mg-protein, which was consistent with the genome sequence analysis. This Sod-Cat dependent detoxification system could be responsible for the higher O2 tolerance of Scalindua than other freshwater anammox species lacking the Sod activity.
  • 安定同位体を用いた窒素循環に関わる微生物代謝活性評価               
    押木守
    バイオサイエンスとインダストリー, 81, 3, 200, 204, 2023年05月, [査読有り], [招待有り], [筆頭著者, 責任著者]
  • Multivariate regression modeling in integrative analysis via sparse regularization               
    Shuichi Kawano, Toshikazu Fukushima, Junichi Nakagawa, Mamoru Oshiki
    arXiv (preprint), 2023年04月
  • Growth of nitrite-oxidizing Nitrospira and ammonia-oxidizing Nitrosomonas in marine recirculating trickling biofilter reactors.
    Mamoru Oshiki, Hirotoshi Netsu, Kyohei Kuroda, Takashi Narihiro, Naoki Fujii, Tomonori Kindaichi, Yoshiyuki Suzuki, Takahiro Watari, Masashi Hatamoto, Takashi Yamaguchi, Nobuo Araki, Satoshi Okabe
    Environmental microbiology, 24, 8, 3735, 3750, 2022年06月07日, [査読有り], [筆頭著者, 責任著者], [国際誌]
    英語, 研究論文(学術雑誌), Aerobic ammonia and nitrite oxidation reactions are fundamental biogeochemical reactions contributing to the global nitrogen cycle. Although aerobic nitrite oxidation yields 4.8-folds less Gibbs free energy (∆Gr ) than aerobic ammonia oxidation in the NH4 + -feeding marine recirculating trickling biofilter reactors operated in the present study, nitrite-oxidizing and not ammonia-oxidizing Nitrospira (sublineage IV) outnumbered ammonia-oxidizing Nitrosomonas (relative abundance; 53.8% and 7.59% respectively). CO2 assimilation efficiencies during ammonia or nitrite oxidation were 0.077 μmol-14 CO2 /μmol-NH3 and 0.053-0.054 μmol-14 CO2 /μmol-NO2 - respectively, and the difference between ammonia and nitrite oxidation was much smaller than the difference of ∆Gr . Free-energy efficiency of nitrite oxidation was higher than ammonia oxidation (31%-32% and 13% respectively), and high CO2 assimilation and free-energy efficiencies were a determinant for the dominance of Nitrospira over Nitrosomonas. Washout of Nitrospira and Nitrosomonas from the trickling biofilter reactors was also examined by quantitative PCR assay. Normalized copy numbers of Nitrosomonas amoA were 1.5- to 1.7-folds greater than Nitrospira nxrB and 16S rRNA gene in the reactor effluents. Nitrosomonas was more susceptible for washout than Nitrospira in the trickling biofilter reactors, which was another determinant for the dominance of Nitrospira in the trickling biofilter reactors.
  • NH2OH disproportionation mediated by anaerobic ammonium-oxidizing (anammox) bacteria
    Mamoru Oshiki, Lin Gao, Lei Zhang, Satoshi Okabe
    Microbes Environ., 37, 2, ME21092, n/a, Japanese Society of Microbial Ecology, 2022年04月, [査読有り], [筆頭著者, 責任著者]
    研究論文(学術雑誌)
  • N2O reduction by Gemmatimonas aurantiaca and potential involvement of Gemmatimonadetes bacteria in N2O reduction in agricultural soils
    Mamoru Oshiki, Yuka Toyama, Toshikazu Suenaga, Akihiko Terada, Yasuhiro Kasahara, Takashi Yamaguchi, Nobuo Araki
    Microbes Environ., 37, 2, ME21090, n/a, Japanese Society of Microbial Ecology, 2022年04月, [査読有り], [筆頭著者, 責任著者]
    研究論文(学術雑誌)
  • Determination of 15 N/14N of ammonium, nitrite, nitrate, hydroxylamine, and hydrazine using colorimetric reagents and matrix-assisted laser desorption ionization–time-of-flight mass spectrometry (MALDI-TOF MS)
    Mamoru Oshiki, Komei Nagai, Satoshi Ishii, Yoshiyuki Suzuki, Nobuo Saito, Takashi Yamaguchi, Nobuo Araki, Satoshi Okabe
    Appl. Environ. Microbiol., 88, 9, e0241621, American Society for Microbiology, 2022年03月14日, [査読有り], [筆頭著者, 責任著者], [国際誌]
    英語, 研究論文(学術雑誌), 15 N/ 14 N analysis for aqueous N species is a powerful tool for obtaining insights into the global N cycle, but the procedure is cumbersome and laborious. The combined use of colorimetric reagents and MALDI-TOF MS, designated color MALDI-TOF MS, enabled us to determine the 15 N atom% of common aqueous N species without laborious sample preparation and chromatographic separation steps; for instance, the 15 N atom% of NO 2 can be determined from >1,000 liquid samples daily at <$1 (U.S.) per 384 samples for routine analysis.
  • Full-scale application of a down-flow hanging sponge reactor combined with a primary sedimentation basin for domestic sewage treatment
    Tsutomu Okubo, Tadashi Tagawa, Masanobu Takahashi, Akinori Iguchi, Mamoru Oshiki, Nobuo Araki, Kengo Kubota, Ahmed Tawfik, Shigeki Uemura, Hideki Harada
    Bioprocess Biosyst. Eng., 45, 4, 701, 709, Springer Science and Business Media LLC, 2022年01月31日, [査読有り]
    研究論文(学術雑誌)
  • Metagenomic Analysis of Five Phylogenetically Distant Anammox Bacterial Enrichment Cultures.
    Mamoru Oshiki, Yoshihiro Takaki, Miho Hirai, Takuro Nunoura, Atsushi Kamigaito, Satoshi Okabe
    Microbes and environments, 37, 3, 2022年, [査読有り], [筆頭著者, 責任著者], [国内誌]
    英語, 研究論文(学術雑誌), Anaerobic ammonium-oxidizing (anammox) bacteria are slow-growing and fastidious bacteria, and limited numbers of enrichment cultures have been established. A metagenomic ana-lysis of our 5 established anammox bacterial enrichment cultures was performed in the present study. Fourteen high-quality metagenome-assembled genomes (MAGs) were obtained, including those of 5 anammox Planctomycetota (Candidatus Brocadia, Ca. Kuenenia, Ca. Jettenia, and Ca. Scalindua), 4 Bacteroidota, and 3 Chloroflexota. Based on the gene sets of metabolic pathways involved in the degradation of polymeric substances found in Chloroflexota and Bacteroidota MAGs, they are expected to be scavengers of extracellular polymeric substances and cell debris.
  • Endpoint recombinase polymerase amplification (RPA) assay for enumeration of thiocyanate-degrading bacteria
    Mamoru Oshiki, Toshikazu Fukushima, Shuichi Kawano, Junichi Nakagawa
    Microbes Environ., 37, 1, ME21073, n/a, Japanese Society of Microbial Ecology, 2022年, [査読有り], [筆頭著者, 責任著者]
    研究論文(学術雑誌)
  • 海洋性Anammox細菌の脱アンモニウム能に及ぼすpHの影響:至適pH制御による脱アンモニウム能の回復               
    田中孝国, 岡悟史, 押木守, 金田一智規, 新田見匡
    工業用水, 669, 51, 55, 2021年11月, [査読有り]
  • 異なるpH条件下における 海洋性Anammox細菌の脱アンモニウム能               
    田中孝国, 岡悟史, 田中昭雄, 押木守, 金田一智規, 新田見匡
    工業用水, 667, 76, 82, 2021年07月, [査読有り]
  • Glycogen metabolism of the anammox bacterium "Candidatus Brocadia sinica".
    Satoshi Okabe, Amrini Amalia Shafdar, Kanae Kobayashi, Lei Zhang, Mamoru Oshiki
    ISME J., 15, 5, 1287, 1301, 2021年05月, [査読有り], [最終著者], [国際誌]
    英語, 研究論文(学術雑誌), Presence of glycogen granules in anaerobic ammonium-oxidizing (anammox) bacteria has been reported so far. However, very little is known about their glycogen metabolism and the exact roles. Here, we studied the glycogen metabolism in "Ca. Brocadia sinica" growing in continuous retentostat cultures with bicarbonate as a carbon source. The effect of the culture growth phase was investigated. During the growing phase, intracellular glycogen content increased up to 32.6 mg-glucose (g-biomass dry wt)-1 while the specific growth rate and ATP/ADP ratio decreased. The accumulated glycogen begun to decrease at the onset of entering the near-zero growth phase and was consumed rapidly when substrates were depleted. This clearly indicates that glycogen was synthesized and utilized as an energy storage. The proteomic analysis revealed that "Ca. B. sinica" synthesized glycogen via three known glycogen biosynthesis pathways and simultaneously degraded during the progress of active anammox, implying that glycogen is being continuously recycled. When cells were starved, a part of stored glycogen was converted to trehalose, a potential stress protectant. This suggests that glycogen serves at least as a primary carbon source of trehalose synthesis for survival. This study provides the first physiological evidence of glycogen metabolism in anammox bacteria and its significance in survival under natural substrate-limited habitat.
  • Complete genome sequence of Thiohalobacter sp. strain COW1, isolated from activated sludge treating coke oven wastewater
    Kentaro Miyazaki, Hikaru Suenaga, Mamoru Oshiki, Shuichi Kawano, Toshikazu Fukushima
    Microbiol. Resour. Announc., 10, 7, e00013-21, American Society for Microbiology, 2021年02月18日, [査読有り], [国際誌]
    英語, 研究論文(学術雑誌), ABSTRACT
    A thiocyanate-degrading bacterium, Thiohalobacter sp. strain COW1, was isolated from activated sludge treating coke oven wastewater, and the complete genome sequence was determined. COW1 contained a single circular chromosome (3.23 Mb; G+C content, 63.4%) in which 2,788 protein-coding genes, 39 tRNA genes, and 3 rRNA genes were identified.
  • 海洋に流出した窒素除去を目的とした海洋性anammox細菌の研究
    田中考国, 押木守, 金田一智規, 新田見匡
    分離技術, 51, 1, 32, 41, 2021年02月, [査読有り]
  • Draft genome sequence of Cytophagales sp. strain WSM2-2, isolated from garden soil
    Masataka Aoki, Mamoru Oshiki, Mizuki Iwagaki, Masashi Hatamoto, Takashi Yamaguchi, Nobuo Araki
    Microbiol. Resour. Announc., 10, 2, e01238-20, American Society for Microbiology, 2021年01月14日, [査読有り]
    研究論文(学術雑誌), ABSTRACT
    We report the draft genome sequence of Cytophagales sp. strain WSM2-2, isolated from garden soil. A 5.5-Mb genome sequence comprising four contigs was successfully obtained using Illumina NovaSeq and MinION sequencers. This draft genome sequence will contribute to the genomic knowledge of the bacterial order Cytophagales.
  • Draft genome sequence of Enterobacter sp. AS-1, a potential eurytrophic recombination host
    Yuki Iwasaki, Yuya Itoiri, Sota Ihara, Hironaga Akita, Mamoru Oshiki, Zen-ichiro Kimura
    J. Genomics, 9, 6, 9, Ivyspring International Publisher, 2021年, [査読有り], [国際誌]
    英語, 研究論文(学術雑誌), Strain AS-1 was isolated from laboratory-scale activated sludge collected in Japan. This strain not only grows on rich medium, including R2A medium, but also forms colonies on medium lacking organic matter other than agar (water agar), indicating it could be used as a eurytrophic recombinant host in material production processes. Here, we present a draft genome sequence of Enterobacter sp. AS-1, which consists of a total of 24 contigs containing 5,207,146 bp, with a GC content of 55.64%, and comprising 4,921 predicted coding sequences. Based on 16S rRNA gene sequence analysis, strain AS-1 was designated as Enterobacter sp. AS-1.
  • Cell-density dependent anammox activity of Candidatus Brocadia sinica regulated by N-acyl homoserine lactone-mediated quorum sensing
    Mamoru Oshiki, Haruna Hiraizumi, Hisashi Satoh, Satoshi Okabe
    Microbes Environ., 35, 4, ME20086, 2020年08月, [査読有り], [筆頭著者, 責任著者], [国内誌]
    英語, 研究論文(学術雑誌), The activity of anaerobic ammonia-oxidizing (anammox) bacteria is considered to depend on cell density; however, this has not yet been confirmed due to the fastidious nature of anammox bacteria (e.g., slow growth, oxygen sensitivity, and rigid aggregate formation). In the present study, the cell density-dependent occurrence of anammox activity (14-15N2 gas production rate) was investigated using planktonic enrichment cultures of Candidatus Brocadia sinica. This activity was detectable when the density of cells was higher than 107‍ ‍cells‍ ‍mL-1 and became stronger with increases in cell density. At the cell densities, the transcription of the BROSI_A1042 and BROSI_A3652 genes, which are potentially involved in the biosynthesis and reception of N-acyl homoserine lactone (AHL), was detectable in Brocadia sinica cells. The presence of AHL molecules in the MBR culture of B. sinica was confirmed by an AHL reporter assay and gas chromatography mass spectrometry analysis. The exogenous addition of the MBR culture extract and AHL molecules (a cocktail of C6, C8, C10, and C12-homoserine lactones) increased the specific 14-15N2 production rate of B. sinica. These results suggest that the specific anammox activity of B. sinica is regulated by AHL-mediated quorum sensing.
