Analysis of natural reservoir of Escherichia albertii, an emerging zoonotic pathogen

• Project/Area Number: 20K06396
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 42020:Veterinary medical science-related
• Research Institution: Osaka Metropolitan University (2022-2023); Osaka Prefecture University (2020-2021)
• Principal Investigator: 日根野谷 淳 大阪公立大学, 大学院獣医学研究科, 准教授 (20548490)
• Co-Investigator(Kenkyū-buntansha): 山崎 伸二 大阪公立大学, 生命環境科学研究科, 教授 (70221653)
• Project Period (FY): 2020-04-01 – 2024-03-31
• Project Status: Completed (Fiscal Year 2023)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2022: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2021: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2020: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
• Keywords: Escherichia albertii / アライグマ / 自然宿主 / Eacdt / stx2 / 志賀毒素 / 野生動物 / アルバーティ細菌 / 新興人獣共通感染症菌

Outline of Research at the Start

本研究では、新興人獣共通感染症細菌Escherichia albertiiの自然界での動態およびヒトへの感染経路の解明、延いては本菌の感染予防法の構築に寄与すべく、野生動物、特にアライグマが自然宿主である可能性について分子疫学研究により評価する。具体的には、①アライグマにおけるE. albertiiの保菌調査を行う。また、保菌率とアライグマの個体情報（季節性、地域性、性差、年齢）との関係性を解析する。②アライグマ分離株と、ヒト臨床分離株の細菌学的特徴を比較解析する。③高病原性が予想される株は動物感染モデルでの評価も加えて、それらの病原性も調べる。

Outline of Annual Research Achievements

新興食中毒細菌であるEscherichia albertiiによる食中毒が世界的に報告される中、特に我が国においては集団食中毒もこれまで度々発生している。本菌は食用動物の他、様々な野生動物からも分離報告があり、本菌もまた人獣共通感染症細菌であることがわかってきたが、自然宿主については未だ同定されていなかった。本研究では、本菌の自然宿主を同定することを目的とした野生動物の保菌調査を行い、アライグマが本菌を高率に保菌することを見出した。最終年度は、分離株の性状解析、特に遺伝学的解析を行った。実施期間中にアライグマ286個体から1,362の分離株を得ることができた。個体内の多様性を簡易PCRにより調べた結果、個体内で同一の遺伝子型が多数見つかり、分離株は最終的に340菌株となった。パルスフィールドゲル電気泳動による解析の結果、340株は119種のPFGEパターンに分けられ、アライグマが遺伝学的に多様なE. albertiを保菌していることが明らかになった。一方で、個体間で同一のパターンを示す菌株も存在し、中には年を越えても維持されるクローンが存在することも明らかになった。更に、保有する臨床分離株（50株）も含めて比較すると、類似したPFGEパターンを示す例が認められた。更に、より識別能が高い方法として全ゲノムシークエンス取得し、SNPsによる菌株の多型解析を行ったところ、アライグマ由来株と患者由来株とで系統的に非常に近いことを示す結果が得られた。これらの結果は、アライグマは自然宿主としてE. albertiiを保菌・維持しており、何らかの経路を介してヒトへの感染源となっている可能性があることを示していると考えられた。

