In vitro excystation of Cryptosporidium parvum.
ABSTRACT Protocols for in vitro excystation of oocysts of Cryptosporidium parvum, including different chemical pre-incubation steps, were compared to examine some of the biochemical triggers involved in excystation and to define an in vitro excystation protocol of a reproducibly high efficiency. Pre-incubation steps which increased the permeability of the oocysts were found to enhance excystation dynamics and pre-treatment of oocysts with saliva was found to decrease the permeability and reduce excystation. Although excystation was maximal after incubation for 4 h, sporozoites tended to lyse over this period, and maximum sporozoite recovery occurred after 30 min. The results obtained are discussed in relation to excystation protocols adopted by different research groups and a number of recommendations are given for in vitro excystation of C. parvum oocysts.
- Research Journal of Parasitology. 01/2009; 4(2):63-69.
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ABSTRACT: The fluorogenic vital dyes assay utilizing 4′,6′-diamidino-2-phenyl-indole (DAPI) and propidium iodide (PI) can be used for determining the viability of individual Cryptosporidium parvum oocysts isolated from environmental samples; however this assay can be affected by the pH of the oocyst suspending medium, resulting in formation of fluorescent yellow DAPI crystals at neutral or alkaline pH. Interference from DAPI crystals can lead to oocyst occlusion, making their viability assessment difficult or subjective. When the oocyst suspending medium is rinsed with HBSS adjusted to pH 4, DAPI crystallization is reduced or prevented without affecting oocyst staining characteristics with these two dyes or with fluorescein isothiocyanate conjugated anti-Cryptosporidium monoclonal antibody.Water Research 01/1999; 33(13):3037-3039. · 5.32 Impact Factor