  • N2O production using native nos-deficient denitrifying bacterial strains screened by a genome mining approach               
    Mamoru Oshiki, Miho Ishimaru, Masashi Hatamoto, Takashi Yamaguchi, Nobuo Araki, Satoshi Okabe
    Bioresour. Technol. Rep., 11, 100529, 2020年08月, [査読有り], [筆頭著者, 責任著者]
  • Formation of denitrifying granules in an upflow sludge blanket reactor with municipal sewage and sodium nitrate feeding
    Takahiro Watari, Wilasinee Kotcharoen, Takanori Omine, Masashi Hatamoto, Nobuo Araki, Mamoru Oshiki, Kazuhisa Mimura, Akihiro Nagano, Takashi Yamaguchi
    Environ. Technol. Innov., 19, 100861, 100861, Elsevier BV, 2020年08月, [査読有り]
    研究論文(学術雑誌)
  • 異なる温度条件下における海洋性Anammox細菌の脱アンモニウム能               
    田中孝国, 柴田真輝, フーレイチア, 田中昭雄, 押木守, 金田一智規, 新田見匡
    工業用水, 661, 69, 75, 2020年07月, [査読有り]
  • Biosynthesis of hydrazine from ammonium and hydroxylamine using an anaerobic ammonium oxidizing bacterium
    Mamoru Oshiki, Koshiro Yamada, Itsuki Kato, Kento Okoshi, Toshiro Imai, Takashi Yamaguchi, Nobuo Araki
    Biotechnology Letters, 42, 6, 979, 985, Springer Science and Business Media {LLC}, 2020年06月17日, [査読有り], [筆頭著者, 責任著者]
    英語, 研究論文(学術雑誌)
  • Simple and reliable enumeration of Escherichia coli concentrations in wastewater samples by measuring β-D-glucuronidase (GUS) activities via a microplate reader
    Hisashi Satoh, Kai Kikuchi, Yutaka Katayose, Shu Tsuda, Reiko Hirano, Yuga Hirakata, Masaaki Kitajima, Satoshi Ishii, Mamoru Oshiki, Masashi Hatamoto, Masahiro Takahashi, Satoshi Okabe
    Sci. Total Environ., 715, 136928, 2020年05月01日, [査読有り]
    研究論文(学術雑誌), © 2020 Elsevier B.V. Monitoring of Escherichia coli concentrations at wastewater treatment plants (WWTPs) is important to ensure process performance and protect public health. However, conventional E. coli enumeration methods are complicated and time- and labor-consuming. Here, we report a novel simple and reliable method based on β-D-glucuronidase (GUS) activity assay to enumerate E. coli concentrations in wastewater (WW) samples. An aliquot (20 μL) of the medium with fluorogenic enzyme substrate for E. coli and 180 μL of a WW sample were added to one well of a 96-well microplate. The microplate was placed in a microplate reader at 37 °C. To this end, the fluorescence intensity of a fluorogenic enzyme substrate for E. coli was measured every 10 min over 3 h to determine GUS activity. The linear increase in the fluorescence intensity representing the GUS activities showed a positive correlation with E. coli concentrations in wastewater samples. However, the correlation equations were specific to WWTPs, which could be due to the difference in the E. coli population structures among WWTPs. We observed that the wastewater matrix is not a limitation to measure the GUS activity, and a WWTP-specific correlation equation can be used as a calibration curve to estimate the E. coli concentrations in the samples collected from that site. A comparison of the results with those of culture-dependent Colilert method proved that the current method is simple and useful for the enumeration of E. coli concentrations in wastewater samples reliably.
  • Food selectivity of anaerobic protists and direct evidence for methane production using carbon from prey bacteria by endosymbiotic methanogen.
    Yuga Hirakata, Masashi Hatamoto, Mamoru Oshiki, Takahiro Watari, Nobuo Araki, Takashi Yamaguchi
    ISME J., 14, 7, 1873, 1885, 2020年04月27日, [査読有り], [国際誌]
    英語, 研究論文(学術雑誌), Anaerobic protists are major predators of prokaryotes in anaerobic ecosystems. However, little is known about the predation behavior of anaerobic protists because almost none have been cultured. In particular, these characteristics of anaerobic protists in the phyla Metamonada and Cercozoa have not been reported previously. In this study, we isolated three anaerobic protists, Cyclidium sp., Trichomitus sp., and Paracercomonas sp., from anaerobic granular sludge in an up-flow anaerobic sludge blanket reactor used to treat domestic sewage. Ingestion and digestion of food bacteria by anaerobic protists with or without endosymbiotic methanogens were demonstrated using tracer experiments with green fluorescent protein and a stable carbon isotope. These tracer experiments also demonstrated that Cyclidium sp. supplied CO2 and hydrogen to endosymbiotic methanogens. While Cyclidium sp. and Trichomitus sp. ingested both Gram-negative and -positive bacteria, Paracercomonas sp. could only take up Gram-negative bacteria. Archaeal cells such as Methanobacterium beijingense and Methanospirillum hungatei did not support the growth of these protists. Metabolite patterns of all three protists differed and were influenced by food bacterial species. These reported growth rates, ingestion rates, food selectivity, and metabolite patterns provide important insights into the ecological roles of these protists in anaerobic ecosystems.
  • Total ammonia nitrogen (TAN) removal performance of a recirculating down-hanging sponge (DHS) reactor operated at 10 to 20ºC with activated carbon               
    Mamoru Oshiki, Takashi Aizuka, Hirotoshi Netsu, Satoshi Oomori, Akihiro Nagano, Takashi Yamaguchi, Nobuo Araki
    Aquaculture, 520, 734963, 2020年01月, [査読有り], [筆頭著者, 責任著者]
  • Draft genome sequence of Deinococcus sp. KR-1, a potential strain for palladium-leaching
    Hironaga Akita, Yuya Itoiri, Akio Kumagai, Noriyo Takeda, Akinori Matsushika, Mamoru Oshiki, Zen-ichiro Kimura
    J. Genomics, 8, 21, 24, Ivyspring International Publisher, 2020年01月, [査読有り]
    研究論文(学術雑誌)
  • Temporal variation of eukaryotic community structures in UASB reactor treating domestic sewage as revealed by 18S rRNA gene sequencing
    Yuga Hirakata, Masashi Hatamoto, Mamoru Oshiki, Takahiro Watari, Kyohei Kuroda, Nobuo Araki, Takashi Yamaguchi
    Sci. Rep., 9, 1, 12783, 12783, 2019年12月01日, [査読有り]
    英語, 研究論文(学術雑誌), © 2019, The Author(s). Eukaryotes are important components of ecosystems in wastewater treatment processes. However, little is known about eukaryotic community in anaerobic wastewater treatment systems. In this study, eukaryotic communities in an up flow anaerobic sludge blanket (UASB) reactor treating domestic sewage during two years of operation were investigated using V4 and V9 regions of 18S rRNA gene for amplicon sequencing. In addition, activated sludge and influent sewage samples were also analyzed and used as the references for aerobic eukaryotic community to characterize anaerobic eukaryotes. The amplicon sequence V4 and V9 libraries detected different taxonomic groups, especially from the UASB samples, suggesting that commonly used V4 and V9 primer pairs could produce a bias for eukaryotic communities analysis. Eukaryotic community structures in the UASB reactor were influenced by the immigration of eukaryotes via influent sewage but were clearly different from the influent sewage and activated sludge. Multivariate statistics indicated that protist genera Cyclidium, Platyophrya and Subulatomonas correlated with chemical oxygen demand and suspended solid concentration, and could be used as bioindicators of treatment performance. Uncultured eukaryotes groups were dominant in the UASB reactor, and their physiological roles need to be examined to understand their contributions to anaerobic processes in future studies.
  • Draft Genome Sequence of the Plant Growth-Promoting Streptomyces sp. Strain 6-11-2
    Ayman Ztouti, Natsumi Saito, Junna Sasaki, Takashi Yamaguchi, Nobuo Araki, Mamoru Oshiki
    Microbiology Resource Announcements, 8, e00877-19, 2019年11月21日, [査読有り], [最終著者, 責任著者]
    研究論文(学術雑誌)
  • Thiocyanate degradation by a highly enriched culture of the neutrophilic halophile Thiohalobacter sp. strain FOKN1 from activated sludge and genomic insights into thiocyanate metabolism
    Oshiki M, Fukushima T, Kawano S, Kasahara Y, Nakagawa J
    Microbes Environ., 34, 4, 402, 412, Japanese Society of Microbial Ecology, 2019年10月, [査読有り], [筆頭著者, 責任著者]
    研究論文(学術雑誌)
  • Dual nitrogen and oxygen isotope fractionation during anaerobic ammonium oxidation by anammox bacteria
    Kanae Kobayashi, Akiko Makabe, Midori Yano, Mamoru Oshiki, Tomonori Kindaichi, Karen L. Casciotti, Satoshi Okabe
    ISME J., 13, 10, 2426, 2436, Springer Science and Business Media LLC, 2019年10月, [査読有り]
    研究論文(学術雑誌)
  • 異なる塩分濃度環境下における海洋性Anammox細菌の脱アンモニウム能
    田中 孝国, 半田 佳幹, フーレイチア, 田中 昭雄, 押木 守, 金田一 智規, 新田見 匡
    工業用水 = Journal of industrial water, 656, 656, 77, 84, 日本工業用水協会, 2019年09月, [査読有り]
    日本語, 研究論文(学術雑誌)
  • Investigation of prospective factors that control Kouleothrix (Type 1851) filamentous bacterial abundance and their correlation with sludge settleability in full-scale wastewater treatment plants
    Nittami, T, Shoji, T, Koshiba, Y, Noguchi, M, Oshiki, M, Kuroda, M, Kindaichi, T, Fukuda, J, Kurisu F
    Process Saf. Environ., 124, 137, 142, 2019年04月, [査読有り]
    研究論文(学術雑誌), © 2019 Institution of Chemical Engineers Kouleothrix (Type 1851) filamentous bacteria associated with bulking incidents were investigated for their quantitative correlations with sludge settleability in municipal wastewater treatment plants (WWTPs) around Japan. Factors affecting their abundance were also explored by statistical analyses with the aim of controlling bulking by suppressing their growth. Real-time quantitative PCR assays showed Kouleothrix spp. mainly ranged from 104−105 copies ng-DNA−1 in WWTPs. They were not always abundant in every WWTP, but their abundance (>105 copies ng-DNA−1) almost always coincided with low activated sludge settleability. Their abundance in nitrogen removal processes was always low (1.08 − 3.83 × 104 copies ng-DNA−1). Denitrification process at the feeding stage may be a useful selection pressure for Kouleothrix spp., leading to their suppression. A stepwise multiple regression analysis revealed that the increase of dissolved oxygen (DO) and pH in an aeration tank have a negative influence, while NH4+ increase in the influent has a positive influence on the Kouleothrix spp. abundance. The three-variable model was significant and accounted for approximately 45% of the variance in the dependent variable (i.e., Kouleothrix spp.). The DO increase had the greatest influence on the Kouleothrix spp. decrease, and thus, the DO control may be a promising way to suppress their growth in WWTPs.
  • 都市下水処理UASBの上昇線流速が槽内の原生動物に及ぼす影響
    吉田 力, 佐藤 剛, 押木 守, 荒木 信夫, 平片 悠河, 幡本 将史, 山口 隆司
    土木学会論文集G(環境), 75, 7, III_19, III_24, 公益社団法人 土木学会, 2019年, [査読有り]
    日本語, 研究論文(学術雑誌),

    本研究では実都市下水を供給したUASBリアクター内の上昇線流速を段階的に変化させ, 槽内に出現する嫌気性原生動物の細胞数と原生動物叢への影響を解析した. 原生動物の細胞数は槽内の上昇線流速が2.4-14.4m/dayの範囲では0-3970cells/mLで変化し,上昇線流速の増加に伴って増加する傾向が見られた.22.4m/dayの上昇線流速では原生動物が汚泥床外へ流亡し,細胞数は100cells/mLまで急激に減少した.UASB内に存在した嫌気性原生動物はMetopus palaeformis,Metopus contortus,Chaenomorpha sp.,Plagiopyla sp.が優占種であり,平均して全18SrRNA遺伝子の約7割を占めた.線流速が大きい条件でUASB内の原生動物の優占種は短期間で変化し,その変化は同一条件の運転期間においても観察された.

  • Health impact of agricultural drainage water for farmers in the West Nile Delta               
    Okubo, T, Iguchi, A, Tanaka, S, Uchida, S, Tagawa, T, Oshiki, M, Araki, A, Tawfik, A, Takahashi, M, Kubota, K, Harada, H, Uemura, S
    Int. J. Environ. Res, 13, 319, 325, 2019年, [査読有り]
    英語, 研究論文(学術雑誌)
  • Synergistic inhibition of anaerobic ammonium oxidation (anammox) activity by phenol and thiocyanate
    Mamoru Oshiki, Yoshiko Masuda, Takashi Yamaguchi, Nobuo Araki
    Chemosphere, 213, 498, 506, Elsevier {BV}, 2018年12月, [査読有り], [筆頭著者, 責任著者]
    研究論文(学術雑誌)
  • Nitric oxide production from nitrite reduction and hydroxylamine oxidation by copper-containing dissimilatory nitrite reductase (nirK) from the aerobic ammonia-oxidizing archaeon, Nitrososphaera viennensis
    Kobayashi S, Hira D, Yoshida K, Toyofuku M, Shida Y, Ogasawara W, Yamaguchi T, Araki N, Oshiki M
    Microbes Environ., 33, 4, 428, 434, Japanese Society of Microbial Ecology, 2018年10月, [査読有り], [最終著者, 責任著者]
    研究論文(学術雑誌)
  • 窒素を食べる微生物たち
    押木守
    生物工学会誌, 96, 7, 408, 2018年07月25日, [筆頭著者, 責任著者]
    日本語
  • Ureolytic prokaryotes in soil: community abundance and diversity
    Mamoru Oshiki, Mitsuru Araki, Yuga Hirakata, Masashi Hatamoto, Takashi Yamaguchi, Nobuo Araki
    Microbes Environ., 33, 2, 230, 233, JAPANESE SOC MICROBIAL ECOLOGY, DEPT BIORESOURCE SCIENCE, 2018年07月, [査読有り], [筆頭著者, 責任著者]
    英語, 研究論文(学術雑誌), Although the turnover of urea is a crucial process in nitrogen transformation in soil, limited information is currently available on the abundance and diversity of ureolytic prokaryotes. The abundance and diversity of the soil 16S rRNA gene and ureC (encoding a urease catalytic subunit) were examined in seven soil types using quantitative PCR and amplicon sequencing with Illumina MiSeq. The amplicon sequencing of ureC revealed that the ureolytic community was composed of phylogenctically varied prokaryotes, and we detected 363 to 1,685 species-level ureC operational taxonomic units (OTUs) per soil sample, whereas 5,984 OTUs were site-specific OTUs found in only one of the seven soil types.
