Cryptosporidium Oocyst Purification Using Discontinuous Gradient Centrifugation

  • Michael J. ArrowoodEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2052)


Many laboratory studies in cryptosporidial research require a source of purified oocysts. Sources can include experimentally infected laboratory animals or from samples collected from naturally infected animals and from clinical cases of human cryptosporidiosis. Purification of oocysts can be accomplished with readily available laboratory equipment including tabletop centrifuges and microcentrifuges. Following purification, oocysts can be stored in antibiotic-supplemented buffers or in 2.5% aqueous potassium dichromate for over 6 months. Ultimately, oocyst viability and infectivity decline to less than 10% after 1 year, so if isolates are expected to be maintained, serial passage in a suitable host at ≤6-month intervals is recommended. Oocysts purified as described in this chapter are suitable for animal infection studies, cell culture studies, and a wide range of molecular biological studies, environmental studies, drug testing, and disinfection studies.


Cryptosporidium Oocyst Purification Isolation Sucrose Sheather’s solution Cesium chloride Centrifugation Gradient 



Disclosures: Use of trade names is for identification only and does not imply endorsement by the Public Health Service or by the U.S. Department of Health and Human Services. The findings and conclusions in this report are those of the author and do not necessarily represent the official position of the Centers for Disease Control and Prevention.


  1. 1.
    Arrowood MJ, Hurd MR, Mead JR (1995) A new method for evaluating experimental cryptosporidial parasite loads using immunofluorescent flow cytometry. J Parasitol 81(3):404–409CrossRefGoogle Scholar
  2. 2.
    Arrowood MJ, Donaldson K (1996) Improved purification methods for calf-derived Cryptosporidium parvum oocysts using discontinuous sucrose and cesium chloride gradients. J Eukaryot Microbiol 43(5):89CrossRefGoogle Scholar
  3. 3.
    Arrowood MJ, Sterling CR (1987) Isolation of Cryptosporidium oocysts and sporozoites using discontinuous sucrose and isopycnic Percoll gradients. J Parasitol 73:314–319CrossRefGoogle Scholar
  4. 4.
    Anderson BC (1985) Moist heat inactivation of Cryptosporidium sp. Am J Public Health 75:1433–1434CrossRefGoogle Scholar
  5. 5.
    Fujino T, Matsui T, Kobayashi F, Haruki K, Yoshino Y, Kajima J, Tsuji M (2002) The effect of heating against Cryptosporidium oocysts. J Vet Med Sci 64(3):199–200CrossRefGoogle Scholar
  6. 6.
    Harp JA, Fayer R, Pesch BA, Jackson GJ (1996) Effect of pasteurization on infectivity of Cryptosporidium parvum oocysts in water and milk. Appl Environ Microbiol 62(8):2866–2868CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Division of Foodborne, Waterborne, and Environmental Diseases, National Center for Emerging and Zoonotic Infectious DiseasesCenters for Disease Control and PreventionAtlantaUSA

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