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Real-time and on-site monitoring of bacterial cells in freshwater environments by using a microfluidic system

Research Project

Project/Area Number 21256002
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section海外学術
Research Field Environmental pharmacy
Research InstitutionOsaka University

Principal Investigator

NASU Masao  大阪大学, 大学院・薬学研究科, 教授 (90218040)

Co-Investigator(Kenkyū-buntansha) TANI Katsuji  大阪大谷大学, 薬学部, 教授 (50217113)
KAWAI Mako  姫路獨協大学, 薬学部, 准教授 (40533922)
YAMAGUCHI Nobuyasu  大阪大学, 大学院・薬学研究科, 准教授 (20252702)
BABA Takashi  大阪大学, 大学院・薬学研究科, 助教 (20423121)
Co-Investigator(Renkei-kenkyūsha) KAWABATA Zen'ichiro  総合地球環境学研究所, 研究部, 教授 (80108456)
IWAMOTO Tomotada  神戸市環境保健研究所, 微生物部, 副部長 (70416402)
ICHIJO Tomoaki  大阪大学, 大学院・薬学研究科, 特任研究員 (20513899)
Research Collaborator 陳 功祥  中国, 浙江大学医学部, 教授
HIEN Lai Thuy  ベトナム, 科学技術アカデミー, 准教授
MUANGKAEW Kanya  タイ, 科学技術省, 研究員
Project Period (FY) 2009 – 2012
Project Status Completed (Fiscal Year 2012)
Budget Amount *help
¥48,100,000 (Direct Cost: ¥37,000,000、Indirect Cost: ¥11,100,000)
Fiscal Year 2011: ¥16,250,000 (Direct Cost: ¥12,500,000、Indirect Cost: ¥3,750,000)
Fiscal Year 2010: ¥18,200,000 (Direct Cost: ¥14,000,000、Indirect Cost: ¥4,200,000)
Fiscal Year 2009: ¥13,650,000 (Direct Cost: ¥10,500,000、Indirect Cost: ¥3,150,000)
Keywords水環境 / on-site モニタリング / 細菌 / アジア / マイクロ流路デバイス / 衛生 / モニタリング / 病原細菌 / オンサイトモニタリング / 蛍光染色
Research Abstract

“Real-time” and “on-site” microbiological methods are required for prevention of waterborne diseases. We fabricated a portable microfluidic system (36 cm × 54 cm × 23 cm, 15 kg) for rapid monitoring of bacterial cells in freshwater at a density in the order of 104- 106/ml.A microfluidic device (size: 48 mm × 23 mm) was designed and fabricated using polydimethylsiloxane (PDMS) and a thin glass sheet. Escherichia coli O157:H7 and Legionella pneumophila cells were spiked in purified freshwater (101- 106cells/ml). Low numbers of targeted cells were collected on a filter and then resuspended in purified freshwater for a 100 to 1,000 -fold concentration. Fluorescent dye was used for nucleic acid stain and direct counting of total bacterial cells. Fluorescent antibody was used for specific detection of E. coli O157:H7 and L. pneumophila cells in the samples. The portable microfluidic system was used for the detection of fluorescently stained cells flowing i n the microchannel, and number of the cells in each sample was determined using original image analysis software. Bacterial cells in the same samples were counted by fluorescence microscopy for comparison. The numbers of bacterial cells determined using the microfluidic system was highly correlated with the microscopic counts. We could count targeted cells in the samples within 2 hours with this microfluidic system. The microfluidic device and portable counting system fabricated in this study will contribute to the microbiological quality control of freshwater.

Report

(5 results)
  • 2012 Final Research Report ( PDF )
  • 2011 Annual Research Report
  • 2010 Annual Research Report
  • 2009 Annual Research Report
  • Products Report
  • Research Products

    (49 results)

All 2013 2012 2011 2010 2009 Other

All Journal Article (17 results) (of which Peer Reviewed: 17 results) Presentation (32 results)

  • [Journal Article] 16S ribosomal RNA gene-based phylogenetic analysis of abundant bacteria in river, canal and potable water in Bangkok2013

    • Author(s)
      Nobuyasu Yamaguchi, Takahiro Nishiguchi, Fuangfa Utrarachkij, Orasa Suthienkul, Masao Nasu.
    • Journal Title

      Thailand. Biol. Pharm. Bull.

