Tracking of antibiotic resistant bacteria in water environment by using genetic and chemical tracers

• Project/Area Number: 19H02272
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 22060:Environmental systems for civil engineering-related
• Research Institution: Kanazawa University
• Principal Investigator: Honda Ryo 金沢大学, 地球社会基盤学系, 教授 (40422456)
• Co-Investigator(Kenkyū-buntansha): 渡部 徹 山形大学, 農学部, 教授 (10302192) ; 西山 正晃 山形大学, 農学部, 准教授 (10802928) ; 原 宏江 金沢大学, 地球社会基盤学系, 助教 (70823524) ; 松浦 哲久 金沢大学, 地球社会基盤学系, 准教授 (90771585) ; 田中 宏明 京都大学, 工学研究科, 教授 (70344017)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000); Fiscal Year 2021: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000); Fiscal Year 2020: ¥6,760,000 (Direct Cost: ¥5,200,000、Indirect Cost: ¥1,560,000); Fiscal Year 2019: ¥7,150,000 (Direct Cost: ¥5,500,000、Indirect Cost: ¥1,650,000)
• Keywords: 薬剤耐性 / 雨天時下水越流水 / 都市河川 / 微生物群集 / 薬剤耐性遺伝子 / 微生物起源追跡 / 薬剤耐性菌 / 残留医薬品類 / 排出源追跡 / 水環境 / 合流式下水道

Outline of Research at the Start

環境中の薬剤耐性菌の多様な排出源からの拡散動態を把握することを目的として，
（１）耐性遺伝子と微量化学物質を用いた排出源追跡手法の開発，
（２）開発した新規手法の適用による薬剤耐性菌の排出源からの拡散動態の解析，を行う。
薬剤耐性遺伝子の配列相同性に基づく進化的プロファイルを用いて，自然由来と流域間の広域移動による長い時間スケールでの薬剤耐性菌の拡散を把握する手法と，微量化学物質マーカーによる分解や沈降を考慮した，より短い時間スケールでの拡散動態を把握する手法を開発し，その手法を実際の流域に適用して，各流域における多様な排出源からの薬剤耐性菌の拡散動態の時間的・空間的スケールを明らかにする。

Outline of Final Research Achievements

A survey of the target urban river revealed that the proportion of antimicrobial resistance genes increased uniformly from upstream to downstream during outflow of combined sewage overflow during wet weather. The proportion of mobile genetic elements, which are associated with spread of antimicrobial resistance among different species by horizontal gene transfer, also increased during wet weather. The in vitro experiment revealed that β-lactamase-producing genes can be horizontally transmitted to other bacteria in the sediment environment. These results indicate that the release of combined sewage overflow during wet weather not only releases a large quantity of antimicrobial resistance genes, but also possibly increase the chance in spread of antimicrobial resistance by horizontal gene transfer.

Academic Significance and Societal Importance of the Research Achievements

本研究の成果により，合流式下水道雨天時越流水が河川環境中における薬剤耐性の拡散動態に与えている影響が明らかになった。都市下水は，環境中に拡散する薬剤耐性の主要な排出源であり，その拡散の低減において，雨天時下水越流水の制御が重要であることが明らかになった。また，環境中における遺伝子水平伝搬による細菌間での薬剤耐性の拡散について，底泥環境下で一定の割合で起こりうることが示唆され，雨天時下水越流水によって，水平伝搬の要因となる遺伝子の放流量も増えることが明らかになった。以上のことは，ワンヘルスの考えのもとで環境中の薬剤耐性の拡散を抑制するために重要な知見である。

