Flow cytometry with fluorescence activated cell sorting (FACS) was used to purify Cryptosporidium (oo)cysts and Giardia cysts from water. With this purification step Cryptosporidium and Giardia were found in a higher percentage of the samples and significantly higher Giardia concentrations were detected in these positive samples. Because FACS removed most of the debris from the concentrated water sample, the microscopic preparations could be examined more rapidly for the presence of (oo)cysts and the morphological characteristics of the (oo)cysts could be interpreted more unambiguously than with the conventional immunofluorescence microscopy method. The use of FACS made it possible to apply PI-staining on environmental samples to determine the fraction of dead (oo)cysts. Sample processing did not appear to influence the PI-staining characteristics of the Cryptosporidium (oo)cysts, but did increase the percentage of PI-positive Giardia cysts. This suggests that this protocol can be used for determining the percentage of dead Cryptosporidium oocysts in environmental samples. Analysis of environmental samples suggests that reservoir storage increases the percentage of PI-positive (dead) oocysts.
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Research Article|
December 01 1998
Improved detection and vital staining of Cryptosporidium and Giardia with flow cytometry Available to Purchase
G. J. Medema;
G. J. Medema
*Microbiological Laboratory for Health Protection, RIVM, P.O. Box 1, 3720 BA Bilthoven, The Netherlands
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F. M. Schets;
F. M. Schets
*Microbiological Laboratory for Health Protection, RIVM, P.O. Box 1, 3720 BA Bilthoven, The Netherlands
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H. Ketelaars;
H. Ketelaars
**Water Storage Company Brabantse Biesbosch, Werkendam, The Netherlands
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G. Boschman
G. Boschman
***Becton Dickinson Immunocytometry Systems Europe, Erembodegem, Aalst, Belgium
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Water Sci Technol (1998) 38 (12): 61–65.
Citation
G. J. Medema, F. M. Schets, H. Ketelaars, G. Boschman; Improved detection and vital staining of Cryptosporidium and Giardia with flow cytometry. Water Sci Technol 1 December 1998; 38 (12): 61–65. doi: https://doi.org/10.2166/wst.1998.0501
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