      Volume: 36 Pages: 872-876

    • URL

      https://www.jstage.jst.go.jp/article/bpb/36/5/36_b13-00012/_pdf

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] 16S ribosomal RNA gene-based phylogenetic analysis of abundant bacteria in river, canal and potable water in Bangkok, Thailand2013

    • Author(s)
      Nobuyasu Yamaguchi
    • Journal Title

      Biol. Pharm. Bull.

      Volume: 36 Pages: 872-876

    • NAID

      130003361421

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Microchip-Based Terminal Restriction Fragment Length Polymorphism for On-Site Analysis of Bacterial Communities in Freshwater2013

    • Author(s)
      Nobuyasu Yamaguchi, Syuhei Matsukawa, Yoko Shintome, Tomoaki Ichijo, Masao Nasu.
    • Journal Title

      Biological and Pharmaceutical Bulletin

      Volume: 36 Issue: 8 Pages: 1305-1309

    • DOI

      10.1248/bpb.b13-00118

    • NAID

      130003361493

    • ISSN
      0918-6158, 1347-5215
    • Related Report
      Products Report
    • Peer Reviewed
  • [Journal Article] Selective enumeration of viable Enterobacteriaceae and active Pseudomonas spp. in milk within 7h by multicolor fluorescence in situ hybridization following microcolony formation.2012

    • Author(s)
      Nobuyasu Yamaguchi, Akiko Kitaguchi, Masao Nasu
    • Journal Title

      J. Biosci. Bioeng.

      Volume: 113 Issue: 6 Pages: 746-750

    • DOI

      10.1016/j.jbiosc.2012.01.009

    • Related Report
      2012 Final Research Report 2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A ten years (2000-2009) surveillance of resistant Enterobacteriaceae in Zhejiang Province, China2012

    • Author(s)
      Rong Zhang, Tomoaki Ichijo, Yan-Yan Hu, Hong-Wei Zhou, Nobuyasu Yamaguchi, Masao Nasu, Gong-Xiang Chen
    • Journal Title

      Microb. Ecol. Health Dis.

      Volume: 23 Issue: 0 Pages: 11609-11618

    • DOI

      10.3402/mehd.v23i0.11609

    • Related Report
      2012 Final Research Report 2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] High Prevalence of <i>qnr</i> and <i>aac(6&prime;)-Ib-cr</i> Genes in Both Water-Borne Environmental Bacteria and Clinical Isolates of <i>Citrobacter freundii</i> in China2012

    • Author(s)
      Rong Zhang, Tomoaki Ichijo, Yong-Lu Huang, Jia-Chang Cai, Hong-Wei Zhou, Nobuyasu Yamaguchi, Masao Nasu, Gong-Xiang Chen.
    • Journal Title

      Microbes and Environments

      Volume: 27 Issue: 2 Pages: 158-163

    • DOI

      10.1264/jsme2.ME11308

    • NAID

      130001396931

    • ISSN
      1342-6311, 1347-4405
    • Related Report
      2012 Final Research Report 2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Rapid Enumeration of Active <i>Legionella pneumophila</i> in Freshwater Environments by the Microcolony Method Combined with Direct Fluorescent Antibody Staining2012

    • Author(s)
      Takashi Baba, Naoko Inoue, Nobuyasu Yamaguchi, Masao Nasu.
    • Journal Title

      Microbes and Environments

      Volume: 27 Issue: 3 Pages: 324-326

    • DOI

      10.1264/jsme2.ME11324

    • NAID

      10030869731

    • ISSN
      1342-6311, 1347-4405
    • Related Report
      2012 Final Research Report 2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Rapid, semiautomated quantification of bacterial cells in freshwater by using a microfluidic device for on-chip staining and counting.2011

    • Author(s)
      Nobuyasu Yamaguchi, Masashi Torii, Yuko Uebayashi, Masao Nasu.
    • Journal Title

      Appl. Environ. Microbiol.

      Volume: 77 Issue: 4 Pages: 1536-1539

    • DOI

      10.1128/aem.01765-10

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Rapid enumeration of low numbers of moulds in tea based drinks using an automated system. Int2011

    • Author(s)
      Nobuyasu Yamaguchi, Kouichi Tanaka, Takashi Baba, Norihide Amano, Masao Nasu.
    • Journal Title

      J. Food Microbiol.

      Volume: 145 Issue: 1 Pages: 365-369

    • DOI

      10.1016/j.ijfoodmicro.2011.01.012

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Rapid, semiautomated quantification of bacterial cells in freshwater by using a microfluidic device for on-chip staining and counting2011

    • Author(s)
      Yamaguchi, N., et al.
    • Journal Title

      Appl.Environ.Microbiol.

