Abel Oliva SPI Sociedade Portuguesa de Inova

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					Biosensor / Biomolecular
 Diagnostics Laboratory

   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
 Project Acronyme: ADDAV    Contract No: ICA4-CT2000-300028
                   INCO 2 PROGRAMME - EU


              Abel Gonzalez Oliva
                      oliva@itqb.unl.pt
              Parceiros no ADDAV
1. Prof. J.S. Ahmed (Co-ordinator), Research Center Bortstel,
   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.
         Objectivos do ADDAV

Os principais objectivos de este
projecto de investigação foram:
                                   Merogony /

Desenvolvimento de métodos de
diagnóstico e vacinas, para
espécies altamente patogénicas
de Theileria sp. que afectam
pequenos ruminantes na China.
Zona endémica de Theileria sp. na RP China




        Geographic distribution of Theileria sp. (China) in the northwest part of the
             People’s Republic of China. Endemic area surrounded in black.
             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
                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
                           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.
                              Artigos científicos

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). 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. 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. 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 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. Helder J. Cruz, Carla C. Rosa, Abel G. Oliva
(2002) Parasitol Res 88: S4-S.
       Presentações em Conferências Int.

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. 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). 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 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.
        Presentações em Conferências Int.

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

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 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 Oliva, A. 2nd.Internation Conference on Ticks and Tick-
borne diseases (ICTTD). Shanghai, China, September 11th- 15th, 2000.
Lanzhou Veterinary Research Institute (LIVRI)