  • 不織布に塗布した海洋性Anammox細菌を用いた反応装置の立ち上げ方法の検討
    田中 孝国, フーレイチア, 田中 昭雄, 押木 守, 金田一 智規, 新田見 匡
    工業用水 = Journal of industrial water, 648, 648, 39, 43, 日本工業用水協会, 2018年05月, [査読有り]
    日本語, 研究論文(学術雑誌)
  • Experimental evidence for in situ nitric oxide production in anaerobic ammonia-oxidizing bacterial granules
    Rathnayake, Rathnayake M. L. D., Oshiki, Mamoru, Ishii, Satoshi, Segawa, Takahiro, Satoh, Hisashi, Okabe, Satoshi
    Environ. Sci. Technol., 52, 10, 5744, 5752, AMER CHEMICAL SOC, 2018年05月, [査読有り]
    英語, 研究論文(学術雑誌), Although nitric oxide (NO) emissions from anaerobic ammonium oxidation (anammox)-based processes were reported previously, the NO production pathways are poorly understood. Here, we investigated the NO production pathways in anammox granules in detail by combining N-15-stable isotope tracer experiments with various inhibitors, microsensor measurements, and transcriptome analysis for key genes of NO2 reduction. NO was emitted from the anammox granules, which account for 0.07% of the N-2 emission. N-15-stable isotope-tracer experiments indicated that most of the N-2 was produced by anammox bacteria, whereas NO was produced from NO2- reduction by anammox and denitrifying bacteria. The NO emission rate was highest at pH 8.0 and accelerated by increasing NH4+ and NO2- concentrations in the culture media. The microsensor analyses showed the in situ NO production rate was highest in the outer layer of the anammox granule where anammox activity was also highest. The detected in situ NO concentrations of up to 2.7 mu M were significantly above physiological thresholds known to affect a wide range of microorganisms present in wastewater. Hence, NO likely plays pivotal roles in the microbial interactions in anammox granules, which needs to be further investigated.
  • Microfluidic PCR amplification and MiSeq amplicon sequencing techniques for high-throughput detection and genotyping of human pathogenic RNA viruses in human feces, sewage, and oysters
    Mamoru Oshiki, Takayuki Miura, Shinobu Kazama, Takahiro Segawa, Satoshi Ishii, Masashi Hatamoto, Takashi Yamaguchi, Kengo Kubota, Akinori Iguchi, Tadashi Tagawa, Tsutomu Okubo, Shigeki Uemura, Hideki Harada, Naohiro Kobayashi, Nobuo Araki, Daisuke Sano
    Front. Microbiol., 9, 830, Frontiers Media S.A., 2018年04月27日, [査読有り], [筆頭著者, 責任著者]
    英語, 研究論文(学術雑誌), Detection and genotyping of pathogenic RNA viruses in human and environmental samples are useful for monitoring the circulation and prevalence of these pathogens, whereas a conventional PCR assay followed by Sanger sequencing is time-consuming and laborious. The present study aimed to develop a high-throughput detection-and-genotyping tool for 11 human RNA viruses [Aichi virus
    astrovirus
    enterovirus
    norovirus genogroup I (GI), GII, and GIV
    hepatitis A virus
    hepatitis E virus
    rotavirus
    sapovirus
    and human parechovirus] using a microfluidic device and next-generation sequencer. Microfluidic nested PCR was carried out on a 48.48 Access Array chip, and the amplicons were recovered and used for MiSeq sequencing (Illumina, Tokyo, Japan)
    genotyping was conducted by homology searching and phylogenetic analysis of the obtained sequence reads. The detection limit of the 11 tested viruses ranged from 100 to 103 copies/μL in cDNA sample, corresponding to 101-104 copies/mL-sewage, 105-108 copies/g-human feces, and 102-105 copies/g-digestive tissues of oyster. The developed assay was successfully applied for simultaneous detection and genotyping of RNA viruses to samples of human feces, sewage, and artificially contaminated oysters. Microfluidic nested PCR followed by MiSeq sequencing enables efficient tracking of the fate of multiple RNA viruses in various environments, which is essential for a better understanding of the circulation of human pathogenic RNA viruses in the human population.
  • Nitrogen cycle evaluation (NiCE) chip for simultaneous analysis of multiple N cycle associated genes
    Mamoru Oshiki, Takahiro Segawa, Satoshi Ishii
    Appl. Environ. Microbiol., 84, 8, e02615-17, American Society for Microbiology, 2018年04月01日, [査読有り], [筆頭著者]
    英語, 研究論文(学術雑誌), Various microorganisms play key roles in the nitrogen (N) cycle. Quantitative PCR (qPCR) and PCR amplicon sequencing of N cycle functional genes allow us to analyze the abundance and diversity of microbes responsible for N-transforming reactions in various environmental samples. However, analysis of multiple target genes can be cumbersome and expensive. PCR-independent analysis, such as metagenomics and metatranscriptomics, is useful but expensive, especially when we analyze multiple samples and try to detect N cycle functional genes present at a relatively low abundance. Here, we present the application of microfluidic qPCR chip technology to simultaneously quantify and prepare amplicon sequence libraries for multiple N cycle functional genes as well as taxon-specific 16S rRNA gene markers for many samples. This approach, named the nitrogen cycle evaluation (NiCE) chip, was evaluated by using DNA from pure and artificially mixed bacterial cultures and by comparing the results with those obtained by conventional qPCR and amplicon sequencing methods. Quantitative results obtained by the NiCE chip were comparable to those obtained by conventional qPCR. In addition, the NiCE chip was successfully applied to examine the abundance and diversity of N cycle functional genes in wastewater samples. Although nonspecific amplification was detected on the NiCE chip, this can be overcome by optimizing the primer sequences in the future. As the NiCE chip can provide a high-throughput format to quantify and prepare sequence libraries for multiple N cycle functional genes, this tool should advance our ability to explore N cycling in various samples.
  • エジプトの高校生を対象とした「水」に関する環境教育プログラムの開発と実践 (下水道協会誌論文集)
    大久保 努, 上村 繁樹, 井口 晃徳, 多川 正, 押木 守, 久保田 健吾, 荒木 信夫, 原田 秀樹, Tawfik Ahmed, 市村 禎二郎
    下水道協会誌 = Journal of Japan Sewage Works Association, 55, 665, 76, 82, 日本下水道協会, 2018年03月, [査読有り]
    日本語
  • 菌叢解析再訪 : 変遷と展望 (特集 環境分野に於ける菌叢解析 : 分子生物学が汚泥に光を当てる)
    伊藤 司, 金田一 智則, 押木 守, 岡部 聡
    EICA : journal of EICA : 環境システム計測制御学会誌, 22, 4, 21, 26, 環境システム計測制御学会, 2018年, [査読有り]
    日本語
  • 最初沈殿池+DHSシステムに付加する下水処理水の灌漑利用のための消毒システムの開発
    景政 柊蘭, 長町 晃宏, 井口 晃徳, 久保田 健吾, 高橋 優信, 原田秀樹, 押木守, 荒木信夫, 大久保努, 上村 繁樹, 多川 正
    土木学会論文集G, 74, 7, 333, 340, 公益社団法人 土木学会, 2018年, [査読有り]
    日本語, 研究論文(学術雑誌), 水不足が深刻な乾燥地域開発途上国であるエジプトは,灌漑用水として下水処理水の再利用を推奨しているが,衛生学的な利用基準を満たしていない.最初沈殿池にDown-flow Hanging Sponge (DHS)を組み合わせた下水処理システム(初沈+DHSシステム)において,HRTを0.8~4.8時間の間変化させ,糞便性大腸菌群の低減を確認した結果,全てのHRTにおいてエジプトの灌漑用水基準(5×103 MPN/100mL)を達成することは困難であった.DHSの後段処理法として銅イオンによる殺菌処理を検討した結果,銅イオン濃度 2 mg/L,静置時間を2時間以上確保することでDHS処理水の衛生学的安全性を確保できると考えられた.また,銅イオンDHS処理水によるトマトの生長阻害及び葉への銅イオン蓄積は確認されなかった.
  • Microbial competition among anammox bacteria in nitrite-limited bioreactors
    Lei Zhang, Yuko Narita, Lin Gao, Muhammad Ali, Mamoru Oshiki, Satoshi Ishii, Satoshi Okabe
    Water Res., 125, 249, 258, PERGAMON-ELSEVIER SCIENCE LTD, 2017年11月, [査読有り]
    英語, 研究論文(学術雑誌), Phylogenetically diverse anammox bacteria have been detected in most of anoxic natural and engineered ecosystems and thus regarded as key players in the global nitrogen cycle. However, ecological niche differentiation of anammox bacteria remains unresolved despite its ecological and practical importance. In this study, the microbial competitions for a common substrate (nitrite) among three anammox species (Le. "Candidatus Brocadia sinica", "Candidatus Jettenia caeni" and "Candidatus Kuenenia stuttgartiensis") were systematically investigated in nitrite-limited gel-immobilized column reactors (GICR) and membrane bioreactors (MBRs) under different nitrogen loading rates (NLRs). 16 S rRNA gene-based population dynamics revealed that "Ca. J. caeni" could proliferate only at low NLRs, whereas "Ca. B. sinica" out competed other two species at higher NLRs in both types of reactors. Furthermore, FISH analysis revealed that "Ca. J. caeni" was mainly present as spherical microclusters at the inner part (low NO2- environment), whereas "Ca. B. sinica" was present throughout the gel beads and granules. This spatial distribution supports the outcomes of the competition experiments. However, the successful competition of "Ca. J. caeni" at low NLR could not be explained with the Monod model probably due to inaccuracy of kinetic parameters such as half saturation constant (K-s) for nitrite and a difference in the maintenance rate (m). In addition, the growth of "Ca. K. stuttgartiensis" could not be observed in any experimental conditions, suggesting possible unknown factor(s) is missing. Taken together, NLR was one of factors determining ecological niche differentiation of "Ca. B. sinica" and "Ca. J. caeni". (C) 2017 Elsevier Ltd. All rights reserved.
  • Genetic diversity of marine anaerobic ammonium-oxidizing bacteria as revealed by genomic and proteomic analyses of "Candidatus Scalindua japonica'
    Mamoru Oshiki, Keisuke Mizuto, Zen-ichiro Kimura, Tomonori Kindaichi, Hisashi Satoh, Satoshi Okabe
    Environ. Microbiol. Rep., 9, 5, 550, 561, WILEY, 2017年10月, [査読有り], [筆頭著者]
    英語, 研究論文(学術雑誌), Anaerobic ammonium-oxidizing (anammox) bacteria affiliated with the genus Candidatus Scalindua' are responsible for significant nitrogen loss in oceans, and thus their ecophysiology is of great interest. Here, we enriched a marine anammox bacterium, Ca. S. japonica' from a Hiroshima bay sediment in Japan, and comparative genomic and proteomic analyses of Ca. S. japonica' were conducted. Sequence of the 4.81-Mb genome containing 4019 coding regions of genes (CDSs) composed of 47 contigs was determined. In the proteome, 1762 out of 4019 CDSs in the Ca. S. japonica' genome were detected. Based on the genomic and proteomic data, the core anammox process and carbon fixation of Ca. S. japonica' were further investigated. Additionally, the present study provides the first detailed insights into the genetic background responsible for iron acquisition and menaquinone biosynthesis in anammox bacterial cells. Comparative analysis of the Ca. Scalindua' genomes revealed that the 1502 genes found in the Ca. S. japonica' genome were not present in the Ca. S. profunda' and Ca. S. rubra' genomes, showing a high genomic diversity. This result may reflect a high phylogenetic diversity of the genus Ca. Scalindua'.
  • Draft genome sequence of Thiohalobacter thiocyanaticus strain FOKN1, a neutrophilic halophile capable of thiocyanate degradation
    Mamoru Oshiki, Toshikazu Fukushima, Shuichi Kawano, Junichi Nakagawa
    Genome Announc., 5, 32, e00799-17, American Society for Microbiology, 2017年08月01日, [査読有り], [筆頭著者, 責任著者]
    英語, 研究論文(学術雑誌), A draft genome sequence of a neutrophilic halophile capable of thiocyanate degradation, Thiohalobacter thiocyanaticus FOKN1, was determined using a PacBio RSII sequencer. A 3.23-Mb circular genome sequence was assembled, in which 3,026 gene-coding sequences, 45 tRNAs, and 1 rrn operon were annotated.