Research Products

• [Int'l Joint Research] UPES(インド)
• [Int'l Joint Research] ルフナ大学(スリランカ)
• [Int'l Joint Research] カセサート大学/ガジャマンガラ大学タンヤブリ校(タイ)
• [Journal Article] MOLECULAR CHARACTERIZATION OF EXTENDED-SPECTRUM BETA-LACTAMASE PRODUCING GENES OF ENTEROBACTERIACEAE ISOLATED FROM MUNCIPAL AND HOSPITAL WASTEWATERS (2021)
  • Author(s): MORI Masaya、NISHIYAMA Masateru、SHIBUKI Rio、BABA Hiroaki、KANAMORI Hajime、WATANABE Toru
  • Journal Title: Journal of Japan Society of Civil Engineers, Ser. G (Environmental Research) Volume: 77 Issue: 7 Pages: III_199-III_207
  • DOI: 10.2208/jscejer.77.7_III_199
  • ISSN: 2185-6648
  • Peer Reviewed
• [Journal Article] Fate and seasonal change of Escherichia coli resistant to different antibiotic classes at each stage of conventional activated sludge process (2020)
  • Author(s): Honda Ryo、Tachi Chihiro、Noguchi Mana、Yamamoto-Ikemoto Ryoko、Watanabe Toru
  • Journal Title: Journal of Water and Health Volume: 18 Issue: 6 Pages: 879-889
  • DOI: 10.2166/wh.2020.013
• [Journal Article] Prevalence of antibiotic resistance in the tropical rivers of Sri Lanka and India (2020)
  • Author(s): Kumar Manish、Sulfikar、Chaminda Tushara、Patel Arbind K.、Sewwandi Himaya、Mazumder Payal、Joshi Madhvi、Honda Ryo
  • Journal Title: Environmental Research Volume: 188 Pages: 109765-109765
  • DOI: 10.1016/j.envres.2020.109765
• [Journal Article] Estimated discharge of antibiotic-resistant bacteria from combined sewer overflows of urban sewage system (2020)
  • Author(s): Honda Ryo、Tachi Chihiro、Yasuda Keisuke、Hirata Tatsuki、Noguchi Mana、Hara-Yamamura Hiroe、Yamamoto-Ikemoto Ryoko、Watanabe Toru
  • Journal Title: npj Clean Water Volume: 3 Issue: 1 Pages: 15-15
  • DOI: 10.1038/s41545-020-0059-5
• [Journal Article] Seasonality impels the antibiotic resistance in Kelani River of the emerging economy of Sri Lanka (2020)
  • Author(s): Kumar Manish、Chaminda G. G. Tushara、Honda Ryo
  • Journal Title: npj Clean Water Volume: 3 Issue: 1 Pages: 12-12
  • DOI: 10.1038/s41545-020-0058-6
• [Journal Article] Treatment enhances the prevalence of antibiotic-resistant bacteria and antibiotic resistance genes in the wastewater of Sri Lanka, and India (2020)
  • Author(s): Kumar Manish、Ram Bhagwana、Sewwandi Himaya、Sulfikar、Honda Ryo、Chaminda Tushara
  • Journal Title: Environmental Research Volume: 183 Pages: 109179-109179
  • DOI: 10.1016/j.envres.2020.109179
• [Journal Article] Concurrence of antibiotic resistant bacteria (ARB), viruses, pharmaceuticals and personal care products (PPCPs) in ambient waters of Guwahati, India: Urban vulnerability and resilience perspective (2019)
  • Author(s): Kumar Manish、Ram Bhagwana、Honda Ryo、Poopipattana Chomphunut、Canh Vu Duc、Chaminda Tushara、Furumai Hiroaki
  • Journal Title: Science of The Total Environment Volume: 693 Pages: 133640-133640
  • DOI: 10.1016/j.scitotenv.2019.133640
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] A Review on Antibiotic Resistance Gene (ARG) Occurrence and Detection in WWTP in Ishikawa, Japan and Colombo, Sri Lanka (2019)
  • Author(s): Sulfikar、Sovannlaksmy Sorn、Honda Ryo、Chaminda Tushara、Kumar Manish
  • Journal Title: Emerging Issues in the Water Environment during Anthropocene: A South East Asian Perspective Volume: 1 Pages: 1-14
  • DOI: 10.1007/978-981-32-9771-5_1
  • ISBN: 9789813297708, 9789813297715
  • Peer Reviewed / Int'l Joint Research
