Reactive dynamics between plasma and microorganism

2012 Fiscal Year Annual Research Report

• Project Area: Creation of Science of Plasma Nano-Interface Interactions
• Project/Area Number: 21110006
• Research Institution: Meijo University
• Principal Investigator: 伊藤 昌文 名城大学, 理工学部, 教授 (10232472)
• Co-Investigator(Kenkyū-buntansha): 太田 貴之 名城大学, 理工学部, 准教授 (10379612) ; 竹田 圭吾 名古屋大学, 工学(系)研究科(研究院), 助教 (00377863)
• Project Period (FY): 2009-04-01 – 2014-03-31
• Keywords: プラズマ / 殺菌 / 蛍光 / 分光 / カビ / レーザ誘起蛍光法

Outline of Annual Research Achievements

ラジカルと植物性菌細胞の階層的反応，特にナノ界面で顕在化するプラズマと細胞との相互作用反応の解明を目的とした．殺菌に大きな影響を及ぼすOラジカルなどの反応性酸素種(reactive oxygen species : ROS)の効果に着目し大気圧酸素ラジカル源を開発し，中性ラジカルの効果を検証した．
真空紫外吸収分光法を用いて基底状態酸素原子と励起状態酸素分子の密度測定とその殺菌効率を評価した．照射距離を変化させラジカルによる殺菌を行った結果，照射距離に依存して殺菌効率は変化した．基底状態酸素原子は寿命が0.5ms以下と短いこと、ミドリカビ胞子の殺菌においては効率が高い殺菌因子の一つであることが示された．
ラジカルと菌の階層構造部位とのナノ界面時空間相互反応ダイナミクスを，種々の手法を用いて解析を試みた．具体的にはSEMを用いて細胞外側組織の形状変化を，共焦点レーザ走査型蛍光顕微鏡を用いて，ラジカルと細胞外部組織の反応を検証した．さらにリアルタイムESR測定システムを構築し，細胞内フリーラジカルの同定とその変化を明らかにした．基底状態酸素原子は細胞表面の構造を大きく破壊することなく細胞内部へ浸透する．その結果，細胞膜や細胞質に存在する細胞小器官へ作用し，細胞内部から生体機能を失活させることが示唆された．また，ESR測定により細胞内のフリーラジカルの存在が明らかになった．このラジカルはベンゾキノンであると考えられる．照射時間の増加とともに，ベンゾキノンの信号が減少した．キノンは抗酸化作用をもつといわれており，ラジカル照射によって抗酸化作用が失われ，脂質過酸化反応等の酸化が促進されたされた結果，細胞死に至ったと推測される．

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

交付申請書に記載した①気相及び液相ラジカル密度測定と殺菌効果，②共焦点レーザ蛍光顕微鏡による２次元反応ダイナミクスモニタリング，③ナノ界面形状及び分子構造解析のそれぞれについて，概要に記述したようにおおむね達成している．

Strategy for Future Research Activity

今後は，さらにミクロな領域，すなわちナノ界面での細胞組織の変化に着目し，ラジカルとナノ界面（細胞壁、細胞膜）や細胞内部の変化の解析を行い，プラズマとカビ細胞との相互作用を解明していく．

Research Products

• [Journal Article] Optical-Fiber Type Broadband Cavity Ring-Down Spectroscopy Using Wavelength-Tunable Ultra-Short Pulsed Light (2013)
  • Author(s): T. Hiraoka, T. Ohta, M. Ito, N. Nishizawa, M. Hori
  • Journal Title: Japanese Journal of Applied Physics Volume: 52 Pages: 040201-01-03
  • DOI: http://dx.doi.org/10.7567/JJAP.52.040201
  • Peer Reviewed
• [Journal Article] Inactivation Process of Penicillium digitatum Spores Treated with Non-equilibrium Atmospheric Pressure Plasma (2013)
  • Author(s): H. Hashizume, T. Ohta, T. Mori, S. Iseki, M. Hori, and M. Ito
  • Journal Title: Japanese Journal of Applied Physics Volume: 52 Pages: 056202-01-04
  • DOI: http://dx.doi.org/10.7567/JJAP.52.056202
  • Peer Reviewed
• [Journal Article] Plasma agriculture based on quantitative monitoring of reactions between fungal cells and atmospheric-pressure plasmas (2012)
  • Author(s): M. Ito, T. Ohta, K. Takeda
  • Journal Title: MRS Online Proceedings Library Volume: 1469 Pages: ww02-07
  • DOI: http://dx.doi.org/10.1557/opl.2012.1008
  • Peer Reviewed
• [Journal Article] Electron Spin Resonance (ESR) Observation of Radicals on Biological Organism Interacted with Plasmas (2012)
  • Author(s): K. Ishikawa, H. Moriyama, K. Tamiya, H. Hashizume, T. Ohta, M. Ito, S. Iseki, H. Tanaka, K. Takeda, H. Kondo, M. Sekine, M. Hori
