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Comparison of Cryoprotectants in the Preservation of Theileria parva Sporozoites Using an In Vitro Infectivity Assay

Tropical Animal Health and Production volume 33, pages 29–41 (2001)Cite this article

Abstract

An in vitro infectivity assay was used to examine five cryoprotectants for their suitability for preserving Theileria parva sporozoites. All five were capable of preserving T. parva sporozoites through freezing, the optimal concentrations being 7.5% for glycerol, 5% for dimethyl sulphoxide (DMSO), poly (vinylpyrrolidone) (PVP) and poly(ethylene glycol) (PEG), and 2.5% for hydroxyethyl starch (HES). When the five cryoprotectants were compared at their optimal concentrations, using a modification of the standard method of stabilate preparation, glycerol was significantly better than the others (p<0.05). Measurement of the effects of each cryoprotectant on the osmolality of the media revealed that glycerol and DMSO elevated the osmolality significantly (p<0.05). Resuscitation of glycerol-preserved sporozoites required the presence of glycerol in the diluent to maintain infectivity. Studies on the effects of equilibration time in glycerol on the infectivity of sporozoites showed that those frozen immediately after mixing (2 min) were as infective as those frozen after 60 min of equilibration.

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REFERENCES

  • Brown, C.G.D., 1979. Propagation of Theileria. In: K. Maramorosch and H. Hirumi (eds), Practical Tissue Culture Application, (Academic Press, New York), 223-254

    Google Scholar 

  • Brown, C.G.D., Stagg, D.A., Purnell, R.E., Kanhai, G.K. and Payne, R.C., 1973. Infection and transformation of bovine lymphoid cells in vitro by infective particles of Theileria parva. Nature, 245, 101-103

    Google Scholar 

  • Büscher, G. and 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, 93-100

    Google Scholar 

  • Callow, L.L. and Farrant, J., 1973. Cryopreservation of the promastigote form of Leishmania tropica var major at different cooling rates. International Journal for Parasitology, 3, 77-88

    Google Scholar 

  • Cunningham, M.P., Lumsden, W.H.R. and Webber, W.A.F., 1963. Preservation of viable trypanosomes in lymph tubes at low temperatures. Experimental Parasitology, 14, 280-284

    Google Scholar 

  • Cunningham, M.P., Brown, C.G.D., Burridge, M.J. and Purnell, R.E., 1973. Cryopreservation of infective particles of Theileria parva. International Journal for Parasitology, 3, 583-587

    Google Scholar 

  • Cunningham, M.P., Brown, C.G.D., Burridge, M.J., Musoke, A.J., Purnell, R.E., Radley, D.E. and Sempembwa, C., 1974. East Coast fever: titration in cattle of suspensions of Theileria parva derived from ticks. British Veterinary Journal, 13, 336-345

    Google Scholar 

  • Dalgliesh, R.J., 1972. Theoretical and practical aspects of freezing parasitic protozoa. AustralianVeterinary Journal, 48, 233-239

    Google Scholar 

  • Dalgliesh, R.J., Jorgensen, W.K. and de Vos, A.J., 1990. Australian frozen vaccines for the control of babesiosis and anaplasmosis in cattle-a review. Tropical Animal Health and Production, 22, 44-52

    Google Scholar 

  • Hentrich, B., Bose, R. and Doherr, M., 1994. Cryopreservation of Babesia caballi cultures. International Journal for Parasitology, 24, 253-254

    Google Scholar 

  • Hernandez, F.J., 1987. Investigation on various aspects of cryopreservation in cattle and rabbit embryos, (PhD thesis, University of Hanover)

  • Karber, G., 1931. Beitrag zur kollektiven behandlung pharmakologischer Reinhenversuche. Archiv für Experimentelle Pathologie und Pharmakologie, 162, 480-487

    Google Scholar 

  • Kimbita, E.N. and Silayo, R.S., 1997. Use of in vitro infectivity assay in comparison with histological techniques in the study of Theileria parva sporozoite maturation. Veterinary Parasitology, 70, 83-97