  • Improvement of a phosphate ion-selective microsensor using bis(dibromophenylstannyl)methane as a carrier
    Hisashi Satoh, Yuji Miyazaki, Shou Taniuchi, Mamoru Oshiki, Rathnayake M, L. D. Rathnayake, Masahiro Takahashi, Satoshi Okabe
    Anal. Sci., 33, 7, 825, 830, JAPAN SOC ANALYTICAL CHEMISTRY, 2017年07月, [査読有り]
    英語, 研究論文(学術雑誌), An ionophore-doped sensing membrane phosphate (PO4) microsensor based on bis(dibromophenylstannyl)methane (Bis microsensor) is described. The Bis microsensor showed a Nernstian response. The response of the Bis microsensor was log-linear down to a monohydrogen phosphate ion (HPO42-) concentration of 0.5 mu M (corresponding to 1.0 mu M of orthophosphate at pH 7.2), whereas the detection limit of PO4-microsensors based on trialkyl/aryltin chloride was 50 mu M of HPO42-. The Bis microsensor showed excellent selectivity for HPO42- against nitrite, nitrate, chloride, bicarbonate and sulfate, as compared with PO4 microsensors based on trialkyl/aryltin chloride. Dissolved oxygen, which is known to interfere with the response of a previously developed cobalt-based potentiometric solid-state PO4 microsensor, had no effect on the response of the ionophore-doped sensing membrane-type microsensors described herein. Only OH- (i.e., pH) interfered with the ionophore-doped sensing membrane-type microsensors.
  • Maximum specific growth rate of anammox bacteria revisited
    Lei Zhang, Yuko Narita, Lin Gao, Muhammad Ali, Mamoru Oshiki, Satoshi Okabe
    Water Res., 116, 296, 303, PERGAMON-ELSEVIER SCIENCE LTD, 2017年06月, [査読有り]
    英語, 研究論文(学術雑誌), Anammox bacteria have long been considered to be slow-growing bacteria. However, it has recently been reported that they could grow much faster than previously thought when they were cultivated in a membrane bioreactor (MBR) with a step-wise decrease in the solid retention time (SRT). Here, we reevaluated the maximum specific growth rates (mu(max)) of three phylogenetically distant anammox bacterial species (i.e. "Ca. Brocadia sinica", "Ca. Jettenia caeni" and "Ca. Scalindua sp.") by directly measuring 16S rRNA gene copy numbers using newly developed quantitative polymerase chain reaction (qPCR) assays. When free-living planktonic "Ca. B. sinica" and "Ca. J. caeni" cells were immobilized in polyvinyl alcohol (PVA) and sodium alginate (SA) gel beads and cultivated in an up-flow column reactor with high substrate loading rates at 37 degrees C, the mu(max) were determined to be 033 +/- 0.02 d(-1) and 0.18 d(-1) (corresponding doubling time of 2.1 day and 3.9 day) from the exponential increases in 16S rRNA genes copy numbers, respectively. These values were faster than the fastest growth rates reported for these species so far. The cultivation of anammox bacteria in gel beads was achieved less than one month without special cultivation method and selection pressure, and the exponential increase in 16S rRNA gene numbers was directly measured by qPCR with high reproducibility; therefore, the resulting mu(max) values were considered accurate. Taken together, the fast growth is, therefore, considered to be an intrinsic kinetic property of anammox bacteria. (C) 2017 Elsevier Ltd. All rights reserved.
  • Removal of human pathogenic viruses in a down-flow hanging sponge (DHS) reactor treating municipal wastewater and health risks associated with utilization of the effluent for agricultural irrigation
    Naohiro Kobayashi, Mamoru Oshiki, Toshihiro Ito, Takahiro Segawa, Masashi Hatamoto, Tsuyoshi Kato, Takashi Yamaguchi, Kengo Kubota, Masanobu Takahashi, Akinori Iguchi, Tadashi Tagawa, Tsutomu Okubo, Shigeki Uemura, Hideki Harada, Toshiki Motoyama, Nobuo Araki, Daisuke Sano
    Water Res., 110, 389, 398, PERGAMON-ELSEVIER SCIENCE LTD, 2017年03月, [査読有り], [責任著者]
    英語, 研究論文(学術雑誌), A down-flow hanging sponge (DHS) reactor has been developed as a cost-effective wastewater treatment system that is adaptable to local conditions in low-income countries. A pilot-scale DHS reactor previously demonstrated stable reduction efficiencies for chemical oxygen demand (COD) and ammonium nitrogen over a year at ambient temperature, but the pathogen reduction efficiency of the DHS reactor has yet to be investigated. In the present study, the reduction efficiency of a pilot-scale DHS reactor fed with municipal wastewater was investigated for 10 types of human pathogenic viruses (norovirus GI, GII and GIV, aichivirus, astrovirus, enterovirus, hepatitis A and E viruses, rotavirus, and sapovirus). DHS influent and effluent were collected weekly or biweekly for 337 days, and concentrations of viral genomes were determined by microfluidic quantitative PCR. Aichivirus, norovirus GI and GII, enterovirus, and sapovirus were frequently detected in DHS influent, and the logio reduction (LR) of these viruses ranged from 1.5 to 3.7. The LR values for aichivirus and norovirus Gil were also calculated using a Bayesian estimation model, and the average LR (standard deviation) values for aichivirus and norovirus GII were estimated to be 1.4 (1.5) and 1.8 (2.5), respectively. Quantitative microbial risk assessment was conducted to calculate a threshold reduction level for norovirus Gil that would be required for the use of DHS effluent for agricultural irrigation, and it was found that LRs of 2.6 and 3.7 for norovirus GII in the DHS effluent were required in order to not exceed the tolerable burden of disease at 10(-4) and 10(-6) disability-adjusted life years loss per person per year, respectively, for 95% of the exposed population during wastewater reuse for irrigation. (C) 2016 Elsevier Ltd. All rights reserved.
  • Eukaryotic community in UASB reactor treating domestic sewage based on 18S rRNA gene sequencing
    Y. Hirakata, M. Hatamoto, M. Oshiki, N. Araki, T. Yamaguchi
    Lecture Notes in Civil Engineering, 4, 218, 224, 2017年, [査読有り]
    英語, 論文集(書籍)内論文, © Springer International Publishing AG 2017. Microbial eukaryotes play important roles in sewage treatment systems. In an up-flow anaerobic sludge blanket (UASB) reactor fed with domestic sewage, anaerobic bacteria, archaea, and microbial eukaryotes (protist and fungi) coexist. To date, bacterial and archaeal communities in the UASB reactor have been widely studied. However, little is known about the eukaryotic community structure and function in most of anaerobic treatment systems. In this study, we analyzed eukaryotic community in the UASB reactor treating domestic sewage over 2 years operational period based on 18S rRNA gene sequences. In addition, multivariate statistics were applied to elucidate the correlation between eukaryotic community and operational conditions of the UASB reactor. The dominant protist groups observed were from phylum Ciliophora, Apicomplexa, Perkisea and Amoebozoa in the UASB reactor. Followings were flagellate protist such as Cercozoa, Sulcozoa, Bicosoecida, Choanomonada, Dinoflagellata and Metamonada. The result showed protist phylum and water temperature were not correlated. Only phylum Sulcozoa positively correlated with COD and suspended solid (SS), whereas other protist showed low correlation. The dominant fungi groups were LKM11, LKM15 and phylum Ascomycota. Uncultured LKM15 correlated with sulfide whereas phylum Discicristoidea and Chytridiomycota showed negative correlation. Phylum Ascomycota seemed most abundant when sulfide was low, but their association with environmental variables remained unclear. These results suggested that some protist and fungi groups could be used as indicator of environmental parameters in the UASB reactor.
  • Ecology and physiology of anaerobic ammonium oxidizing bacteria
    Mamoru Oshiki, Hisashi Satoh, Satoshi Okabe
    Environ. Microbiol., 18, 9, 2784, 2796, WILEY-BLACKWELL, 2016年09月, [査読有り], [筆頭著者]
    英語, Anaerobic ammonium oxidation (anammox) is a microbial process in which NH4+ is oxidized to N-2 gas with NO2- as an electron acceptor. The anammox process is mediated by bacterial members affiliated with the phylum Planctomycetes, which are ubiquitously detected from anoxic natural and man-made ecosystems and a key player in the global nitrogen cycle. In the past two decades, phylogenetically different anammox bacteria have been recognized in natural and synthetic ecosystems (i.e. 'Candidatus Kuenenia', 'Candidatus Brocadia', 'Candidatus Jettenia', 'Candidatus Anammoxoglobus' and 'Candidatus Scalindua' genera), and the geographic distributions of these anammox bacteria indicate that they have genus-specific or species-specific habitats. Recently, we revealed the physiological characteristics of 'Ca. Jettenia' in addition to 'Ca. Kuenenia', 'Ca. Brocadia' and 'Ca. Scalindua', and, as a result, it is possible to compare the physiological characteristics of the anammox bacteria and discuss their niche partitioning. Therefore, we summarize the current knowledge of anammox bacterial ecology and physiology in this review to assess the potential ecological niche partitioning of anammox bacteria in natural and synthetic ecosystems.
  • Hydroxylamine-dependent anaerobic ammonium oxidation (anammox) by "Candidatus Brocadia sinica"
    Mamoru Oshiki, Muhammad Ali, Kaori Shinyako-Hata, Hisashi Satoh, Satoshi Okabe
    Environ. Microbiol., 18, 9, 3133, 3143, WILEY-BLACKWELL, 2016年09月, [査読有り], [筆頭著者]
    英語, 研究論文(学術雑誌), Although metabolic pathways and associated enzymes of anaerobic ammonium oxidation (anammox) of 'Ca. Kuenenia stuttgartiensis' have been studied, those of other anammox bacteria are still poorly understood. NO2- reduction to NO is considered to be the first step in the anammox metabolism of 'Ca. K. stuttgartiensis', however, 'Ca. Brocadia' lacks the genes that encode canonical NO-forming nitrite reductases (NirS or NirK) in its genome, which is different from 'Ca. K. stuttgartiensis'. Here, we studied the anammox metabolism of 'Ca. Brocadia sinica'. N-15-tracer experiments demonstrated that 'Ca. B. sinica' cells could reduce NO2- to NH2OH, instead of NO, with as yet unidentified nitrite reductase(s). Furthermore, N2H4 synthesis, downstream reaction of NO2- reduction, was investigated using a purified 'Ca. B. sinica' hydrazine synthase (Hzs) and intact cells. Both the 'Ca. B. sinica' Hzs and cells utilized NH2OH and NH1+, but not NO and NH1+, for N2H4 synthesis and further oxidized N2H4 to N-2 gas. Taken together, the metabolic pathway of 'Ca. B. sinica' is NH2OH-dependent and different from the one of 'Ca. K. stuttgartiensis', indicating metabolic diversity of anammox bacteria.
  • Effects of predation by protists on prokaryotic community function, structure, and diversity in anaerobic granular sludge
    Yuga Hirakata, Mamoru Oshiki, Kyohei Kuroda, Masashi Hatamoto, Kengo Kubota, Takashi Yamaguchi, Hideki Harada, Nobuo Araki
    Microbes Environ., 31, 3, 279, 287, JAPANESE SOC MICROBIAL ECOLOGY, DEPT BIORESOURCE SCIENCE, 2016年09月, [査読有り], [責任著者]
    英語, 研究論文(学術雑誌), Predation by protists is top-down pressure that regulates prokaryotic abundance, community function, structure, and diversity in natural and artificial ecosystems. Although the effects of predation by protists have been studied in aerobic ecosystems, they are poorly understood in anoxic environments. We herein studied the influence of predation by Metopus and Caenomorpha ciliates-ciliates frequently found in anoxic ecosystems-on prokaryotic community function, structure, and diversity. Metopus and Caenomorpha ciliates were cocultivated with prokaryotic assemblages (i.e., anaerobic granular sludge) in an up-flow anaerobic sludge blanket (UASB) reactor for 171 d. Predation by these ciliates increased the methanogenic activities of granular sludge, which constituted 155% of those found in a UASB reactor without the ciliates (i.e., control reactor). Sequencing of 16S rRNA gene amplicons using Illumina MiSeq revealed that the prokaryotic community in the UASB reactor with the ciliates was more diverse than that in the control reactor; 2,885-3,190 and 2,387-2,426 operational taxonomic units (>97% sequence similarities), respectively. The effects of predation by protists in anaerobic engineered systems have mostly been overlooked, and our results show that the influence of predation by protists needs to be examined and considered in the future for a better understanding of prokaryotic community structure and function.
  • 一次沈殿+DHSシステムによる衛生指標微生物の処理性能評価
    長町 晃宏, 井口 晃徳, 瀬戸 雄太, 久保田 健吾, 押木 守, 荒木 信夫, 大久保 努, 上村 繁樹, 高橋 優信, 原田 秀樹, 多川 正
    土木学会論文集G(環境), 72, 7, III_187, III_195, 公益社団法人 土木学会, 2016年, [査読有り]
    日本語, 活性汚泥法の代替下水処理技術として一次沈殿処理の後段処理にDown-flow Hanging Sponge (DHS)を組み合わせた低コスト・低エネルギー消費型下水処理システム(一次沈殿+DHSシステム)を考案し,1,000日以上に及ぶ連続通水実験を実施した.その結果,衛生指標微生物としての大腸菌群及び糞便性大腸菌群の対数除去率は,DHSのHRTが0.8時間の時にはそれぞれ1.45 log,1.88 logと低かった.この時のBOD,SSの除去率は80%以下で,硝化反応も見られなかった.しかしDHSのHRTを3.2時間とすることで,大腸菌群及び糞便性大腸菌群の対数除去率は,2.73 log,3.15 logを示し,BOD,SSの除去率もそれぞれ90%以上を達成した.同一下水を処理している活性汚泥法による大腸菌群及び糞便性大腸菌群の対数除去率がそれぞれ2.55 log,2.19 logであり,BOD,SS除去率もほぼ同等だった.以上より,一次沈殿+DHSシステムは,HRTを管理することで活性汚泥法とほぼ同等の衛生指標微生物除去性能を得ることができる.
