1. Abel Gonzalez Oliva oliva@itqb.unl.pt oliva@itqb.unl.pt Biosensor / Biomolecular Diagnostics Laboratory Project Acronyme: ADDAV Contract No: ICA4-CT2000-300028 INCO 2 PROGRAMME- EU Project Acronyme: ADDAV Contract No: ICA4-CT2000-300028 INCO 2 PROGRAMME - EU Molecular and immunological characterization of merozoite antigens and their encoding genes of a Theileria species highly pathogenic for small ruminants in China: application for the development of diagnostics and vaccine 2001 - 2005

2. 1.Prof. J.S. Ahmed (Co-ordinator), Research Center Bortstel, Borstel, Germany. Borstel, Germany. 2. Prof. Hong Yin, Lanzhou Veterinary Research Institute (LIVRI), Chinese Academy of Agricultural Sciences, Xujiaping 11, Yanchangbu, Lanzhou, Gansu, 730046, PR of China. 3. Prof. J. Wu Animal Quarantine Institule, Ministry of Agriculture (AQIMA), 369 Nanjing Road, Qingdao 266032, PR of China. 4. Prof. Z. Yuan, Institute of Animal Husbandry of Gannan Tibet Nationality Autonomous Prefecture, Gansu Province, PR China. 5. Prof. Abel G. Oliva, Instituto de Biologia Experimental e Technologica (IBET) Biosensor/Biomolecular Diagnostics Laboratory - Apartado 12, 2781-901 Oeiras, Portugal. Parceiros no ADDAV

3. Objectivos do ADDAV Os principais objectivos de este projecto de investigação foram: Desenvolvimento de métodos de diagnóstico e vacinas, para espécies altamente patogénicas de Theileria sp. que afectam pequenos ruminantes na China.

4. Geographic distribution of Theileria sp. (China) in the northwest part of the Peoples Republic of China. Endemic area surrounded in black. Zona endémica de Theileria sp. na RP China

5. Tarefas do IBET no ADDAV Developed tasks: Participation in the infection process and in serum/blood collection Infected blood characterisation by PCR and Reverse line blot (RLB) technique. Merozoites isolation and purification studies as well as identification of potential antigens. Antigens characterisation by mass spectrometry Establishment of In vitro cultivation of Theileria sp. (China) merozoites in erythrocytes

6. Treino de pessoal Coolaboração na preparação do Workshop in Biochemical & Molecular Biology Techniques para estudantes Chineses. Lanzhou - 12-26 June 2002 Joana Miranda (PhD Student) tem realizado numerosas visitas ao Lanzhou Veterinary Research Institute – Lanzhou, República Popular da China, onde desenvolveu trabalho experimental relativo a parasita Theileria sp., sob a supervisão do Prof. Yin Hong), nomeadamente para recolha de amostras de soro e sangue ovino, isolamento de merozoites, purificação de RNA e produção de cDNA, e desenvolvimento e validação de teste de ELISA Junho 2002 e Abril-Maio 2005

7. Workshop Workshop Establishment of tick colonies and tissue cultures of Theileria annulata for the immunisation against Tropical Theilerosis. CMDT & IBET, Lisbon, Portugal, from the 31st March until the 10th of April, 2003.

