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Morphological changes and viability of Cryptosporidium parvum sporozoites after excystation in cell-free culture media

Published online by Cambridge University Press:  27 August 2010

M. MATSUBAYASHI ,
H. ANDO ,
I. KIMATA ,
H. NAKAGAWA ,
M. FURUYA ,
H. TANI  and
K. SASAI
M. MATSUBAYASHI
Affiliation:
Department of Food and Nutrition, Osaka Yuhigaoka Gakuen Junior College, Osaka, Osaka 543-0073, Japan
H. ANDO
Affiliation:
Department of Veterinary Internal Medicine, Division of Veterinary Science, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, Osaka 598-8531, Japan
I. KIMATA
Affiliation:
Department of Parasitology, Graduate School of Medicine, Osaka City University, Osaka, Osaka 545-8585, Japan
H. NAKAGAWA
Affiliation:
Department of Central Laboratory, Graduate School of Medicine, Osaka City University, Osaka, Osaka 545-8585, Japan
M. FURUYA
Affiliation:
Department of Veterinary Internal Medicine, Division of Veterinary Science, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, Osaka 598-8531, Japan
H. TANI
Affiliation:
Department of Veterinary Internal Medicine, Division of Veterinary Science, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, Osaka 598-8531, Japan
K. SASAI*
Affiliation:
Department of Veterinary Internal Medicine, Division of Veterinary Science, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, Osaka 598-8531, Japan
*
*Corresponding author: Department of Veterinary Internal Medicine, Division of Veterinary Science, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, Osaka 598-8531, Japan. Tel: +81 72 463 5385. Fax: +81 72 463 5387. E-mail: [email protected].
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Summary

Cryptosporidium parvum, belonging to the phylum Apicomplexa, is a major cause of waterborne gastroenteritis throughout the world. The sporozoites are thought to invade host enterocytes using an active process termed gliding motility. However, the biological and morphological changes within the sporozoites during this process are not fully understood. In the present study, excysted sporozoites of C. parvum were analysed ultrastructurally in vitro and their viability was evaluated using fluorescent dyes. The sporozoites excysted from oocysts changed morphologically from banana-shaped to rod-shaped and finally to a rounded shape, in culture media in 3 h. Transmission microscopy revealed that the distance between the apical end and the nucleus was markedly reduced, dense granules were present close to the rhoptry in the apical region, amylopectin granules were absent, and membranes of round sporozoites were less clear. A fluorescent assay showed that the rate of survival decreased from 89% to 56% at 0–3 h (84·3% for banana-shaped and 49·2% for rod-shaped sporozoites). Therefore, post-excysted sporozoites in vitro underwent morphological changes and a rapid loss of viability. This staining method is useful, inexpensive and provides an alternative to more costly and intensive flow cytometric assays or infectivity assays with host cells in vitro.

Keywords

Cryptosporidium parvumexcystationmorphological changessporozoitesviability
Type
Research Article
Information
Parasitology , Volume 137 , Issue 13 , November 2010 , pp. 1861 - 1866
Copyright
Copyright © Cambridge University Press 2010

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