Environmental adaptation mechanisms of Clostridium difficile (CD)

• Project/Area Number: 17K08822
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Bacteriology (including mycology)
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: SAITO Ryoichi 東京医科歯科大学, 大学院医歯学総合研究科, 准教授 (00581969)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2019: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2017: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: クロストリジウム・ディフィシル / デオキシコール酸 / 芽胞 / 毒素 / ラクトバシラス属 / 環境適応 / 緊縮応答 / トランスクリプトーム解析 / 細胞毒性試験 / 増殖 / トキシン / トランスクリプトーム

Outline of Final Research Achievements

Clostridium difficile causes a spectrum of symptoms, ranging from mild diarrhea to severe pseudomembranous colitis and even death. C. difficile infection (CDI) is a well-known cause of health-care-associated infectious diarrhea related to the disruption of the indigenous intestinal microbiota due to prolonged drug treatment. We found that: DCA inhibited efficient C. difficile vegetative growth in a dose-dependent manner; DCA regulated toxin production under vegetative growth conditions by repressing tcdB expression; DCA reduced C. difficile sporulation efficiency via spo0A downregulation; and DCA downregulated spore coat-related genes. Furthermore, we also demonstrated that Lactobacillus leichmunnii strain ATCC 7830 impacted the C. difficile gene expressions such as toxin production, amino acid metabolism, and membrane transport.

Academic Significance and Societal Importance of the Research Achievements

CD感染症は先進国に加え、近年は途上国でも増加が認められるため、世界的に注視すべき重要な感染症である。本研究により、CDの環境適応機構で最も重要な発芽・芽胞形成に関わる生理学的機構や臨床にも有益な毒素産生機構の基盤情報を提供できた。これらの成果は、上記の機構に関わる主要タンパク質の機能解析等を通して、CD感染症の新たな治療戦略や感染対策についても貢献できる可能性を示す。

Research Products

• [Journal Article] Epidemiology and virulence-associated genes of Clostridioides difficile isolates and factors associated with toxin EIA results at a university hospital in Japan (2020)
  • Author(s): Okada Yuta、Yagihara Yuka、Wakabayashi Yoshitaka、Igawa Gene、Saito Ryoichi、Higurashi Yoshimi、Ikeda Mahoko、Tatsuno Keita、Okugawa Shu、Moriya Kyoji
  • Journal Title: Access Microbiology Volume: 2 Issue: 2 Pages: 1-9
  • DOI: 10.1099/acmi.0.000086
  • Peer Reviewed / Open Access
• [Journal Article] Long-term observation of antimicrobial susceptibility and molecular characterisation of Campylobacter jejuni isolated in a Japanese general hospital 2000?2017 (2019)
  • Author(s): Yamada Kageto、Saito Ryoichi、Muto Saori、Sasaki Masakazu、Murakami Hinako、Aoki Kotaro、Ishii Yoshikazu、Tateda Kazuhiro
  • Journal Title: Journal of Global Antimicrobial Resistance Volume: 18 Pages: 59-63
  • DOI: 10.1016/j.jgar.2019.02.001
  • Peer Reviewed
• [Journal Article] Infective endocarditis caused by Cardiobacterium valvarum (2019)
  • Author(s): Washio Yohei、Sakamoto Shun-ichiro、Saito Ryoichi、Nei Takahito、Morishima Masayo、Shinoyama Akihiro、Tashiro Ayaka、Sugimoto Ryoji
  • Journal Title: Access Microbiology Volume: 1 Issue: 8
  • DOI: 10.1099/acmi.0.000040
  • Peer Reviewed / Open Access
• [Journal Article] Whole-genome analysis of EC129, an NDM-5-, CTX-M-14-, OXA-10- and MCR-1-co-producing Escherichia coli ST167 strain isolated from Japan (2019)
  • Author(s): Nukui Yoko, Ayibieke Alafate, Taniguchi Makoto, Aiso Yoshibumi, Shibuya Yuka, Sonobe Kazunari, Nakajima Jun. Kanehira Saki, Hadano Yoshiro, Tohda Shuji, Koike Ryuji, Saito Ryoichi