      Volume: 77 Pages: 1536-1539

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Rapid enumeration of low numbers of moulds in tea based drinks using an automated system2011

    • Author(s)
      Yamaguchi, N., et al.
    • Journal Title

      Int.J.Food Microbiol.

      Volume: 145 Pages: 365-369

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Staphylococcus epidermidis forms floating micro-colonies in platelet concentrates at the early stage of contamination.2009

    • Author(s)
      Nobuyasu Yamaguchi, Yasuo Motoyama, Mami Matsumoto, Tomoaki Ichijo, Hideto Nagumo, Noboru Kagami, Yoshihiko Tani, Masahiro Satake, Masao Nasu.
    • Journal Title

      J. Health Sci.

      Volume: 55 Pages: 726-731

    • NAID

      130000127960

    • URL

      https://www.jstage.jst.go.jp/article/jhs/55/5/55_5_726/_pdf

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Rapid on-chip flow cytometric detection of Listeria monocytogenes in milk2009

    • Author(s)
      Nobuyasu Yamaguchi, Masafumi Ikeda, Masao Nasu.
    • Journal Title

      J. Health Sci.

      Volume: 55 Pages: 851-856

    • NAID

      130000127978

    • URL

      https://www.jstage.jst.go.jp/article/jhs/55/5/55_5_851/_pdf

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Rapid detection of starved Escherichia coh with respiratory activity in potable water by signal-amplified in situ hybridization following formazan reduction2009

    • Author(s)
      Yamaguchi N., et al.
    • Journal Title

      Microbes and Environments 24

      Pages: 286-290

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Staphylococcus epidermidis forms floating micro-colonies in platelet concentrates at the early stage of contamination2009

    • Author(s)
      Yamaguchi N., et al.
    • Journal Title

      J.Health Sci. 55

      Pages: 726-731

    • NAID

      130000127960

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Rapid on-chip flow cytometric detection of Listeria monocytogenes in milk2009

    • Author(s)
      Yamaguchi N., et al.
    • Journal Title

      J.Health Sci. 55

      Pages: 851-856

    • NAID

      130000127978

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Microchip-based terminal restriction fragment length polymorphism (T -RFLP) for on-site analysis of bacterial communities in freshwater.

    • Author(s)
      Nobuyasu Yamaguchi, Syuhei Matsukawa, Yoko Shintome, Tomoaki Ichijo, Masao Nasu.
    • Journal Title

      Biol. Pharm. Bull.

      Volume: (印刷中)

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Presentation] マイクロ流路システムによる冷却塔水中の Legionella pneumophila のオンサイトモニタリング2012

    • Author(s)
      山口 進康,他
    • Organizer
      フォーラム 2012:衛生薬学・環境トキシコロジー
    • Place of Presentation
      名古屋
    • Year and Date
      2012-10-26
    • Related Report
      2012 Final Research Report
  • [Presentation] マイクロ流路システムによる水環境中の Legionella pneumophila の迅速モニタリング.2012

    • Author(s)
      山口 進康,他
    • Organizer
      日本微生物生態学会第28回大会
    • Place of Presentation
      豊橋
    • Year and Date
      2012-09-21
    • Related Report
      2012 Final Research Report
  • [Presentation] Rapid monitoring of Legionella pneumophila in cooling tower water with a microfluidic system.2012

    • Author(s)
      N. Yamaguchi, et al
    • Organizer
      ASM 2012 General Meeting
    • Place of Presentation
      サンフランシスコ,アメリカ
    • Year and Date
      2012-06-18
    • Related Report
      2012 Final Research Report
  • [Presentation] 冷却塔水中の Legionella pneumophila のマイクロ流路システムによる迅速モニタリング2012

    • Author(s)
      山口 進康,他
    • Organizer
      日本薬学会第132 年会
    • Place of Presentation
      札幌
    • Year and Date
      2012-03-31
    • Related Report
      2012 Final Research Report
  • [Presentation] 冷却塔水中のレジオネラのマイクロ流路システムによる on-site モニタリング2012

    • Author(s)
      山口 進康,他
    • Organizer
      第85回日本細菌学会総会
    • Place of Presentation
      長崎
    • Year and Date
      2012-03-29
    • Related Report
      2012 Final Research Report
  • [Presentation] Rapid monitoring of Legionella pneumophila in cooling tower qater with a microfluidic system2012

