Skip to main content
SpringerLink
Log in

Flowcytometric assessment of the effect of drugs on Giardia lamblia trophozoites in vitro

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

The effect of albendazole, vincristine and metronidazole on the adherence, morphology, cell cycle and viability of Giardia lamblia trophozoites was evaluated. Albendazole rendered the trophozoites unable to attach which is a prerequisite for establishment of Giardia infection. The adherence assay showed that albendazole caused the detachment of trophozoites at a significantly lower concentration (0.1 μg/ml) as compared to metronidazole and vincristine (40 and 60 μg/ml, respectively) in the time period of 9 h. However, at the higher concentration (10 μg/ml) albendazole caused the similar effect within the time period of 3 h. The microscopic studies revealed that albendazole brought about gross morphological changes in G. lamblia, i.e., rounding of trophozoites and disruption of ventral adhesive disc along with vacuolation in cytoplasm. Metronidazole caused the ballooning of cytoplasm and induced the most lethal effect on trophozoites (8.17% viability) as compared to vincristine and albendazole (72.13 and 65.49% viability) as shown by flowcytometric analysis. Further, the flowcytometric findings indicated that the three drugs led to the arrest of growth of trophozoites at different stages of cell cycle. Albendazole and vincristine interfered with the formation of spindle/basal body microtubules thereby arresting the cell cycle in G2 + M phase whereas metronidazole arrested the growth in S-phase by inhibiting the DNA segregation and cell division. Hence, flowcytometry has been usefully employed to study the effect of drugs on viability and DNA of G. lamblia trophozoites in the present study. (Mol Cell Biochem 265: 151–160, 2004)

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

  1. Webster LT: Drugs used in chemotherapy of protozoal infections. In: AG Gilman, TW Rall, AS Nies, P Taylor (eds). Goodman and Gilmans 'Pharmacological Basis of Therapeutics,' 8th edn. Pergamon Press, New York, 1990, pp 978–1007

    Google Scholar 

  2. Gardner TB, Hill DR: Treatment of giardiasis. Clin Microbiol Rev 14(1): 114–128, 2001

    Google Scholar 

  3. Upcroft P, Upcroft JA: Drug targets and mechanisms of resistance in the anaerobic protozoa. Clin Microbiol Rev 14(1): 150–164, 2001

    Google Scholar 

  4. Vanden Bossche H: Pharmacology of anthelmintics. In: H Vanden Bossche, D Theimnpont, PG Janssens (eds). Chemotherapy of Gas-trointestinal Helminths. Springer-Verlag, Berlin, 1985, pp 125–171

    Google Scholar 

  5. Meloni BP, Thompson RCA, Reynoldson JA, Seville P: Albendazole, a more effective antigiardial agent in vitro than metronidazole or tinida-zole. Trans Roy Soc Trop Med Hyg 81: 637–640, 1990

    Google Scholar 

  6. Liu LX, Weller PF: Antiparasitic drugs. N Engl J Med 334: 1178–1184, 1996

    Google Scholar 

  7. Harris JC, Plummer S, Lloyd D: Antigiardial drugs. Appl Microbiol Biotechnol 57: 614–619, 2001

    Google Scholar 

  8. Crouch AA, Seow WK, Thong YH: Effect of 23 chemotherapeutic agents on the adherence and growth of Giardia lamblia in vitro.Trans Roy Soc Trop Med Hyg 72: 431–432, 1986

    Google Scholar 

  9. Edind TD, Hang TI, Chakraborty PR: Activity of the anthelmintic ben-zimidazoles against Giardia lamblia in vitro.JInf Dis 162: 408–411, 1990

    Google Scholar 

  10. Morgan UM, Reynoldson JA, Thompson RCA: Activities of several benzimadazoles and tubulin inhibitors against Giardia spp. in vitro. Antimicrob Agents Chemoth 37: 328–331, 1993

    Google Scholar 

  11. Campanati L, Monteiro-Leal LH: The effects of the antiprotozoal drugs metronidazole and furazolidone on trophozoites of Giardia lamblia (P1 strain). Parasitol Res 88(1): 80–85, 2002

    Google Scholar 

  12. Byers CD: Clinical applications of flowcytometry. Clin Lab Sci 6(3): 174–176, 1993

    Google Scholar 

  13. Goetzman EA: Flowcytometry: Basic concepts and clinical applications in Immunodiagnostics. Clin Lab Sci 6(3): 177–182, 1993

    Google Scholar 

  14. Azas N, Giorgio CD, Delmas F, Gasquet M, Timon-David P: Leishmania infantum promastigotes: Flowcytometry as a possible tool for assessing the effects of drugs on cellular functions. Exp Parasitol 87: 1–7, 1997

    Google Scholar 

  15. Fouchet P, Jayat C, Hechard Y, Ratinaud M, Frelat G: Recent advances of flowcytometry in fundamental and applied microbiology. Biol Cell 78: 95–109, 1993

