Metagenomic analysis of uncultured Anaplasmataceae pathogen from a single tick in Japan

• Project/Area Number: 26460532
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Bacteriology (including mycology)
• Research Institution: University of Shizuoka
• Principal Investigator: Ohashi Norio 静岡県立大学, 食品栄養科学部, 教授 (10169039)
• Co-Investigator(Kenkyū-buntansha): 吉川 悠子 日本獣医生命科学大学, 獣医学部, 助教 (00580523)
• Project Period (FY): 2014-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: メタゲノム / 細菌 / 新興感染症 / Anaplasma / マダニ媒介 / ゲノム / マダニ媒介性 / Neoehrlichia

Outline of Final Research Achievements

Human Granulocytic Anaplasmosis (HGA) is an emerging infectious disease and caused by Anaplasma phagocytophilum, that infects primarily neutrophils. In Japan, we do not have any Japanese A. phagocytophilum isolate, because its isolation from patients is extremely difficult. In this study, therefore we attempted the metagenomic analysis to obtain the genome sequence of uncultured A. phagocytophilum from a single Ixodes persulcatus tick using several next-generation and third-generation sequencers such as ION PGM System, MiSeq, HiSeq, and MinION. Eventually, we successfully obtained almost full-length genome sequence with 1. 48 Mb of the uncultured A. phagocytophilum in Japan using the whole-genome amplification DNA from salivary glands of the single tick. Thus, we believe that the genome sequence ofJapanese A. phagocytophilum obtained in this study will contribute the development of molecular-based HGA-diagnostic tool for public health significance.

Research Products

• [Journal Article] Evaluation of Diagnostic Assay for Rickettsioses Using Duplex Real-Time PCR in Multiple Laboratories in Japan (2018)
  • Author(s): Kawamori F, Shimazu Y, Sato H, Monma N, Ikegaya A, Yamamoto S, Fujita H, Morita H, Tamaki Y, Takamoto N, Su H, Shimada S, Shimamura Y, Masuda S, Ando S, Ohashi N
  • Journal Title: Japanese Journal of Infectious Diseases Volume: 71 Issue: 4 Pages: 267-273
  • DOI: 10.7883/yoken.JJID.2017.447
  • NAID: 130007418376
  • ISSN: 1344-6304, 1884-2836
  • Year and Date: 2018-07-31
  • Peer Reviewed / Open Access
• [Journal Article] Case of Human Infection with <i>Anaplasma phagocytophilum</i> in Inner Mongolia, China (2018)
  • Author(s): Gaowa, Wulantuya, Yin X, Cao M, Guo S, Ding C, Lu Y, Luo J, Kawabata H, Ando S, Su H, Shimada M, Takamoto N, Shimamura Y, Masuda S, Ohashi N
  • Journal Title: Japanese Journal of Infectious Diseases Volume: 71 Issue: 2 Pages: 155-157
  • DOI: 10.7883/yoken.JJID.2017.450
  • NAID: 130006517093
  • ISSN: 1344-6304, 1884-2836
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Case report: Clinical features of a case of suspected Borrelia miyamotoi disease in Hokkaido, Japan (2017)
  • Author(s): Yamano K, Ito T, Kiyanagi K, Yamazaki H, Sugawara M, Saito T, Ohashi N, Zamoto-Niikura A, Sato K, Kawabata H.
  • Journal Title: Am. J. Trop. Med. Hyg. Volume: 97 Issue: 1 Pages: 84-87
  • DOI: 10.4269/ajtmh.16-0699
  • Peer Reviewed
• [Journal Article] Interaction between various apple procyanidin and staphylococcal enterotoxin A and their inhibitory effects on toxin activity. (2017)
  • Author(s): Shimamura Y, Hirai C, Sugiyama Y, Utsumi M, Yanagida A, Murata M, Ohashi N, Masuda, S.
