Mouse Models for Use in Cryptosporidium Infection Studies and Quantification of Parasite Burden Using Flow Cytometry, qPCR, and Histopathology

Methods Mol Biol. 2020:2052:229-251. doi: 10.1007/978-1-4939-9748-0_14.

Abstract

Cryptosporidiosis threatens life of young children in developing countries and newborn calves around the world. No vaccine or therapy can prevent or cure this diarrhea-inducing enteric disease caused by Cryptosporidium spp. protozoan parasites. There is an essential need to discover new therapeutic drugs efficient in reducing parasite burden in infected individuals. Research therefore relies on reliable small animal models of cryptosporidiosis. Here, we present excellent mouse models which can efficiently mimic pathogenesis of human and bovine cryptosporidiosis. We also describe methods to purify C. parvum oocysts from stool and intestine of infected mice to facilitate oocyst quantification. Moreover, we present protocols using flow cytometry, quantitative polymerase chain reaction, and histopathology to accurately quantify parasite burden in stool or intestine samples.

Keywords: Cryptosporidium parvum; Flow cytometry; Histopathology; IL-12 knock-out mice; Interferon gamma knock-out mice; Interferon gamma receptor knock-out mice; Oocysts; Quantitative polymerase chain reaction; SCID mice.

MeSH terms

  • Animals
  • Cryptosporidiosis / parasitology*
  • Cryptosporidiosis / pathology
  • Cryptosporidium
  • Cryptosporidium parvum / growth & development
  • Cryptosporidium parvum / isolation & purification*
  • Disease Models, Animal
  • Feces / parasitology
  • Flow Cytometry / methods
  • Ileum / cytology
  • Ileum / parasitology
  • Ileum / pathology
  • Interferon gamma Receptor
  • Interleukin-12 / genetics
  • Mice
  • Mice, Knockout
  • Mice, SCID
  • Oocysts / growth & development
  • Oocysts / isolation & purification*
  • Real-Time Polymerase Chain Reaction / methods
  • Receptors, Interferon / genetics
  • Workflow

Substances

  • Receptors, Interferon
  • Interleukin-12