  • Journal Title: Journal of Global Antimicrobial Resistance Volume: 18 Pages: 148-150
  • DOI: 10.1016/j.jgar.2019.07.001
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Gene Composition as a Potential Barrier to Large Recombinations in the Bacterial Pathogen Klebsiella pneumoniae (2019)
  • Author(s): Comandatore Francesco、Sassera Davide、Bayliss Sion C、Scaltriti Erika、Gaiarsa Stefano、Cao Xiaoli、Gales Ana C、Saito Ryoichi、Pongolini Stefano、Brisse Sylvain、Feil Edward J、Bandi Claudio
  • Journal Title: Genome Biology and Evolution Volume: 11 Issue: 11 Pages: 3240-3251
  • DOI: 10.1093/gbe/evz236
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Molecular epidemiologic and clinical analysis of Helicobacter cinaedi bacteremia in Japan (2019)
  • Author(s): Nukui Yoko, Chino Takashi, Tani Chihiro, Sonobe Kazunari, Aiso Yoshibumi, Tohda Shuji, Koike Ryuji, Saito Ryoichi
  • Journal Title: Helicobacter Volume: 25 Issue: 1 Pages: 23-27
  • DOI: 10.1111/hel.12675
  • Peer Reviewed / Open Access
• [Journal Article] Hypervirulent clade 2, ribotype 019/sequence type 67 Clostridioides difficile strain from Japan (2019)
  • Author(s): Saito Ryoichi, Usui Yukino, Ayibieke Alafate, Nakajima Jun, Prah Isaac, Sonobe Kazunari, Aiso Yoshibumi, Ito Shiori, Itsui Yasuhiro, Hadano Yoshiro, Nukui Yoko, Koike Ryuji, Tohda Shuji
  • Journal Title: Gut Pathogens Volume: 11 Issue: 1 Pages: 54-61
  • DOI: 10.1186/s13099-019-0336-3
  • Peer Reviewed / Open Access
• [Journal Article] Molecular characterisation of the NDM-1-encoding plasmid p2189-NDM in an Escherichia coli ST410 clinical isolate from Ghana (2018)
  • Author(s): Ayibieke Alafate、Sato Wakana、Mahazu Samiratu、Prah Isaac、Addow-Thompson John、Ohashi Mitsuko、Suzuki Toshihiko、Iwanaga Shiroh、Ablordey Anthony、Saito Ryoichi
  • Journal Title: PLOS ONE Volume: 13 Issue: 12 Pages: 0209623-0209623
  • DOI: 10.1371/journal.pone.0209623
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Novel mechanism responsible for high-level macrolide resistance in <em>Moraxella catarrhalis</em> (2018)
  • Author(s): Kasai Ayako、Ohta Ayaka、Maeda Yuina、Yamada Kageto、Tao Kazuyuki、Saito Ryoichi
  • Journal Title: Infection and Drug Resistance Volume: Volume 11 Pages: 2137-2140
  • DOI: 10.2147/idr.s181714
  • Peer Reviewed / Open Access
• [Journal Article] Clinical and microbiological characteristics and occurrence of <em>Klebsiella pneumoniae</em> infection in Japan (2018)
  • Author(s): Ikeda Mahoko、Mizoguchi Miyuki、Oshida Yukie、Tatsuno Keita、Saito Ryoichi、Okazaki Mitsuhiro、Okugawa Shu、Moriya Kyoji
  • Journal Title: International Journal of General Medicine Volume: Volume 11 Pages: 293-299
  • DOI: 10.2147/ijgm.s166940
  • Peer Reviewed / Open Access
• [Journal Article] RelA/DTD-mediated regulation of spore formation and toxin production by Clostridium perfringens type A strain SM101 (2018)
  • Author(s): Saito Ryoichi、Talukdar Prabhat K.、Alanazi Saud S.、Sarker Mahfuzur R.
  • Journal Title: Microbiology Volume: 164 Issue: 5 Pages: 835-847
  • DOI: 10.1099/mic.0.000655
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] The inhibitory effects of essential oil constituents against germination, outgrowth and vegetative growth of spores of Clostridium perfringens type A in laboratory medium and chicken meat (2018)
  • Author(s): Alanazi Saud、Alnoman Maryam、Banawas Saeed、Saito Ryoichi、Sarker Mahfuzur R.