• [Presentation] in vitro伝達実験を用いたバンコマイシン耐性遺伝子のグラム陽性細菌に対する伝播ポテンシャルの評価 (2022)
  • Author(s): 西山正晃，馬場啓聡，金森肇，渡部徹
  • Organizer: 第56回日本水環境学会年会
• [Presentation] Global comparison of antimicrobial resistance genes in wastewater treatment plants based on metagenomic analysis (2021)
  • Author(s): Yuta Morinaga, Hiroe Hara-Yamamura, Norihisa Matsuura, Toru Watanabe, Ryo Honda
  • Organizer: The 14th Annual Conference on Challenges in Environmental Science and Engineering
  • Int'l Joint Research
• [Presentation] Profiling and international comparison of antibiotic resistome in activated sludge by metagenomic data analysis (2021)
  • Author(s): Yuta Morinaga , Than Van Huy, Sawako Asakura, Sorn Sovannlaksmy, Hiroe Hara-Yamamura, Norihisa Matsuura, Toru Watanabe, Ryo Honda
  • Organizer: The 9th IWA Microbial Ecology and Water Engineering (MEWE) Specialist Conference
  • Int'l Joint Research
• [Presentation] 都市下水と病院排水中の腸内細菌科細菌が保有するESBL産生遺伝子の特徴 (2021)
  • Author(s): 西山正晃，馬場啓聡，金森肇，渡部徹
  • Organizer: 土木学会東北支部技術研究発表会
• [Presentation] Removal Mechanisms of Antibiotic Resistance Genes in Membrane Bioreactor (2020)
  • Author(s): Rongxuan Wang, Norihisa Matsuura, Hiroe Hara-Yamamura, Ryo Honda
  • Organizer: nternational Conference on Challenges in Environmental Science & Engineering
  • Int'l Joint Research
• [Presentation] Fate of Antibiotic Resistant Bacteria from Wastewater to Water Environment: From Asian Perspectives (2020)
  • Author(s): Ryo Honda
  • Organizer: 6th International Forum on Sustainable Future in Asia, National Institute for Environmental Studies, Japan
  • Int'l Joint Research / Invited
• [Presentation] Fate and Control of Antibiotic Resistant Bacteria from Domestic Wastewate (2019)
  • Author(s): Ryo Honda
  • Organizer: Pre-conference Workshop on Antibiotic Resistance in Wastewater in IWA-MEWE2019
  • Int'l Joint Research
• [Presentation] Assessment of elevated risk by antibiotic resistance in indirect reuse of treated livestock wastewater for irrigation (2019)
  • Author(s): Ryo Honda, Lin Mengyu, Tatsuki Hirata, Hiroe Hara-Yamamura, Ryoko Yamamoto-Ikemoto, Toru Watanab
  • Organizer: 12th IWA International Conference on Water Reclamation and Reuse
  • Int'l Joint Research
• [Presentation] Transition of antibiotic resistome in wastewater treatment process (2019)
  • Author(s): Ryo Honda, Norihisa Matsuura, Sorn Sovannlaksmy, Yalkhin Masakke, Hiroe Hara-Yamamura, Toru Watanabe
  • Organizer: IWA Microbial Ecology and Water Engineering 2019
  • Int'l Joint Research
• [Presentation] Aeration induces antibiotic resistance gene expression in wastewater treatment plant. (2019)
  • Author(s): Sulfikar, Sorn Sovannlaksmy, Hiroe Hara-Yamamura, Ryoko Yamamoto-Ikemoto, Toru Watanabe, Ryo Honda
  • Organizer: IWA Microbial Ecology and Water Engineering 2019
  • Int'l Joint Research
• [Book] Springer Nature, Singapore (2020)
  • Author(s): Manish Kumar, Daniel D Snow, Ryo Honda, Santanu Mukherjee
  • Total Pages: 436
  • Publisher: Contaminants in Drinking and Wastewater Sources.
  • ISBN: 9789811545993
• [Book] Emerging Issues in the Water Environment during Anthropocene: A South East Asian Perspective (2019)
  • Author(s): Manish Kumar, Daniel D. Snow, Ryo Honda
  • Total Pages: 365
  • Publisher: Springer Nature, Singapore
  • ISBN: 9789813297708
• [Funded Workshop] Pre-conference Workshop on Antibiotic Resistance in Wastewater in IWA-MEWE2019 (2019)