  • Journal Title: MRS Online Proceedings Library Volume: 1469 Pages: ww02-06
  • DOI: http://dx.doi.org/10.1557/opl.2012.928
  • Peer Reviewed
• [Journal Article] Real-time in situ electron spin resonance measurements on fungal spores of Penicillium digitatum during exposure of oxygen plasmas (2012)
  • Author(s): K. Ishikawa, H. Mizuno, H. Tanaka, K. Tamiya, H. Hashizume, T. Ohta, M. Ito, S. Iseki, K. Takeda, H. Kondo, M. Sekine, M. Hori
  • Journal Title: APPLIED PHYSICS LETTERS Volume: 101 Pages: 013704-01-04
  • DOI: http://dx.doi.org/10.1063/1.4733387
  • Peer Reviewed
• [Presentation] 酸素ラジカル処理による出芽酵母の増殖効果 (2013)
  • Author(s): 橋爪博司、 鈴木実、 太田貴之、 堀 勝、 伊藤昌文
  • Organizer: 第60回応用物理学会春季学術講演会
  • Place of Presentation: 神奈川工科大学
  • Year and Date: 2013-03-27 – 2013-03-30
• [Presentation] ミドリカビ胞子からの電子スピン共鳴信号の同定(3) (2013)
  • Author(s): 水野寛子、 石川健治、 田中宏昌、 橋爪博司、 太田貴之、 伊藤昌文、 竹田圭吾、 近藤博基、 関根誠、 堀 勝
  • Organizer: 第60回応用物理学会春季学術講演会
  • Place of Presentation: 神奈川工科大学
  • Year and Date: 2013-03-27 – 2013-03-30
• [Presentation] 酸素ラジカル殺菌中のミドリカビ胞子の形態変化 (2013)
  • Author(s): 太田貴之、 橋爪博司、 杉原慶紀、 竹田圭吾、 石川健治、 堀 勝、 伊藤昌文
  • Organizer: 第60回応用物理学会春季学術講演会
  • Place of Presentation: 神奈川工科大学
  • Year and Date: 2013-03-27 – 2013-03-30
• [Presentation] Effects of reactive oxygen species on inactivation of Penicillium digitatum spores dependent on exposure distance using an atmospheric-pressure oxygen radical source (2013)
  • Author(s): H. Hashizume, T. Ohta, F. Jia, K. Takeda, K. Ishikawa, M. Hori,M. Ito
  • Organizer: The 6th International Conference on PLAsma-Nano Technology Science(IC-PLANTS 2013)
  • Place of Presentation: Gifu, Japan
  • Year and Date: 2013-02-02 – 2013-02-03
• [Presentation] Raman spectra variation of microorganisms treated by atmospheric-pressure oxygen-radical source (2013)
  • Author(s): H. Kato, H. Hashizume, T. Ohta, M. Hiramatsu, M. Hori, M. Ito
  • Organizer: The 6th International Conference on PLAsma-Nano Technology Science(IC-PLANTS 2013)
  • Place of Presentation: Gifu, Japan
  • Year and Date: 2013-02-02 – 2013-02-03
• [Presentation] Measurement of species in non-equilibrium atmospheric pressure plasma using near infrared absorption spectroscopy with supercontinuum light (2013)
  • Author(s): M. Hori, K. Takeda, T. Ohta, M. Ito, N. Nishizawa
  • Organizer: 5th International Symposium on Advanced Plasma Science and its Applications for Nitrides and Nanomaterials
  • Place of Presentation: Aichi, Japan
  • Year and Date: 2013-01-28 – 2013-02-01
• [Presentation] Dependence of exposure distance on inactivation of P. digitatum spores using low-temperature atmospheric pressure radical sourc (2013)
  • Author(s): M. Ito, H. Hashizume, T. Ohta, F. Jia, K. Takeda, K. Ishikawa, M. Hori
  • Organizer: 5th International Symposium on Advanced Plasma Science and its Applications for Nitrides and Nanomaterials
  • Place of Presentation: Aichi, Japan
  • Year and Date: 2013-01-28 – 2013-02-01