  • Journal Title: Food Microbiology Volume: 73 Pages: 311-318
  • DOI: 10.1016/j.fm.2018.02.003
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Antimicrobial susceptibility to β-lactam antibiotics and production of BRO β-lactamase in clinical isolates of Moraxella catarrhalis from a Japanese hospital (2017)
  • Author(s): Yamada Kageto、Arai Katsumi、Saito Ryoichi
  • Journal Title: Journal of Microbiology, Immunology and Infection Volume: 50 Issue: 3 Pages: 386-389
  • DOI: 10.1016/j.jmii.2016.08.003
  • Peer Reviewed
• [Journal Article] Molecular Characterization of Fluoroquinolone-Resistant Moraxella catarrhalis Variants Generated In Vitro by Stepwise Selection (2017)
  • Author(s): Yamada Kageto、Saito Ryoichi、Muto Saori、Kashiwa Machiko、Tamamori Yoshiko、Fujisaki Shingo
  • Journal Title: Antimicrobial Agents and Chemotherapy Volume: 61 Issue: 10
  • DOI: 10.1128/aac.01336-17
  • Peer Reviewed
• [Journal Article] 表現型からみるcarbapenemase-producing Enterobacteriaceaeの網羅的スクリーニング法 (2017)
  • Author(s): 山田景土、長野則之、齋藤良一
  • Journal Title: 日本臨床微生物学雑誌 Volume: 27 Pages: 267-274
  • NAID: 40021347014
  • Peer Reviewed
• [Journal Article] Evaluation of the modified carbapenem inactivation method and sodium mercaptoacetate-combination method for the detection of metallo-b-lactamase production by carbapenemase-producing Enterobacteriaceae. (2016)
  • Author(s): Yamada K, Kashiwa M, Arai K, Nagano N, Saito R.
  • Journal Title: Journal of Microbiological Methods Volume: 132 Pages: 112-115
  • DOI: 10.1016/j.mimet.2016.11.013
  • Peer Reviewed
• [Presentation] Virulence Profile of Diarrhegenic E. coli from the Western region of Ghana (2020)
  • Author(s): Isaac Prah, Alafate Ayibieke, Atsushi Iguchi Samiratu Mahazu, Wakana Sato, Toshihiko Suzuki, Shoji Yamaoka, Shiroh Iwanaga, Anthony Ablordey, Ryoichi Saito
  • Organizer: 第93回日本細菌学会総会
• [Presentation] Epidemiological characteristics of bloodstream S. epidermidis isolates from two hospitals in Tokyo (2020)
  • Author(s): Alafate Ayibieke, Kageto Yamada, Yoshibumi Aiso, Yoshiro Hadano, Yoko Nukui, Ryuji Koike, Shuji Tohda, Ryoichi Saito
  • Organizer: 第93回日本細菌学会総会
• [Presentation] 本邦初のNDM-5メタロ-β-ラクターマーゼ産生大腸菌ST167による感染事例 (2019)
  • Author(s): 貫井陽子, 羽田野義郎, 齋藤良一, 相曽啓史, 東田修二, 小池竜司
  • Organizer: 第93回日本感染症学会学術講演会
• [Presentation] Antimicrobial susceptibility and extended-spectrum β-lactamase profiles of Escherichia coli and Klebsiella spp. isolated from Ghana (2019)
  • Author(s): Wakana Sato, Samiratu Mahazu, Isaac Prah, John Addow-Thompson, Alafate Ayibieke, Shiroh Iwanaga, Anthony Ablordy, Ryoichi Saito
  • Organizer: The 23rd IFCC-EFLM European Congress of Clinical Chemistry and Laboratory Medicine
  • Int'l Joint Research
• [Presentation] Impact of deoxycholate on the growth, toxin production, and spore formation of Clostridioides difficile (2019)
  • Author(s): Yukino Usui, Yuko Kamiichi, Shu Okugawa, Ryoichi Saito
  • Organizer: The 23rd IFCC-EFLM European Congress of Clinical Chemistry and Laboratory Medicine
  • Int'l Joint Research
• [Presentation] Antibiotic resistance among Gram-negative glucose non-fermenting bacilli from Ghana (2019)
