Skip to main content
SpringerLink
Log in

Trypanosoma congolense: an in vitro assay to distinguish drug-resistant from drug-sensitive populations

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

Abstract

An in vitro assay to distinguish drug-resistant from drug-sensitive populations ofTrypanosoma congolense has been developed. The incorporation of radiolabelled hypoxanthine by procyclic trypanosomes in vitro was measured after 48 h exposure to different concentrations of trypanocides. In the presence of either isometamidium chloride (Samorin) or diminazene aceturate (Berenil), the ability of procyclics of a drug-sensitive stock (TREU 1627) to incorporate hypoxanthine at 28° C was impaired to a much greater extent than that of procyclics of a drug-resistant stock (TREU 1467), when compared with control organisms grown in the absence of drugs. Serum from a rabbit given 1 mg/kg Samorin also inhibited incorporation of radiolabel in TREU 1627 procyclics more severely than in TREU 1467 procyclics, although the difference between stocks was not substantial. When used with cultured bloodstream forms maintained at 35° C, the assay could distinguish the stocks in the presence of Samorin, but no difference was detected between the populations in their incorporation of hypoxanthine after exposure to Berenil.

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

  • Borowy NK, Fink E, Hirumi H (1985a)Trypanosoma brucei: Five commonly used trypanocides assayed in vitro with a mammalian feeder layer system for cultivation of bloodstream forms. Exp Parasitol 60:323–330

    PubMed  Google Scholar 

  • Borowy NK, Fink E, Waithaka HK, Hirumi H (1985b) An assay for screening drugs against animal infective bloodstream forms ofT. b. brucei in vitro. Drugs Exp Clin Res 11:155–161

    PubMed  Google Scholar 

  • Brown HC, Ross CA, Holmes PH, Luckins AG, Taylor AM (1987) Adaptation ofTrypanosoma congolense stocks to in vitro culture does not change their sensitivity to isometamidium. Acta Trop 44:373–374

    PubMed  Google Scholar 

  • Brun R, Kunz C (1989) In vitro drug sensitivity test forTrypanosoma brucei subgroup bloodstream trypomastigotes. Acta Trop 46:361–368

    PubMed  Google Scholar 

  • Chulay JD, Haynes JM, Giggs CL (1983)Plasmodium falciparum assessment of in vitro growth using [3H]-hypoxanthine. Exp Parasitol 55:138–146

    PubMed  Google Scholar 

  • Desjardins RE, Casero RA Jr, Willet GP, Childs GP, Canfield CJ (1980)Trypanosoma rhodesiense: semiautomated microtesting for quantitation of antitrypanosomal activity in vitro. Exp Parasitol 50:260–271

    PubMed  Google Scholar 

  • Fairclough R (1963) Observations of the use of Berenil against trypanosomiasis in cattle in Kenya. Vet Rec 75:1107–1112

    Google Scholar 

  • Fish WR, Marr JJ, Berens RL (1982a) Purine metabolism inTrypanosoma brucei gambiense. Biochim Biophys Acta 714:422–428

    PubMed  Google Scholar 

  • Fish WR, Looker DL, Marr JJ, Berens RL (1982b) Purine metabolism in the bloodstream forms ofTrypanosoma gambiense andTrypanosoma rhodesiense. Biochim Biophys Acta 719:223–231

    PubMed  Google Scholar 

  • Gray AR, Roberts CJ (1971) The stability of resistance to diminazene aceturate and quinapyramine sulphate in a strain ofTrypanosoma vivax during cyclical transmission through antelope. Parasitology 63:163–168

    PubMed  Google Scholar 

  • Gray MA, Ross CA, Taylor AM, Tetley L, Luckins AG (1985) In vitro cultivation ofTrypanosoma congolense: production of infective forms from metacyclic trypanosomes cultured on bovine endothelial cell monolayers. Acta Trop 42:99–110

    PubMed  Google Scholar 

  • Irvin AD, Young ER (1977) Possible in vitro test for screening drugs for activity againstBabesia and other blood protozoa. Nature 269:407–409

    PubMed  Google Scholar 

  • Kellner H-M, Eckert HG, Voltz MH (1985) Studies in cattle on the disposition of the anti-trypanosomal drug diminazene diaceturate (Berenil). Tropenmed Parasitol 36:199–204