• [Presentation] Real-time in-situ monitoring of fluorescent images of P. digitatum spores during oxygen radical treatment using confocal laser microscopy (2013)
  • Author(s): H. Hashizume, T. Towatari, H. Ohashi, T. Ohta, M. Hori, M. Ito
  • Organizer: 5th International Symposium on Advanced Plasma Science and its Applications for Nitrides and Nanomaterials
  • Place of Presentation: Aichi, Japan
  • Year and Date: 2013-01-28 – 2013-02-01
• [Presentation] Electron spin resonance study of plasma-biological surface interaction for food hygiene (2013)
  • Author(s): K. Ishikawa, H. Mizuno, H. Tanaka, H. Hashizume, T. Ohta, M. Ito, K. Takeda, H. Kondo, M. Sekine, M. Hori
  • Organizer: 5th International Symposium on Advanced Plasma Science and its Applications for Nitrides and Nanomaterials
  • Place of Presentation: Aichi, Japan
  • Year and Date: 2013-01-28 – 2013-02-01
• [Presentation] Plasma-biological surface interaction investigated by electron spin resonance (2013)
  • Author(s): H. Mizuno, K. Ishikawa, H. Tanaka, H. Hashizume, T. Ohta, M. Ito, K. Takeda, H. Kondo, M. Sekine, M. Hori
  • Organizer: 5th International Symposium on Advanced Plasma Science and its Applications for Nitrides and Nanomaterials
  • Place of Presentation: Aichi, Japan
  • Year and Date: 2013-01-28 – 2013-02-01
• [Presentation] Raman spectroscopy of Penicillium digitatum spores treated by atmospheric-pressure oxygen-radical source (2013)
  • Author(s): H. Kato, H. Hashizume, T. Ohta, M. Hiramatsu, M. Ito, M. Hori
  • Organizer: 5th International Symposium on Advanced Plasma Science and its Applications for Nitrides and Nanomaterials
  • Place of Presentation: Aichi, Japan
  • Year and Date: 2013-01-28 – 2013-02-01
• [Presentation] Measurement of singlet oxygen molecule densities in the inactivation process of P. digitatum spores using low-temperature atmospheric pressure radical source (2013)
  • Author(s): H. Hashizume, T. Ohta, M. Ito, F. Jia, K. Takeda, K. Ishikawa, M. Hori
  • Organizer: 5th International Symposium on Advanced Plasma Science and its Applications for Nitrides and Nanomaterials
  • Place of Presentation: Aichi, Japan
  • Year and Date: 2013-01-28 – 2013-02-01
• [Presentation] Inactivation efficiencies of P. digitatum spores dependent on exposure distances using an atmospheric-pressure oxygen radical source (2013)
  • Author(s): H. Hashizume, T. Ohta, F. Jia, K. Takeda, K. Ishikawa, M. Hori,M. Ito
  • Organizer: 2nd International Workshop on Advanced Plasma Science
  • Place of Presentation: Aichi, Japan
  • Year and Date: 2013-01-28 – 2013-01-28
  • Invited
• [Presentation] O(3Pj)とO2（1Δg）密度測定によるミドリカビ胞子の殺菌因子の評価 (2013)
  • Author(s): 橋爪博司、太田貴之、賈鳳東、竹田圭吾、石川健治、堀勝、伊藤昌文
  • Organizer: 第30回プラズマプロセシング研究会（SPP-30）
  • Place of Presentation: アクトシティ浜松
  • Year and Date: 2013-01-21 – 2013-01-23
• [Presentation] 大気圧酸素ラジカル源によって処理された微生物のラマン分光 (2013)
  • Author(s): 加藤寛人、橋爪博司、太田貴之、平松美根男、堀勝、伊藤昌文
  • Organizer: 第30回プラズマプロセシング研究会（SPP-30）