    • Author(s)
      N. Yamaguchi
    • Organizer
      American Society for Microbiology 2012 General Meeting
    • Place of Presentation
      San Francisco
    • Related Report
      2011 Annual Research Report
  • [Presentation] マイクロ流路システムによる水環境中のLegionella pneumophilaの迅速モニタリング2012

    • Author(s)
      山口 進康
    • Organizer
      日本微生物生態学会第28回大会
    • Place of Presentation
      豊橋
    • Related Report
      2011 Annual Research Report
  • [Presentation] マイクロ流路システムによる冷却塔水中のLegionella pneumophila のオンサイトモニタリング2012

    • Author(s)
      山口 進康
    • Organizer
      フォーラム2012:衛生薬学・環境トキシコロジー
    • Place of Presentation
      名古屋
    • Related Report
      2011 Annual Research Report
  • [Presentation] 冷却塔水中の Legionella pneumophila のマイクロ流路システムによるモニタリング2011

    • Author(s)
      山口 進康,他
    • Organizer
      フォーラム 2011:衛生薬学・環境トキシコロジー
    • Place of Presentation
      金沢
    • Year and Date
      2011-10-28
    • Related Report
      2012 Final Research Report
  • [Presentation] 冷却塔水中のレジオネラのマイクロ流路システムによる迅速モニタリング2011

    • Author(s)
      山口 進康,他
    • Organizer
      第61回日本薬学会近畿支部大会
    • Place of Presentation
      神戸
    • Year and Date
      2011-10-22
    • Related Report
      2012 Final Research Report
  • [Presentation] マイクロ流路システムによる水環境中の Legionella pneumophila の迅速検出2011

    • Author(s)
      山口 進康,他
    • Organizer
      日本微生物生態学会第27回大会
    • Place of Presentation
      京都
    • Year and Date
      2011-10-09
    • Related Report
      2012 Final Research Report
  • [Presentation] マイクロ流路システムによる水環境中の細菌の迅速モニタリング2011

    • Author(s)
      山口 進康,他
    • Organizer
      日本宇宙生物科学会第25回大会
    • Place of Presentation
      横浜
    • Year and Date
      2011-09-30
    • Related Report
      2012 Final Research Report
  • [Presentation] 水環境中のレジオネラのオンサイトモニタリング2011

    • Author(s)
      山口 進康,他
    • Organizer
      日本温泉科学会第64回大会
    • Place of Presentation
      有馬
    • Year and Date
      2011-09-08
    • Related Report
      2012 Final Research Report
  • [Presentation] Abundance of Legionella pneumophila in cooling tower water determined by using a microfluidic device.2011

    • Author(s)
      N. Yamaguchi, et al
    • Organizer
      IUMS2011
    • Place of Presentation
      札幌
    • Year and Date
      2011-09-08
    • Related Report
      2012 Final Research Report
  • [Presentation] Automated staining and counting of bacterial cells in freshwater environments by using a microfluidic device.2011

    • Author(s)
      N. Yamaguchi, et al
    • Organizer
      Water Convention 2011
    • Place of Presentation
      シンガポール
    • Year and Date
      2011-07-05
    • Related Report
      2012 Final Research Report
  • [Presentation] Rapid enumeration of Legionella pneumophila in cooling tower water by using a microfluidic system.2011

    • Author(s)
      N. Yamaguchi, et al
    • Organizer
      ASM 2011 General Meeting
    • Place of Presentation
      ニューオーリンズ,アメリカ
    • Year and Date
      2011-05-23
    • Related Report
      2012 Final Research Report
  • [Presentation] Rapid monitoring of bacterial cells in freshwater by using a microfluidic system.2011

    • Author(s)
      N. Yamaguchi, et al
    • Organizer
      7th International Microbial Space Workshop
    • Place of Presentation
      クレルモン・フェラン,フランス
    • Year and Date
      2011-05-19
    • Related Report
      2012 Final Research Report
  • [Presentation] マイクロ流路システムを用いた水環境中の細菌の迅速モニタリング.2011

    • Author(s)
      山口 進康,他
    • Organizer
      日本薬学会第131 年会
    • Place of Presentation
      静岡
    • Year and Date
      2011-03-30
    • Related Report
      2012 Final Research Report
  • [Presentation] マイクロ流路システムを用いた水環境中の細菌の迅速モニタリング2011

    • Author(s)
      山口進康, 他
    • Organizer
      日本薬学会第131年会
    • Place of Presentation
      静岡
    • Year and Date
      2011-03-30
    • Related Report
      2010 Annual Research Report
  • [Presentation] Rapid monitoring of Legionella pneumophila in aquatic environment by using a microfluidic device.2010