  • 下水再生処理におけるヒト腸管系ウイルスの目標除去効率の算定法
    伊藤寿宏, 押木守, 小林直央, 加藤毅, 瀬川高弘, 幡本将史, 山口隆司, 原田秀樹, 北島正章, 岡部聡, 佐野大輔
    土木学会論文集 G(環境)(Web), 72, 7, III_305‐III_313(J‐STAGE), III_313, 公益社団法人 土木学会, 2016年, [査読有り]
    日本語, 本研究では、流入下水中のヒト腸管系ウイルス濃度分布、及びWHOが推奨する許容年間疾病負荷(10<sup>-6</sup> disability-adjusted life years per person per year)に基づいて、定量的微生物リスク評価(quantitative microbial risk assessment: QMRA)の手法を用いて下水再生水の利用用途ごとにウイルス除去効率の目標値を算出する手法を構築・提案する。代表的なヒト腸管系ウイルスとしてノロウイルスに着目し、流入下水中のノロウイルス濃度モニタリングデータを使用することで、6種類の下水再生水利用シナリオにおけるノロウイルスの下水再生水中許容濃度及び除去効率の目標値を試算した。本研究で提案した方法により算出したヒト腸管系ウイルス除去効率目標値を使用する際には、下水再生システム稼動後においても未処理下水中のヒト腸管系ウイルス濃度をモニタリングすることが求められる。また、用量反応モデル情報の更新と、下水再生利用が行われる地域の状況に基づいた曝露シナリオとパラメータの更新についても継続して取り組むことが重要である。
  • Draft genome sequence of an anaerobic ammonium-oxidizing bacterium, "Candidatus Brocadia sinica"
    Mamoru Oshiki, Kaori Shinyako-Hata, Hisashi Satoh, Satoshi Okabe
    Genome Announc., 3, 2, e00267-15, American Society for Microbiology, 2016年, [査読有り], [筆頭著者]
    英語, 研究論文(学術雑誌), A draft genome sequence of an anaerobic ammonium-oxidizing (anammox) bacterium, "Candidatus Brocadia sinica," was determined by pyrosequencing and by screening a fosmid library. A 4.07-Mb genome sequence comprising 3 contigs was assembled, in which 3,912 gene-coding regions, 47 tRNAs, and a single rrn operon were annotated.
  • High-cell-density cultivation of Nitrosomonas europaea in a membrane bioreactor for performing protein purification and characterization studies
    Mamoru Oshiki, Ryota Takagi, Masashi Hatamoto, Takashi Yamaguchi, Nobuo Araki
    J. Gen. Appl. Microbiol., 62, 6, 330, 333, MICROBIOL RES FOUNDATION, 2016年, [査読有り], [筆頭著者, 責任著者]
    英語, 研究論文(学術雑誌)
  • Effects of dissolved oxygen and pH on nitrous oxide production rates in autotrophic partial nitrification granules
    Rathnayake M, L, D. Rathnayake, Mamoru Oshiki, Satoshi Ishii, Takahiro Segawa, Hisashi Satoh, Satoshi Okabe
    Bioresour. Technol., 197, 15, 22, ELSEVIER SCI LTD, 2015年12月, [査読有り]
    英語, 研究論文(学術雑誌), The effects of dissolved oxygen (DO) and pH on nitrous oxide (N2O) production rates and pathways in autotrophic partial nitrification (PN) granules were investigated at the granular level. N2O was primarily produced by betaproteobacterial ammonia-oxidizing bacteria, mainly Nitrosomonas europaea, in the oxic surface layer (<200 mu m) of the autotrophic PN granules. N2O production increased with increasing bulk DO concentration owing to activation of the ammonia (i.e., hydroxylamine) oxidation in this layer. The highest N2O emissions were observed at pH 7.5, although the ammonia oxidation rate was unchanged between pH 6.5 and 8.5. Overall, the results of this study suggest that in situ analyses of PN granules are essential to gaining insight into N2O emission mechanisms in a granule. (C) 2015 Elsevier Ltd. All rights reserved.
  • Identification and detection of prokaryotic symbionts in the ciliate Metopus from anaerobic granular sludge
    Yuga Hirakata, Mamoru Oshiki, Kyohei Kuroda, Masashi Hatamoto, Kengo Kubota, Takashi Yamaguchi, Hideki Harada, Nobuo Araki
    Microbes Environ., 30, 4, 335, 338, JAPANESE SOC MICROBIAL ECOLOGY, DEPT BIORESOURCE SCIENCE, 2015年12月, [査読有り], [責任著者]
    英語, 研究論文(学術雑誌), The aim of the present study was to investigate the prokaryotic community structure of the anaerobic ciliate, Metopus sp. using rRNA sequencing, fluorescence in situ hybridization ( FISH), and transmission electron microscopy (TEM). Metopus sp. was physically separated from anaerobic granular sludge in a domestic wastewater treatment plant and anoxically cultivated for 7 d. 16S rRNA gene sequences from the prokaryotes Methanoregula boonei and Clostridium aminobutyricum were abundantly detected in Metopus ciliates. The FISH analysis using the oligonucleotide probes Mg1200b and Cla568 demonstrated that these prokaryotes were localized within Metopus cells. These results identify M. boonei- and C. aminobutyricum-like prokaryotes as novel endosymbionts of Metopus ciliates.
  • Rapid and successful start-up of anammox process by immobilizing the minimal quantity of biomass in PVA-SA gel beads
    Muhammad Ali, Mamoru Oshiki, Lashitha Rathnayake, Satoshi Ishii, Hisashi Satoh, Satoshi Okabe
    Water Res., 79, 147, 157, PERGAMON-ELSEVIER SCIENCE LTD, 2015年08月, [査読有り]
    英語, 研究論文(学術雑誌), Rapid start-up of anaerobic ammonium oxidation (anammox) process in up-flow column reactors was successfully achieved by immobilizing minimal quantity of biomass in polyvinyl alcohol (PVA)-sodium alginate (SA) gel beads. The changes in the reactor performance (i.e., nitrogen removal rate; NRR) were monitored with time. The results demonstrate that the reactor containing the immobilized biomass concentration of 0.33 g-VSS L-1 achieved NRR of 10.8 kg-N m(-3) d(-1) after 35-day operation, whereas the reactor containing the granular biomass of 2.5 g-VSS L-1 could achieve only NRR of 3.5 kg-N m(-3) d(-1). This indicates that the gel immobilization method requires much lower seeding biomass for start-up of anammox reactor. To explain the better performance of the immobilized biomass, the biological and physicochemical properties of the immobilized biomass were characterized and compared with the naturally aggregated granular biomass. Effective diffusion coefficient (De) in the immobilized biomass was directly determined by microelectrodes and found to be three times higher than one in the granular biomass. High anammox activity (i.e., NH4+ and NO2- consumption rates) was evenly detected throughout the gel beads by microelectrodes due to faster and deeper substrate transport. In contrast, anammox activity was localized in the outer layers of the granular biomass, indicating that the inner biomass could not contribute to the nitrogen removal. This difference was in good agreement with the spatial distribution of microbes analysed by fluorescence in situ hybridization (FISH). Based on these results, PVA-SA gel immobilization is an efficient strategy to initiate anammox reactors with minimal quantity of anammox biomass. (C) 2015 Elsevier Ltd. All rights reserved.
  • Physiological characterization of anaerobic ammonium oxidizing bacterium 'Candidatus Jettenia caeni'.
    Ali M, Oshiki M, Awata T, Isobe K, Kimura Z, Yoshikawa H, Hira D, Kindaichi T, Satoh H, Fujii T, Okabe S
    Environ. Microbiol., 17, 6, 2172, 2189, WILEY-BLACKWELL, 2015年06月, [査読有り]
    英語, 研究論文(学術雑誌), To date, six candidate genera of anaerobic ammonium-oxidizing (anammox) bacteria have been identified, and numerous studies have been conducted to understand their ecophysiology. In this study, we examined the physiological characteristics of an anammox bacterium in the genus CandidatusJettenia'. Planctomycete KSU-1 was found to be a mesophilic (20-42.5 degrees C) and neutrophilic (pH 6.5-8.5) bacterium with a maximum growth rate of 0.0020h(-1). Planctomycete KSU-1 cells showed typical physiological and structural features of anammox bacteria; i.e. N-29(2) gas production by coupling of (NH4+)-N-15 and (NO2-)-N-14, accumulation of hydrazine with the consumption of hydroxylamine and the presence of anammoxosome. In addition, the cells were capable of respiratory ammonification with oxidation of acetate. Notably, the cells contained menaquinone-7 as a dominant respiratory quinone. Proteomic analysis was performed to examine underlying core metabolisms, and high expressions of hydrazine synthase, hydrazine dehydrogenase, hydroxylamine dehydrogenase, nitrite/nitrate oxidoreductase and carbon monoxide dehydrogenase/acetyl-CoA synthase were detected. These proteins require iron or copper as a metal cofactor, and both were dominant in planctomycete KSU-1 cells. On the basis of these experimental results, we proposed the name Ca. Jettenia caeni' sp. nov. for the bacterial clade of the planctomycete KSU-1.
  • 嫌気性アンモニア酸化(anammox)細菌の廃水処理への適用 (特集総説 極限生物たちが切り拓く未来の環境バイオテクノロジー)
    押木 守, 佐藤 久, 岡部 聡
    Journal of environmental biotechnology, 14, 1, 21, 29, 環境バイオテクノロジー学会, 2014年10月, [査読有り], [筆頭著者]
    日本語
  • Simple, rapid and effective preservation and reactivation of anaerobic ammonium oxidizing bacterium "Candidatus Brocadia sinica"
    Muhammad Ali, Mamoru Oshiki, Satoshi Okabe
    Water Res., 57, 215, 222, PERGAMON-ELSEVIER SCIENCE LTD, 2014年06月, [査読有り]
    英語, 研究論文(学術雑誌), It is still the biggest challenge to secure enough seeding biomass for rapid start-up of fullscale (anaerobic ammonium oxidation) anammox processes due to slow growth. Preservation of active anammox biomass could be one of the solutions. In this study, biomass of anammox bacterium, "Candidatus Brocadia sinica", immersed in various nutrient media were preserved at 80 C, 4 C and room temperature. After 45, 90 and 150 days of preservation, specific anammox activity (SAA) of the preserved anammox biomass was determined by measuring (29)N2 production rate and transcription levels of hzsA gene encoding hydrazine synthase alpha subunit. Storage in nutrient medium containing 3 mM of molybdate at room temperature with periodical (every 45 days) supply of Nlit and NO2- was proved to be the most effective storage technique for "Ca. Brocadia sinica" biomass. Using this preservation condition, 96, 92 and 65% of the initial SAA was sustained after 45, 90 and 150 days of storage, respectively. Transcription levels of hzsA gene in biomass correlated with the SAA (R-2 = 0.83), indicating it can be used as a genetic marker to evaluate the anammox activity of preserved biomass. Furthermore, the 90-day-stored biomass was successfully reactivated by immobilizing in polyvinyl alcohol (6%, w/v) and sodium alginate (2%, w/v) gel and then inoculated to up-flow column reactors. Total nitrogen removal rates rapidly increased to 7 kg-N m(-3) d(-1) within 35 days of operation. Based on these results, the room temperature preservation with molybdate addition is simple, cost-effective and feasible at a practical scale, which will accelerate the practical use of anammox process for wastewater treatment. (C) 2014 Elsevier Ltd. All rights reserved.
  • Source identification of nitrous oxide on autotrophic partial nitrification in a granular sludge reactor
    R. M. L, D. Rathnayake, Y. Song, A. Tumendelger, M. Oshiki, S. Ishii, H. Satoh, S. Toyoda, N. Yoshida, S. Okabe
    Water Res., 47, 19, 7078, 7086, PERGAMON-ELSEVIER SCIENCE LTD, 2013年12月, [査読有り]
    英語, 研究論文(学術雑誌), Emission of nitrous oxide (N2O) during biological wastewater treatment is of growing concern since N2O is a major stratospheric ozone-depleting substance and an important greenhouse gas. The emission of N2O from a lab-scale granular sequencing batch reactor (SBR) for partial nitrification (PN) treating synthetic wastewater without organic carbon was therefore determined in this study, because PN process is known to produce more N2O than conventional nitrification processes. The average N2O emission rate from the SBR was 0.32 +/- 0.17 mgN L-1 h(-1), corresponding to the average emission of N2O of 0.8 +/- 0.4% of the incoming nitrogen load (1.5 +/- 0.8% of the converted NI-14"). Analysis of dynamic concentration profiles during one cycle of the SBR operation demonstrated that N2O concentration in off-gas was the highest just after starting aeration whereas N2O concentration in effluent was gradually increased in the initial 40 min of the aeration period and was decreased thereafter. Isotopomer analysis was conducted to identify the main N2O production pathway in the reactor during one cycle. The hydroxylamine (NH2OH) oxidation pathway accounted for 65% of the total N2O production in the initial phase during one cycle, whereas contribution of the NO reduction pathway to N2O production was comparable with that of the NH2OH oxidation pathway in the latter phase. In addition, spatial distributions of bacteria and their activities in single microbial granules taken from the reactor were determined with microsensors and by in situ hybridization. Partial nitrification occurred mainly in the oxic surface layer of the granules and ammonia-oxidizing bacteria were abundant in this layer. N2O production was also found mainly in the oxic surface layer. Based on these results, although N2O was produced mainly via NH2OH oxidation pathway in the autotrophic partial nitrification reactor, N2O production mechanisms were complex and could involve multiple N2O production pathways. (C) 2013 Elsevier Ltd. All rights reserved.
