Morphological changes and viability of Cryptosporidium parvum sporozoites after excystation in cell-free culture media.
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.
Matsubayashi
M
M
Department of Food and Nutrition, Osaka Yuhigaoka Gakuen Junior College, Osaka, Japan.
Ando
H
H
Kimata
I
I
Nakagawa
H
H
Furuya
M
M
Tani
H
H
Sasai
K
K
D000818:Animals; D016785:Cryptosporidium parvum / Q000254:growth & development* / Q000502:physiology / Q000648:ultrastructure*; D003470:Culture Media; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D046529:Microscopy, Electron, Transmission; D033661:Oocysts / Q000502:physiology / Q000648:ultrastructure; D034101:Sporozoites / Q000254:growth & development / Q000502:physiology / Q000648:ultrastructure
D003470:Culture Media; D005456:Fluorescent Dyes