  • Place of Presentation: アクトシティ浜松
  • Year and Date: 2013-01-21 – 2013-01-23
• [Presentation] 酸素ラジカル処理されたミドリカビ胞子の蛍光観察 (2013)
  • Author(s): 橋爪博司、 太田貴之、 竹田圭吾、 石川健治、 堀 勝、 伊藤昌文
  • Organizer: 第30回プラズマプロセシング研究会（SPP-30）
  • Place of Presentation: アクトシティ浜松
  • Year and Date: 2013-01-21 – 2013-01-23
• [Presentation] Plasma-Biological Surface Interaction for Food Hygiene: Real-time in situ Electron Spin Resonance Measurements (2012)
  • Author(s): K. Ishikawa, H. Mizuno, H. Tanaka, H. Hashizume, T. Ohta, M. Ito, K. Takeda, H. Kondo, M. Sekine, M. Hori
  • Organizer: 34th International Symposium on Dry Process
  • Place of Presentation: the University of Tokyo, Japan
  • Year and Date: 2012-11-15 – 2012-11-16
• [Presentation] ミドリカビ胞子細菌における活性酸素の定量的解析 (2012)
  • Author(s): 橋爪博司、太田貴之、伊藤昌文、賈鳳東、竹田圭吾、石川健治、堀勝
  • Organizer: 第５回プラズマ技術産業応用センター（PLACIA）＆プラズマが拓くものづくり研究会(PLAM)国際シンポジウム
  • Place of Presentation: 愛知県名古屋市サイエンス交流プラザ
  • Year and Date: 2012-10-24 – 2012-10-24
• [Presentation] Measurement of activated species generated by 60 Hz excited atmospheric pressure Ar plasma in atmospheric gas (2012)
  • Author(s): K. Takeda, J. Jolibois, K. Ishikawa, H. Tanaka, H. Kano, M. Sekine, M. Hori
  • Organizer: 65th Annual Gaseous Electronics Conference
  • Place of Presentation: Dallas, USA
  • Year and Date: 2012-10-22 – 2012-10-26
• [Presentation] Absolute Density and Translational Temperature of Atomic Oxygen Radical in Surface Wave Oxygen Plasma with High Rare Gas Dilution (2012)
  • Author(s): K. Takeda, M. Hori
  • Organizer: 11th APCPST (Asia Pacific Conference on Plasma Science and Technology) and 25th SPSM (Symposium on Plasma Science for Materials)
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 2012-10-02 – 2012-10-05
• [Presentation] Real-Time Electron Spin Resonance Study on Fungal Spores during Inactivation (2012)
  • Author(s): K. Ishikawa, H. Mizuno, H. Tanaka, H. Hashizume, T. Ohta, M. Ito, K. Takeda
  • Organizer: 11th APCPST (Asia Pacific Conference on Plasma Science and Technology) and 25th SPSM (Symposium on Plasma Science for Materials)
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 2012-10-02 – 2012-10-05
• [Presentation] Disinfection of Fungal Spores by Atmospheric Pressure Radical Source (2012)
  • Author(s): H. Hashizume, S. Nagano, T. Ohta, M. Ito, K. Takeda, K. Ishikawa, M. Hori
  • Organizer: 11th APCPST (Asia Pacific Conference on Plasma Science and Technology) and 25th SPSM (Symposium on Plasma Science for Materials)
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 2012-10-02 – 2012-10-05
• [Presentation] 大気圧酸素ラジカル源を用いたミドリカビ胞子の殺菌 (2012)
  • Author(s): 長野晋、加藤寛人、橋爪博司、太田貴之、伊藤昌文、竹田圭吾、石川健治、堀勝
  • Organizer: 平成24年度電気関係学会 東海支部連合大会
  • Place of Presentation: 豊橋技術科学大学
  • Year and Date: 2012-09-24 – 2012-09-25
• [Presentation] 大気圧プラズマ処理によるミドリカビ胞子のラマンスペクトルの変化 (2012)
  • Author(s): 加藤寛人、橋爪博司、太田貴之、平松美根男、伊藤昌文、堀勝
  • Organizer: 平成24年度電気関係学会 東海支部連合大会
  • Place of Presentation: 豊橋技術科学大学