    Google Scholar 

  16. Leonce S, Burbridge M: Flowcytometry: Auseful technique in the study of multi-drug resistance. Biol Cell 78: 63–68, 1993

    Google Scholar 

  17. Diamond LS, Harlow D, Cunnick CC: Anewmedium for the cultivation of Entamoeba histolytica and other entamoebae. Trans Roy Soc Trop Med Hyg 72: 431–432, 1978

    Google Scholar 

  18. Ormerod MG: Further applications to cell biology. In: MG Ormerod (ed). Flowcytometry-A Practical Approach. IRL Press, Oxford Uni-versity Press, England, 1990, pp 265–266

    Google Scholar 

  19. Reynoldsen JA, Thompson RCA, Meloni BP: The mode of action of benzimidazoles against Giardia and their chemotherpeutic potential against Giardia and other parasitic protozoa. In: GH Coombs, MJ North (eds). Biochemical Protozoology. Taylor & Francis, London, United Kingdom, 1991, pp 587–593

    Google Scholar 

  20. Lacey E: The biochemistry of anthelminitic resistance. In: N Anderson, P Waller (eds). Resistance in Nematodes to Anthelminitic Drugs. CSIRO, Glebe, NSW, 1985, pp 69–78

  21. Sangster NC, Prichard RK, Lacey E: Tubulin and benzimidazole re-sistance in Trichostrongylus colubriformis (Nematoda). J Parasitol 71: 644–651, 1985

    Google Scholar 

  22. Lacey E, Prichard RK: Interactions of benzimidazoles (BZ) with tubulin from BZ-sensitive and BZ-resistant isolates of Haemonchus contortus. Mol Biochem Parasitol 19: 171–181, 1986

    Google Scholar 

  23. Lacey E: Role of the cytoskeletal protein tubulin in the mode of action and mechanism of drug resistance to benzimidazoles. Int J Parasitol 18: 855–936, 1988

    Google Scholar 

  24. Lacey E: Mode of action of benzimidazoles. Parasitol Today 6: 112–115, 1990.

    Google Scholar 

  25. Samuelson J: Why metronidazole is active against both bacteria and parasites. Antimicro Agents Chemother 43: 1533–1541, 1999

    Google Scholar 

  26. Townson SM, Hanson GR, Upcroft JA, Upcroft P: A purified ferredoxin from Giardia duodenalis. Eur J Biochem 220: 439–446, 1994

    Google Scholar 

  27. Upcroft J, Upcroft P: My favorite cell: Giardia. Bioassays 20: 256–263, 1998

    Google Scholar 

  28. Venkatesan P: Albendazole. Antimicrob Chemother 41: 145–147, 1998

    Google Scholar 

  29. McCracken RO, Stillwell WH: A possible mode of action of benzimi-dazole antihelmintics. Int J Parasitol 21: 99–104, 1991

    Google Scholar 

  30. Erlandsen SL, Meyer EA: In: SE Erlandsen, EA Meyer (eds). Giardia and Giardiasis. Plenum Press, New York, 1984

    Google Scholar 

  31. Clark JT, Holberton DV: Triton-labile antigens in flagella isolated from Giardia lamblia.Parasitol Res 74: 415–423, 1988

    Google Scholar 

  32. Tracy JW, Webster LT: Drugs used in the chemotherapy of protozoal infections. In: JG Hardman, LE Limbird (eds). The Pharmacological Basis of Therapeutics, 9th edn. McGraw-Hill, New York, NY, 1996, pp 987–1008

    Google Scholar 

  33. Muller M: Mode of action of metronidazole on anaerobic bacteria and protozoa. Surgery 93: 165–171, 1983

    Google Scholar 

  34. Edwards DI: Reduction of nitroimidazoles in vitro and DNA damage. Biochem Pharmacol 35: 53–58, 1986

    Google Scholar 

  35. Edwards DI: Nitroimidazole drugs-Action and resistence mechanisms. I. Mechanisms of action. J Antimicrob Chemother 31: 9–20, 1993

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Indian Council of Medical Research, New Delhi

    Harpreet Sandhu & Nirmal Kumar Ganguly

  2. Department of Parasitology, Postgraduate Institute of Medical Education and Research, Chandigarh, India

    Ramesh Chander Mahajan

Authors
  1. Harpreet Sandhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ramesh Chander Mahajan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Nirmal Kumar Ganguly
    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

Sandhu, H., Mahajan, R.C. & Ganguly, N.K. Flowcytometric assessment of the effect of drugs on Giardia lamblia trophozoites in vitro . Mol Cell Biochem 265, 151–160 (2004). https://doi.org/10.1023/B:MCBI.0000044392.01704.5f

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/B:MCBI.0000044392.01704.5f

Search

Navigation