Comparison of in vitro growth characteristics of Cryptosporidium hominis (IdA15G1) and Cryptosporidium parvum (Iowa-IIaA17G2R1 and IIaA18G3R1)

Abstract

Cryptosporidium is a major cause of diarrhoeal disease and mortality in young children in resource-poor countries, for which no vaccines or adequate therapeutic options are available. Infection in humans is primarily caused by two species: C. hominis and C. parvum. Despite C. hominis being the dominant species infecting humans in most countries, very little is known about its growth characteristics and life cycle in vitro, given that the majority of our knowledge of the in vitro development of Cryptosporidium has been based on C. parvum. In the present study, the growth and development of two C. parvum isolates (subtypes Iowa-IIaA17G2R1 and IIaA18G3R1) and one C. hominis isolate (subtype IdA15G1) in HCT-8 cells were examined and compared at 24 h and 48 h using morphological data acquired with scanning electron microscopy. Our data indicated no significant differences in the proportion of meronts or merozoites between species or subtypes at either time-point. Sexual development was observed at the 48-h time-point across both species through observations of both microgamonts and macrogamonts, with a higher frequency of macrogamont observations in C. hominis (IdA15G1) cultures at 48-h post-infection compared to both C. parvum subtypes. This corresponded to differences in the proportion of trophozoites observed at the same time point. No differences in proportion of microgamonts were observed between the three subtypes, which were rarely observed across all cultures. In summary, our data indicate that asexual development of C. hominis is similar to that of C. parvum, while sexual development is accelerated in C. hominis. This study provides new insights into differences in the in vitro growth characteristics of C. hominis when compared to C. parvum, which will facilitate our understanding of the sexual development of both species.

A pumpless and tubeless microfluidic device enables extendedin vitrodevelopment ofCryptosporidium parvum

“…Scanning electron micrographs demonstrated a large amount of heterogeneity of both asexual and sexual developmental stages after two days of infection under fluid shear stress conditions. Under static conditions, in vitro development of C. parvum has been previously observed to be largely asexual at the same time-point (35,41).…”

“…The C. parvum (Iowa-IIaA17G2R1) oocysts were obtained from BioPoint Pty Ltd (Sydney, Australia) and the subtype was confirmed using a nested PCR targeting the 60-kilodalton glycoprotein (gp60) locus described previously (33,34), with minor modifications to reagent and primer concentrations reported elsewhere (35). The C. parvum oocysts underwent excystation pre-treatment as described previously (36).…”