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In vitro encystation and excystation of Blastocystis ratti

Published online by Cambridge University Press:  01 February 1999

X. Q. CHEN ,
M. SINGH ,
J. HOWE ,
L. C. HO ,
S. W. TAN  and
E. H. YAP
X. Q. CHEN
Affiliation:
Department of Microbiology, Faculty of Medicine, National University of Singapore, 10 Kent Ridge Crescent, Singapore 119260
M. SINGH
Affiliation:
Department of Microbiology, Faculty of Medicine, National University of Singapore, 10 Kent Ridge Crescent, Singapore 119260
J. HOWE
Affiliation:
Department of Microbiology, Faculty of Medicine, National University of Singapore, 10 Kent Ridge Crescent, Singapore 119260
L. C. HO
Affiliation:
Department of Microbiology, Faculty of Medicine, National University of Singapore, 10 Kent Ridge Crescent, Singapore 119260
S. W. TAN
Affiliation:
Department of Microbiology, Faculty of Medicine, National University of Singapore, 10 Kent Ridge Crescent, Singapore 119260
E. H. YAP
Affiliation:
Department of Microbiology, Faculty of Medicine, National University of Singapore, 10 Kent Ridge Crescent, Singapore 119260
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Abstract

Cysts of Blastocystis ratti were produced in vitro by culturing the parasite in Iscove's modified Dulbecco's medium (IMDM) with increasing concentrations of horse serum. Yields up to 3×106 cysts/ml of culture medium were obtained after 72 h. Encystation efficiency was time, strain and inoculum size dependent. A viability of >70% was determined by flow cytometry employing fluorescein diacetate and propidium iodide staining. The presence of chitin as a cyst wall component was demonstrated by Calcofluor White M2R staining with which cystic stages showed blue fluorescence. The changes in morphology during excystation were examined by transmission electron microscopy. The cyst enlarged in size and some vacuoles appeared within the condensed cytoplasm. The vacuoles were full of inclusions and small glycogen aggregates. Coalescence of the vacuoles led to central body formation. Glycogen deposits were prominent throughout the excystation process. Some cysts divided by binary fission before the completion of the excystation.

Keywords

Blastocystis rattiencystationexcystation
Type
Research Article
Information
Parasitology , Volume 118 , Issue 2 , February 1999 , pp. 151 - 160
Copyright
1999 Cambridge University Press

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