Development of comprehensive quantification method for pathogenic bacteria that contributes to the clarification of the occurrence of pathogenic bacteria in the aquatic environment

• Project/Area Number: 25630226
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Civil and environmental engineering
• Research Institution: Kitasato University
• Principal Investigator: INOUE Daisuke 北里大学, 医療衛生学部, 准教授 (70448091)
• Co-Investigator(Kenkyū-buntansha): SEI Kazunari 北里大学, 医療衛生学部, 教授 (80324177) ; IWASHITA Masato 北里大学, 医療衛生学部, 助教 (00137895)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2013: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
• Keywords: 病原性細菌 / 水環境 / 網羅的定量解析 / DNAマイクロアレイ / DNAインターカレーター処理 / 網羅的解析 / DNAマイクロアレイ法

Outline of Final Research Achievements

This study aimed to develop a comprehensive quantification method for over 900 species of pathogenic bacteria and to clarify the occurrence of those pathogenic bacteria in the aquatic environment using the developed method. To these aims, this study tried to develop a selective detection method of viable bacteria in water using DNA intercalating treatment, to develop a quantitative DNA microarray method to simultaneously quantify over 900 pathogenic bacterial species, and to understand the occurrence of pathogenic bacteria in aquatic environments in Japan and other countries. Results of this study clarified the DNA intercalating treatment conditions useful for various bacterial species to some extent, and showed some important hints for the establishment of a quantitative DNA microarray method. In addition, this study could grasp the occurrence of over 900 pathogenic bacterial species in the Sagami River basin and in water samples collected from the Kathmandu valley, Nepal.

Research Products

• [Journal Article] High-throughput DNA microarray detection of pathogenic bacteria in shallow well groundwater in the Kathmandu Valley, Nepal (2015)
  • Author(s): Daisuke Inoue, Takuji Hinoura, Noriko Suzuki, Junqin Pang, Rabin Malla, Sadhana Shrestha, Saroj Kumar Chapagain, Hiroaki Matsuzawa, Takashi Nakamura, Yasuhiro Tanaka, Michihiko Ike, Kei Nishida, Kazunari Sei
  • Journal Title: Current Microbiology Volume: 70 Issue: 1 Pages: 43-50
  • DOI: 10.1007/s00284-014-0681-x
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] ネパール・カトマンズ盆地の各種水試料中における病原性細菌汚染の実態調査 (2015)
  • Author(s): 吉永隼人, 井上大介, Bikash Malla, 田中靖浩, Jeevan B. Sherchand, 原本英司, 清和成
  • Organizer: 日本水処理生物学会第52回大会
  • Place of Presentation: 福岡県北九州市（北九州国際会議場）
  • Year and Date: 2015-11-11
• [Presentation] Monitoring of bacterial pathogens in Sagami River basin for the grasp of the river water pollution by bacterial pathogens (2015)
  • Author(s): Kazunari Sei, Hayato Yoshinaga, Tomoharu Kamibayashi, Kaori Miyauchi, Takuji Hinoura, Daisuke Inoue
  • Organizer: 18th International Symposium on Health-Related Water Microbiology (WaterMicro2015)
  • Place of Presentation: Lisbon, Portugal（Faculdade de Medicina Dentaria）
  • Year and Date: 2015-09-13
  • Int'l Joint Research
• [Presentation] Comprehensive analysis of pathogenic bacteria occurred in jar water, community well groundwater and river water in the Kathmandu Valley, Nepal (2015)
  • Author(s): Hayato Yoshinaga, Daisuke Inoue, Bikash Malla, Rajani Ghaju, Dinesh Bhandari, Sarmila Tandukar, Yasuhiro Tanaka, Jeevan B. Sherchand, Eiji Haramoto, Kazunari Sei
  • Organizer: Water and Environment Technology Conference 2015
  • Place of Presentation: 東京都千代田区（日本大学駿河台キャンパス）
  • Year and Date: 2015-08-05
  • Int'l Joint Research
• [Presentation] DNAマイクロアレイを用いた相模川水系の病原性細菌汚染の調査 (2014)
  • Author(s): 吉永隼人, 上林智遥, 宮内華織, 日野浦拓之, 井上大介, 清和成
  • Organizer: 日本水処理生物学会第51回大会
  • Place of Presentation: 山梨県甲府市（山梨県JA会館）
  • Year and Date: 2014-11-12 – 2014-11-14
• [Presentation] 様々な細菌種の生菌選択検出に適用可能なDNAインターカレーター処理法の開発 (2014)
  • Author(s): 井上大介, 上林智遥, 吉永隼人, 清和成
  • Organizer: 第48回日本水環境学会年会
  • Place of Presentation: 東北大学川内北キャンパス（宮城県仙台市）
• [Presentation] Comprehensive detection of pathogenic bacteria in shallow groundwater in Kathmandu Valley, Nepal, using DNA microarray (2013)
  • Author(s): Takuji Hinoura, Noriko Suzuki, Junqin Pang, Rabin Malla, Shrestha Malla, Saroj K. Chapagain, Hiroaki Matsuzawa, Yasuhiro Tanaka, Daisuke Inoue, Michihiko Ike, Kei Nishida, Kazunari Sei
  • Organizer: 6th Advanced Engineering Technology for Environment and Energy－Environment, Energy and Sustainable Development
  • Place of Presentation: 大阪大学吹田キャンパス（大阪府吹田市）
• [Presentation] DNA microarray analysis of bacterial pathogens in shallow well groundwater in the Kathmandu Valley, Nepal (2013)
  • Author(s): Kazunari Sei, Takuji Hinoura, Kaori Miyauchi, Saroj K. Chapagain, Hiroaki Matsuzawa, Takashi Nakamura, Yasuhiro Tanaka, Daisuke Inoue, Kei Nishida
  • Organizer: 17th International Symposium on Health-Related Water Microbiology
  • Place of Presentation: Majestic Palace Hotel (Florianopolis, Brazil)
• [Presentation] DNAマイクロアレイを用いた相模原水系における病原性細菌分布の網羅的解析 (2013)
  • Author(s): 井上大介, 上林智遥, 吉永隼人, 宮内華織, 日野浦拓之, 清和成
  • Organizer: 日本水処理生物学会第50回大会
  • Place of Presentation: 神戸市水道局たちばな職員研修センター（兵庫県神戸市）