  • Year and Date: 2012-09-24 – 2012-09-25
• [Presentation] Oxidative Inactivation Mechanism of Fungal Spores Employing Atmospheric-pressure Oxygen Radical Source (2012)
  • Author(s): T. Ohta, H. Hashizume, M. Ito, K. Takeda, K. Ishikawa, M. Hori
  • Organizer: International Union of Materials Research Societies - Internet Conference on Electronic Materials (IUMRS-ICEM 2012)
  • Place of Presentation: Yokohama, Japan
  • Year and Date: 2012-09-23 – 2012-09-28
• [Presentation] Real-time / In-situ Electron Spin Resonance Analysis of Surface Reactions on Organic / Biological Materials Interacted with Plasmas (2012)
  • Author(s): K. Ishikawa, H. Moriyama, N. Sumi, K. Tamiya, H. Hashizume, T. Ohta, M. Ito, A. Kono, H. Horibe, H. Tanaka, K. Takeda, H. Kondo, M. Sekine, M. Hori
  • Organizer: International Union of Materials Research Societies - Internet Conference on Electronic Materials (IUMRS-ICEM 2012)
  • Place of Presentation: Yokohama, Japan
  • Year and Date: 2012-09-23 – 2012-09-28
  • Invited
• [Presentation] Inactivation Mechanism of Fungal Spores by Atmospheric Pressure Plasma (2012)
  • Author(s): T. Ohta, H. Hashizume, M. Ito
  • Organizer: Ninth International Conference on Flow Dynamics
  • Place of Presentation: Sendai, Japan
  • Year and Date: 2012-09-19 – 2012-09-21
  • Invited
• [Presentation] 酸素ラジカル照射によるミドリカビ胞子のラマンスペクトルの変化 (2012)
  • Author(s): 加藤寛人，橋爪博司，太田貴之，平松美根男，伊藤昌文，堀勝
  • Organizer: 平成24年秋季第73回応用物理学会学術講演会
  • Place of Presentation: 愛媛大学
  • Year and Date: 2012-09-11 – 2012-09-14
• [Presentation] 活性酸素種の密度測定によるミドリカビ胞子殺菌の定量的評価の検討 (2012)
  • Author(s): 橋爪博司，太田貴之，伊藤昌文，賈鳳東，竹田圭吾，石川健治，堀勝
  • Organizer: 平成24年秋季第73回応用物理学会学術講演会
  • Place of Presentation: 愛媛大学
  • Year and Date: 2012-09-11 – 2012-09-14
• [Presentation] Atmospheric Pressure Plasma Processes and their Diagnostics for Life Sciences (2012)
  • Author(s): M. Hori, M. Tanaka, K. Ishikawa, H. Kondo, K. Takeda, M. Sekine, F. Kikkawa, K. Nakamura, M. Hayashi, H. Kajiyama, M. Mizuno, H. Ohta, M. Ito, H. Kano
  • Organizer: The 2nd International Symposium for Plasma Biosciences
  • Place of Presentation: Seoul, Korea
  • Year and Date: 2012-08-12 – 2012-08-14
  • Invited
• [Presentation] Oxidative Inactivation of Fungal Spores by Atmospheric Pressure Oxygen Radical Source (2012)
  • Author(s): T. Ohta, H. Hashizume, M. Ito, K. Takeda, K. Ishikawa, M. Hori
  • Organizer: The 9th International Bioelectrics Symposium (BIOELECTRICS 2012)
  • Place of Presentation: Kumamoto, Japan
  • Year and Date: 2012-08-05 – 2012-08-08
• [Presentation] Inactivation Efficiencies of Reactive Oxygen Species on Spores by Atmospheric-Pressure O2/Ar Plasma Inactivation of Penicillium digitatum (2012)
  • Author(s): H. Hashizume, T. Ohta, M. Ito, F. Jia, K. Takeda, K. Ishikawa, M. Hori
  • Organizer: 2012 IEEE 39th International Conference on Plasma Sciences (ICOPS)
  • Place of Presentation: Edinburgh, UK
  • Year and Date: 2012-07-08 – 2012-07-13