8. Establishment of continuous in vitro culture of erythrocytic stages of a Theileria species highly pathogenic for small ruminants in China. Establishment of continuous in vitro culture of erythrocytic stages of a Theileria species highly pathogenic for small ruminants in China. Miranda J., Nascimento E., Cruz H., Zweigarth E., Oliva A. (2006) Vet Parasit. (submitted) Identification of homologous genes of T. annulata proteins in the genome of Theileria sp. (China). Identification of homologous genes of T. annulata proteins in the genome of Theileria sp. (China). Miranda J., Bakheit M., Schneider I.., Haller D., Ahmed J., Hong Y., Oliva A., Seitzer U. Ann NYAcad Sciences (submitted) Development of a recombinant indirect ELISA for the diagnosis of Theileria sp. (China) infection in small ruminants. Development of a recombinant indirect ELISA for the diagnosis of Theileria sp. (China) infection in small ruminants. Miranda J, Bakheit MA, Liu Z, Yin H, Yongjuan M, Guo S, Beyer D, Oliva A, Ahmed JS, Seitzer U (2006) - Parasitol Res (in press) Identification and characterization of merozoite antigens of a Theileria species highly pathogenic for small ruminants in China Identification and characterization of merozoite antigens of a Theileria species highly pathogenic for small ruminants in China. Miranda J, Nascimento E, Cruz H, Hong Y, A. Coelho, Ahmed J, Oliva A (2006) Journal of Veterinary Pathology Ann NYAcad Sci (in press). Identification of antigenic proteins of a Theileria species pathogenic for small ruminants in China: characterization of the immune response and assessment of diagnostic potential Identification of antigenic proteins of a Theileria species pathogenic for small ruminants in China: characterization of the immune response and assessment of diagnostic potential J.Miranda, B.Stumme, D. Beyer, H. Cruz, A.G. Oliva, D. Wicklein, H. Yin, L. Jianxun, J.S. Ahmed, U. Seitzer. (2004) Ann NYAcad Sci, 1026:1-4 Immunosensors for diagnostic applications Immunosensors for diagnostic applications. Helder J. Cruz, Carla C. Rosa, Abel G. Oliva (2002) Parasitol Res 88: S4-S. Artigos científicos

9. Presentações em Conferências Int. Establishment of Continuous in vitro Culture of Erythrocytic Stages of a Theileria species Highly Pathogenic for Small Ruminants in China. Establishment of Continuous in vitro Culture of Erythrocytic Stages of a Theileria species Highly Pathogenic for Small Ruminants in China. JMiranda, ENascimento, HCruz, YHong, EZweygarth, AOliva June 26 -July 1st 2005, Hanoi/Vietnam. Identification and characterization of merozoite antigens of Theileria species 2 highly pathogenic for small ruminants in China. Identification and characterization of merozoite antigens of Theileria species 2 highly pathogenic for small ruminants in China. Miranda, J.; Nascimento, E.; Cruz, H.; HYin.; Ahmed, J.; Oliva A. June 26 -July 1st 2005, Hanoi/Vietnam. Identification of homologous genes of T. annulata proteins in the genome of Theileria sp. (China). Identification of homologous genes of T. annulata proteins in the genome of Theileria sp. (China). JMiranda, MBakheit, ISchneider, DHaller, JAhmed, HYin, Abel G. Oliva, Ulrike Seitzer June 26 -July 1st 2005, Hanoi/Vietnam. Identification of antigenic proteins of a Theileria species pathogenic for small ruminants in China: characterisation of the immune response and assessment of diagnosis potential Identification of antigenic proteins of a Theileria species pathogenic for small ruminants in China: characterisation of the immune response and assessment of diagnosis potential JMiranda, BStumme, DBeyer, HCruz, AOliva, DWicklein, HYin, LJianxun, JAhmed, USeitzer (2003) 7th Biennal Conference STVM (Society for Tropical Veterniary Medicine) 22nd – 28th June 2003 Foz do Iguaçu, Brasil.

10. Identification and characterization of different Theileria china antigen/sera preparations from small ruminants Identification and characterization of different Theileria china antigen/sera preparations from small ruminants Helder J.S. Cruz, Joana Miranda, Abel G. Oliva (2002) Central Asia and European Union Commission International Conference on Ticks and tick-borne Diseases, Tashkent, Republic of Uzbekistan, 13-15 Mai. 2002. Antigen characterization and purification of Theileria china infected sera from small ruminants Antigen characterization and purification of Theileria china infected sera from small ruminants Helder J.S. Cruz, Joana Miranda, Luo JX, Yin Hong and Abel G. Oliva (2001) INCO- DEV Meeting: Hypodermosis and Theileria diseases International Conference, Lanzhou, Republic of China, 16th -19th Oct. 2001. Immunosensors for diagnostic Immunosensors for diagnostic Oliva, A. 2nd.Internation Conference on Ticks and Tick- borne diseases (ICTTD). Shanghai, China, September 11th- 15th, 2000. Presentações em Conferências Int. Identification and characterization of merozoite antigens of Theileria species highly pathogenic for small ruminants in China. Identification and characterization of merozoite antigens of Theileria species highly pathogenic for small ruminants in China. (oral presentation) J. Miranda, E. Nascimento, H. Cruz, H. Yin, A. Oliva. EMOP XI, Valencia, Spain, 2004

11. Lanzhou Veterinary Research Institute (LIVRI)