Potential of symbionts in controlling arthropod-borne diseases

• Project/Area Number: 19H03118
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 42020:Veterinary medical science-related
• Research Institution: Hokkaido University
• Principal Investigator: Nakao Ryo 北海道大学, 獣医学研究院, 准教授 (50633955)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥17,290,000 (Direct Cost: ¥13,300,000、Indirect Cost: ¥3,990,000); Fiscal Year 2021: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2020: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000); Fiscal Year 2019: ¥6,630,000 (Direct Cost: ¥5,100,000、Indirect Cost: ¥1,530,000)
• Keywords: 吸血性節足動物 / 共生微生物 / マイクロビオーム / マダニ / Coxiella / Francisella / Occidentia / PNA / Gregarina / パラトランスジェネシス / 微生物叢

Outline of Research at the Start

昆虫（蚊やハエ）やダニなどの吸血性節足動物はヒトや動物に重篤な疾病を引き起こす様々な病原体を媒介する。一方で、節足動物は病原体以外にも、ヒトや動物に病気を起こさない多様な微生物を体内に保有し、共生微生物を含めた超個体として存在している。本研究では、３者（節足動物―病原体―共生微生物）の共生状態を維持するために重要な節足動物因子および共生微生物因子を特定することで、節足動物媒介性疾病の新たな制御法を開発することを目的とする。

Outline of Final Research Achievements

Blood-feeding arthropods such as ticks and mosquitoes can carry pathogens to humans and animals. They also harbor a variety of seemingly non-pathogenic microorganisms; however, their diversity and roles have not yet been clearly understood. This study disclosed bacterial microbiomes of ticks from domestic and foreign origins by sequencing 16S ribosomal RNA gene (rDNA). The dominant taxa found in tick bacterial microbiome, including Coxiella, Spiroplasma, and Rickettsia, were further genetically characterized to understand their transmission manners in ticks. The method to explore the tick eukaryotic microbiome was developed by designing a peptide nucleic acid (PNA) clamp method which suppresses amplification of tick 18S rDNA, leading to the first detection of Gregarina in ticks. Collectively, these results provide fundamental information to develop control strategies for vector-borne diseases.

Academic Significance and Societal Importance of the Research Achievements

国内外のマダニが保有する微生物叢が明らかになり、Coxiella、Spiroplasma、Rickettsia等の優占細菌群の存在を突き止めた。それら優占細菌群のマダニ体内での役割を解析することで、マダニやマダニ媒介性病原体の制御法開発に向けた研究展開が可能となった。マダニの真核生物叢の解析手法を世界で初めて開発した。その手法を論文として公開したことで、他の研究者も容易に真核生物叢解析が可能となり、これまで全く注目されてこなかったマダニ保有真核生物叢の情報集積が期待できる。これまでに認知されていない病原体や新規微生物の発見につながる可能性があり、学術的、社会的意義は大きい。