• [Presentation] Electron Spin Resonance (ESR) study of Radicals on Biological Organism Created by Interaction with Plasma (2012)
  • Author(s): K. Ishikawa, H. Tanaka, H. Moriyama, S. Iseki, K. Takeda, S. Tajima, H. Kondo, M. Sekine, M. Hori, K. Tamiya, H. Hashizume, T. Ohta, M. Ito, H. Kano
  • Organizer: 2012 IEEE 39th International Conference on Plasma Sciences (ICOPS)
  • Place of Presentation: Edinburgh, UK
  • Year and Date: 2012-07-08 – 2012-07-13
• [Presentation] Inactivation of Penicillium digitatum spores by reactive oxygen radicals employing atmospheric-pressure oxygen radical source (2012)
  • Author(s): T. Ohta, H. Hashizume, M. lto, F. Jia, K. Takeda, K. Ishikawa, M. Hori
  • Organizer: 4th international conference on plasma medicine (ICPM4)
  • Place of Presentation: Orl#233;ans, France
  • Year and Date: 2012-06-17 – 2012-06-21
• [Presentation] Diagnostics of Atomic Oxygen in O2/Rare Gas Mixture Plasma with Vacuum Ultraviolet Laser Absorption Spectroscopy (2012)
  • Author(s): K. Takeda, M. Hori
  • Organizer: 5th international workshop on plasma spectroscopy
  • Place of Presentation: Riviera, France
  • Year and Date: 2012-05-13 – 2012-05-16
• [Presentation] Oxidation Process of Penicillium digitatum Spores Using Atmospheric-pressure Plasma Observed by Fluorescent Microscopy (2012)
  • Author(s): H. Hashizume, T. Ohta, S. Iseki, T. Mori, M. Ito, M. Hori
  • Organizer: 2012 MRS Spring Meeting Exhibition
  • Place of Presentation: San Francisco, USA
  • Year and Date: 2012-04-09 – 2012-04-13
• [Presentation] Electron Spin Resonance (ESR) Observation of Radicals on Biological Organism Interacted with Plasmas (2012)
  • Author(s): K. Ishikawa, S. Iseki, H. Moriyama, H. Tanaka, H. Hashizume, T. Ohta, M. Ito, K. Takeda, H. Kondo, M. Sekine, M. Hori
  • Organizer: 2012 MRS Spring Meeting Exhibition
  • Place of Presentation: San Francisco, USA
  • Year and Date: 2012-04-09 – 2012-04-13
• [Presentation] Plasma Agriculture Based on Quantitative Monitoring of Reactions between Fungal Cells and Atmospheric-pressure Plasmas (2012)
  • Author(s): M. Ito, T. Ohta, K. Takeda
  • Organizer: 2012 MRS Spring Meeting Exhibition
  • Place of Presentation: San Francisco, USA
  • Year and Date: 2012-04-09 – 2012-04-13
  • Invited
• [Presentation] Terahertz Time-domain Spectroscopic Sensing of Penicillium digitatum Inactivated by Non-equilibrium Atmospheric Pressure Plasmas (2012)
  • Author(s): T. Hiraoka, N. Ebizuka, K. Takeda, T. Ohta, K. Ishikawa, M. Ito, K. Kawase, M. Sekine, M. Hori
  • Organizer: 2012 MRS Spring Meeting Exhibition
  • Place of Presentation: San Francisco, USA
  • Year and Date: 2012-04-09 – 2012-04-13
• [Patent(Industrial Property Rights)] 酵母の増殖方法および酵母の生菌数の制御方法 (2013)
  • Inventor(s): 伊藤昌文, 太田貴之, 橋爪博司, 堀 勝
  • Industrial Property Rights Holder: 名城大学， 名古屋大学
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2013-45836
  • Filing Date: 2013-03-07