  • Cultivation of planktonic anaerobic ammonium oxidation (anammox) bacteria using membrane bioreactor
    Oshiki, Mamoru, Awata, Takanori, Kindaichi, Tomonori, Satoh, Hisashi, Okabe, Satoshi
    Microbes Environ., 28, 4, 436, 443, JAPANESE SOC MICROBIAL ECOLOGY, DEPT BIORESOURCE SCIENCE, 2013年12月, [査読有り], [筆頭著者]
    英語, 研究論文(学術雑誌), Enrichment cultures of anaerobic ammonium oxidation (anammox) bacteria as planktonic cell suspensions are essential for studying their ecophysiology and biochemistry, while their cultivation is still laborious. The present study aimed to cultivate two phylogenetically distinct anammox bacteria, "Candidatus Brocadia sinica" and "Ca. Scalindua sp." in the form of planktonic cells using membrane bioreactors (MBRs). The MBRs were continuously operated for more than 250 d with nitrogen loading rates of 0.48-1.02 and 0.004-0.09 kgN m(-3) d(-1) for "Ca. Brocadia sinica" and "Ca. Scalindua sp.", respectively. Planktonic anammox bacterial cells were successfully enriched (>90%) in the MBRs, which was confirmed by fluorescence in-situ hybridization and 16S rRNA gene sequencing analysis. The decay rate and half-saturation constant for NO2- of "Ca. Brocadia sinica" were determined to be 0.0029-0.0081 d(-1) and 0.47 mgN L-1, respectively, using enriched planktonic cells. The present study demonstrated that MBR enables the culture of planktonic anammox bacterial cells, which are suitable for studying their ecophysiology and biochemistry.
  • Nitrate-dependent ferrous iron oxidation by anaerobic ammonium oxidation (anammox) bacteria
    M. Oshiki, S. Ishii, K. Yoshida, N. Fujii, M. Ishiguro, Hisashi Satoh, S. Okabe
    Appl. Environ. Microbiol., 79, 13, 4087, 4093, 2013年07月, [査読有り], [筆頭著者]
    英語, 研究論文(学術雑誌), We examined nitrate-dependent Fe2+ oxidation mediated by anaerobic ammonium oxidation (anammox) bacteria. Enrichment cultures of "Candidatus Brocadia sinica" anaerobically oxidized Fe2+ and reduced NO3- to nitrogen gasat rates of 3.7±0.2 and 1.3±0.1 (mean±standard deviation [SD]) nmol mg protein-1 min-1, respectively (37°C and pH 7.3). This nitrate reduction rate is an order of magnitude lower than the anammox activity of "Ca. Brocadia sinica" (10 to 75 nmol NH4+ mg protein-1 min-1). A 15N tracer experiment demonstrated that coupling of nitrate-dependent Fe2+ oxidation and the anammox reaction was responsible for producing nitrogen gas from NO3- by "Ca. Brocadia sinica." The activities of nitrate-dependent Fe2+ oxidation were dependent on temperature and pH, and thehighest activities were seen at temperatures of 30 to 45°C and pHs ranging from 5.9 to 9.8. The mean half-saturation constant for NO3 -±SD of "Ca. Brocadia sinica" was determined to be 51±21 μM. Nitrate-dependent Fe2+ oxidation was further demonstrated by another anammox bacterium, "Candidatus Scalindua sp.," whose rates of Fe2+ oxidation and NO3- reduction were 4.7±0.59 and 1.45±0.05 nmol mg protein-1 min-1, respectively (20°C and pH 7.3). Co-occurrence of nitrate-dependent Fe2+ oxidation and the anammox reaction decreased the molar ratios of consumed NO2 - to consumed NH4 + (δNO2-/δNH4 +) and produced NO3- to consumed NH4 + (δNO3 -/δNH4 +). These reactions are preferable to the application of anammox processes for wastewater treatment. © 2013, American Society for Microbiology.
  • Physiological characterization of an anaerobic ammonium-oxidizing bacterium belonging to the "Candidatus Scalindua" group
    Takanori Awata, Mamoru Oshiki, Tomonori Kindaichi, Noriatsu Ozaki, Akiyoshi Ohashi, Satoshi Okabe
    Appl. Environ. Microbiol., 79, 13, 4145, 4148, AMER SOC MICROBIOLOGY, 2013年07月, [査読有り]
    英語, 研究論文(学術雑誌), The phylogenetic affiliation and physiological characteristics (e.g., K-s and maximum specific growth rate [mu(max)]) of an anaerobic ammonium oxidation (anammox) bacterium, "Candidatus Scalindua sp.," enriched from the marine sediment of Hiroshima Bay, Japan, were investigated. "Candidatus Scalindua sp." exhibits higher affinity for nitrite and a lower growth rate and yield than the known anammox species.
  • Microbial community composition of polyhydroxyalkanoate-accumulating organisms in full-scale wastewater treatment plants operated in fully aerobic mode
    Mamoru Oshiki, Motoharu Onuki, Hiroyasu Satoh, Takashi Mino
    Microbes Environ., 28, 1, 96, 104, JAPANESE SOC MICROBIAL ECOLOGY, DEPT BIORESOURCE SCIENCE, 2013年03月, [査読有り], [筆頭著者, 責任著者]
    英語, 研究論文(学術雑誌), The removal of biodegradable organic matter is one of the most important objectives in biological wastewater treatments. Polyhydroxyalkanoate (PHA)-accumulating organisms (PHAAOs) significantly contribute to the removal of biodegradable organic matter; however, their microbial community composition is mostly unknown. In the present study, the microbial community composition of PHAAOs was investigated at 8 full-scale wastewater treatment plants (WWTPs), operated in fully aerobic mode, by fluorescence in situ hybridization (FISH) analysis and post-FISH Nile blue A (NBA) staining techniques. Our results demonstrated that 1) PHAAOs were in the range of 11-18% in the total number of cells, and 2) the microbial community composition of PHAAOs was similar at the bacterial domain/phylum/class/order level among the 8 full-scale WWTPs, and dominant PHAAOs were members of the class Alphaproteobacteria and Betaproteobacteria. The microbial community composition of alpha- and beta-proteobacterial PHAAOs was examined by 16S rRNA gene clone library analysis and further by applying a set of newly designed oligonucleotide probes targeting 16S rRNA gene sequences of alpha- or beta-proteobacterial PHAAOs. The results demonstrated that the microbial community composition of PHAAOs differed in the class Alphaproteobacteria and Betaproteobacteria, which possibly resulted in a different PHA accumulation capacity among the WWTPs (8.5-38.2 mg-C g-VSS-1 h(-1)). The present study extended the knowledge of the microbial diversity of PHAAOs in full-scale WWTPs operated in fully aerobic mode.
  • Revealing microbial community structures in large- and small-scale activated sludge systems by barcoded pyrosequencing of 16S rRNA gene
    Purnika Damindi Ranasinghe, Hiroyasu Satoh, Mamoru Oshiki, Kenshiro Oshima, Wataru Suda, Masahira Hattori, Takashi Mino
    Water Sci. Technol., 66, 10, 2155, 2161, IWA PUBLISHING, 2012年, [査読有り]
    英語, 研究論文(学術雑誌), The diversity of bacterial groups in activated sludge from large- and small-scale wastewater treatment plants was explored by barcoded pyrosequencing of 16S rRNA gene. Activated sludge samples (three small and 17 large scale) were collected from 12 wastewater treatment plants to clarify precise taxonomy and relative abundances. DNA was extracted, and amplified by 4 base barcoded 27f/519r primer set. The 454 Titanium (Roche) pyrosequences were obtained and analyses performed by Quantitative Insight Into Microbial Ecology (QIIME) with around 100,000 reads. Sequence statistics were computed, while constructing a phylogenetic tree and heatmap. Computed results explained total microbial diversity at phylum and class level and resolution was further extended to Operational Taxonomic Unit (OTU) based taxonomic assignment for investigating community distribution based on individual sample. Composition of sequence reads were compared and microbial community structures for large- and small-scale treatment plants were identified as major phyla (Proteobacteria and Bacteroidetes) and classes (Betaproteobacteria and Bacteroidetes). Also, family level breakdowns were explained and differences in family Nitrospiraceae and phylum Actinobacteria found at their species level were also illustrated. Thus, the pyrosequencing method provides high resolution insight into microbial community structures in activated sludge that might have been unnoticed with conventional approaches.
  • N2O emission from a partial nitrification-anammox process and identification of a key biological process of N2O emission from anammox granules
    Satoshi Okabe, Mamoru Oshiki, Yoshitaka Takahashi, Hisashi Satoh
    Water Res., 45, 19, 6461, 6470, PERGAMON-ELSEVIER SCIENCE LTD, 2011年12月, [査読有り]
    英語, 研究論文(学術雑誌), Emission of nitrous oxide (N2O) during biological wastewater treatment is of growing concern. The emission of N2O from a lab-scale two-reactor partial nitrification (PN)-anammox reactor was therefore determined in this study. The average emission of N2O from the PN and anammox process was 4.0 +/- 1.5% (9.6 +/- 3.2% of the removed nitrogen) and 0.1 +/- 0.07% (0.14 +/- 0.09% of the removed nitrogen) of the incoming nitrogen load, respectively. Thus, a larger part (97.5%) of N2O was emitted from the PN reactor. The total amount of N2O emission from the PN reactor was correlated to nitrite (NO2-) concentration in the PN effluent rather than DO concentration. In addition, further studies were performed to indentify a key biological process that is responsible for N2O emission from the anammox process (i.e., granules). In order to characterize N2O emission from the anammox granules, the in situ N2O production rate was determined by using microelectrodes for the first time, which was related to the spatial organization of microbial community of the granule as determined by fluorescence in situ hybridization (FISH). Microelectrode measurement revealed that the active N2O production zone was located in the inner part of the anammox granule, whereas the active ammonium consumption zone was located above the N2O production zone. Anammox bacteria were present throughout the granule, whereas ammonium-oxidizing bacteria (AOB) were restricted to only the granule surface. In addition, addition of penicillin G that inhibits most of the heterotrophic denitrifiers and AOB completely inhibited N2O production in batch experiments. Based on these results obtained, denitrification by putative heterotrophic denitrifiers present in the inner part of the granule was considered the most probable cause of N2O emission from the anammox reactor (i.e., granules). (C) 2011 Elsevier Ltd. All rights reserved.
  • Development of long-term stable partial nitrification and subsequent anammox process
    Satoshi Okabe, Mamoru Oshiki, Yoshitaka Takahashi, Hisashi Satoh
    Bioresour. Technol., 102, 13, 6801, 6807, 2011年07月, [査読有り]
    英語, 研究論文(学術雑誌), The partial nitrification reactor was successfully started up and operated stably for more than 250days with a maximum nitrite production rate of 1.12kg-Nm-3day-1. The important factors for successful partial nitrification were high ammonium loading rate (>
    1.0kg-Nm-3day-1) and relatively high pH (ca. 8.0), giving high free ammonia concentrations (>
    10mg NH3-NL-1). In addition, the air flow rate must be controlled at the ratio of air flow rate to ammonium loading rate below 0.1 (mair3day-1)/(kg-Nm-3day-1). After the establishment of stable partial nitrification, the effluent NO2--N/NH4+-N ratio and effluent NO3--N concentration were 1.20±0.33 and 1.2±1.0mg-NL-1, respectively, which was then fed into an granular-sludge anammox reactor. Consistent nitrogen removal was achieved for more than 250days with a maximum nitrogen removal rate of 15.0kg-TNm-3day-1. © 2011 Elsevier Ltd.
  • Physiological characteristics of the anaerobic ammonium-oxidizing bacterium "Candidatus Brocadia sinica"
    Mamoru Oshiki, Masaki Shimokawa, Naoki Fujii, Hisashi Satoh, Satoshi Okabe
    Microbiology, 157, 6, 1706, 1713, SOC GENERAL MICROBIOLOGY, 2011年06月, [査読有り], [筆頭著者]
    英語, 研究論文(学術雑誌), The present study investigated the phylogenetic affiliation and physiological characteristics of bacteria responsible for anaerobic ammonium oxidization (anammox); these bacteria were enriched in an anammox reactor with a nitrogen removal rate of 26.0 kg N m(-3) day(-1). The anammox bacteria were identified as representing 'Candidatus Brocadia sinica' on the basis of phylogenetic analysis of rRNA operon sequences. Physiological characteristics examined were growth rate, kinetics of ammonium oxidation and nitrite reduction, temperature, pH and inhibition of anammox. The maximum specific growth rate (mu(max).) was 0.0041 h(-1), corresponding to a doubling time of 7 days. The half-saturation constants (K(s)) for ammonium and nitrite of 'Ca. B. sinica' were 28 +/- 4 and 86 +/- 4 mu M, respectively, higher than those of 'Candidatus Brocadia anammoxidans' and 'Candidatus Kuenenia stuttgartiensis'. The temperature and pH ranges of anammox activity were 25-45 degrees C and pH 6.5-8.8, respectively. Anammox activity was inhibited in the presence of nitrite (50% inhibition at 16 mM), ethanol (91% at 1 mM) and methanol (86% at 1 mM). Anammox activities were 80 and 70% of baseline in the presence of 20 mM phosphorus and 3% salinity, respectively. The yield of biomass and dissolved organic carbon production in the culture supernatant were 0.062 and 0.005 mol C (mol NH(4)(+))(-1), respectively. This study compared physiological differences between three anammox bacterial enrichment cultures to provide a better understanding of anammox niche specificity in natural and man-made ecosystems.
  • 微生物の多様性―「遺伝子の多様性」/「細胞の多様性」から見た生物多様性 バイオフィルム
    押木守, 岡部聡
    生物の科学 遺伝, 65, 3, 48, 54, エヌ・ティー・エス, 2011年05月01日, [筆頭著者]
    日本語
  • Rapid quantification of polyhydroxyalkanoates (PHA) concentration in activated sludge with the fluorescent dye Nile blue A
    M. Oshiki, H. Satoh, T. Mino
    Water Sci. Technol., 64, 3, 747, 753, IWA PUBLISHING, 2011年, [査読有り], [筆頭著者, 責任著者]
    英語, 研究論文(学術雑誌), The present study was conducted (1) to develop a rapid quantification method of polyhydroxyalkanoates (PHA) concentration in activated sludge by Nile blue A staining and fluorescence measurement and (2) to perform on-line monitoring of PHA concentrations in activated sludge. Activated sludge samples collected from laboratory scale sequencing batch reactors and full-scale wastewater treatment plants were stained with Nile blue A and their fluorescence intensities were determined. There was a high correlation (R(2)>0.97) between the fluorescence intensities of Nile blue A and PHA concentrations in activated sludge determined by gas chromatography. The Nile blue A staining and fluorescence measurement method allows us to determine PHA concentrations in activated sludge within only five minutes and up to 96 samples can be measured at once by using microplate reader. On-line monitoring of PHA concentrations in activated sludge was achieved by using a fluorometer equipped with a flow cell and the time point at which PHA concentration in activated sludge reached the maximum level could be identified. In addition, we examined the influence of pH, floc size and co-existing chemicals in activated sludge suspension on the fluorescence intensities of Nile blue A.
