Surveillance and Control of Animal Protozoan Diseases

OIE Collaborating Centre Reports

Activities in 2012

Title of Collaborating Centre: / Surveillance and Control of Animal Protozoan Diseases
Address of Collaborating Centre / National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Nishi 2-13, Inada-cho Obihiro, Hokkaido 080-8555, JAPAN
Tel.: / +81-155-49-5641
Fax: / +81-155-49-5643
e-mail address: /
website: / http://www.obihiro.ac.jp/~protozoa
Name of Director of Institute (Responsible Official): / Prof. Hiroshi Suzuki, Ph.D.
Name (including Title and Position) of Head of the Collaborating Centre (formally OIE Contact Point): / Prof. Ikuo Igarashi, D.V.M., Ph.D.
Name (including Title and Position) of writer of this report
(if different from above)


Summary of activities specifically related to the mandate of
OIE Collaborating Centres

1. Activities as a centre of research, expertise, standardisation and dissemination of techniques within the remit of the mandate given by the OIE

Since the National Research Center for Protozoan Diseases was designated as an OIE Collaborating Centre in May 2008, we had established the OIE Reference Centre laboratory that prevents biological hazards caused by imported biological samples with BSL2 level. At this laboratory, sample preparations including serum isolation and DNA extraction can be performed serological tests such as for ELISA, IFAT, immunochromatographic test and molecular tests such as molecular cloning, PCR/LAMP and DNA sequencing for diagnostics and epidemiological surveillances.

2. Proposal or development of any procedure that will facilitate harmonisation of international regulations applicable to the surveillance and control of animal diseases, food safety or animal welfare

Molecular diagnostics such as PCR and Loop-mediated isothermal amplification (LAMP) and serological methods including ELISA and an immunochormatographic test (ICT) for specific detection of Trypanosoma, Babesia, Theileria, Toxoplasma and Neopspora have been developed. Evaluation and standardization of these methods for field application are ongoing.

Novel anti-protozoan potential drugs have evaluated for different protozoan parasites and further evaluation of these drugs have done for Babesia of treatment of protozoan diseases in the fields.

Epidemiological survey on tick-vectors that are responsible for the transmission of the protozoan diseases were done for the control of tick-born protozoan diseases in the future.

3. Networking

a) Maintenance of a network with other OIE Collaborating Centres designated for the same specialty, and

None

b) Should the need arise, maintenance of a network with Collaborating Centres in other disciplines

There were some disagreements in the sensitivity of IFAT for equine piroplasmosis among the different laboratories in the world. One possibility is that different antigen sources may contribute these differences. Therefore, the standardisation of antigen sources will be necessary in the future.

4. Placement of expert consultants at the disposal of the OIE

Dr. N. Inoue (OIE RL expert for Surra) participated in the Meeting of the Group on Non Tsetse Transmitted Animal Trypanosomoses (NTTAT) (Chairperson: Dr. Louis Touratier), which was held on 20 May, 2012 at the Hotel Ampere, Paris.

5. Provision of scientific and technical training, within the remit of the mandate given by the OIE, to personnel from OIE Member Countries

1. Advanced Research Course on International Animal Health was held at The National Research Center for Protozoan Diseases, Obihiro, Japan, financed by Japan International Cooperation Agency (JICA). Five young scientists from various countries (Argentina, Mongolia, Thailand and Zambia) have been trained for basic molecular biology and molecular diagnostics for 10 months (Oct. 2011-Aug. 2012). Another 8 young scientists from developing countries (Indonesia, Mongolia, Thailand, Uganda and Zambia) are being trained for basic molecular biology and molecular diagnostics for 10 months (Oct. 2012-Aug. 2013).

2. Thirteen foreign (Afghanistan, Benin, China, Egypt, Kenya, Mongolia, Philippines, Sri Lanka, Thailand and Viet Nam) and ten Japanese graduate students were trained by the global COE program ("Frontier Program for Animal Global Health and Hygiene"). Eleven foreign from different countries (Syria, Bolivia, Burkina Faso, China, Egypt and Kenya) and six Japanese postdoctoral fellows f have been worked on protozoan diseases.

6. Organisation of scientific meetings on behalf of the OIE

International Workshop on molecular diagnosis of Equine Piroplasmosis was held at National Research Center on Equine, Hisar, India in cooperation with National Research Center for Protozoan Diseases, Obihiro, Japan during November 29 to December 8 inviting 11 participants (Afghanistan, Bangladesh, Bhutan, India, Nepal and Sri Lanka) sponsored under OIE Twining Project.