  • Journal Title: Toxins Volume: 9(8) Issue: 8 Pages: 243-243
  • DOI: 10.3390/toxins9080243
  • Peer Reviewed / Open Access
• [Journal Article] Amelioration of Citrobacter rodentium proliferation in early stage of infection in mice by pre-treatment with Lactobacillus brevis KB290 and verification using in vivo bioluminescence imaging (2017)
  • Author(s): Waki N, Kuwabara Y, Yoshikawa Y, Suganuma H, Koide H, Oku N, Ohashi N
  • Journal Title: FEMS Microbiol. Lett. Volume: 364 Pages: fnw254-fnw254
  • DOI: 10.1093/femsle/fnw254
  • Peer Reviewed
• [Journal Article] Detection and characterization of emerging relapsing fever pathogen, Borrelia miyamotoi, from Ixodes ricinus tick on rural Trakya (Thrace) region of north western Turkey (2016)
  • Author(s): Sakakibara Y, Sen E, Sato K, Kawabata H, Ohashi N, Masuzawa T
  • Journal Title: Vector Borne Zoonotic Dis. Volume: 16 Issue: 12 Pages: 797-799
  • DOI: 10.1089/vbz.2016.2012
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Prevalence of Borrelia miyamotoi in Ixodes persulcatus in Irkutsk City and its neighboring territories, Russia. (2016)
  • Author(s): Khasnatinov, M. A., Danchinova, G. A., Takano, A., Kawabata, H., Ohashi, N., Masuzawa, T.
  • Journal Title: Ticks Tick Borne Dis. Volume: 7 Issue: 2 Pages: 394-397
  • DOI: 10.1016/j.ttbdis.2015.12.016
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Molecular and serological survey of Rickettsiales bacteria in wild sika deer (Cervus nippon nippon) in Shizuoka prefecture, Japan: High prevalence of Anaplasma species (2015)
  • Author(s): Wu, D., Wuritu, Yoshikawa, Y., Gaowa, Kawamori, F., Ikegaya, A., Ohtake, M., Ohashi, M., Shimada, M., Takada, A., Iwai, K., and Ohashi, N.
  • Journal Title: Jpn. J. Infect. Dis. Volume: Advance Publication Issue: 5 Pages: 434-437
  • DOI: 10.7883/yoken.jjid.2015.003
  • NAID: 130005098926
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Anaplasma phagocytophilum antibodies in humans, Japan, 2010-2011. (2014)
  • Author(s): Gaowa, Yoshikawa, Y., Ohashi, N., Wu, D., Kawamori, F., Ikegaya, A., Watanabe, T., Saitoh, K., Takechi, D., Murakami, Y., Shichi, D., Aso, K., and Ando, S.
  • Journal Title: Emerging Infectious Diseases Volume: 20 Issue: 3 Pages: 508-509
  • DOI: 10.3201/eid2003.131337
  • Peer Reviewed / Open Access
• [Presentation] マダニ内在性細菌群のメタ16S解析から見るCoxiellaとRickettsia (2018)
  • Author(s): 佐藤綾香，高本直矢，高野愛，大石沙織，阿部冬樹，神田隆，平良雅克，藤田博己，島村裕子，大橋典男
  • Organizer: 第91回日本細菌学会総会（福岡）
• [Presentation] アナプラズマ症 (2017)
  • Author(s): 大橋典男
  • Organizer: 第90回日本細菌学会総会 （シンポジウム）
  • Place of Presentation: 仙台
  • Year and Date: 2017-03-19
• [Presentation] マダニ内在性細菌群のRickettsiaは異常な生殖系統の宿主で消えていく (2017)
  • Author(s): 高本直矢, 田井仁, 佐藤寛子, 川森文彦, 池ヶ谷朝香, 高田伸弘, 大橋典男
  • Organizer: 第90回日本細菌学会総会
  • Place of Presentation: 仙台
