Comparison of viability assays for Cryptosporidium parvum oocysts after disinfection.
In order to test various viability assays for Cryptosporidium parvum oocysts were used to infect HCT-8 cells in vitro or baby mice. Infected cells were either stained with fluorescent anti-Cryptosporidium-antibody or lysed and subjected to C. parvum-specific PCR after 48 h. Titrations with infective oocysts were performed and compared to oocysts disinfected with Neopredisan for 2 h at varying concentrations. Caecal smears and histological sections from infected animals were examined in parallel. The number of foci of parasite development in vitro after immunofluorescent staining correlated well with the infection dose. PCR was less quantifiable and the results were not always reproducible, especially when low infection doses were used. Disinfection resulted in a dose-dependent reduction of oocyst infectivity when compared to the controls in all three assays. The infection of cells cultured in vitro with oocysts of C. parvum provides a suitable tool for the estimation of viability after treatment with chemical disinfectants. Immunofluorescence is easy to perform and gives quantitative results, while PCR-based detection of parasite DNA, although possible, requires the use of more sophisticated tools for quantification.
Joachim
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Institute for Parasitology, Faculty of Veterinary Medicine, University of Leipzig, An den TierKliniken 33, Leipzig 04103, Germany. ajoachim@vetmed.uni-leipzig.de
Eckert
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E
Petry
F
F
Bialek
R
R
Daugschies
A
A
D000818:Animals; D002460:Cell Line; D048429:Cell Size / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003408:Cresols / Q000494:pharmacology*; D016785:Cryptosporidium parvum / Q000166:cytology / Q000187:drug effects* / Q000235:genetics; D016054:DNA, Protozoan / Q000032:analysis; D004203:Disinfection; D004305:Dose-Response Relationship, Drug; D005455:Fluorescent Antibody Technique; D005659:Fungicides, Industrial / Q000494:pharmacology*; D051379:Mice; D033661:Oocysts / Q000166:cytology* / Q000187:drug effects*; D016133:Polymerase Chain Reaction
D003408:Cresols; D016054:DNA, Protozoan; D005659:Fungicides, Industrial; C006984:chlorocresol