Skip to main content

Methods for Giardia Culture, Cryopreservation, Encystation, and Excystation In Vitro

  • Chapter
Giardia

Abstract

The main objective of this chapter is to present the most widely accepted methods for routine culturing, encysting, and excysting Giardia lamblia in vitro. In addition, we include basic methods to cryopreserve and recover Giardia trophozoites. Encystation and excystation methods described were originally optimized for strain WB clone C6 (ATCC 50803; the first genome strain), and can serve as a foundation for other strains or species of Giardia. Throughout the chapter, we highlight important factors and procedural details that should assist one to improve or maintain high efficiencies of Giardia growth and differentiation. We also include “recipe” cards for all media and solutions used for protocols detailed in the chapter for ease of use in laboratory settings. Although we have attempted to summarize the literature, we only have direct experience with the strain and procedures used in our laboratory and cannot critically evaluate others’ methods.

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

Access this chapter

Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 169.00
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 219.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  • Adam RD (1991) The biology of Giardia spp. Microbiol Rev 55: 706–732

    PubMed  CAS  Google Scholar 

  • Adam RD (2001) Biology of Giardia lamblia. Clin Microbiol Rev 14: 447–475

    Article  PubMed  CAS  Google Scholar 

  • Arguello-Garcia R, Bazan-Tejeda ML, and Ortega-Pierres G (2009) Encystation commitment in Giardia duodenalis: a long and winding road. Parasite 16: 247–258

    Article  PubMed  CAS  Google Scholar 

  • Bernander R, Palm JE, and Svard SG (2001) Genome ploidy in different stages of the Giardia lamblia life cycle. Cell Microbiol 3: 55–62

    Article  PubMed  CAS  Google Scholar 

  • Bingham AK and Meyer EA (1979) Giardia excystation can be induced in vitro in acidic solutions. Nature 277: 301–302

    Article  PubMed  CAS  Google Scholar 

  • Bingham AK, Jarroll EL Jr, Meyer EA, and Radulescu S (1979) Giardia sp.: physical factors of excystation in vitro, and excystation vs eosin exclusion as determinants of viability. Exp Parasitol 47: 284–291

    Article  PubMed  CAS  Google Scholar 

  • Boucher SE and Gillin FD (1990) Excystation of in vitro-derived Giardia lamblia cysts. Infect Immun 58: 3516–3522

    PubMed  CAS  Google Scholar 

  • Carranza PG and Lujan HD (2010) New insights regarding the biology of Giardia lamblia. Microbes Infect 12: 71–80

    Article  PubMed  CAS  Google Scholar 

  • Clark CG and Diamond LS (2002) Methods for cultivation of luminal parasitic protists of clinical importance. Clin Microbiol Rev 15: 329–341

    Article  PubMed  Google Scholar 

  • Diamond LS (1964) Freeze-preservation of protozoa. Cryobiology 1: 95–102

    Article  PubMed  CAS  Google Scholar 

  • Diamond LS, Harlow DR, and Cunnick CC (1978) A new medium for the axenic cultivation of Entamoeba histolytica and other Entamoeba. Trans R Soc Trop Med Hyg 72: 431–432

    Article  PubMed  CAS  Google Scholar 

  • Eichinger D (2001) Encystation in parasitic protozoa. Curr Opin Microbiol 4: 421–426

    Article  PubMed  CAS  Google Scholar 

  • Feely DE, Gardner MD, and Hardin EL (1991) Excystation of Giardia muris induced by a phosphate-bicarbonate medium: localization of acid phosphatase. J Parasitol 77: 441–448

    Article  PubMed  CAS  Google Scholar 

  • Gault MJ, Reiner DS, and Gillin FD (1985) Tolerance of axenically cultured Entamoeba histolytica and Giardia lamblia to a variety of antimicrobial agents. Trans R Soc Trop Med Hyg 79: 60–62

    Article  PubMed  CAS  Google Scholar 

  • Gillin FD, Reiner DS, Gault MJ, et al. (1987) Encystation and expression of cyst antigens by Giardia lamblia in vitro. Science 235: 1040–1043

    Article  PubMed  CAS  Google Scholar 

  • Gillin FD, Reiner DS, and Boucher SE (1988) Small-intestinal factors promote encystation of Giardia lamblia in vitro. Infect Immun 56: 705–707

    PubMed  CAS  Google Scholar 

  • Gillin FD, Boucher SE, Rossi SS, and Reiner DS (1989) Giardia lamblia: the roles of bile, lactic acid, and pH in the completion of the life cycle in vitro. Exp Parasitol 69: 164–174

    Article  PubMed  CAS  Google Scholar 

  • Hautus MA, Kortbeek LM, Vetter JC, and Laarman JJ (1988) In vitro excystation and subsequent axenic growth of Giardia lamblia. Trans R Soc Trop Med Hyg 82: 858–861

    Article  PubMed  CAS  Google Scholar 

  • Kane AV, Ward HD, Keusch GT, and Pereira ME (1991) In vitro encystation of Giardia lamblia: large-scale production of in vitro cysts and strain and clone differences in encystation efficiency. J Parasitol 77: 974–981

    Article  PubMed  CAS  Google Scholar 

  • Karapetyan A (1962) In vitro cultivation of Giardia duodenalis. J Parasitol 48: 337–340

    Article  PubMed  CAS  Google Scholar 

  • Kaul D, Rani R, and Sehgal R (2001) Receptor-Ck regulates giardia encystation process. Mol Cell Biochem 225: 167–169