  • Year and Date: 2017-03-19
• [Presentation] RF6/A 感染細胞におけるAnaplasma phagocytophilumのP44外膜蛋白質群の発現解析 (2017)
  • Author(s): 島田雅彦, 蘇泓如, 大橋典男
  • Organizer: 第90回日本細菌学会総会
  • Place of Presentation: 仙台
  • Year and Date: 2017-03-19
• [Presentation] P44-specific peptide antigens for serodiagnosis of human granulocytic anaplasmosis (2017)
  • Author(s): 蘇泓如, 伊藤圭祐, 河原崎泰昌, 島田雅彦, 大橋典男
  • Organizer: 第90回日本細菌学会総会（仙台）
  • Place of Presentation: 仙台
  • Year and Date: 2017-03-19
• [Presentation] Anaplasma phagocytophilum in Japan (2017)
  • Author(s): 1.Ohashi N, Gaowa, Wuritu, Wu D, Kawamori F, Su H, Shimada M, Takamoto N, Tai H, Ando S
  • Organizer: International Congress on Rickettsiae and other Intracellular Bacteria (Marseille, France)
  • Int'l Joint Research
• [Presentation] Rickettsia sp. LONの細胞内増殖評価法の検討 (2017)
  • Author(s): 1.田井仁，高本直矢，藤田博己，島田雅彦，蘇泓如，大橋典男
  • Organizer: 第24回リケッチア研究会 (東京)
• [Presentation] 単為生殖がもたらすダニ内在性リケッチアへの影響 (2017)
  • Author(s): 2.高本直矢，田井仁，佐藤寛子，平良雅克，高野愛，高田伸弘，川森文彦，池ヶ谷朝香，大石沙織，阿部冬樹，神田隆，藤田博己，大橋典男
  • Organizer: 第24回リケッチア研究会 (東京)
• [Presentation] マダニ内に存在する細菌叢のメタ16S解析 (2017)
  • Author(s): 3.佐藤綾香，高本直矢，笹原榛華，松﨑光ノ介，高野愛，大石沙織，阿部冬樹，神田隆，平良雅克，藤田博己，島村裕子，増田修一，大橋典男
  • Organizer: 第24回リケッチア研究会 (東京)
• [Presentation] アナプラズマ感染における異なる宿主細胞のlive imaging (2016)
  • Author(s): 島田雅彦, 蘇泓如, 大橋典男
  • Organizer: 第23回リケッチア研究会
  • Place of Presentation: 東京
  • Year and Date: 2016-12-03
• [Presentation] マダニから抽出したRNAの紅斑熱群リケッチア検査への活用について (2016)
  • Author(s): 池ヶ谷朝香, 原稔美, 酒井悠希子, 阿部冬樹, 佐原啓二, 大橋典男
  • Organizer: 第23回リケッチア研究会
  • Place of Presentation: 東京
  • Year and Date: 2016-12-03
• [Presentation] つつが虫病および紅斑熱の迅速診断法の検討 (2016)
  • Author(s): 川森文彦, 池ヶ谷朝香, 佐原啓二, 安藤秀二, 大橋典男
  • Organizer: 第23回リケッチア研究会
  • Place of Presentation: 東京
  • Year and Date: 2016-12-03
• [Presentation] フタトゲチマダニの両性/単為生殖系統における共生細菌群の解析 (2016)
  • Author(s): 高本直矢, 田井仁, 佐藤寛子, 川森文彦, 池ヶ谷朝香, 高田伸弘, 大橋典男
  • Organizer: 第23回リケッチア研究会
  • Place of Presentation: 東京
  • Year and Date: 2016-12-03
• [Presentation] マダニゲノムとアマプラズマ細菌を見る (2016)
  • Author(s): 大橋典男，高本直矢，田井仁
  • Organizer: 第24回ダニと疾患のインタフェースに関するセミナー（SADI）鹿児島大会
  • Place of Presentation: 指宿市
  • Year and Date: 2016-05-27
• [Presentation] ペプチドアレイを用いたアナプラズマ細菌P44抗原のエピトープマッピング (2016)
  • Author(s): 蘇泓如，呉東興，伊藤圭祐，河原崎泰昌，大橋典男
  • Organizer: 第24回ダニと疾患のインタフェースに関するセミナー（SADI）鹿児島大会
  • Place of Presentation: 指宿市
  • Year and Date: 2016-05-27
• [Presentation] Anaplasma phagocytophilum specific P44 epitopes identified by peptide array analysis (2016)
  • Author(s): 蘇泓如, 呉東興, 伊藤圭祐, 河原崎泰昌, 大橋典男
  • Organizer: 第89回日本細菌学会総会
  • Place of Presentation: 大阪
  • Year and Date: 2016-03-23
• [Presentation] Whole genome shotgun sequencing of Ixodes persulcatus infected with Anaplasma phagocytophilum T162 (cutting-edge report) (2015)
  • Author(s): Ohashi, N., Takamoto, N., Wu, D., Tai H.
