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Host serum modifies the drug susceptibility of Babesia bovis in vitro

Published online by Cambridge University Press:  02 December 2004

S. BORK
Affiliation:
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan
M. OKAMURA
Affiliation:
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan
T. MATSUO
Affiliation:
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan
S. KUMAR
Affiliation:
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan
N. YOKOYAMA
Affiliation:
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan
I. IGARASHI
Affiliation:
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan

Abstract

Babesia parasites generally require a defined percentage of serum in the culture medium for their in vitro growth. In this study, we attempted to culture Babesia bovis in a serum-free condition. The growth pattern and morphology of B. bovis in serum-free (plain) GIT medium were unaltered as compared to those of the standard growth condition containing 40% bovine serum in M199. When exposed to the test drugs, the parasite in plain GIT medium showed clearly lower IC50 values than those in 40% serum-containing GIT medium, indicating that several serum components may interfere with the drug bio-availability. Therefore, the serum-free culture system is useful for standardizing drug test protocols and understanding the roles of serum factors in the drug test.

Type
Research Article
Copyright
© 2004 Cambridge University Press

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References

REFERENCES

ASAHI, H. & KANAZAWA, T. ( 1994). Continuous cultivation of intraerythrocytic Plasmodium falciparum in a serum-free medium with the use of a growth-promoting factor. Parasitology 109, 397401.CrossRefGoogle Scholar
AVARZED, A., IGARASHI, I., KANEMURA, T., HIRUMI, K., OMATA, Y., SAITO, A., OYAMADA, T., NAGASAWA, H., TOYODA, Y. & SUZUKI, N. ( 1997). Improved in vitro cultivation of Babesia caballi. Journal of Veterinary Medical Science 59, 479481.CrossRefGoogle Scholar
BORK, S., YOKOYAMA, N., MATSUO, T., CLAVERIA, F. G., FUJISAKI, K. & IGARASHI, I. ( 2003 a). Growth inhibitory effects of triclosan on equine and bovine parasites. American Journal of Tropical Medicine and Hygiene 68, 334340.Google Scholar
BORK, S., YOKOYAMA, N., MATSUO, T., CLAVERIA, F. G., FUJISAKI, K. & IGARASHI, I. ( 2003 b). Clotrimazole, ketoconazole, and clodinafop-propargyl as potent growth inhibitors of equine Babesia parasites during in vitro culture. Journal of Parasitology 89, 604606.Google Scholar
BORK, S., YOKOYAMA, N., MATSUO, T., CLAVERIA, F. G., FUJISAKI, K. & IGARASHI, I. ( 2003 c). Clotrimazole, ketoconazole, and clodinafop-propargyl inhibit the in vitro growth of Babesia bigemina and Babesia bovis (Phylum Apicomplexa). Parasitology 127, 15.Google Scholar
BORK, S., YOKOYAMA, N., IKEHARA, Y., KUMAR, S., SUGIMOTO, C. & IGARASHI, I. ( 2004). Growth inhibitory effect of heparin on Babesia parasites. Antimicrobial Agents and Chemotherapy 48, 236241.CrossRefGoogle Scholar
GOFF, W. L. & YUNKER, C. E. ( 1986). Babesia bovis: increased percentage parasitized erythrocytes in cultures and assessment of growth by incorporation of [3H] hypoxanthine. Experimental Parasitology 62, 202210.CrossRefGoogle Scholar
GORENFLOT, A., BRASSEUR, P., PRECIGOUT, E., L'HOSTIS, M., MARCHAND, A. & SCHRÉVEL, J. ( 1991). Cytological and immunological responses to Babesia divergens in differential hosts: ox, gerbil, man. Parasitology Research 77, 312.CrossRefGoogle Scholar
GRANDE, N., PRECIGOUT, E., ANCELIN, M. L., MOUBRI, K., CARCY, B., LEMESTRE, J. L., VIAL, H. & GORENFLOT, A. ( 1997). Continuous in vitro culture of Babesia divergens in a serum-free medium. Parasitology 115, 8189.CrossRefGoogle Scholar
HOLMAN, P., FRERICHS, W. M., CHIEVES, L. & WAGNER, G. G. ( 1993). Culture confirmation of the carrier status of Babesia caballi-infected horses. Journal of Clinical Microbiology 31, 689701.Google Scholar
HOLMAN, P., FRERICHS, W. M., OLSON, D. & WAGNER, G. G. ( 1994). Babesia equi erythrocytic stage continuously cultured in an enriched medium. Journal of Parasitology 80, 232236.CrossRefGoogle Scholar
IKADAI, H., MARTIN, M. D., NAGASAWA, H., FUJISAKI, K., SUZUKI, N., MIKAMI, T., KUDO, N., OYAMADA, T. & IGARASHI, I. ( 2001). Analysis of a growth-promoting factor for Babesia caballi cultivation. Journal of Parasitology 87, 14841486.CrossRefGoogle Scholar
JACKSON, L. A., WALDRON, S. J., WEIER, H. M., NICOLL, C. L. & COOKE, B. M. ( 2001). Babesia bovis: culture of laboratory-adapted parasite lines and clinical isolates in a chemically defined medium. Experimental Parasitology 99, 168174.CrossRefGoogle Scholar
JAMES, M. A., LEVY, M. G. & RISTIC, M. ( 1981). Isolation and partial characterization of culture-derived soluble Babesia bovis antigens. Infection and Immunity 31, 358361.Google Scholar
KUTTLER, K. L. & YOUNG, A. ( 1984). Chemotherapy of babesiosis. In Malaria and Babesiosis (ed. Ristic, M., Thomas, A. & Kreier, J. P.), pp. 151172. Martinus Nijhoff, Boston, MA.CrossRef
LEVY, M. G. & RISTIC, M. ( 1980). Babesia bovis: continuous cultivation in a microaerophilous stationary phase culture. Science 27, 12181220.CrossRefGoogle Scholar
LEVY, M. G., ERP, E. & RISTIC, M. ( 1981). Cultivation of Babesia. In Babesiosis (ed. Ristic, M. & Kreier, J. P.), pp. 207223. Academic Press, New York.
NEVES, L. ( 1991). Babesia bovis: cultivation techniques and immunochemical studies. M.V.Sc. thesis, University of Liverpool.
OFULLA, A. O., OREGA, A. S., GITHURE, J. I., BURANS, J. P., ALEMAN, G. M., JOHNSON, A. J. & MARTIN, S. K. ( 1994). Determination of fifty percent inhibitory concentrations (IC50) of antimalarial drugs against Plasmodium falciparum parasites in a serum-free medium. Antimicrobial Agents and Chemotherapy 51, 214218.CrossRefGoogle Scholar
SCHRÉVEL, J., GRELLIER, P. & RIGOMER, D. ( 1992). New approaches in in vitro cultures of Plasmodium falciparum and Babesia divergens by using serum-free medium based on human high density lipoproteins. Memorias do Instituto Oswaldo Cruz 87 (Suppl. III), 7175.CrossRefGoogle Scholar
VEGA, C. A., BUENING, G. M., GREEN, T. J. & CARSON, C. A. ( 1985). In vitro cultivation of Babesia bigemina. American Journal of Veterinary Research 46, 416420.Google Scholar
ZWEYGARTH, E., VAN NIEKERK, C. J. & DE WAAL, D. T. ( 1999). Continuous in vitro cultivation of Babesia caballi in serum-free medium. Parasitology Research 85, 413416.CrossRefGoogle Scholar