  • Author(s): Alafate Ayibieke, Ayumi Kobayashi, Samiratu Mahazu, Wakana Sato, Isaac Prah, John Addow-Thompson, Shiroh Iwanaga, Anthony Ablordy, Ryoichi Saito
  • Organizer: The 23rd IFCC-EFLM European Congress of Clinical Chemistry and Laboratory Medicine
  • Int'l Joint Research
• [Presentation] Clinical features, molecular epidemiology, and virulence-associated genes of Clostridioides difficile isolates in patients at a University Hospital in Japan (2019)
  • Author(s): Yuta Okada, Yuka Yagihara, Yoshitaka Wakabayashi, Gene Igawa, Ryoichi Saito, Yoshimi Higurashi, Mahoko Ikeda, Keita Tatsuno, Shu Okugawa, Kyoji Moriya
  • Organizer: ASM Microbe 2019
  • Int'l Joint Research
• [Presentation] Molecular Epidemiological Analysis of Multidrug-Resistant Pseudomonas aeruginosa that Caused an Outbreak in an Intensive Care Unit in Japan (2019)
  • Author(s): Y. Nukui, Y. Maki, Y. Hadano, Y. Aiso, J. Nakajima, K. Sonobe, R. Saito, S. Tohda, R. Koike
  • Organizer: ASM Microbe 2019
  • Int'l Joint Research
• [Presentation] 感染症の病原体核酸検査の現状と将来展望－細菌の遺伝子型解析による院内感染制御－ (2019)
  • Author(s): 齋藤良一
  • Organizer: 日本臨床検査医学会 第75回関東・甲信越支部例会
  • Invited
• [Presentation] ガーナ共和国のフルオロキノロン系薬耐性、基質特異性拡張型β-ラクタマーゼ産生Escherichia coliに関する分子疫学研究 (2019)
  • Author(s): 岩澤茉莉絵, 佐藤和佳菜, Alafate Ayibieke, Isaac Prah, Samiratu Mahazu, 岩永史朗, Anthony Ablordey, 齋藤良一
  • Organizer: 第14回日本臨床検査学教育学会学術大会
• [Presentation] デオキシコール酸に対するClostridioides difficileの代謝調節 (2019)
  • Author(s): 臼井雪乃, 上市裕子,奥川 周, 東田修二, 齋藤良一
  • Organizer: 第49回日本嫌気性菌感染症学会総会・学術集会
• [Presentation] 血液培養陽性例へのASによりみえてくるものは (2019)
  • Author(s): 相曽啓史, 渋谷有香, 丸谷美保子, 鳥羽美貴子, 森美奈子, 齋藤良一, 羽田野義郎, 貫井陽子, 高橋弘充
  • Organizer: 第34回日本環境感染学会総会・学術集会
• [Presentation] Emergence of an NDM-1-producing Escherichia coli from Ghana (2019)
  • Author(s): Alafate Ayibieke, Wakana Sato, Samiratu Mahazu, Mitsuko Ohhashi, Toshihiko Suzuki, Shiroh Iwanaga, Anthony Ablordey, Ryoichi Saito
  • Organizer: 第30回日本臨床微生物学会総会・学術集会
• [Presentation] Moraxella catarrhalis同定法の評価 (2019)
  • Author(s): 武藤沙起里, 山田景土, 西山宏幸, 齋藤良一
  • Organizer: 第30回日本臨床微生物学会総会・学術集会
• [Presentation] ガーナ共和国の臨床分離株における腸内細菌科細菌の薬剤感受性成績 (2019)
  • Author(s): 佐藤和佳菜, Alafate Ayibieke, 小林あゆみ, 谷千尋, 日暮芳己, 佐藤智明, 森屋恭爾, 鈴木敏彦, 大橋光子, 岩永史朗, 齋藤良一
  • Organizer: 第30回日本臨床微生物学会総会・学術集会
• [Presentation] デオキシコール酸がClostridioides difficileの増殖能、毒素産生能および芽胞形成能に与える影響 (2019)
  • Author(s): 臼井雪乃, 上市裕子,内田のぞみ, 奥川 周, 園部一成, 東田修二, 齋藤良一
  • Organizer: 第30回日本臨床微生物学会総会・学術集会
• [Presentation] Moraxella catarrhalisのβ-ラクタマーゼ阻害剤配合薬耐性に関わる分子機構 (2019)
  • Author(s): 太田彩花, 大塚 武, 長谷川瑛美, 笠井彩子, 山田景土, 山口佳宏, 和知野純一, 木村幸司, 荒川 宜親, 齋藤良一
  • Organizer: 第30回日本臨床微生物学会総会・学術集会
• [Presentation] Cardiobacterium valvarumによる感染性心内膜炎の一例 (2018)
  • Author(s): 武井美澪, 鷲尾洋平, 篠山明宏, 井上淳, 遠藤康実, 根井貴仁, 齋藤良一
  • Organizer: 第14回東京都医学検査学会
• [Presentation] アモキシリン/クラブラン酸耐性Moraxella catarrhalisが有する耐性機構の解析 (2018)
  • Author(s): 太田彩花, 大塚 武, 長谷川瑛美, 笠井彩子, 山田景土, 山口佳宏, 和知野純一, 木村幸司, 齋藤良一
  • Organizer: 第55回関甲信支部・首都圏支部医学検査学会
• [Presentation] ガーナ共和国の臨床分離株における薬剤感受性成績 (2018)
  • Author(s): 佐藤和佳菜, Alafate Ayibieke, 小林あゆみ, 谷千尋, 齋藤良一
  • Organizer: 第55回関甲信支部・首都圏支部医学検査学会
• [Presentation] Clostridium difficileの増殖、毒素産生および芽胞形成に対する胆汁酸の影響 (2018)
  • Author(s): 臼井雪乃, 上市裕子, 内田のぞみ, 奥川 周, 園部一成, 東田修二, 齋藤良一
  • Organizer: 第55回関甲信支部・首都圏支部医学検査学会