    Article  PubMed  CAS  Google Scholar 

  • Keister DB (1983) Axenic culture of Giardia lamblia in TYI-S-33 medium supplemented with bile. Trans R Soc Trop Med Hyg 77: 487–488

    Article  PubMed  CAS  Google Scholar 

  • Lujan HD, Mowatt MR, Byrd LG, and Nash TE (1996) Cholesterol starvation induces differentiation of the intestinal parasite Giardia lamblia. Proc Natl Acad Sci USA 93: 7628–7633

    Article  PubMed  CAS  Google Scholar 

  • Lujan HD, Mowatt MR, and Nash TE (1997) Mechanisms of Giardia lamblia differentiation into cysts. Microbiol Mol Biol Rev 61: 294–304

    PubMed  CAS  Google Scholar 

  • Meyer EA (1970) Isolation and axenic cultivation of Giardia trophozoites from the rabbit, chinchilla, and cat. Exp Parasitol 27: 179–183

    Article  PubMed  CAS  Google Scholar 

  • Meyer EA (1976) Giardia lamblia: isolation and axenic cultivation. Exp Parasitol 39: 101–105

    Article  PubMed  CAS  Google Scholar 

  • Meyer EA and Chadd JA (1967) Preservation of Giardia trophozoites by freezing. J Parasitol 53: 1108–1109

    Article  PubMed  CAS  Google Scholar 

  • Oberhuber G, Kastner N, and Stolte M (1997) Giardiasis: a histologic analysis of 567 cases. Scand J Gastroenterol 32: 48–51

    Article  PubMed  CAS  Google Scholar 

  • Palm D, Weiland M, McArthur AG, et al. (2005) Developmental changes in the adhesive disk during Giardia differentiation. Mol Biochem Parasitol 141: 199–207

    Article  PubMed  CAS  Google Scholar 

  • Phillips RE, Boreham PF, and Shepherd RW (1984) Cryopreservation of viable Giardia intestinalis trophozoites. Trans R Soc Trop Med Hyg 78: 604–606

    Article  PubMed  CAS  Google Scholar 

  • Reiner DS, Douglas H, and Gillin FD (1989) Identification and localization of cyst-specific antigens of Giardia lamblia. Infect Immun 57: 963–968

    PubMed  CAS  Google Scholar 

  • Reiner DS, Hetsko ML, Das S, et al. (1993) Giardia lamblia: absence of cyst antigens and reduced secretory vesicle for mation and bile salt uptake in an encystation-deficient subline. Exp Parasitol 77: 461–472

    Article  PubMed  CAS  Google Scholar 

  • Reiner DS, Hetsko ML, and Gillin FD (1995) A lipoprotein-cholesterol-albumin serum substitute stimulates Giardia lamblia encystation vesicle formation. J Eukaryot Microbiol 42: 622–627

    Article  PubMed  CAS  Google Scholar 

  • Rice EW and Schaefer FW 3rd (1981) Improved in vitro excystation procedure for Giardia lamblia cysts. J Clin Microbiol 14: 709–710

    PubMed  CAS  Google Scholar 

  • Schaefer FW 3rd, Rice EW, and Hoff JC (1984) Factors promoting in vitro excystation of Giardia muris cysts. Trans R Soc Trop Med Hyg 78: 795–800

    Article  PubMed  Google Scholar 

  • Schupp DG and Erlandsen SL (1987) Determination of Giardia muris cyst viability by differential interference contrast, phase, or brightfield microscopy. J Parasitol 73: 723–729

    Article  PubMed  CAS  Google Scholar 

  • Schupp DG, Januschka MM, Sherlock LA, et al. (1988) Production of viable Giardia cysts in vitro: determination by fluorogenic dye staining, excystation, and animal infectivity in the mouse and Mongolian gerbil. Gastroenterology 95: 1–10

    PubMed  CAS  Google Scholar 

  • Stefanic S, Morf L, Kulangara C, et al. (2009) Neogenesis and maturation of transient Golgi-like cisternae in a simple eukaryote. J Cell Sci 122: 2846–2856

    Article  PubMed  CAS  Google Scholar 

  • Sun CH, McCaffery JM, Reiner DS, and Gillin FD (2003) Mining the Giardia lamblia genome for new cyst wall proteins. J Biol Chem 278: 21701–21708

    Article  PubMed  CAS  Google Scholar 

  • Visvesvara GS (1980) Axenic growth of Giardia lamblia in Diamond’s TPS-1 medium. Trans R Soc Trop Med Hyg 74: 213–215

    Article  PubMed  CAS  Google Scholar 

  • Warhurst DC and Wright SG (1979) Cryopreservation of Giardia intestinalis. Trans R Soc Trop Med Hyg 73: 601

    Article  PubMed  CAS  Google Scholar 

  • Wieder SC, Keister DB, and Reiner DS (1983) Mass cultivation of Giardia lamblia in a serum-free medium. J Parasitol 69: 1181–1182

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2011 Springer-Verlag/Wien

About this chapter

Cite this chapter

Davids, B.J., Gillin, F.D. (2011). Methods for Giardia Culture, Cryopreservation, Encystation, and Excystation In Vitro . In: Luján, H.D., Svärd, S. (eds) Giardia. Springer, Vienna. https://doi.org/10.1007/978-3-7091-0198-8_23

Download citation

Publish with us

Policies and ethics