Research Products

• [Journal Article] Seasonality of detection rate of <i>Escherichia albertii</i> in wild raccoons (<i>Procyon lotor</i>) in Osaka, Japan (2024)
  • Author(s): XU Bingting、HATANAKA Noritoshi、AWASTHI Sharda Prasad、HINENOYA Atsushi、YAMASAKI Shinji
  • Journal Title: Journal of Veterinary Medical Science Volume: 86 Issue: 2 Pages: 180-183
  • DOI: 10.1292/jvms.23-0372
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed / Open Access
• [Journal Article] Endotoxin-Free Stx2B-C-CPE Vaccine and Its Optimized Adjuvant Regimen for Preventing Food Poisoning (2023)
  • Author(s): Hosomi Koji、Shimoyama Atsushi、Hinenoya Atsushi、Hatanaka Noritoshi、Noguchi Takafumi、Ebina Hirotaka、Tojima Yoko、Furuta Mari、Kondoh Masuo、Kiyono Hiroshi、Yamasaki Shinji、Fukase Koichi、Kunisawa Jun
  • Journal Title: Frontiers in Bioscience-Landmark Volume: 28 Issue: 1 Pages: 15-15
  • DOI: 10.31083/j.fbl2801015
  • Peer Reviewed / Open Access
• [Journal Article] Critical role of the RpoE stress response pathway in polymyxin resistance of<i>Escherichia coli</i> (2023)
  • Author(s): Zeng Ximin、Hinenoya Atsushi、Guan Ziqiang、Xu Fuzhou、Lin Jun
  • Journal Title: Journal of Antimicrobial Chemotherapy Volume: 78 Issue: 3 Pages: 732-746
  • DOI: 10.1093/jac/dkad003
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Detection, Isolation, and Molecular Characterization of <i>Escherichia albertii</i> from Wild Birds in West Japan (2022)
  • Author(s): Hinenoya Atsushi, Awasthi Sharda Prasad, Yasuda Noritomo, Nagano Keigo, Hassan Jayedul, Takehira Keiji, Hatanaka Noritoshi, Saito Shun, Watabe Takashi, Yoshizawa Miki, Inoue Haruna, Yamasaki Shinji.
  • Journal Title: Japanese Journal of Infectious Diseases Volume: 75 Issue: 2 Pages: 156-163
  • DOI: 10.7883/yoken.JJID.2021.355
  • NAID: 130008080699
  • ISSN: 1344-6304, 1884-2836
  • Year and Date: 2022-03-31
  • Peer Reviewed
• [Journal Article] Isolation and characterization of <i>Escherichia albertii</i> from wild and safeguarded animals in Okayama Prefecture and its prefectural borders, Japan (2022)
  • Author(s): NAKA Atsushi、HINENOYA Atsushi、AWASTHI Sharda Prasad、YAMASAKI Shinji
  • Journal Title: Journal of Veterinary Medical Science Volume: 84 Issue: 9 Pages: 1299-1306
  • DOI: 10.1292/jvms.22-0213
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed / Open Access
• [Journal Article] Longitudinal surveillance and comparative characterization of Escherichia albertii in wild raccoons in the United States (2022)
  • Author(s): Hinenoya Atsushi、Wang Huiwen、Patrick Erin M.、Zeng Ximin、Cao Liu、Li Xing-Ping、Lindsey Rebecca L.、Gillespie Barbara、He Qiang、Yamasaki Shinji、Lin Jun
  • Journal Title: Microbiological Research Volume: 262 Pages: 127109-127109
  • DOI: 10.1016/j.micres.2022.127109
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Piperine, an active ingredient of white pepper, suppresses the growth of multidrug‐resistant toxigenic Vibrio cholerae and other pathogenic bacteria. (2022)
  • Author(s): Manjunath G.B., Awasthi S.P., Zahid M.S.H., Hatanaka N., Hinenoya A., Iwaoka E., Aoki S., Ramamurthy T., Yamasaki S.
  • Journal Title: Letters in Applied Microbiology Volume: 74 Issue: 4 Pages: 472-481
  • DOI: 10.1111/lam.13646
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Identification of a recently dominant sublineage in Salmonella 4,[5],12:i:- sequence type 34 isolated from food animals in Japan. (2021)
  • Author(s): Arai Nobuo, Sekizuka Tsuyoshi, Tamamura-Andoh Yukino, Barco Lisa, Hinenoya Atsushi, Yamasaki Shinji, Iwata Taketoshi, Watanabe-Yanai Ayako, Kuroda Makoto, Akiba Masato, Kusumoto Masahiro.
  • Journal Title: Frontiers in Microbiology Volume: 12 Pages: 690947-690947
  • DOI: 10.3389/fmicb.2021.690947
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Critical role of 3′-downstream region of pmrB in polymyxin resistance in Escherichia coli BL21(DE3). (2021)
  • Author(s): Xu Fuzhou, Hinenoya Atsushi, Zeng Ximin, Li Xing-Ping, Guan Ziqiang, Lin Jun.
  • Journal Title: Microorganisms Volume: 9 Issue: 3 Pages: 655-655
  • DOI: 10.3390/microorganisms9030655
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Prevalence of mobile colistin resistance (mcr) genes in extended-spectrum β-lactamase-producing Escherichia coli isolated from retail raw foods in Nha Trang, Vietnam. (2021)
  • Author(s): Le Phong Quoc, Awasthi Sharda Prasad, Hatanaka Noritoshi, Hinenoya Atsushi, Hassan Jayedul, Ombarak Rabee Alhossiny, Iguchi Atsushi, Tran Nga Thuy Thi, Dao Khanh Van Thi, Vien Mai Quang, Le Huy Xuan, Do Hung Thai, Yamamoto Yoshimasa, Yamasaki Shinji.