• [Presentation] Molecular epidemiological analysis of Clostridium difficile that caused an outbreak in a Japanese hospital (2018)
  • Author(s): Y. Nukui, Y. Aiso, T. Chino, R. Saito, C. Tani, J. Nakajima, K. Sonobe, S. Tohda, R. Koike
  • Organizer: ASM Microbe 2018
  • Int'l Joint Research
• [Presentation] Genetic profile in the development of macrolide resistance in Moraxella catarrhalis (2018)
  • Author(s): Ayaka Ohta, Ayako Kasai, Yuina Maeda, Kageto Yamada, Ryoichi Saito
  • Organizer: ASM Microbe 2018
  • Int'l Joint Research
• [Presentation] Dissemination of qac genes among Staphylococcus epidermidis isolates from a University teaching hospital in Tokyo (2018)
  • Author(s): Alafate Ayibieke, Miyuki Mizoguchi, Kageto Yamada, Tomoaki Sato, Chino Takashi, Yoshibumi Aiso, Toshihide Fujie, Yoko Nukui, Kyoji Moriya, Ryuji Koike, Shuji Tohda, Ryoichi Saito
  • Organizer: ASM Microbe 2018
  • Int'l Joint Research
• [Presentation] First clinical isolation of NDM-1 carbapenemase-producing Escherichia coli in Ghana (2018)
  • Author(s): Alafate Ayibieke, Wakana Sato, Samiratu Mahazu, Isaac Prah, John Thompson, Chihiro Tani, Mitsuko Ohashi, Shiroh Iwanaga, Toshihiko Suzuki, Anthony Ablordey, Ryoichi Saito
  • Organizer: ASM Microbe 2018
  • Int'l Joint Research
• [Presentation] RelA/DTD-mediated regulation of spore formation and toxin production by Clostridium perfringens type A strain SM101 (2018)
  • Author(s): Ryoichi Saito, Prabhat K. Talukdar, Saud S. Alanazi, Mahfuzur R. Sarker
  • Organizer: ASM Microbe 2018
  • Int'l Joint Research
• [Presentation] The inhibitory effects of essential oil constituents against germination, outgrowth and vegetative growth of spores of Clostridium perfringens type A in laboratory medium and chicken meat (2018)
  • Author(s): Saud Alanazi, Maryam Alnoman, Saeed Banawas, Ryoichi Saito, Mahfuzur R. Sarker
  • Organizer: ASM Microbe 2018
  • Int'l Joint Research
• [Presentation] CDアウトブレイクの分子疫学解析 (2018)
  • Author(s): 貫井陽子, 相曽啓史, 齋藤良一, 谷千尋, 東田修二, 小池竜司
  • Organizer: 第92回日本感染症学会学術講演会
• [Presentation] Amoxicillin/clavulanic acid低感受性Moraxella catarrhalisが分離された一例 (2018)
  • Author(s): 大塚 武、静野健一、太田彩花、笠井彩子、山田景土、齋藤良一
  • Organizer: 第67回日本医学検査学会
• [Presentation] Carbapenemase-producing Enterobacteriaceaeスクリーニング法の比較検討 Modified-Hodge test vs Carba NP test vs carbapenem inactivation method (2017)
  • Author(s): 山田景土, 柏 真知子, 長野則之, 齋藤良一
  • Organizer: 第29回日本臨床微生物学会総会・学術集会
• [Presentation] Helicobacter cinaedi菌血症の臨床的・分子疫学的解析 (2017)
  • Author(s): 貫井陽子, 齋藤良一, 谷 千尋, 相曽啓史, 藤江俊秀, 東田修二, 小池竜司, 茅野崇
  • Organizer: 第91回日本感染症学会総会・学術講演会
• [Presentation] 細菌の薬剤耐性化 (2017)
  • Author(s): 齋藤良一
  • Organizer: 第12回日本臨床検査学教育学会学術大会
  • Invited
• [Presentation] The Clostridium perfringens RelA and DTD control sporulation (2017)
  • Author(s): Ryoichi Saito、Mahfuzur R. Sarker
  • Organizer: The 29th World Congress of World Association of Societies of Pathology and Laboratory Medicine
  • Int'l Joint Research
• [Presentation] Donor of mosaic pena gene of ceftriaxone resistant neisseria gonorrhoeae fc428 and gu140106 (2017)
  • Author(s): Makoto Ohnishi, Gene Igawa, Misato Dorin, Ken Shimuta, Ryoichi Saito, Shu-Ichi Nakayama
  • Organizer: STI and HIV 2017 World Congress
  • Int'l Joint Research