    Google Scholar 

  • Leef, J.L., Strome, C.P.A. and Beaudoin, R.L., 1979. Low temperature preservation of sporozoites of Plasmodium berghei. Bulletin of the World Health Organization, 57, 87-91

    Google Scholar 

  • Lin, D.B., Su, K.E. and Yu, J.C., 1995. Studies on cryopreservation of Toxoplasma gondii and its antigenicity in mice. American Journal of Tropical Medicine and Hygiene, 53, 392-396

    Google Scholar 

  • Lovelock, J.E., 1953. The mechanism of protective action of glycerol against haemolysis by freezing and thawing. Biochimica et Biophysica Acta, 11, 28-36

    Google Scholar 

  • Mangold, A.J., Aguirre, D.H. and Guglielmone, A.A., 1990. Post thawing viability of vaccine for bovine babesiosis and anaplasmosis cryopreserved with glycerol.Veterinary Parasitology, 37, 301-306

    Google Scholar 

  • Norton, C.C. and Joyner, L.P., 1968. The freeze preservation of coccidia. Research in Veterinary Science, 9, 598-600

    Google Scholar 

  • Palmer, D.A., Buening, G.M. and Carson, C.A., 1982. Cryopreservation of Babesia bovis for in vitro cultivation. Parasitology, 84, 567-572

    Google Scholar 

  • Purnell, R.E., Brown, C.G.D., Cunningham, M.P., Burridge, M.J., Kirimi, I.M. and Ledger, M.A., 1973. East Coast fever: correlation between morphology and infectivity of Theileria parva developing in its tick vector. Parasitology, 66, 539-544

    Google Scholar 

  • Radley, D.E., Brown, C.G.D., Burridge, M.J., Cunningham, M.P., Kirimi, I.M., Purnell, R.E. and Young, A.S., 1975. East Coast fever: 1. Chemoprophylactic immunization of cattle against Theileria parva (Muguga) and five theilerial strains. Veterinary Parasitology, 1, 35-41

    Google Scholar 

  • Rasheda, M., Qazi, M.H. and Mazar, R., 1995. Purification and preservation of sporozoites of Eimeria garnhami. Sahad Journal of Agriculture, 11, 189-193

    Google Scholar 

  • Snedecor, G.W. and Cochran, W.G., 1989. Statistical Methods, 8th edn, (Iowa State University Press, Ames, IA)

    Google Scholar 

  • Solis, J., 1972. Turkey Leucocytozoon infection. 2. Cryobiological preservation of Leucocytozoon smithi sporozoites. Poultry Science, 51, 1747-1752

    Google Scholar 

  • Standfast, N.F. and Jorgensen, W.K., 1997. Comparison of the infectivity of Babesia bovis, Babesia bigemina and Anaplasma centrale for cattle after cryopreservation in either dimethyl sulphoxide (DMSO) or polyvinylpyrrolidone (PVP). Australian Veterinary Journal, 75, 62-63

    Google Scholar 

  • Wilde, J.K.H., Brown, C.G.D., Hulliger, L., Gall, D. and Macleod, W.G., 1968. East Coast Fever: experiments with tissues of infected ticks. British Veterinary Journal, 124, 196-208

    Google Scholar 

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Authors and Affiliations

  1. International Livestock Research Institute, Nairobi, Kenya; and

    E.N. Kimbita, R.S. Silayo & T.T. Dolan

  2. Department of Veterinary Microbiology and Parasitology, Sokoine University of Agriculture, Morogoro, Tanzania

    E.N. Kimbita & R.S. Silayo

  3. Livestock Services Ltd, Nairobi, Kenya

    T.T. Dolan

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  1. E.N. Kimbita
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  2. R.S. Silayo
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  3. T.T. Dolan
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Kimbita, E., Silayo, R. & Dolan, T. Comparison of Cryoprotectants in the Preservation of Theileria parva Sporozoites Using an In Vitro Infectivity Assay. Tropical Animal Health and Production 33, 29–41 (2001). https://doi.org/10.1023/A:1010329409637

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  • DOI: https://doi.org/10.1023/A:1010329409637

  • cryoprotectants
  • equilibration
  • infectivity
  • osmolality
  • sporozoites
  • Theileria parva

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