Research Products

• [Int'l Joint Research] サウスバリー大学/マンスーラ大学(エジプト)
• [Int'l Joint Research] リロングウェ農業天然資源大学(マラウイ)
• [Int'l Joint Research] ザンビア大学(ザンビア)
• [Int'l Joint Research] クイーンズランド大学(オーストラリア)
• [Int'l Joint Research] リバプール大学(英国)
• [Int'l Joint Research] ベルリン自由大学(ドイツ)
• [Int'l Joint Research] ザンビア大学(ザンビア)
• [Int'l Joint Research] リロングウェ農業・ 自然資源大学(マラウイ)
• [Int'l Joint Research] ザンビア大学(ザンビア)
• [Int'l Joint Research] ハルツーム大学(スーダン)
• [Int'l Joint Research] ガブリエル・レネ・モレノ国立自治大学(ボリビア)
• [Int'l Joint Research] ガーナ大学(ガーナ)
• [Int'l Joint Research] リロングウェ農業・ 自然資源大学(マラウイ)
• [Int'l Joint Research]
• [Journal Article] Reconstruction of mitochondrial genomes from raw sequencing data provides insights on the phylogeny of Ixodes ticks and cautions for species misidentification (2022)
  • Author(s): Mohamed Wessam Mohamed Ahmed、Moustafa Mohamed Abdallah Mohamed、Kelava Samuel、Barker Dayana、Matsuno Keita、Nonaka Nariaki、Shao Renfu、Mans Ben J.、Barker Stephen C.、Nakao Ryo
  • Journal Title: Ticks and Tick-borne Diseases Volume: 13 Issue: 1 Pages: 101832-101832
  • DOI: 10.1016/j.ttbdis.2021.101832
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Description of the female, nymph and larva and mitochondrial genome, and redescription of the male of Ixodes barkeri Barker, 2019 (Acari: Ixodidae), from the short-beaked echidna, Tachyglossus aculeatus, with a consideration of the most suitable subgenus for this tick (2022)
  • Author(s): Barker Dayana、Kelava Samuel、Shao Renfu、Seeman Owen D.、Jones Malcolm K.、Nakao Ryo、Barker Stephen C.、Apanaskevich Dmitry A.
  • Journal Title: Parasites & Vectors Volume: 15 Issue: 1 Pages: 117-117
  • DOI: 10.1186/s13071-022-05165-2
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] High infection rate of tick-borne protozoan and rickettsial pathogens of cattle in Malawi and the development of a multiplex PCR for Babesia and Theileria species identification (2022)
  • Author(s): Chatanga Elisha、Maganga Emmanuel、Mohamed Wessam Mohamed Ahmed、Ogata Shohei、Pandey Gita Sadaula、Abdelbaset Abdelbaset Eweda、Hayashida Kyoko、Sugimoto Chihiro、Katakura Ken、Nonaka Nariaki、Nakao Ryo
  • Journal Title: Acta Tropica Volume: 231 Pages: 106413-106413
  • DOI: 10.1016/j.actatropica.2022.106413
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Rediscovery of Ixodes confusus in Australia with the first description of the male from Australia, a redescription of the female and the mitochondrial (mt) genomes of five species of Ixodes (2022)
  • Author(s): Barker Dayana、Kelava Samuel、Seeman Owen D.、Shao Renfu、Seaniger James R.、Jones Malcolm K.、Apanaskevich Maria A.、Nakao Ryo、Apanaskevich Dmitry A.、Barker Stephen C.
  • Journal Title: International Journal for Parasitology: Parasites and Wildlife Volume: 18 Pages: 1-11
  • DOI: 10.1016/j.ijppaw.2022.03.006
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Screening of tick-borne pathogens in argasid ticks in Zambia: Expansion of the geographic distribution of Rickettsia lusitaniae and Rickettsia hoogstraalii and detection of putative novel Anaplasma species. (2021)
  • Author(s): Qiu Y, Simuunza M, Kajihara M, Chambaro H, Harima H, Eto Y, Simulundu E, Squarre D, Torii S, Takada A, Hangmbe BM, Sawa H, Sugimoto C, Nakao R
  • Journal Title: Ticks Tick Borne Dis Volume: 12 Issue: 4 Pages: 101720-101720
  • DOI: 10.1016/j.ttbdis.2021.101720
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Applications of Blocker Nucleic Acids and Non-Metazoan PCR Improves the Discovery of the Eukaryotic Microbiome in Ticks (2021)
  • Author(s): Taya Yurie、Kinoshita Gohta、Mohamed Wessam Mohamed Ahmed、Moustafa Mohamed Abdallah Mohamed、Ogata Shohei、Chatanga Elisha、Ohari Yuma、Kusakisako Kodai、Matsuno Keita、Nonaka Nariaki、Nakao Ryo
  • Journal Title: Microorganisms Volume: 9 Issue: 5 Pages: 1051-1051
