Bioimaging analysis on erythrocyte invasion mechanisms of Babesia bovis

• Project/Area Number: 25850199
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Veterinary medical science
• Research Institution: Nagasaki University
• Principal Investigator: ASADA Masahito 長崎大学, 熱帯医学研究所, 助教 (40587028)
• Research Collaborator: YAHATA Kazuhide 長崎大学, 熱帯医学研究所, 助教 ; HAKIMI Hassan 長崎大学, 熱帯医学研究所, 特任研究員 ; YOKOYAMA Naoaki 帯広畜産大学, 原虫病研究センター, 教授 ; IGARASHI Ikuo 帯広畜産大学, 原虫病研究センター, 教授 ; KANEKO Osamu 長崎大学, 熱帯医学研究所, 教授 ; SUAREZ E. Carlos ワシントン州立大学, 獣医学部, 准教授 ; KAWAZU Shin-ichiro 帯広畜産大学, 原虫病研究センター, 教授
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: バベシア / ウシ / 赤血球侵入 / 滑走運動 / 遺伝子組換え / イメージング / バベシア原虫 / 遺伝子改変 / 獣医寄生虫 / 原虫病学 / 獣医寄生虫学 / バイオイメージング

Outline of Final Research Achievements

Babesia bovis is an intraerythrocytic apicomplexan parasite, which is transmitted by ticks and causes the disease bovine babesiosis in cattle. In this study, we developed transfection system on B. bovis by using double selection with WR99210 and blasticidin-S, which enables us to perform double transfection in the parasite. Then, we analyzed gliding motility of B. bovis merozoite by disrupting TRAP gene families. Moreover, we analyzed the egress mechanism of the parasite. We found egress was artificially induced in vitro using calcium ionophore A23187 and thapsigargin which increase Ca(2+) concentration in the cytosol of the parasite cells. These finding will contribute to develop new therapeutic strategies to bovine babesiosis.