  • Journal Title: International Journal of Food Microbiology Volume: 346 Pages: 109164-109164
  • DOI: 10.1016/j.ijfoodmicro.2021.109164
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Prevalence, serovar, and antimicrobial resistance of nontyphoidal Salmonella in vegetable, fruit, and water samples in Ho Chi Minh City, Vietnam. (2021)
  • Author(s): Nguyen Dao Thi Anh, Awasthi Sharda Prasad, Hoang Phuong Hoai, Nguyen Phuc Do, Jayedul Hassan, Hatanaka Noritoshi, Hinenoya Atsushi, Van Dang Chinh, Faruque Shah M., Yamasaki Shinji.
  • Journal Title: Foodborne Pathogens and Disease Volume: 18 Issue: 5 Pages: 354-363
  • DOI: 10.1089/fpd.2020.2891
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Prevalence, O-genotype and Shiga toxin (Stx) 2 subtype of Stx-producing <i>Escherichia coli</i> strains isolated from Argentinean beef cattle (2021)
  • Author(s): OKUNO Kentaro, AWASTHI Sharda Prasad, KOPPRIO German A., IGUCHI Atsushi, HATANAKA Noritoshi, HINENOYA Atsushi, LARA Ruben Jose, YAMASAKI Shinji.
  • Journal Title: Journal of Veterinary Medical Science Volume: 83 Issue: 4 Pages: 630-636
  • DOI: 10.1292/jvms.21-0002
  • NAID: 130008023264
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Isolation and characterization of Escherichia albertii in poultry at the pre‐harvest level. (2021)
  • Author(s): Hinenoya Atsushi、Li Xing‐Ping、Zeng Ximin、Sahin Orhan、Moxley Rodney A.、Logue Catherine M.、Gillespie Barbara、Yamasaki Shinji、Lin Jun
  • Journal Title: Zoonoses and Public Health Volume: 68 Issue: 3 Pages: 213-225
  • DOI: 10.1111/zph.12812
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Quantification of Vibrio cholerae cholix exotoxin by sandwich bead-ELISA (2021)
  • Author(s): Awasthi Sharda Prasad、Chowdhury Nityananda、Hatanaka Noritoshi、Hinenoya Atsushi、Ramamurthy Thandavarayan、Asakura Masahiro、Yamasaki Shinji
  • Journal Title: Journal of Medical Microbiology Volume: 70 Issue: 4 Pages: 001311-001311
  • DOI: 10.1099/jmm.0.001311
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Accurate identification of Escherichia albertii by matrix-assisted laser desorption ionization-time of flight mass spectrometry (2020)
  • Author(s): Hatanaka Noritoshi、Awasthi Sharda Prasad、Hinenoya Atsushi、Ueda Osamu、Yamasaki Shinji
  • Journal Title: Journal of Microbiological Methods Volume: 173 Pages: 105916-105916
  • DOI: 10.1016/j.mimet.2020.105916
  • Peer Reviewed
• [Journal Article] Prevalence of Escherichia albertii in Raccoons (Procyon lotor), Japan (2020)
  • Author(s): Hinenoya Atsushi、Nagano Keigo、Awasthi Sharda P.、Hatanaka Noritoshi、Yamasaki Shinji
  • Journal Title: Emerging Infectious Diseases Volume: 26 Issue: 6 Pages: 1304-1307
  • DOI: 10.3201/eid2606.191436
  • Peer Reviewed
• [Journal Article] Development of XRM-MacConkey agar selective medium for the isolation of Escherichia albertii (2020)
  • Author(s): Hinenoya Atsushi、Nagano Keigo、Okuno Kentaro、Nagita Akira、Hatanaka Noritoshi、Awasthi Sharda Prasad、Yamasaki Shinji
  • Journal Title: Diagnostic Microbiology and Infectious Disease Volume: 97 Issue: 1 Pages: 115006-115006
  • DOI: 10.1016/j.diagmicrobio.2020.115006
  • Peer Reviewed
• [Journal Article] Chlorous acid is a more potent antibacterial agent than sodium hypochlorite against Campylobacter (2020)
  • Author(s): Hatanaka Noritoshi、Awasthi Sharda Prasad、Goda Hisataka、Kawata Hiroyuki、Uchino Yuzuru、Kubo Takahiro、Aoki Shigeru、Hinenoya Atsushi、Yamasaki Shinji
  • Journal Title: Food Control Volume: 111 Pages: 107046-107046
  • DOI: 10.1016/j.foodcont.2019.107046
  • Peer Reviewed
• [Presentation] Distribution of Escherichia albertii, an emerging enteropathogen, in wild raccoons in Japan (2023)
  • Author(s): Atsushi Hinenoya, Bingting Xu, Moeko Yamazaki, Sharda Prasad Awasthi, Noritoshi Hatanaka, Shinji Yamasaki
  • Organizer: 2023 International Conference on Emerging Infectious Diseases in the Pacific Rim, Joint Panel Conference on Cholera and Other Bacterial Enteric Infections
  • Int'l Joint Research
• [Presentation] Within-host diversity of Escherichia albertii in wild raccoons (2023)
  • Author(s): Atsushi Hinenoya, Moeko Yamazaki, Bingting Xu, Sharda Prasad Awasthi ,Noritoshi Hatanaka, Shinji Yamasaki
  • Organizer: 第96回日本細菌学会総会
• [Presentation] Survey of Escherichia albertii in wild birds in Japan (2021)
  • Author(s): Atsushi Hinenoya, Sharada Prasad Awasthi, Noritomo Yasuda, Keigo Nagano, Jayedul Hassan, Keiji Takehira, Noritoshi Hatanaka, Haruna Inoue, Shinji Yamasaki
  • Organizer: 第94回日本細菌学会総会
• [Presentation] 新興人獣共通感染症細菌Escherichia albertiiの検査法開発およびその応用 (2021)
  • Author(s): 日根野谷 淳
  • Organizer: 第94回日本細菌学会総会
  • Invited