  • DOI: 10.3390/microorganisms9051051
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Molecular Detection and Genotyping of Coxiella-Like Endosymbionts in Ticks Collected from Animals and Vegetation in Zambia (2021)
  • Author(s): Kobayashi Toshiya、Chatanga Elisha、Qiu Yongjin、Simuunza Martin、Kajihara Masahiro、Hang’ombe Bernard Mudenda、Eto Yoshiki、Saasa Ngonda、Mori-Kajihara Akina、Simulundu Edgar、Takada Ayato、Sawa Hirofumi、Katakura Ken、Nonaka Nariaki、Nakao Ryo
  • Journal Title: Pathogens Volume: 10 Issue: 6 Pages: 779-779
  • DOI: 10.3390/pathogens10060779
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] BoLA-DRB3 gene haplotypes show divergence in native Sudanese cattle from taurine and indicine breeds (2021)
  • Author(s): Salim Bashir、Takeshima Shin-nosuke、Nakao Ryo、Moustafa Mohamed A. M.、Ahmed Mohamed-Khair A.、Kambal Sumaya、Mwacharo Joram M.、Alkhaibari Abeer M.、Giovambattista Guillermo
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 17202-17202
  • DOI: 10.1038/s41598-021-96330-7
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Molecular Survey of Babesia and Anaplasma Infection in Cattle in Bolivia (2021)
  • Author(s): Ogata Shohei、Pereira Juan Antonio Cristian、Jhonny Loza Vega Ariel、Carolina Herbas Perez Gladys、Matsuno Keita、Orba Yasuko、Sawa Hirofumi、Kawamori Fumihiko、Nonaka Nariaki、Nakao Ryo
  • Journal Title: Veterinary Sciences Volume: 8 Issue: 9 Pages: 188-188
  • DOI: 10.3390/vetsci8090188
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Comparing the gut microbiome along the gastrointestinal tract of three sympatric species of wild rodents (2021)
  • Author(s): Anders Jason L.、Moustafa Mohamed Abdallah Mohamed、Mohamed Wessam Mohamed Ahmed、Hayakawa Takashi、Nakao Ryo、Koizumi Itsuro
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 19929-19929
  • DOI: 10.1038/s41598-021-99379-6
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Emergence of the invasive malaria vector Anopheles stephensi in Khartoum State, Central Sudan (2021)
  • Author(s): Ahmed Ayman、Khogali Rua、Elnour Mohammed-Ahmed B.、Nakao Ryo、Salim Bashir
  • Journal Title: Parasites & Vectors Volume: 14 Issue: 1 Pages: 511-511
  • DOI: 10.1186/s13071-021-05026-4
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Complete Genome Sequence of Leptospira kobayashii Strain E30, Isolated from Soil in Japan (2021)
  • Author(s): Nakao Ryo、Masuzawa Toshiyuki、Nakamura Shuichi、Koizumi Nobuo
  • Journal Title: Microbiology Resource Announcements Volume: 10 Issue: 45
  • DOI: 10.1128/mra.00907-21
  • Peer Reviewed / Open Access
• [Journal Article] Amblyomma testudinarium infestation on a brown bear (Ursus arctos yesoensis) captured in Hokkaido, a northern island of Japan. (2021)
  • Author(s): Nakao R, Shinjo K, Sakiyama T, Ogata S, Kusakisako K, Kinoshita G, Naguib D, Chatanga E, Mohamed WMA, Moustafa MAM, Matsuno K, Ito T, Nonaka N, Sashika M, Tsubota T, Shimozuru M
  • Journal Title: Parasitol Int Volume: 80 Pages: 102209-102209
  • DOI: 10.1016/j.parint.2020.102209
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Molecular identification and genetic characterization of tick-borne pathogens in sheep and goats at two farms in the central and southern regions of Malawi. (2021)
  • Author(s): Chatanga E, Kainga H, Maganga E, Hayashida K, Katakura K, Sugimoto C, Nonaka N, Nakao R
  • Journal Title: Ticks Tick Borne Dis Volume: 12 Issue: 2 Pages: 101629-101629
  • DOI: 10.1016/j.ttbdis.2020.101629
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Anthropogenic interferences lead to gut microbiome dysbiosis in Asian elephants and may alter adaptation processes to surrounding environments. (2021)
  • Author(s): Moustafa MAM, Chel HM, Thu MJ, Bawm S, Htun LL, Win MM, Oo ZM, Ohsawa N, Lahdenper M, Mohamed WMA, Ito K, Nonaka N, Nakao R, Katakura K