    Google Scholar 

  • Klatt P, Hajdu P (1976) Pharmacokinetic investigations on diminazene and rolitetracycline in comparison to a combination of both. Vet Rec 99:372–374

    PubMed  Google Scholar 

  • Kratzer RD, Turkoson PK, Karanja WM, Ondiek FO (1989) Studies in cattle on the pharmacokinetics and bioavailability of the anti-trypanosomal drug isometamidium. Proceedings of the 20th International Scientific Council for Trypanosomiasis Research and Control, OAU/ISCTRC, Nairobi (in press)

  • Kupper W, Wolters M (1983) Observations on drug resistance inTrypanosoma (Nannomonas) congolense andTrypanosoma (Duttonella) vivax in cattle at a feedlot in the Northern Ivory Coast. Tropenmed Parasitol 34:203–205

    PubMed  Google Scholar 

  • Mbwambo HA, Mella PNP, Lekaki KA (1988) Berenil (diminazene aceturate)-resistantTrypanosoma congolense in cattle under natural tsetse challenge at Kibaha, Tanzania. Acta Trop 45:239–244

    PubMed  Google Scholar 

  • North TW, Wyler DJ (1987) DNA synthesis in promastigotes ofLeishmania major andLeishmania donovani. Mol Biochem Parasitol 22:215–221

    PubMed  Google Scholar 

  • Ochieng A, Gray MA (1989) Establishment of cultures ofTrypanosoma congolense from infected cattle. KEMRI/KETRI Rep (in press)

  • Owolabi OA, Molyneux DH (1988) Effects of trypanocides onHerpetosoma in a mammalian feeder layer cell culture. Vet Parasitol 30:107–112

    PubMed  Google Scholar 

  • Peregrine AS (1987) Factors influencing the duration of isometamidium prophylaxis against bovine trypanosomiasis. PhD Thesis, Unversity of Glasgow

  • Perschke H, Vollner L (1985) Determination of the trypanocidal drugs homidium, isometamidium and quinapyramine in bovine serum or plasma using HPLC. Acta Trop 42:209–216

    PubMed  Google Scholar 

  • Pinder M, Authie E (1984) The appearance of isometamidium resistantTrypanosoma congolense in West Africa. Acta Trop 41:247–252

    PubMed  Google Scholar 

  • Sones KR, Njogu AJ, Holmes PH (1988) Assessment of sensitivity ofTrypanosoma congolense to isometamidium chloride: a comparison of tests using cattle and mice. Acta Trop 45:153–164

    PubMed  Google Scholar 

  • Van Hoeve K, Cunningham MP (1964) Prophylactic activity of Berenil against trypanosomes in treated cattle. Vet Rec 76:260

    Google Scholar 

  • Waithaka HK, Borowy NK, Gettinby G, Scott J, Hirumi H (1987) Trypanocidal drug screening onTrypanosoma (Duttonella) vivax in vitro. Proceedings of the 19th International Scientific Council for Trypanosomiasis Research and Control. OAU/ISCTRC, Nairobi, pp 296–300

  • Whitelaw DD, Bell IR, Holmes PH, Moloo SK, Hirumi H, Urquart GM, Murray M (1986) Isometamidium chloride prophylaxis againstTrypanosoma congolense challenge and the development of immune responses in Boran cattle. Vet Rec 118:722–726

    PubMed  Google Scholar 

  • Williamson J (1980a) Chemoresistance in trypanosomes. Report on the expert consultation on research on trypanosomiases. Food and Agriculture Organisation of the United Nations, Rome, pp 84–89

    Google Scholar 

  • Williamson J (1980b) Present situation of research for new trypanocidal drugs. Report on the expert consultation on research on trypanosomiases. Food and Agriculture Organisation of the United Nations Rome, pp 90–96

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Centre for Tropical Veterinary Medicine, Easter Bush, EH25 9RG, Roslin, Midlothian, UK

    C. A. Ross & A. M. Taylor

Authors
  1. C. A. Ross
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. M. Taylor
    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

Ross, C.A., Taylor, A.M. Trypanosoma congolense: an in vitro assay to distinguish drug-resistant from drug-sensitive populations. Parasitol Res 76, 326–331 (1990). https://doi.org/10.1007/BF00928187

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00928187

Keywords

Search

Navigation