7. Coordination of scientific and technical studies in collaboration with other laboratories, organisations or collaborating centres

1. Collaboration, training and technology transfer to the National Research Institute for Equines, India is ongoing under the OIE Twinning Program (Equine Piroplasmosis).

2. Molecular epidemiological survey of bovine piroplasmosis in Viet Nam was carried out in collaboration with Hue University (Prof. T. B. Lan).

3. Molecular epidemiological survey of human and animal African trypanosomoses was carried out with University of Zambia (Dr. Boniface Namangala), and Hokkaido University (Prof. Chihiro Sugimoto).

4. Epidemiological studies on animal babesiosis and theileriosis in China were carried out in collaboration with Yanbian University (Prof. S. Zhang).

5. Epidemiological studies on equine piroplasmosis in Mongolia were carried out in collaboration with the Institute of Veterinary Medicine (Dr. B. Battsetseg).

6. Epidemiological studies on bovine piroplasmosis in Thailand were carried out in collaboration with Kasetsart University (Dr. T. Inpankaew and Prof. S. Jittapalapong).

7. Molecular epidemiological survey of bovine babesiosis in Syria was carried out in collaboration with Al-Baath University (Prof. D. Tabbaa).

8. Research project on molecular epidemiology on ovine babesiosis in Turkey is ongoing in collaboration with Selcuk University (Prof. F. Sevinc).

9. Development of a gene konck-out/complementatin system based in WR99210/dhfr and blasticidin/bsd selection markers for functional gene characterization in Babesia bovis has started with Agricultural Research Service, United States Department of Agriculture（ARS-USDA）, (Dr. Carlos E. Suarez).

10. Research project on molecular epidemiology on bovine piroplasmosis was done in collaboration with Veterinary Research Institute in Sri Lanka (Dr. T. Sivakumar).

11. Several research projects on malaria, ticks, bovine piroplasmosis, anti-Babesia drugs, Toxoplasma, African trypanosomiasis were carried out with Jichi Medical College, Kagoshima University, Kanazawa University, Kitazato University, Gifu University and The University of Tokyo.

8. Publication and dissemination of any information within the remit of the mandate given by the OIE that may be useful to Member Countries of the OIE

1.  Aboulaila M, Munkhjargal T, Sivakumar T, Ueno A, Nakano Y, Yokoyama M, Yoshinari T, Nagano D, Katayama K, El-Bahy N, Yokoyama N, Igarashi I. 2012. Apicoplast-targeting antibacterials inhibit the growth of Babesia parasites. Antimicrob Agents Chemother., 56:3196-3206.

2.  Aboulaila M, Terkawi MA, Seuseu FJ, Ota N, de Macedo AC, Yokoyama N, Xuan X, Igarashi I. 2012. Expression and immunological characterization of the heat shock protein-70 homologue from Babesia bigemina. Vet Parasitol., 190:401-410.

3.  Altangerel K, Sivakumar T, Battsetseg B, Battur B, Ueno A, Igarashi I, Yokoyama N. 2012. Phylogenetic relationships of Mongolian Babesia bovis isolates based on the merozoite surface antigen (MSA)-1, MSA-2b, and MSA-2c genes. Vet Parasitol., 184: 309-319.

4.  Aung, K. M., Boldbaatar, D., Umemiya-Shirafuji, R., Liao, M., Xuan, X., Suzuki, H., Galay, R. L., Tanaka, T. and Fujisaki, K. 2012. Scavenger receptor mediates systemic RNA interference in ticks. PLOS ONE, 7:e33504.

5.  Asada M, Goto Y, Yahata K, Yokoyama N, Kawai S, Inoue N, Kaneko O, Kawazu S. 2012. Gliding motility of Babesia bovis merozoites visualized by time-lapse video microscopy. PLOS ONE. 7(4):e35227.

6.  Asada, M., Tanaka, M., Goto, Y., Yokoyama, N., Inoue, N., and Kawazu, S. 2012. Stable expression of green fluorescent protein and targeted disruption of thioredoxin peroxidase-1 gene in Babesia bovis with the WR99210/dhfr selection system. Mol. Biochem. Parasitol., 181:162-170.

7.  Badolo A, Okado K, Guelbeogo WM, Aonuma H, Bando H, Fukumoto S, Sagnon N, Kana H. 2012. Development of an allele-specific, loop-mediated, isothermal amplification method (AS-LAMP) to detect the L1014F kdr-w mutation in Anopheles gambiae s. l. Malar J. 2012. 11: 227.

