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In vitro titration of Theileria parva tick derived stabilates

Published online by Cambridge University Press:  01 March 2004

T. MARCOTTY ,
N. SPEYBROECK ,
D. BERKVENS ,
G. CHAKA ,
R. BESA ,
M. MADDER ,
T. DOLAN ,
B. LOSSON  and
J. BRANDT
T. MARCOTTY
Affiliation:
Veterinary Department, Institute of Tropical Medicine, Nationalestraat 155, 2000 Antwerp, Belgium
N. SPEYBROECK
Affiliation:
Veterinary Department, Institute of Tropical Medicine, Nationalestraat 155, 2000 Antwerp, Belgium
D. BERKVENS
Affiliation:
Veterinary Department, Institute of Tropical Medicine, Nationalestraat 155, 2000 Antwerp, Belgium
G. CHAKA
Affiliation:
Provincial Veterinary Office, PO Box 510155 Chipata, Zambia
R. BESA
Affiliation:
Provincial Veterinary Office, PO Box 510155 Chipata, Zambia
M. MADDER
Affiliation:
Veterinary Department, Institute of Tropical Medicine, Nationalestraat 155, 2000 Antwerp, Belgium
T. DOLAN
Affiliation:
Livestock Services, PO Box 24437, 00502 Karen, Nairobi, Kenya
B. LOSSON
Affiliation:
Faculty of Veterinary Medicine, University of Liège, B42, 4000 Liège, Belgium
J. BRANDT
Affiliation:
Veterinary Department, Institute of Tropical Medicine, Nationalestraat 155, 2000 Antwerp, Belgium
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Abstract

Immunization against the protozoan Theileria parva by infection and treatment has proved to be very efficient for the control of East Coast fever, an acute and often-fatal lymphoproliferative tick-borne disease of cattle in Eastern, Central and Southern Africa. The immunizing dose of live T. parva sporozoites used in this method is usually determined by in vivo titration. An alternative in vitro method of quantification of sporozoites in whole tick-derived stabilates is proposed. The method consists of incubating serially diluted T. parva stabilates with bovine peripheral blood lymphocytes, the host cell that is infected naturally. Allowing the cultures to incubate undisturbed for the full cultivation period (10 days) reduced the variability among replicate titrations. Fungal contaminations were avoided by centrifuging stabilates at 400 g prior to the incubation, which did not precipitate sporozoites significantly. Fungistatics, Nystatin and Flucytosine, did not appear to interfere with the in vitro development of T. parva but their effect on fungal growth was limited. In vitro titration data were compared to in vivo infection data for 2 stabilates. In vitro titration of T. parva sporozoites should allow more ethical and efficient research on the preparation and storage of T. parva tick-derived stabilates.

Keywords

Theileria parvaimmunizationtitrationin vitro
Type
Research Article
Information
Parasitology , Volume 128 , Issue 2 , February 2004 , pp. 131 - 137
Copyright
2004 Cambridge University Press

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