Genomic and virulence analysis of in vitro cultured Cryptosporidium parvum

PLoS Pathog. 2024 Feb 28;20(2):e1011992. doi: 10.1371/journal.ppat.1011992. eCollection 2024 Feb.

Abstract

Recent advances in the in vitro cultivation of Cryptosporidium parvum using hollow fiber bioreactor technology (HFB) have permitted continuous growth of parasites that complete all life cycle stages. The method provides access to all stages of the parasite and provides a method for non-animal production of oocysts for use in clinical trials. Here we examined the effect of long-term (>20 months) in vitro culture on virulence-factors, genome conservation, and in vivo pathogenicity of the host by in vitro cultured parasites. We find low-level sequence variation that is consistent with that observed in calf-passaged parasites. Further using a calf model infection, oocysts obtained from the HFB caused diarrhea of the same volume, duration and oocyst shedding intensity as in vivo passaged parasites.

MeSH terms

  • Animals
  • Cryptosporidiosis* / parasitology
  • Cryptosporidium parvum* / genetics
  • Cryptosporidium*
  • Feces
  • Genomics
  • Oocysts
  • Virulence

Grants and funding

Financial support was provided by The Bill and Melinda Gates Foundation awards OPP1117675 (N.Y.), OPP1151701 (J.C.K), Investment 44418 from the Bill and Melinda Gates Foundation (M.W.R.). Investment GH VAP NG-ID20 from Bill and Melinda Gates Foundation (NY). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. M.M., R.P.B., and D.S., received salary support from the BMGF.