8.  Cao S, Aboge GO, Terkawi MA, Yu L, Kamyingkird K, Luo Y, Li Y, Goo YK, Yamagishi J, Nishikawa Y, Yokoyama N, Suzuki H, Igarashi I, Maeda R, Inpankaew T, Jittapalapong S, Xuan X. 2012. Molecular detection and identification of Babesia bovis and Babesia bigemina in cattle in northern Thailand. Parasitol Res., 111:1259-1266.

9.  Chiou SP, Yokoyama N, Igarashi I, Kitoh K, Takashima Y. 2012. Serum of Babesia rodhaini infected mice down regulates catalase activity of healthy erythrocytes. Exp Parasitol. 132(3):327-333.

10.  Enomoto, M., Kawazu, S., Kawai, S., Furuyama, Y., Ikegami, T., Watanabe, J., and Mikoshiba, K. 2012. Blockage of spontaneous Ca2+ oscillation causes cell death in intraerythrocitic Plasmodium falciparum. PLoS ONE 7: e39499.

11.  Galay RL, Maeda H, Aung KM, Umemiya-Shirafuji R, Xuan X, Igarashi I, Tsuji N, Tanaka T, Fujisaki K. 2012. Anti-babesial activity of a potent peptide fragment derived from longicin of Haemaphysalis longicornis. Trop Anim Health Prod. 44: 343-348.

12.  Goo YK, Aboge GO, Terkawi MA, Jia H, Yamagishi J, Sunaga F, Namikawa K, Cha SY, Jang HK, Kim S, Nishikawa Y, Xuan X. 2012. Four promising antigens, BgP32, BgP45, BgP47, and BgP50, for serodiagnosis of Babesia gibsoni infection were classified as B. gibsoni merozoite surface protein family. Parasitol Int. 61:364-368.

13.  Goo YK, Lee N, Alaa Terkawi M, Luo Y, Oluga Aboge G, Nishikawa Y, Suzuki H, Kim S, Xuan X. 2012. Development of a rapid immunochromatographic test using a recombinant thrombospondin-related adhesive protein of Babesia gibsoni. Vet Parasitol., 190:595-598.

14.  Goto, Y., Duthie MS., Nguyen, TT., Asada, M., Kawazu, S., Carter, D., and Inoue, N. 2012. Serological characterizations of tandem repeat proteins for detection of African trypanosome infection in cattle. Parasitol. Int., 60:538-540.

15.  Hakimi H, Asada M, Angeles JM, Kawai S, Inoue N, Kawazu S. 2012. Plasmodium vivax and Plasmodium knowlesi: Cloning, expression and functional analysis of 1-Cys peroxiredoxin. Exp Parasitol.,133:101-105.

16.  Hakimi H, Asada M, Angeles JM, Inoue N, Kawazu S. 2012. Cloning and characterization of Plasmodium vivax thioredoxin peroxidase-1. Parasitol Res., 111:525-529.

17.  Hayashida, K., Hara, Y., Abe, T., Yamasaki, C., Toyoda, A., Kosuge, T., Suzuki, Y., Sato, Y., Kawashima, S., Katayama, T., Wakaguri, H., Inoue, N., Homma, K., Tada-Umezaki, M., Yagi, Y., Fujii, Y., Habara, T., Kanehisa, M., Watanabe, H., Ito, K., Gojobori, T., Sugawara, H., Imanishi, T., Weir, W., Gardner, M., Pain, A., Shiels, B., Hattori, M., Nene, V. and Sugimoto, C. 2012. Comparative genome analysis of three eukaryotic parasites with differing abilities to transform leukocytes reveals key mediators of Theileria-induced leukocyte transformation. MBio 3:e00204-212.

18.  Hiasa J, Kohara J, Nishimura M, Xuan X, Tokimitsu H, Nishikawa Y. 2012. ELISAs based on rNcGRA7 and rNcSAG1 antigens as an indicator of Neospora caninum activation. Vet Parasitol., 187:379-385.

19.  Hiasa J, Nishimura M, Itamoto K, Xuan X, Inokuma H, Nishikawa Y. 2012. ELISAs based on Neospora caninum dense granule protein 7 and profilin for estimating the stage of neosporosis. Clin Vaccine Immunol., 19:411-417.

20.  Ishigami, S. T., Goto, Y., Inoue, N., Kawazu, S., Matsumoto, Y., Imahara, Y., Tarumi, M., Nakai, H., Fusetani., N. and Nakao, Y. 2012. Cristaxenicin A, an antiprotozoal xenicane diterpenoid from the deep sea gorgonian Acanthoprimnoa cristata. J Org Chem., 77: 10962-10966.

