Skip to main content
SpringerLink
Log in

Purification of the erythrocytic stages ofBabesia bigemina from cultures

  • Original Investigations
  • Published:
Parasitology Research Aims and scope Submit manuscript

Abstract

Exposure of erythrocytes infected withBabesia bigemina to glycerol-enhanced osmotic shock yielded preparations containing infected erythrocyte ghosts, free parasites, and some intact erythrocytes. The released parasites were purified and concentrated by centrifugation in Percoll gradient. Recovered free parasites were shown by the fluorescein acetate technique to be metabolically active, but their infectivity in vitro was low. It was demonstrated by electron microscopy that most of the released parasites had intact plasma membranes. There was only slight contamination of the free-parasite preparation with host erythrocyte debris.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Dalgliesh RJ (1972) Effects of low temperature preservation and route of inoculation on infectivity ofBabesia bigemina in blood diluted with glycerol. Res Vet Sci 13: 540–545

    Google Scholar 

  • Figueroa JV, Buening GM, Kinden DA, Green TJ (1990) Identification of common surface antigens amongBabesia bigemina isolates by using monoclonal antibodies. Parasitology 100: 161–175

    Google Scholar 

  • Gravely SM, Kreier JP (1974)Babesia microti (Gray strain): removal from infected hamster erythrocytes by continuous-flow ultrasonication. Trpenmed Parasitol 25: 198–206

    Google Scholar 

  • Hall R, McBride J, Morgan G, Tait A, Zolg JW, Walliker D, Scaife J (1983) Antigens of the erythrocytic stages of the human malaria parasitePlasmodium falciparum detected by monoclonal antibodies. Mol Biochem Parasitol 7: 247–265

    Google Scholar 

  • Langreth SG, Jensen DJ, Reese RT, Trager W (1978) Fine structure of human malaria in vitro. J Protozool 25: 443–452

    Google Scholar 

  • Levy MG, Ristic M (1983) Cultivation and in vitro studies ofBabesia. In: Jensen JB (ed) In vitro cultivation of protozoan parasites. CRC Press, Boca Raton, Florida, pp 221–241

    Google Scholar 

  • Mahoney DF (1967) Bovine babesiosis: preparation and assessment of complement fixing antigens. Exp Parasitol 20: 232–241

    Google Scholar 

  • Mahoney DF (1972) Immune response to hemoprotozoa: II.Babesia spp. In: Soulsby EJL (ed) Immunity to animal parasites. Academic Press, New York, pp 301–341

    Google Scholar 

  • McElwain TF, Perryman LE, Davis WC, McGuire TC (1987) Antibodies define multiple proteins with epitopes exposed on the surface of liveBabesia bigemina merozoites. J Immunol 138: 2298–2304

    Google Scholar 

  • Mishell BB, Shiigi SM, Henry C, Chan EL, North J, Gallily R, Slomich M, Miller K, Marbrook J, Parks D, Good AH (1980) Preparation of mouse cell suspensions. In: Mishell BB, Shiigi SM (eds) Selected methods in cellular immunology. WH Freeman, New York, pp 3–27

    Google Scholar 

  • O'Donoghue PJ, Friedhoff KT, Vizcaino OG, Weyreter H (1985) The detection of IgM and IgG antibodies againstBabesia bigemina in bovine sera using semi-defined antigens in enzyme immunoassays. Vet Parasitol 18: 1–12

    Google Scholar 

  • Rodriguez SD, Buening GM, Carson CA (1986)Babesia bovis purification and concentration of merozoites and infected bovine erythrocytes. Exp Parasitol 61: 236–243

    Google Scholar 

  • Sanders SK, Alexander EL, Braylan RC (1975) A high-yield teennique for preparing cells fixed in suspension for scanning electron microscopy. J Cell Biol 67: 476–480

    Google Scholar 

  • Shimizu S, Suzuki K, Nakamura K, Kadota K, Fujisaki K, Ito S, Minami T (1988) Isolation ofTheileria sergenti piroplasms from infected erythrocytes and development of an enzyme-linked immunosorbent assay for serodiagnosis ofT. sergenti infections. Res Vet Sci 45: 206–212

    Google Scholar 

  • Vega CA, Buening GM, Green TJ, Carson CA (1985a) In vitro cultivation ofBabesia bigemina. Am J Vet Res 46: 416–420

    Google Scholar 

  • Vega CA, Buening GM, Rodriguez SD, Carson CA, McLaughlin K (1985b) Cryopreservation ofBabesia bigemina for in vitro cultivation. Am J Vet Res 46: 421–423

    Google Scholar 

  • Vega CA, Buening GM, Rodriguez SD, Carson CA (1986) Concentration and enzyme content of in vitro-culturedBabesia bigemina-infected erythrocytes. J Protozool 33(4): 514–518

    Google Scholar 

  • Wunderlich F, Schillinger G, Helwig M (1985) Fractionation ofPlasmodium chabaudi-infected erythrocytes into parasited and ghosts. Z Parasitenkd 71: 545–551

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Veterinary Microbiology, College of Veterinary Medicine, University of Missouri-Columbia, 65211, Columbia, MO, USA

    J. V. Figueroa & G. M. Buening

  2. Department of Veterinary Pathology, College of Veterinary Medicine, University of Missouri-Columbia, 65211, Columbia, MO, USA

    D. A. Kinden

Authors
  1. J. V. Figueroa
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. G. M. Buening
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. D. A. Kinden
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Figueroa, J.V., Buening, G.M. & Kinden, D.A. Purification of the erythrocytic stages ofBabesia bigemina from cultures. Parasitol Res 76, 675–680 (1990). https://doi.org/10.1007/BF00931086

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00931086

Keywords

Search

Navigation