  • A great leap forward in microbial ecology
    Satoshi Okabe, Mamoru Oshiki, Yoichi Kamagata, Nobuyasu Yamaguchi, Masanori Toyofuku, Yutaka Yawata, Yosuke Tashiro, Nobuhiko Nomura, Hiroyuki Ohta, Moriya Ohkuma, Akira Hiraishi, Kiwamu Minamisawa
    Microbes Environ., 25, 4, 230, 240, JAPANESE SOC MICROBIAL ECOLOGY, DEPT BIORESOURCE SCIENCE, 2010年12月, [査読有り]
    英語, Ribosomal RNA (rRNA) sequence-based molecular techniques emerged in the late 1980s, which completely changed our general view of microbial life. Coincidentally, the Japanese Society of Microbial Ecology (JSME) was founded, and its official journal "Microbes and Environments (M&E)" was launched, in 1985. Thus, the past 25 years have been an exciting and fruitful period for M&E readers and microbiologists as demonstrated by the numerous excellent papers published in M&E. In this minireview, recent progress made in microbial ecology and related fields is summarized, with a special emphasis on 8 landmark areas; the cultivation of uncultured microbes, in situ methods for the assessment of microorganisms and their activities, biofilms, plant microbiology, chemolithotrophic bacteria in early volcanic environments, symbionts of animals and their ecology, wastewater treatment microbiology, and the biodegradation of hazardous organic compounds.
  • Separation of PHA-accumulating cells in activated sludge based on differences in buoyant density
    Mamoru Oshiki, Motoharu Onuki, Hiroyasu Satoh, Takashi Mino
    J. Gen. Appl. Microbiol., 56, 2, 163, 167, MICROBIOL RES FOUNDATION, 2010年04月, [査読有り], [筆頭著者, 責任著者]
    英語, 研究論文(学術雑誌)
  • Acetate uptake by PHA-accumulating and non-PHA-accumulating organisms in activated sludge from an aerobic sequencing batch reactor fed with acetate
    M. Oshiki, H. Satoh, T. Mino
    Water Sci. Technol., 62, 1, 8, 14, IWA PUBLISHING, 2010年, [査読有り], [筆頭著者, 責任著者]
    英語, 研究論文(学術雑誌), The present study was conducted to evaluate the specific acetate uptake rates of microorganisms with and without polyhydroxyalkanoates (PHA) accumulation. Activated sludge was aerobically incubated with 75 mgCL(-1) radiolabeled or non-labeled acetate, and acetate consumption and PHA accumulation were monitored. Microorganisms were quantified as follows: all microbial cells by DAPI staining, whole acetate utilizing organisms by microautoradiography, and PHA-accumulating organisms by staining with Nile blue A. The abundance of acetate-utilizing organisms without PHA accumulation was also calculated from the outcomes. The estimate of acetate utilized by PHAAOs included both the acetate converted to PHA and that used to supply reducing power and ATP. Acetate utilized by PHAAOs and non-PHAAOs were divided by their respective abundances to obtain their respective specific acetate uptake rates: PHAAOs ranged between 5.3 and 8.0 x 10(-10) mgC cell(-1) h(-1), and non-PHAAOs ranged between 2.8 and 4.2 x 10(-10) mgC cell(-1) h(-1).
  • 活性汚泥プロセスにおける有機性一時貯蔵物質の蓄積
    押木 守, 佐藤 弘泰, 小貫 元治, 味埜 俊
    下水道協会誌, 46, 566, 126, 137, 日本下水道協会, 2009年12月15日, [査読有り], [筆頭著者, 責任著者]
    日本語, 研究論文(学術雑誌)
  • 微生物の持つ有機物貯蔵能力を利用した省エネルギー型廃水処理法の提案               
    押木 守, 佐藤 弘泰, 味埜 俊
    用水と廃水, 51, 41, 49, 2009年, [査読有り], [筆頭著者, 責任著者]
    日本語, 研究論文(学術雑誌)
  • Occurrence of polyhydroxyalkanoate as temporal carbon storage material in activated sludge during the removal of organic pollutants
    Oshiki, M, Yang, Y, Onuki, M, Satoh, H, Peng, Y.-Z, Mino, T
    J. Water Environ. Technol, 6, 2, 77, 83, Japan Society on Water Environment, 2008年, [査読有り], [筆頭著者, 責任著者]
    英語, 研究論文(学術雑誌), The amounts of temporal carbon storage material, polyhydroxyalkanoate (PHA), were quantified in the course of wastewater treatment by different activated sludge reactors operated at Beijing University of Technology, Beijing, China. The studied reactors were five continuous ones and two sequencing batch ones. In all the reactors, PHA was detected, and its contents within activated sludge ranged between 0.2% and 3.2%. Soluble organic matters loaded to the reactors were mainly removed in the first anaerobic or microaerophilic tank and one fifth to all of removed soluble organic matters were tentatively stored as PHA in the studied reactors. It was concluded that not a small part of soluble organic matters in sewage is converted to PHA in the course of their removal by activated sludge.
  • PHA-accumulating microorganisms in full-scale wastewater treatment plants
    Mamoru Oshiki, Motoharu Onuki, Hiroyasu Satoh, Takashi Mino
    Water Sci. Technol., 58, 1, 13, 20, IWA PUBLISHING, 2008年, [査読有り], [筆頭著者, 責任著者]
    英語, 研究論文(学術雑誌), A study was conducted to clarify phylogenetic affiliations of polyhydroxyalkanoate (PHA)-accumulating microorganisms in full-scale activated sludge processes. Activated sludge samples obtained from three full-scale activated sludge processes were aerobically incubated with excess acetate to increase their PHA content. The buoyant density separation method was applied to selectively collect PHA-accumulating cells, which were then analysed by the group-level FISH and the PCR-DGGE-sequencing methods, and possible PHA-accumulating microbial groups were screened. A set of oligonucleotide probes targeting the microbial groups suspected to accumulate PHA was introduced, and seven oligonucleotide probes were newly designed for this purpose. PHA accumulation of probe-positive cells was confirmed by the post-FISH PHA staining method, wherein PHA staining with Nile Blue A (NBA) was applied after FISH. As a result, the following seven bacterial groups were found to have PHA: Dechloromonas, Accumulibacter, Thauera, Zoogloea, Comamonas, Competibacter and a novel cluster in Beta-proteobacteria. Based on the results of the post-FISH PHA staining method, these seven bacterial groups were estimated to account for around four-tenths to two-thirds of total PHA-accumulating microorganisms.
  • 活性汚泥による酢酸摂取へのPHA蓄積細菌の寄与
    押木 守, 小貫 元治, 佐藤 弘泰, 味埜 俊
    環境工学研究論文集, 43, 195, 203, Japan Society of Civil Engineers, 2006年11月17日, [査読有り], [筆頭著者, 責任著者]
    日本語, 研究論文(学術雑誌), 実下水処理場の活性汚泥を用いたバッチ試験を異なる電子受容体存在下で実施し, 活性汚泥による酢酸摂取へPHA蓄積細菌が寄与した割合を評価した. その際にNile blue Aで検出される細菌をPHA蓄積細菌とみなし, 存在割合をNile blue AとDAPIの二重染色法により定量評価した. 酢酸摂取にNile blue A陽性細菌が寄与した割合は2~10割であり, 好気条件で培養した活性汚泥でNile blue A陽性細菌は全菌の1~2割を占めた. さらにFISH法により<I>Candidatus</I> "Accumulibacter phosphatis"と<I>Candidatus</I> "Competibacter phosphatis"の存在割合を定量評価し, これらの細菌がNile blue A陽性細菌の中で1~9割を占めることを明らかにした.
  • 溶存酸素濃度および基質のC/N比が脱窒素汚泥内のnirS mRNA転写量に及ぼす影響
    押木 守, 荒木 信夫, 竹林 賢, 長野 晃弘
    水環境学会誌, 28, 11, 683, 687, 公益社団法人 日本水環境学会, 2005年11月10日, [査読有り], [筆頭著者]
    日本語, 研究論文(学術雑誌), The transcription level of the nitrite reductase gene of denitrifying bacteria in a lab-scale denitrifying reactor was investigated by real-time reverse transcription (RT)-PCR technique as an indicator of in-situ microbial activity. Two types of batch experiment were conducted to clarify the effects of dissolved oxygen (DO) and carbon/nitrogen (C/N) ratio on the relative nirS mRNA level in denitrifying sludge. The reverse transcription of total RNA with the primer, nirS 6R, specific for the nirS gene was suitable for the subsequent quantification of the nirS gene by real-time PCR. The presence of DO and a low C/N ratio in the medium resulted in a rapid decrease in the relative transcription level of nirS mRNA from 105 to 103 copies·ng-1 total RNA within four hours of the batch experiments. The relative transcription remained at the same level after nitrate concentration decreased to a level low enough to reduce denitrification rate. Although there was no relationship between the denitrification rate of the reactor and the relative transcription level of nirS mRNA in the batch experiments, a quick response of transcriptional induction of nirS mRNA corresponding to the changes in DO and C/N ratio was detected.
  • amoA mRNAの転写量に基づいたin-situにおけるアンモニア酸化活性の評価
    押木 守, 荒木 信夫, 山口 隆司, 山崎 慎一, 原田 秀樹
    環境工学研究論文集, 40, 63, 70, Japan Society of Civil Engineers, 2003年11月14日, [査読有り], [筆頭著者]
    日本語, 研究論文(学術雑誌), The relationship between ammonia-oxidizing activities and <I>amoA</I> mRNA (encoding ammonia monooxigenase) levels was investigated by batch experiments with nitrifying biofilms taken from a lab-scale bioreactor. RT-PCR technology was applied to determine copy numbers of <I>amoA</I> mRNA in total RNA extracted from the sludge sample. Ammonia oxidation rates and transcription levels of <I>amoA</I> mRNA were monitored throughout the experiment that ammonia was fed as the concentration rose abruptly up to 20 or 100 mg-N/L whenever ammonia was consumed completely. A large ammonia turnover rate occurred immediately after every ammonia shock load, although the <I>amoA</I> mRNA level was still low. The turnover rate gradually reduced with decreasing ammonia concentration, while the <I>amoA</I> mRNA level increased until several hours after every ammonia shock load. The tendency of the <I>amoA</I> mRNA transcription changed a decrease as ammonia concentration dropped as low as Km value of <I>Nitrosomonas</I>. The induction of <I>amoA</I> mRNA was detected under the presence of low ammonia less than 5 mg-N/L. However the <I>amoA</I> mRNA level was not correlated with the ammonia turnover rate in the range from 5 to 100mg-N/L. It was indicated that the <I>amoA</I> mRNA level in the nitrifying sludge was not directly connected to the <I>in-situ</I> activity of ammonia-oxidizing bacteria.