  • Journal Title: Sci Rep Volume: 11 Issue: 1 Pages: 741-741
  • DOI: 10.1038/s41598-020-80537-1
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Spiroplasma Infection among Ixodid Ticks Exhibits Species Dependence and Suggests a Vertical Pattern of Transmission (2021)
  • Author(s): Ogata Shohei、Mohamed Wessam Mohamed Ahmed、Kusakisako Kodai、Thu May June、Qiu Yongjin、Moustafa Mohamed Abdallah Mohamed、Matsuno Keita、Katakura Ken、Nonaka Nariaki、Nakao Ryo
  • Journal Title: Microorganisms Volume: 9 Issue: 2 Pages: 333-333
  • DOI: 10.3390/microorganisms9020333
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Exploring Prokaryotic and Eukaryotic Microbiomes Helps in Detecting Tick-Borne Infectious Agents in the Blood of Camels (2021)
  • Author(s): Mohamed Wessam Mohamed Ahmed、Ali Alsagher O.、Mahmoud Hassan Y. A. H.、Omar Mosaab A.、Chatanga Elisha、Salim Bashir、Naguib Doaa、Anders Jason L.、Nonaka Nariaki、Moustafa Mohamed Abdallah Mohamed、Nakao Ryo
  • Journal Title: Pathogens Volume: 10 Issue: 3 Pages: 351-351
  • DOI: 10.3390/pathogens10030351
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Phylogenies from mitochondrial genomes of 120 species of ticks: Insights into the evolution of the families of ticks and of the genus Amblyomma. (2020)
  • Author(s): Kelava S, Mans BJ, Shao R, Moustafa MAM, Matsuno K, Takano A, Kawabata H, Sato K, Fujita H, Ze C, Plantard O, Hornok S, Gao S, Barker D, Barker SC, Nakao R
  • Journal Title: Ticks Tick Borne Dis Volume: 12 Issue: 1 Pages: 101577-101577
  • DOI: 10.1016/j.ttbdis.2020.101577
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Molecular detection and characterization of tick-borne hemoparasites and Anaplasmataceae in dogs in major cities of Malawi (2020)
  • Author(s): Chatanga Elisha、Kainga Henson、Razemba Tinotenda、Ssuna Richard、Swennen Lieza、Hayashida Kyoko、Sugimoto Chihiro、Katakura Ken、Nonaka Nariaki、Nakao Ryo
  • Journal Title: Parasitology Research Volume: 120 Issue: 1 Pages: 267-276
  • DOI: 10.1007/s00436-020-06967-y
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Distinct haplotypes and free movement of Aedes aegypti in Port Sudan, Sudan. (2020)
  • Author(s): Elnour MAB, Moustafa MAM, Khogali R, Azrag RS, Alanazi AD, Kheir A, Nakao R, de Meeus T, Salim B
  • Journal Title: J Appl Entomol Volume: 144 Issue: 9 Pages: 817-823
  • DOI: 10.1111/jen.12816
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] First record and analysis of the COI gene of Cobboldia elephantis obtained from a captive Asian elephant from Myanmar. (2020)
  • Author(s): Chel HM, Nakao R, Ohsawa N, Oo ZM, Nonaka N, Katakura K
  • Journal Title: Parasitol Int Volume: 75 Pages: 102035-102035
  • DOI: 10.1016/j.parint.2019.102035
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Bacterial and protozoan pathogens/symbionts in ticks infecting wild grasscutters (Thryonomys swinderianus) in Ghana (2020)
  • Author(s): Adenyo C, Ohya K, Qiu Y, Takashima Y, Ogawa H, Matsumoto T, Thu MJ, Sato K, Kawabata H, Katayama Y, Omatsu T, Mizutani T, Fukushi H, Nonaka N, Katakura K, Inoue-Murayama M, Kayang B, Nakao R
  • Journal Title: Acta Trop Volume: 205 Pages: 105388-105388
  • DOI: 10.1016/j.actatropica.2020.105388
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Characteristics and application of tick cell lines (2019)
  • Author(s): Nakao Ryo
  • Journal Title: Medical Entomology and Zoology Volume: 70 Issue: 4 Pages: 175-179
  • DOI: 10.7601/mez.70.175
  • NAID: 130007772671
  • ISSN: 0424-7086, 2185-5609
  • Year and Date: 2019-12-25
  • Peer Reviewed / Open Access
• [Journal Article] Complete Genome Sequence of Rickettsia asiatica Strain Maytaro1284, a Member of Spotted Fever Group Rickettsiae Isolated from an Ixodes ovatus Tick in Japan. (2019)
