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In vitro titration of Theileria parva tick derived stabilates

Published online by Cambridge University Press:  01 March 2004

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

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.

Type
Research Article
Copyright
2004 Cambridge University Press

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References

REFERENCES

BROWN, C. G. D., STAGG, D. A., PURNELL, R. E., KANHAI, G. K. & PAYNE, R. C. (1973). Infection and transformation of bovine lymphoid cells in vitro by infective particles of Theileria parva. Nature, London 245, 101103.CrossRefGoogle Scholar
BÜSCHER, G. & OTIM, B. (1986). Quantitative studies on Theileria parva in the salivary glands of Rhipicephalus appendiculatus adults: quantitation and prediction of infection. International Journal for Parasitology 16, 93100.CrossRefGoogle Scholar
CLEVES, M. A., GOULD, W. W. & GUTIERREZ, R. G. (2002). An Introduction to Survival Analysis Using Stata. Stata Press, College Station.
COLLET, D. (1991). Modelling Binary Data. Chapman and Hall, London.CrossRef
CUNNINGHAM, M. P., BROWN, C. G. D., BURRIDGE, M. J. & PURNELL, R. E. (1973). Cryopreservation of infective particles of Theileria parva. International Journal for Parasitology 3, 583587.CrossRefGoogle Scholar
DOLAN, T. T., YOUNG, A. S., LOSOS, G. J., McMILLAN, I., MINDER, C. E. & SOULSBY, K. (1984). Dose dependent responses of cattle to Theileria parva stabilate infection. International Journal for Parasitology 14, 8995.CrossRefGoogle Scholar
DORNY, P., SPEYBROECK, N., VERSTRAETE, M., BAEKE, A., DE BECKER, A., BERKVENS, D. & VERCRUYSSE, J. (2002). Serological survey of Toxoplasma gondii, feline immunodeficiency virus and feline leukaemia virus in urban stray cats in Belgium. Veterinary Record 151, 626629.CrossRefGoogle Scholar
GODDEERIS, B. M. & MORRISON, W. I. (1988). Techniques for the generation, cloning, and characterization of bovine cytotoxic T cells specific for the protozoan Theileria parva. Journal of Tissue Culture Methods 11, 101110.CrossRefGoogle Scholar
JARRETT, W. F., CRIGHTON, G. W. & PIRIE, H. M. (1969). Theileria parva: kinetics of replication. Experimental Parasitology 24, 925.CrossRefGoogle Scholar
KANHAI, G. K., PEGRAM, R. G., HARGREAVES, S. K., HOVE, T. & DOLAN, T. T. (1997). Immunisation of cattle in Zimbabwe using Theileria parva (Boleni) without concurrent tetracycline therapy. Tropical Animal Health and Production 2, 9298.CrossRefGoogle Scholar
MARCOTTY, T., BILLIOUW, M., CHAKA, G., BERKVENS, D., LOSSON, B. & BRANDT, J. (2001). Immunisation against East Coast fever by the infection and treatment method: evaluation of the use of ice baths for field delivery and appraisal of an acid formulation of long-acting tetracycline. Veterinary Parasitology 99, 175187.CrossRefGoogle Scholar
MÄSER, P., GRETHER-BÜHLER, Y., KAMINSKY, R. & BRUN, R. (2002). An anti-contamination cocktail for the in vitro isolation and cultivation of parasitic protozoa. Parasitology Research 88, 172174.CrossRefGoogle Scholar
MEHTA, C. R. & PATEL, N. R. (1995). Exact logistic regression: theory and examples. Statistics in Medicine 14, 21432160.CrossRefGoogle Scholar
MUSISI, F. L., QUIROGA, J. C., MUTUGI, J. J., JACOBSEN, J., DE CASTRO, J. J. & DI GUILIO, G. (1992). Immunisation against East Coast Fever: Recent experiences with the trivalent vaccine in East and Central Africa. In Bovine Theileriosis, 3rd Symposium on Tropical and Animal Health and Production (ed. Paling, R. W. ), pp. 2632. Office for International Co-operation, Utrecht.
OLE-MOI YOI, O. K. (1989). Theileria parva: An intracellular protozoan parasite that induces reversible lymphocyte transformation. Experimental Parasitology 69, 204210.CrossRefGoogle Scholar
RADLEY, D. E., BROWN, C. G., BURRIDGE, M. J., CUNNINGHAM, M. P., PEIRCE, M. A. & PURNELL, R. E. (1974). East Coast fever: quantitative studies of Theileria parva in cattle. Experimental Parasitology 36, 278287.CrossRefGoogle Scholar
RADLEY, D. E., BROWN, C. G. D., CUNNINGHAM, M. P., KIMBER, C. D., MUSISI, F. L., PAYNE, R. C., PURNELL, R. E., STAGG, S. M. & YOUNG, A. S. (1975). East Coast fever. 3. Chemoprophilactic immunisation of cattle using oxytetracycline and a combination of theilerial strains. Veterinary Parasitology 1, 5160.Google Scholar
STATACORP (2001). Stata Statistical Software: Release 7.0. Stata Corporation, Texas.
UILENBERG, G. (1999). Immunization against diseases caused by Theileria parva: a review. Tropical Medicine and International Health 4, A12A20.CrossRefGoogle Scholar
WILKIE, G., KIRVAR, E. & BROWN, C. G. D. (2002). Validation of an in vitro method to determine infectivity of cryopreserved sporozoites in stabilates of Theileria spp. Veterinary Parasitology 104, 199209.CrossRefGoogle Scholar