その他活動・業績

講演・口頭発表等

  • Transmission of antibiotic-resistance plasmids in activated sludge               
    Mamoru Oshiki, Kosuke Higuchi, Kiko Ohara, Satoshi Okabe
    The 10th International Water Industry Conference (IWIC2024), EXCO, Daegu, Korea, 2024年11月12日, 口頭発表(招待・特別)
    2024年11月12日 - 2024年11月15日, [招待講演]
  • 亜硝酸還元酵素による亜酸化窒素(N2O)の生成               
    押木 守, 小林 諒斗, 中屋 佑紀, 服部 祥平, 豊田 栄, 佐藤 久, 岡部 聡
    日本微生物生態学会 第36回大会 (11月27日〜11月30日、浜松アクトシティ) 3_31_S-a02 2023年11月, 2023年11月30日, 口頭発表(招待・特別)
    2023年11月28日 - 2023年11月30日, [招待講演]
  • 生物学的廃水処理プロセスにおける薬剤耐性プラスミドの動態               
    押木 守, 樋口 宏介, 小原 紀子, 岡部 聡
    日本微生物生態学会 第36回大会 (11月27日〜11月30日、浜松アクトシティ) 1_31_S-p05 2023年11月, 2023年11月28日, 口頭発表(招待・特別)
    2023年11月28日 - 2023年11月30日, [招待講演]
  • 下廃水処理に関わる 微生物群とその生理について               
    押木守
    生物工学会北日本支部シンポジウム, 2022年12月17日, 口頭発表(招待・特別)
    [招待講演]
  • Insights into the microbial ecology of nitrifying bacteria in marine recirculating trickling biofilter reactors               
    Mamoru Oshiki, Hirotoshi Netsu, Kyohei Kuroda, Takashi Narihiro, Naoki Fujii, Tomonori Kindaichi, Yoshiyuki Suzuki, Takashiro Watari, Masashi Hatamoto, Takashi Yamaguchi, Nobuo Araki, Satoshi Okabe
    12th Asian Symposium on Microbial Ecology, 2022年04月19日
    [招待講演]
  • 嫌気性アンモニウム酸化(anammox)細菌クロニクル; 未知の代謝を解明する               
    押木守
    第四回北関東磐越地区化学技術フォーラム, 2018年
    郡山地域職業訓練センター, [招待講演]
  • Candidatus Brocadia sinicaによる嫌気性アンモニウム酸化反応               
    Oshiki M
    The 8th Asian Symposium on Microbiology Ecology, 2016年
    台北, [招待講演]
  • 未知転じて既知となる: 嫌気性アンモニア酸化細菌の発見がもたらした衝撃               
    押木守
    48回GRL浜松セミナー, 2014年
    静岡大学, [招待講演]
  • 嫌気性アンモニウム酸化細菌による亜硝酸還元               
    押木守
    平成25年度日本水環境学会北海道支部学術講演会, 2013年
    北海道大学, [招待講演]
  • 嫌気性アンモニア酸化(Anammox)細菌の代謝多様性と廃水処理への応用               
    押木守
    第65回日本生物工学会大会, 2013年
    広島国際会議場, [招待講演]
  • 嫌気性アンモニア酸化(アナモックス)細菌のゲノム解析が明らかにする生理学的特性               
    押木守
    第15回水環境学会シンポジウム, 2012年
    佐賀大学, [招待講演], [国内会議]
  • 省エネ・創エネ型下水処理技術の提案               
    Oshiki M
    International Workshop on Water and Wastewater Treatment, 2009年
    台湾, [招待講演], [国際会議]
  • 活性汚泥による有機物除去へのPHA蓄積細菌の寄与               
    第11回水環境学会シンポジウム, 2008年
    関西大学, [招待講演]

所属学協会

  • 現在
    日本微生物生態学会               
  • 現在
    日本水環境学会               
  • 現在
    土木学会               
  • 現在
    日本生物工学会               
  • 現在
    日本農芸化学会               
  • 現在
    日本ゲノム微生物学会               

共同研究・競争的資金等の研究課題

  • 高温多湿な亜熱帯地域に適合するレトロフィット型高効率窒素除去プロセスの国際開発
    科学研究費助成事業
    2024年09月09日 - 2028年03月31日
    押木 守
    日本学術振興会, 国際共同研究加速基金(海外連携研究), 北海道大学, 24KK0196
  • 嫌気性アンモニウム酸化細菌の代謝経路の解明と産業利用にむけた基盤創出
    科学研究費助成事業
    2023年04月 - 2027年03月
    押木 守
    日本学術振興会, 基盤研究(B), 北海道大学, 研究代表者, 23K26807
  • 非持続的窒素循環を革新する新規生物電気化学的窒素変換技術の創出
    科学研究費助成事業
    2023年04月 - 2027年03月
    岡部 聡, 押木 守
    日本学術振興会, 基盤研究(A), 北海道大学, 研究分担者, 23H00192
  • 閉鎖循環式陸上養殖における海洋性アナモックス細菌による革新的窒素除去
    科学研究費助成事業
    2020年10月27日 - 2024年03月31日
    金田一 智規, MOJIRI AMIN, 押木 守, 粟田 貴宣
    本研究は、閉鎖循環式陸上養殖の普及・安定化・低コスト化の上で課題であった窒素排水処理を克服するために、塩分環境下で活性のある海洋性アナモックス細菌を用いた排水処理プロセスを導入した次世代型陸上養殖システムをスウェーデン養殖研究センター(ヨーテボリ大学)と共同で開発することである。具体的には、(1)海洋性アナモックス細菌の大量培養方法の確立、(2)前処理として部分硝化を組み込んだアナモックスリアクターの開発、(3)閉鎖循環式陸上養殖の循環海水を対象とした窒素排水処理の最適化と検証を行う。
    2021年度もコロナ禍により海外渡航が難しいため、ヨーテボリ大学とメールによる複数回の実験計画の打ち合わせ、およびWeb会議によるミーティングを行った。さらに日本国内の研究分担者とはメールを中心に一部対面での実験打ち合わせを行った。
    2021年度は、海洋性アナモックス細菌の培地成分として添加している微量金属元素に着目し、それらの種類および濃度を変えて培養を行い、海洋性アナモックス細菌の活性に及ぼす影響を検討した。まず人工海水として用いているシーライフ中に含まれる微量金属元素の濃度をICP-MSによって明らかにした。その後、シーライフ中の微量金属元素のみで海洋性アナモックス細菌の活性が低下するかどうかを検討した。その際、微量金属元素ができるだけ含まれない海水を作製するために高純度試薬を用いて海水を合成した実験系も構築し、シーライフに微量金属元素を添加した対照系と3つの実験系を比較した。その結果、いずれの実験系においても海洋性アナモックス細菌の活性に差は見られなかった。また、16S rRNA遺伝子に基づく細菌叢解析を行った結果、海洋性アナモックス細菌の構成比は変化しないことがわかった。
    日本学術振興会, 国際共同研究加速基金(国際共同研究強化(B)), 広島大学, 20KK0244
  • 好気性脱窒反応を促進した下水処理場エアレーションタンク単槽での窒素除去技術の開発
    科学研究費助成事業
    2020年04月01日 - 2024年03月31日
    荒木 信夫, 川上 周司, 青木 仁孝, 押木 守, 渡利 高大
    好気性脱窒能力を有する活性汚泥微生物群の多様性を明らかにするため、実都市下水処理を行っている標準活性汚泥処理装置のエアレーションタンクから活性汚泥を採取し、NO3-N(濃度:約30 mg-N/L)を含む培地にてバッチ式の集積培養を繰り返し行った。最終的に得られた集積培養系による12時間の培養期間における全窒素(TN)除去率は約2割、NO3-N除去率は約5割であった。この結果から、培地に含まれるNO3-N(濃度:約30 mg-N/L)の一部は、好気性脱窒反応によって除去されている可能性が示唆された。また、既知の好気性脱窒能を有すると報告されているAcinetobacter属やComamonas属の細菌が本実験で得られた集積培養系においても好気性脱窒反応を担っている可能性が、16S rRNA遺伝子を標的としたアンプリコンシーケンス解析により示された。
    活性汚泥内での原位置(in-situ)での好気性脱窒反応の速度を評価する手法として、15N標識硝酸(15NO3)を用いて15N標識窒素ガス(15-15N2)の生成速度を評価し、好気性脱窒速度を測定する方法を開発した。本年度は好気性脱窒細菌として知られているParacoccus属細菌を用いて、本手法の検証を行なった。その結果、試験に供したParacoccus属細菌が好気条件下で15N標識硝酸から生成した窒素ガス(30N2ガス)を生成すること、また、好気性脱窒活性がDO濃度に強い影響をうけることを明らかにした。
    日本学術振興会, 基盤研究(B), 20H02290
  • 嫌気的硫黄酸化と電子伝達を活性化した生物学的水処理技術の構築
    科学研究費助成事業
    2020年04月01日 - 2023年03月31日
    山口 隆司, 幡本 将史, 押木 守, 渡利 高大
    開発する水処理システムは、システム前段装置として途上国で普及している上昇流嫌気性汚泥床(UASB)、後段装置として申請者らが開発を進めてきた下降流懸架式スポンジ型好気性反応槽(DHS)を組み合わせた構成である。システムの有機物(BOD)分解者としては、メタン生成古細菌、脱窒素細菌、BOD酸化菌に加えて、硫黄酸化還元サイクルの微生物も積極的に活用することとしている。システム前段UASBは、下水を押出流れで供給する形式とし、内部に直接異種間電子伝達促進ゲル担体を投入する。後段DHSは,スポンジ担体利用散水型装置であり、BOD酸化と硝化を図る。硫黄サイクル微生物によるBOD除去は、特に、低温時に有用となる。本システムを通年連続運転し、黄サイクル微生物によるBOD除去および直接異種間電子伝達促進ゲル担体の制御方法を確立していくものである。特に、保持微生物の評価のための研究として下記を進めた。
    ◎ 嫌気的硫黄酸化について高活性な微生物を培養するため連続式培養装置運転。
    ◎ 嫌気的硫黄酸化の発現汚泥を対象に次世代シーケンサーを用いた網羅的解析。
    ◎ 可視化に成功した嫌気的硫黄酸化微生物の候補は、シングルセルゲノム解析により遺伝子レベルでの機能解明を行ための準備。
    当該年度は、未知な事象である「嫌気的硫黄酸化」に関する電子伝達に関わる反応メカニズムの解明と、低温条件下での嫌気的硫黄酸化還元の制御に関する知見収集が大幅に進む実績を得た。
    日本学術振興会, 基盤研究(A), 長岡技術科学大学, 20H00641
  • 新規好気性アンモニア酸化反応の分子メカニズム解明に向けた触媒分子の分離と機能解明               
    日本学術振興会科学研究費補助金 基盤研究C
    2019年04月 - 2022年03月
    押木守
    日本学術振興会, 研究代表者, 競争的資金
  • ゲノムマイニングおよび培養試験から存在が見いだされた新規な亜硝酸還元酵素の正体と機能をつきとめる               
    2018年04月 - 2020年03月
    押木守
    公益財団法人発酵研究所一般研究助成, 研究代表者, 競争的資金
  • 水処理微生物のバイオインフォマティクス               
    2016年07月 - 2019年06月
    福島寿和
    日本製鉄株式会社(旧 新日鐵住金株式会社)・電気通信大学, 競争的資金
  • 分子・物質合成プラットフォーム利用促進事業: 嫌気性アンモニウム酸化(anammox)細菌におけるヒドラジンN2H4分子の細胞イメージング               
    2018年04月 - 2019年03月
    押木守
    千歳科学技術大学, 研究代表者, 競争的資金
  • 窒素循環に新たな1ページを加える第三の窒素ガス生成経路を担う微生物群の同定               
    日本学術振興会科学研究費補助金 若手(B)
    2017年04月 - 2019年03月
    押木守
    日本学術振興会, 研究代表者, 競争的資金
  • エネルギー資源を生み出す排水処理技術の開発:液体燃料ヒドラジンN2H4の生産               
    2017年09月 - 2018年03月
    押木守
    公益財団法人インテリジェント・コスモス学術振興財団, 研究代表者, 競争的資金
  • ヒドラジンをエネルギーとして生産する新規な窒素含有排水処理法の開発               
    第一回イムラ・ジャパン賞
    2017年04月 - 2018年03月
    押木守
    イムラ・ジャパン株式会社, 研究代表者, 競争的資金
  • 膜分離技術を利用した脱塩素呼吸Dehalococcoides属細菌の高密度培養による超高速有機塩素嫌気分解リアクターの構築               
    育英奨学会研究助成金
    2016年04月 - 2017年03月
    押木守
    公益財団法人ユニオンツール, 研究代表者, 競争的資金
  • ナノ流路集積デバイスを用いた病原性微生物・ウイルスの迅速定量               
    2015年04月 - 2016年03月
    押木守
    公益財団法人 内田エネルギー科学振興財団, 研究代表者, 競争的資金
  • 超省エネ都市下水処理の安定性の強化にむけた嫌気性原生動物データベース(PD2)の整備               
    2015年04月 - 2016年03月
    押木守
    公益財団法人 前田記念工学振興財団, 研究代表者, 競争的資金
  • ナノ流路デバイスを用いた病原性微生物の迅速検査技術の開発               
    研究プロジェクト経費若手研究
    2015年04月 - 2016年03月
    押木守
    独立行政法人国立高等専門学校機構, 研究代表者, 競争的資金
  • 鉄還元型嫌気性アンモニア酸化プロセスによる超省エネルギーかつ省コストな窒素除去技術の創出               
    若手研究助成
    2014年04月 - 2016年03月
    押木守
    公益財団法人鉄鋼環境基金, 研究代表者, 競争的資金
  • 都市下水処理UASB内への嫌気性原生動物の高濃度培養技術の開発               
    日本学術振興会科学研究費補助金 基盤研究(B)
    2013年04月 - 2016年03月
    荒木信夫
    日本学術振興会, 競争的資金
  • 超省エネルギー型下水処理プロセスにおける病原性微生物の動態               
    2014年10月 - 2015年03月
    押木守
    高専−長岡技科大共同研究助成, 研究代表者, 競争的資金
  • 水田土壌由来N2Oガスの生成経路および生成細菌の同定               
    2014年07月 - 2015年03月
    押木守
    公益財団法人山口育英奨学会, 研究代表者, 競争的資金
  • アンモニア酸化細菌による亜酸化窒素(N2O)生成機構の解明               
    育英奨学会研究助成金
    2014年04月 - 2015年03月
    押木守
    公益財団法人ユニオンツール, 研究代表者, 競争的資金
  • メタボローム解析に基づくANAMMOXプロセスの高効率・安定化技術の創出               
    日本学術振興会科学研究費補助金 特別研究員奨励費
    2011年04月 - 2013年03月
    押木守
    日本学術振興会, 研究代表者, 競争的資金
  • 有機物の一時貯蔵によって減少する脱窒量の把握とその改善に向けた検討               
    日本学術振興会科学研究費補助金 研究活動スタート支援
    2009年04月 - 2011年03月
    押木守
    日本学術振興会, 研究代表者, 競争的資金
  • 活性汚泥におけるPHA蓄積細菌群集の分子生物学的手法による解析               
    日本学術振興会科学研究費補助金 特別研究員奨励費
    2007年04月 - 2008年03月
    押木守
    日本学術振興会, 研究代表者, 競争的資金

産業財産権

  • 微生物群の特定方法               
    特許権, 福島寿和, 中川淳一, 加藤敏朗, 川野秀一, 押木守, 日本製鉄株式会社
    特願特願2019-105590, 2019年06月05日
    特開2020-036579, 2020年03月12日
    特許7299485, 2023年06月20日
  • 遺伝子又は該遺伝子産物の定量方法及びモニタリング方法               
    特許権, 福島寿和, 中川淳一, 押木守, 川野秀一, 日本製鉄株式会社
    特願特願2020-100233, 2020年06月09日
    特許特開2021-193883, 2021年12月27日

社会貢献活動

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