21.  Kameyama K, Nishimura M, Punsantsogvoo M, Ibrahim HM, Xuan X, Furuoka H, Nishikawa Y. 2012. Immunological characterization of Neospora caninum cyclophilin. Parasitology, 39:294-301.

22.  Kimura, R., Komaki-Yasuda, K., Kawazu, S., and Kano, S. 2012. 2-Cys peroxiredoxin of Plasmodium falciparum is involved in resistance to heat stress of the parasite. Parasitol. Int., 62: 137-143.

23.  Kimura, A., Nishikawa, H., Nomura, N., Mitsuyama, J., Fukumoto, S., Inoue, I., and Kawazu, S. 2012. In vitro and in vivo antimalarial activity of T-2307, a novel arylamidine. Antimicrob. Agents Chemother., 56: 2191-2193.

24.  Kumar S, Yokoyama N, Kim JY, Bork-Mimm S, Inoue N, Xuan X, Igarashi I, Sugimoto C. 2012. Theileria equi merozoite antigen-2 interacts with actin molecule of equine erythrocyte during their asexual development. Exp Parasitol., 132:508-512.

25.  Kume A, Boldbaatar D, Takazawa Y, Umemiya-Shirafuji R, Tanaka T, Fujisaki K. 2012. RNAi of the translation inhibition gene 4E-BP identified from the hard tick, Haemaphysalis longicornis, affectslipid storage during the off-host starvation period of ticks. Parasitol Res., 111: 889-896.

26.  Li Y, Terkawi MA, Nishikawa Y, Aboge GO, Luo Y, Ooka H, Goo YK, Yu L, Cao S, Sun Y, Yamagishi J, Masatani T, Yokoyama N, Igarashi I, Xuan X. 2012. Macrophages are critical for cross-protective immunity conferred by Babesia microti against Babesia rodhaini infection in mice. Infect Immun., 80: 311-320.

27.  Luo Y, Terkawi MA, Jia H, Aboge GO, Goo YK, Cao S, Li Y, Yu L, Ooka H, Kamyingkird K, Masatani T, Zhang S, Nishikawa Y, Igarashi I, Xuan X. 2012. A double antibody sandwich enzyme-linked immunosorbent assay for detection of secreted antigen 1 of Babesia microti using hamster model. Exp Parasitol., 130:178-182.

28.  Mekata, H., Konnai, S., Mingala, C. N., Abes, N. S., Gutierrez, C. A., Dargantes, A. P., Witola, W. H., Inoue, N., Onuma, M., Murata, S. and Ohashi. K. 2012. Kinetics of regulatory dendritic cells in inflammatory responses during Trypanosoma evansi infection. Parasite Immunol., 34:318-329.

29.  Munkhjargal T, Aboulaila M, Terkawi MA, Sivakumar T, Ichikawa M., Davaasuren B, Nyamjargal T, Yokoyama N, Igarashi I. 2012. Inhibitory Effects of Pepstatin A and Mefloquine on the Growth of Babesia Parasites. Am J Trop Med Hyg., 87:681-688.

30.  Munkhjargal T, Aboulaila M, Ueno A, Sivakumar T, Nakano Y, Yokoyama M, Yokoyama N, Igarashi I. 2012. Cloning and characterization of histone deacetylase from Babesia bovis. Vet Parasitol., 190: 423-433.

31.  Nguyen, TT., Goto, Y., Lun, ZR., Kawazu, S., and Inoue, N. 2012. Tandem Repeat Proteins as potential diagnostic antigen for Trypanosoma evansi infection. Parasitol Res., 110:733-739.

32.  Ooka H, Terkawi MA, Shinuo C, Aboge G, Goo YK, Luo Y, Li Y, Nishikawa Y, Igarashi I, Xuan X. 2012. Molecular and immunological characterization of a novel 32-kDa secreted protein of Babesia microti. J Parasitol., 98:1045-1048.

33.  Shimamoto Y, Sasaki M, Ikadai H, Ishizuka M, Yokoyama N, Igarashi I, Hoshi F, Kitamura H. 2012. Down-regulation of hepatic cytochrome P450 3A in mice infected with Babesia microti. J Vet Med Sci., 74: 241-245.

34.  Sivakumar T, Altangerel K, Battsetseg B, Battur B, Aboulaila M, Munkhjargal T, Yoshinari T, Yokoyama N, Igarashi I. 2012. Genetic detection of Babesia bigemina from Mongolian cattle using apical membrane antigen-1 gene-based PCR assay. Vet Parasitol. 187:17-22.