    • Author(s)
      N. Yamaguchi, et al
    • Organizer
      ISME-13
    • Place of Presentation
      シアトル,アメリカ
    • Year and Date
      2010-08-26
    • Related Report
      2012 Final Research Report
  • [Presentation] Rapid monitoring of Legionella pneumophila in aquatic environment by using a microfluidic device2010

    • Author(s)
      Yamaguchi, N., et al.
    • Organizer
      ISME-13
    • Place of Presentation
      シアトル
    • Year and Date
      2010-08-26
    • Related Report
      2010 Annual Research Report
  • [Presentation] Rapid enumeration of Legionella pneumophsla in aquatic environment by using a microfluidic device2010

    • Author(s)
      Yamaguchi, N., et al.
    • Organizer
      ASM 2010 General Meeting
    • Place of Presentation
      サンディエゴ
    • Year and Date
      2010-05-25
    • Related Report
      2010 Annual Research Report
  • [Presentation] Rapid enumeration of Legionella pneumophila in aquatic environment by using a microfluidic device.2010

    • Author(s)
      N. Yamaguchi, et al
    • Organizer
      ASM 2010 General Meeting
    • Place of Presentation
      サンディエゴ,アメリカ
    • Year and Date
      2010-05-15
    • Related Report
      2012 Final Research Report
  • [Presentation] マイクロ流路システムを用いた水環境中の Legionella pneumophilaの迅速モニタリング2010

    • Author(s)
      山口 進康,他
    • Organizer
      日本薬学会第130 年会
    • Place of Presentation
      岡山
    • Year and Date
      2010-03-30
    • Related Report
      2012 Final Research Report
  • [Presentation] マイクロ流路システムを用いた水環境中のLegionella pneumophilaの迅速モニタリング2010

    • Author(s)
      山口進康, 他
    • Organizer
      日本薬学会第130年会
    • Place of Presentation
      岡山
    • Year and Date
      2010-03-30
    • Related Report
      2009 Annual Research Report
  • [Presentation] 冷却塔水中のレジオネラのマイクロ流路デバイスによる迅速モニタリング.2010

    • Author(s)
      山口 進康,他
    • Organizer
      第83回日本細菌学会総会
    • Place of Presentation
      横浜
    • Year and Date
      2010-03-29
    • Related Report
      2012 Final Research Report
  • [Presentation] 冷却塔水中のレジオネラのマイクロ流路デバイスによる迅速モニタリング2010

    • Author(s)
      山口進康, 他
    • Organizer
      第83回日本細菌学会総会
    • Place of Presentation
      横浜
    • Year and Date
      2010-03-29
    • Related Report
      2009 Annual Research Report
  • [Presentation] マイクロ流路デバイスを用いた水環境中のレジオネラの迅速検出2009

    • Author(s)
      山口 進康,他
    • Organizer
      日本微生物生態学会第25回大会
    • Place of Presentation
      広島
    • Year and Date
      2009-11-22
    • Related Report
      2012 Final Research Report 2009 Annual Research Report
  • [Presentation] マイクロ流路デバイスを用いた水環境中のレジオネラの迅速モニタリング2009

    • Author(s)
      山口 進康,他
    • Organizer
      フォーラム 2009:衛生薬学・環境トキシコロジー
    • Place of Presentation
      沖縄
    • Year and Date
      2009-11-05
    • Related Report
      2012 Final Research Report
  • [Presentation] マイクロ流路デバイスを用いた水環境中のレジオネラの迅速モニタリング2009

    • Author(s)
      山口進康, 他
    • Organizer
      フォーラム2009:衛生薬学・環境トキシコロジー
    • Place of Presentation
      沖縄
    • Year and Date
      2009-11-05
    • Related Report
      2009 Annual Research Report
  • [Presentation] Rapid detection of Legionella pneumophila in aquatic environment by using a microfluidic device.2009

    • Author(s)
      N. Yamaguchi, et al.
    • Organizer
      Legionella 2009
    • Place of Presentation
      パリ,フランス
    • Year and Date
      2009-10-16
    • Related Report
      2012 Final Research Report
  • [Presentation] Rapid detection of Legionella pneumophila in aquatic environment by using a microfluidic device2009

    • Author(s)
      Yamaguchi, N., et al.
    • Organizer
      Legionella 2009
    • Place of Presentation
      パリ、フランス
    • Year and Date
      2009-10-16
    • Related Report
      2009 Annual Research Report

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Published: 2009-04-01   Modified: 2016-09-09  

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