  • Organizer: 第22回リケッチア研究会
  • Place of Presentation: 東京
  • Year and Date: 2015-11-28
• [Presentation] アナプラズマ－情報update (2015)
  • Author(s): 大橋典男
  • Organizer: 第22回リケッチア研究会
  • Place of Presentation: 東京
  • Year and Date: 2015-11-28
• [Presentation] 静岡県における日本紅斑熱の疫学的考察 (2015)
  • Author(s): 川森文彦, 池ヶ谷朝香, 荒畑沙織, 酒井悠希子，佐原啓二, 大橋典男
  • Organizer: 第22回リケッチア研究会
  • Place of Presentation: 東京
  • Year and Date: 2015-11-28
• [Presentation] アナプラズマ科細菌感染症に関する最近の話題について (2015)
  • Author(s): 大橋典男
  • Organizer: 第23回ダニと疾患のインタフェースに関するセミナー (SADI) 東日本大震災復興祈念大会
  • Place of Presentation: 名取
  • Year and Date: 2015-06-26
• [Presentation] 野生動物におけるリケッチア目細菌の分子疫学調査 (2015)
  • Author(s): 呉東興, 大橋典男, 烏日図, 高娃, 吉川悠子, 川森文彦, 池ヶ谷朝香
  • Organizer: 第23回ダニと疾患のインタフェースに関するセミナー (SADI) 東日本大震災復興祈念大会
  • Place of Presentation: 名取
  • Year and Date: 2015-06-26
• [Presentation] One-tube nested PCRによるOrientia tsutugamushiの検出 (2015)
  • Author(s): 川森文彦, 池ヶ谷朝香, 荒畑沙織, 佐原啓二, 大橋典男
  • Organizer: 第23回ダニと疾患のインタフェースに関するセミナー (SADI) 東日本大震災復興祈念大会
  • Place of Presentation: 名取
  • Year and Date: 2015-06-26
• [Presentation] 静岡県におけるSFTSウイルスの浸淫実態 (2015)
  • Author(s): 池ヶ谷朝香, 荒畑沙織, 佐原啓二, 川森文彦, 大橋典男
  • Organizer: 第23回ダニと疾患のインタフェースに関するセミナー (SADI) 東日本大震災復興祈念大会
  • Place of Presentation: 名取
  • Year and Date: 2015-06-26
• [Presentation] アナプラズマ症の特異抗体検出による患者探索の現状報告 (2014)
  • Author(s): 呉東興，高娃，吉川悠子，川森文彦，池ヶ谷朝香，川上万里，岸本壽男，森田裕司，中堂園文子，御供田睦代，能勢裕久，池田賢一，増澤俊幸，安藤秀二，大橋典男
  • Organizer: 第21回リケッチア研究会 研究発表会
  • Place of Presentation: 東京
  • Year and Date: 2014-12-20
• [Presentation] Anaplasma phagocytophilum感染患者血清中に存在する抗体の検出法に関する検討 (2014)
  • Author(s): 呉東興，高娃，吉川悠子，川森文彦，川上万理，岸本壽男，森田裕司，増澤俊幸，安藤秀二，大橋典男
  • Organizer: 第97回日本細菌学会関東支部総会
  • Place of Presentation: 東京
  • Year and Date: 2014-10-30 – 2014-10-31
• [Presentation] 日本国内に潜在する新興感染症「アナプラズマ症」 (2014)
  • Author(s): 大橋典男
  • Organizer: 平成26年度中部地区獣医師大会・獣医学術中部地区学会
  • Place of Presentation: 静岡
  • Year and Date: 2014-08-31
  • Invited
• [Presentation] Tick-associated Anaplasmataceae pathogens in Japan (2014)
  • Author(s): Ohashi N., Yoshikawa Y., Gaowa, Wuritu, Wu D., Kawamori F.
  • Organizer: XIV International Congress of Acarology
  • Place of Presentation: Kyoto
  • Year and Date: 2014-07-14 – 2014-07-18
  • Invited
• [Presentation] わが国の新興アナプラズマ症 (2014)
  • Author(s): 川森文彦, 大橋典男
  • Organizer: 衛生微生物技術協議会第35回研究会
  • Place of Presentation: 東京
  • Year and Date: 2014-06-26 – 2014-06-27
  • Invited
• [Book] 臨床医学eBook, Enfection, 感染症診療とダニワールド, アナプラズマ症 (2016)
  • Author(s): 大橋典男
  • Total Pages: 66
  • Publisher: シーニュ社