Establishment of an evaluation system for erythrocyte invasion by bovine Babesia parasite

• Project/Area Number: 15K14862
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Veterinary medical science
• Research Institution: Obihiro University of Agriculture and Veterinary Medicine
• Principal Investigator: Yokoyama Naoaki 帯広畜産大学, 原虫病研究センター, 教授 (80301802)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: バベシア / 赤血球 / 侵入 / 評価系 / ワクチン

Outline of Final Research Achievements

The Babesia bovis culture was subjected to a cold treatment to induce the release of merozoites from infected erythrocytes, and the free merozoites were collected. Next, an erythrocyte invasion assay for Babesia was established using these free merozoites. Subsequently, the erythrocyte invasion assay was used to investigate the role of Babesia bovis BOV57 antigen in the erythrocyte invasion by merozoites. The findings showed that the antibodies specific to BOV57 antigen significantly inhibit the invasion of merozoites into bovine erythrocytes. This observation suggested that the BOV57 antigen plays an important role in the invasion of merozoites into erythrocytes. Therefore, BOV57 is a potential candidate antigen for the development of subunit vaccines against bovine babesiosis.

Research Products

• [Int'l Joint Research] 国立獣医学研究所(スリランカ)
• [Int'l Joint Research] 国立生命科学大学・獣医学研究所(モンゴル)
• [Int'l Joint Research] フエ大学/フエ農林大学(ベトナム)
• [Int'l Joint Research] 国立畜産局/カセタルト大学(タイ)
• [Int'l Joint Research] サダト大学(エジプト)
• [Int'l Joint Research] 獣医学研究所(スリランカ)
• [Int'l Joint Research] フエ大学/フエ農林大学(ベトナム)
• [Int'l Joint Research] モンゴル農業大学(モンゴル)
• [Int'l Joint Research] 国立動物科学研究所(タイ)
• [Journal Article] Babesia bovis BOV57, a Theileria parva P67 homolog, is an invasion-related, neutralization-sensitive antigen (2017)
  • Author(s): Ishizaki Takahiro、Sivakumar Thillaiampalam、Hayashida Kyoko、Takemae Hitoshi、Tuvshintulga Bumduuren、Munkhjargal Tserendorj、Guswanto Azirwan、Igarashi Ikuo、Yokoyama Naoaki
  • Journal Title: Infection, Genetics and Evolution Volume: 54 Pages: 138-145
  • DOI: 10.1016/j.meegid.2017.06.025
  • NAID: 120006411414
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Type-specific PCR assays for Babesia bovis msa-1 genotypes in Asia: Revisiting the genetic diversity in Sri Lanka, Mongolia, and Vietnam. (2016)
  • Author(s): Liyanagunawardena N, Sivakumar T, Kothalawala H, Silva SS, Battsetseg B, Lan DT, Inoue N, Igarashi I, Yokoyama N.
  • Journal Title: Infect Genet Evol. Volume: 37 Pages: 64-69
  • DOI: 10.1016/j.meegid.2015.10.029
  • NAID: 120006390924
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] RBC Invasion and invasion-inhibition assays using free merozoites isolated after cold treatment of Babesia bovis in vitro culture. (2016)
  • Author(s): Ishizaki, T., Sivakumar, T., Hayashida, K., Tuvshintulga, B., Igarashi, I., and Yokoyama N.
  • Journal Title: Exp. Parasitol. Volume: 166 Pages: 10-15
  • DOI: 10.1016/j.exppara.2016.03.010
  • NAID: 120006390946
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Genetic diversity and antigenicity variation of Babesia bovis merozoite surface antigen-1 (MSA-1) in Thailand. (2016)
  • Author(s): Tattiyapong, M., Sivakumar, T., Takemae, H., Simking, P., Jittapalapong, S., Igarashi, I., and Yokoyama, N.
  • Journal Title: Infect. Genet. Evol. Volume: 41 Pages: 255-261
  • DOI: 10.1016/j.meegid.2016.04.021
  • NAID: 120006390947
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Epidemiology of bovine hemoprotozoa parasites in cattle and water buffalo in Vietnam (2016)
  • Author(s): Weerasooriya, G., Sivakumar, T., Lan, D.T., Long, P.T., Takemae, H., Igarashi, I., Inoue, N., and Yokoyama, N.
  • Journal Title: Journal of Veterinary Medical Science Volume: 78 Issue: 8 Pages: 1361-1367
  • DOI: 10.1292/jvms.16-0099
  • NAID: 130005261519
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] A longitudinal study of Babesia and Theileria infections in cattle in Sri Lanka. (2016)
  • Author(s): Sivakumar, T., Kothalawala, H., Weerasooriya, G., Silva, S.S.P., Puvanendiran, S., Munkhjargal, T., Igarashi, I., and Yokoyama, N.
  • Journal Title: Veterinary Parasitology: Regional Studies and Reports Volume: 6 Pages: 20-27