Research Products

• [Journal Article] Establishment of transient and stable transfection systems for Babesia ovata (2016)
  • Author(s): Hakimi H, Yamagishi J, Kegawa Y, Kaneko O, Kawazu SI, Asada M
  • Journal Title: Parasit Vectors Volume: 9 Issue: 1 Pages: 171-171
  • DOI: 10.1186/s13071-016-1439-z
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Asada M, Yahata K, Hakimi H, Yokoyama N, Igarashi I, Kaneko O, Suarez CE, Kawazu SI. Transfection of Babesia bovis by double selection with WR99210 and Blasticidin-S and its application for functional analysis of thioredoxin peroxidase-1 (2015)
  • Author(s): Asada M, Yahata K, Hakimi H, Yokoyama N, Igarashi I, Kaneko O, Suarez CE, Kawazu SI
  • Journal Title: PLOS One Volume: 10(5) Issue: 5 Pages: e0125993-e0125993
  • DOI: 10.1371/journal.pone.0125993
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Calcium ions are involved in egress of Babesia bovis merozoites from bovine erythrocytes (2015)
  • Author(s): Mossad E, Asada M, Nakatani D, Inoue N, Yokoyama N, Kaneko O, Kawazu S.
  • Journal Title: J Vet Med Sci. Volume: ７７（１） Pages: 53-58
  • NAID: 130004693911
  • Peer Reviewed
• [Journal Article] Plasmodium knowlesi thioredoxin peroxidase 1 binds to nucleic acids and has RNA chaperone activity. (2014)
  • Author(s): Hakimi H, Suganuma K, Usui M, Masuda-Suganuma H, Angeles JM, Asada M, Kawai S, Inoue N, Kawazu S.
  • Journal Title: Parasitol Res. Volume: １１３（１） Issue: 11 Pages: 3957-3962
  • DOI: 10.1007/s00436-014-4060-0
  • NAID: 120006986305
  • Peer Reviewed
• [Journal Article] Cloning and characterization of a 2-Cys peroxiredoxin from Babesia gibsoni. (2014)
  • Author(s): Masatani T, Asada M, Ichikawa-Seki M, Usui M, Terkawi MA, Hayashi K, Kawazu S, Xuan X.
  • Journal Title: J Vet Med Sci. Volume: 76-1 Pages: 139-143
  • NAID: 130003362301
  • Peer Reviewed
• [Journal Article] Molecular Characterization of a New Babesia bovis Thrombospondin-Related Anonymous Protein (BbTRAP2) (2013)
  • Author(s): Terkawi MA, Ratthanophart J, Salama A, Aboulaila M, Asada M, Ueno A, Alhasan H, Guswanto A, Masatani T, Yokoyama N, Nishikawa Y, Xuan X, Igarashi I
  • Journal Title: PLoS One Volume: 8-12
  • Peer Reviewed
• [Journal Article] Identification, cloning and characterization of BmP41, a common antigenic protein of Babesia microti. (2013)
  • Author(s): Masatani T, Ooka H, Terkawi MA, Cao S, Luo Y, Asada M, Hayashi K, Nishikawa Y, Xuan X
  • Journal Title: J Vet Med Sci Volume: 75-7 Pages: 967-970
  • NAID: 130003362150
  • Peer Reviewed
• [Presentation] バベシア原虫メロゾイト滑走運動のバイオイメージング研究 (2015)
  • Author(s): 麻田正仁、河津信一郎
  • Organizer: 第48回日本原生生物学会大会
  • Place of Presentation: 国立感染症研究所（東京都・新宿区）
  • Year and Date: 2015-11-06
• [Presentation] Transfection of Babesia bovis by Double Selection with WR99210 and Blasticidin-S and Its Application for Functional Analysis of Thioredoxin Peroxidase-1 (2015)
  • Author(s): Masahito Asada, Kazuhide Yahata, Hassan Hakimi, Naoaki Yokoyama, Ikuo Igarashi, Osamu Kaneko, Carlos E. Suarez, Shin-ichiro Kawazu
  • Organizer: The 14th Awaji International Forum on Infection and Immunity
  • Place of Presentation: 淡路夢舞台国際会議場(兵庫県・淡路市)
  • Year and Date: 2015-09-08
  • Int'l Joint Research
• [Presentation] Establishment of transient and stable transfection system for Babesia ovata (2015)
  • Author(s): Hassan Hakimi、山岸潤也、外川裕人、田中健、金子修、河津信一郎、麻田正仁
  • Organizer: 第158回日本獣医学会
  • Place of Presentation: 北里大学(青森県・十和田市)
  • Year and Date: 2015-09-07
• [Presentation] 原虫とその感染症のゲノム・トランスクリプトーム解析 (2015)
  • Author(s): 田中健、山岸潤也、Hassan Hakimi、麻田正仁、河津信一郎
  • Organizer: 第158回日本獣医学会
  • Place of Presentation: 北里大学（青森県・十和田市）
  • Year and Date: 2015-09-07
• [Presentation] バベシア原虫メロゾイトの宿主赤血球遊出機構 (2015)
  • Author(s): 麻田正仁、Ehab Mossaad、 Hassan Hakimi、矢幡一英、河津信一郎、金子修
  • Organizer: 第13回分子寄生虫学マラリア研究フォーラム合同大会
  • Place of Presentation: 帯広畜産大学(北海道・帯広市）
  • Year and Date: 2015-08-30
• [Presentation] WR99210選択システム及びブラスチシジンS選択システムの併用によるBabesia bovis遺伝子改変技術の確立及びノックアウト遺伝子相補実験への応用 (2015)
  • Author(s): 麻田正仁、矢幡一英、Hassan Hakimi、横山直明、五十嵐郁男、金子修、Carlos Suarez、河津信一郎
  • Organizer: 第84回日本寄生虫学会大会
  • Place of Presentation: 杏林大学 （東京都・三鷹市）
  • Year and Date: 2015-03-21 – 2015-03-22
• [Presentation] Babesia bovisにおけるダブルトランスフェクション法の確立 (2014)
  • Author(s): 麻田正仁、Carlos Suarez、Hassan Hakimi、横山直明、五十嵐郁男、矢幡一英、金子修、河津信一郎
  • Organizer: 第157回日本獣医学会大会
  • Place of Presentation: 北海道大学 (北海道・札幌市)
  • Year and Date: 2014-09-09 – 2014-09-12
• [Presentation] バベシア原虫における遺伝子改変技術の開発 (2014)
  • Author(s): 麻田正仁、矢幡一英、Hassan Hakimi、横山直明、五十嵐郁男、金子修、Carlos Suarez、河津信一郎
  • Organizer: 第12回分子寄生虫学マラリア研究フォーラム合同大会
  • Place of Presentation: 帯広畜産大学 (北海道・帯広市)
  • Year and Date: 2014-08-31 – 2014-09-03
• [Presentation] Gliding motility of Babesia bovis merozoites (2014)
  • Author(s): Asada M., Goto Y., Yahata K., Yokoyama N., Inoue N., Kaneko O., Kawazu SI.
  • Organizer: 第13回国際寄生虫学会(ICOPAXIII)
  • Place of Presentation: Camino Real Hotel （メキシコ・メキシコシティー）
  • Year and Date: 2014-08-10 – 2014-08-15
• [Presentation] Gliding motility of Babesia bovis merozoites visualized by time-lapse video microscopy (2014)
  • Author(s): Masahito Asada, Yasuyuki Goto, Kazuhide Yahata, Naoaki Yokoyama, Noboru Inoue, Osamu Kaneko, Shin-ichiro Kawazu
  • Organizer: 第3回感染症若手フォーラム
  • Place of Presentation: 「やすらぎ伊王島」海の見えるホテル（長崎市）
• [Presentation] バベシア原虫におけるダブルトランスフェクション法の確立 (2014)
  • Author(s): 麻田正仁、Carlos Suarez、横山直明、五十嵐郁男、矢幡一英、金子修、河津信一郎
  • Organizer: 第83回日本寄生虫学会大会
  • Place of Presentation: 愛媛大学城北キャンパス（松山市）
• [Presentation] Study on gliding motility of Babesia bovis merozoites using bioimaging analysis (2014)
  • Author(s): Masahito Asada and Shin-ichiro Kawazu
  • Organizer: 第6回ASEAN熱帯医学寄生虫学会議 シンポジウム
  • Place of Presentation: Intercontinental Kuala Lumpur (マレーシア・クアラルンプール)
  • Invited
• [Presentation] Babesia bovisチオレドキシンペルオキシターゼBbTPx-1ノックアウト原虫は活性窒素種負荷に対する感受性が上昇する (2013)
  • Author(s): 麻田正仁、Carlos Suarez、薄井美帆、後藤康之、横山直明、井上昇、矢幡一英、金子修、河津信一郎
  • Organizer: 第156回日本獣医学会学術集会
  • Place of Presentation: 岐阜大学（岐阜市）