  • Author(s): Thu MJ, Qiu Y, Yamagishi J, Kusakisako K, Ogata S, Moustafa MAM, Isoda N, Sugimoto C, Katakura K, Nonaka N, Nakao R
  • Journal Title: Microbiol Resour Announc Volume: 8(37) Issue: 37 Pages: 37-37
  • DOI: 10.1128/mra.00886-19
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Mutations in the TaPIN1 peptidyl prolyl isomerase gene in Theileria annulata parasites isolated in Sudan (2019)
  • Author(s): Salim Bashir、Chatanga Elisha、Jannot Guillaume、Mossaad Ehab、Nakao Ryo、Weitzman Jonathan B.
  • Journal Title: International Journal for Parasitology: Drugs and Drug Resistance Volume: 11 Pages: 101-105
  • DOI: 10.1016/j.ijpddr.2019.11.001
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] 人工核酸を用いた選択的PCR法によるマダニ保有真核生物叢の解析 (2021)
  • Author(s): 田谷友里恵、木下豪太、Wessam Mohamed Ahmed Mohamed、 Mohamed Abdallah Mohamed Moustafa、尾針由真、松野啓太、野中成晃、中尾亮
  • Organizer: 第90回日本寄生虫学会 第32回日本臨床寄生虫学会 合同大会
• [Presentation] ボリビア多民族国サンタクルス県の牛におけるBabesiaおよびAnaplasmaの感染状況 (2021)
  • Author(s): 小方昌平、Juan Antonio Pereira、松野啓太、大場靖子、澤洋文、川森文彦、野中成晃、中尾亮
  • Organizer: 第164回日本獣医学会学術集会
• [Presentation] Application of selective whole genome amplification (SWGA) for tick mitochondrial genome sequencing (2021)
  • Author(s): Elisha Chatanga, Tateki Matsumoto, Mohamed Abdallah Mohamed Moustafa, Yuma Ohari, Yongjin Qiu, Ken Katakura, Nariaki Nonaka, Ryo Nakao
  • Organizer: 第164回日本獣医学会学術集会
• [Presentation] マダニが保有する真核生物叢解析法の開発 (2021)
  • Author(s): 田谷友里恵、木下豪太、Wessam Mohamed Ahmed Mohamed、 Mohamed Abdallah Mohamed Moustafa、尾針由真、松野啓太、野中成晃、中尾亮
  • Organizer: 第164回日本獣医学会学術集会
• [Presentation] マダニから分離したリケッチエラ共生菌が保有するビタミンB群代謝関連遺伝子群の特定 (2021)
  • Author(s): 大前日希、May June Thu、林直樹、今里裕平、野中成晃、中尾亮
  • Organizer: 日本寄生虫学会・日本衛生動物学会第67回北日本支部合同大会
• [Presentation] Haemaphysalis longicornis が保有する Coxiella 様共生細菌のビタミン B 群およびその補因子の代謝に関わる遺伝子の探索 (2020)
  • Author(s): 草木迫 浩大、山本 佳代子、今里 裕平、山岸 潤也、白藤(梅宮) 梨可、玄 学南、野中 成晃、中尾 亮
  • Organizer: 第89回日本寄生虫学会大会
• [Presentation] マダニ中の多様なフレボウイルスは何者か？ (2019)
  • Author(s): 松野啓太、中尾亮、草木迫浩大、大場靖子、澤洋文
  • Organizer: 第162回日本獣医学会学術集会
• [Presentation] Sequences of two genes encoding Theileria parva CD8+ antigens reveal the presence of Muguga vaccine strain in non-vaccinated cattle in Malawi (2019)
  • Author(s): Elisha Chatanga, Ryo Nakao, Kyoko Hayashida, Bashir Salim, Chihiro Sugimoto, Ken Katakura, Nariaki Nonaka
  • Organizer: 第162回日本獣医学会学術集会
• [Presentation] 2017、2018年に北海道道央地域のヤマトマダニから分離されたダニ媒介性脳炎ウイルスの性状解析 (2019)
  • Author(s): 高橋侑嗣、小林進太郎、石塚万里子、中尾亮、苅和宏明、好井健太朗
  • Organizer: 第162回日本獣医学会学術集会
• [Presentation] マダニ中のフレボウイルスの遺伝子系統解析に基づく性状推定 (2019)
  • Author(s): 松野啓太、中尾亮、梶原将大、下田宙、海老原秀喜、高田礼人、前田健、岡松正敏、迫田義博
  • Organizer: 第162回日本獣医学会学術集会
• [Presentation] マダニが保有するスピロプラズマ属細菌の多様性 (2019)
  • Author(s): 小方昌平、中尾亮、片倉賢、野中成晃
  • Organizer: 第162回日本獣医学会学術集会
• [Presentation] フタトゲチマダニ由来カタラーゼ遺伝子の同定ならびに組換え体を用いた抗酸化活性の検証 (2019)
  • Author(s): 草木迫 浩大、中尾 亮、田仲 哲也、野中 成晃
  • Organizer: 第65回日本寄生虫学会・日本衛生動物学会北日本支部合同大会
• [Presentation] Characterization of tick-borne encephalitis virus isolated from Ixodes ovatus in Central area of Hokkaido in 2017, 2018 (2019)
  • Author(s): 高橋侑嗣、小林進太郎、石塚万里子、中尾亮、苅和宏明、好井健太朗
  • Organizer: 第67回日本ウイルス学会学術集会
• [Presentation] Human borreliosis caused by a novel New World relapsing fever borrelia in the Afrotropic ecozone (2019)
  • Author(s): Yongjin Qiu, Ryo Nakao, Bernard Mudenda Hang’ombe, Kozue Sato, Masahiro Kajihara, Sharon Kanchela, Katendi Changula, Ayato Takada, Hirohumi Sawa, Chihiro Sugimoto, Hiroki Kawabata
  • Organizer: 第60回日本熱帯医学会大会
• [Presentation] Phylogeny and diversity of tick symbionts in Japan (2019)
  • Author(s): Ryo Nakao
  • Organizer: Entomology 2019, the 67th Annual Meeting of the Entomological Society of America
  • Int'l Joint Research / Invited