  • DOI: 10.1016/j.vprsr.2016.11.009
  • NAID: 120006390921
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Calcium ions are involved in egress of <i>Babesia bovis</i> merozoites from bovine erythrocytes (2015)
  • Author(s): Mossaad, E., Asada, M., Nakatani, D., Inoue, N., Yokoyama, N., Kaneko, O., and Kawazu, S.
  • Journal Title: Journal of Veterinary Medical Science Volume: 77 Issue: 1 Pages: 53-58
  • DOI: 10.1292/jvms.14-0391
  • NAID: 130004693911
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] An epidemiological survey of bovine Babesia and Theileria parasites in cattle, buffaloes, and sheep in Egypt. (2015)
  • Author(s): Elsify, A., Sivakumar, T., Nayel, M., Salama, A., Elkhtam, A., Rizk, M., Mosaab, O., Sultan, K., Elsayed, S., Igarashi, I., and Yokoyama, N.
  • Journal Title: Parasitol. Int. Volume: 64 Issue: 1 Pages: 79-85
  • DOI: 10.1016/j.parint.2014.10.002
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Genetic variations in merozoite surface antigen genes of Babesia bovis detected in Vietnamese cattle and water buffaloes. (2015)
  • Author(s): Yokoyama, N., Sivakumar, T., Tuvshintulga, B., Hayashida, K., Igarashi, I., Inoue, N., Long, P.T., and Lan, D.T.
  • Journal Title: Infect. Genet. Evol. Volume: 30 Pages: 288-295
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Optimization of a Fluorescence-Based Assay for Large-Scale Drug Screening against Babesia and Theileria Parasites. (2015)
  • Author(s): Rizk, M.A., El-Sayed, S.A., Terkawi, M.A., Youssef, M.A., El Said el Sel, S., Elsayed, G., El-Khodery, S., El-Ashker, M., Elsify, A., Omar, M., Salama, A., Yokoyama, N., and Igarashi, I.
  • Journal Title: PLoS One. Volume: 10 Issue: 4 Pages: e0125276-e0125276
  • DOI: 10.1371/journal.pone.0125276
  • Peer Reviewed / Open Access / Int'l Joint Research
• [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] Evaluation of inhibitory effect of methylene blue against Babesia and Theileria parasites. (2015)
  • Author(s): Tuvshintulga, B., Sivakumar, T., Salama, A.A., Yokoyama, N., and Igarashi, I.
  • Journal Title: J. Protozool. Res. Volume: 25 Pages: 18-28
  • NAID: 120006391034
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Analysis of the genetic diversity of Babesia bovis msa-1 in Thailand using type-specific polymerase chain reaction. (2015)
  • Author(s): Tattiyapong, M., Sivakumar, T., Takemae, H., Simking, P., Jittapalapong, S., Igarashi, I., and Yokoyama, N.
  • Journal Title: J. Protozool. Res. Volume: 25 Pages: 38-43
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Cocktail of Theileria equi antigens for detecting infection in equines. (2015)
  • Author(s): El-Sayed, S.A.E.-S., Rizk, M.A., Terkawi, M.A., Mousa, A., El-Said, E.-S.E.-S., Elsayed, G., Fouda, M., Yokoyama, N., and Igarashi, I.
  • Journal Title: Asian Pacific Journal of Tropical Biomedicine Volume: 5 Issue: 12 Pages: 977-981
  • DOI: 10.1016/j.apjtb.2015.09.001
  • Peer Reviewed / Int'l Joint Research
• [Presentation] Babesia bovis BOV57, a Theileria parva P67 homolog, is an invasion-related, neutralization-sensitive antigen. (2017)
  • Author(s): 石崎隆弘、ティライアンパラム シバクマール、林田京子、五十嵐郁男、横山直明
  • Organizer: 第160回日本獣医学会学術集会
• [Presentation] スリランカ国の牛におけるバベシア及びタイレリアの分子疫学調査 (2016)
  • Author(s): Sivakumar Thillaiampalam、五十嵐郁男、横山直明
  • Organizer: 第57回日本熱帯医学学会大会
  • Place of Presentation: 一橋大学（東京都千代田区）
  • Year and Date: 2016-11-05
• [Presentation] Effect of cold treatment of Babesia bovis-infected erythrocytes: A novel technique to isolate free, viable, invasion-competent merozites. (2015)
  • Author(s): Yokoyama, N., Ishizaki, T., Sivakumar, T., and Igarashi, I.
  • Organizer: 第56回日本熱帯医学会大会
  • Place of Presentation: 大阪大学（大阪府吹田市）
  • Year and Date: 2015-12-04
• [Presentation] The molecular diagnostic tools for bovine piroplasmosis. (2015)
  • Author(s): Yokoyama, N.
  • Organizer: 57th Annual Convention of the Sri Lanka Veterinary Association
  • Place of Presentation: Oak-Ray and Earl's Regency (Kandy, Sri Lanka)
  • Year and Date: 2015-07-19
  • Int'l Joint Research / Invited
• [Remarks] 帯広畜産大学・原虫病研究センター ホームページ
  • URL: http://www.obihiro.ac.jp/~protozoa/index.html