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High-yield amplification of Cryptosporidium parvum in interferon γ receptor knockout mice

Published online by Cambridge University Press:  31 July 2008

J. von OETTINGEN ,
M. NATH-CHOWDHURY ,
B. J. WARD ,
A. C. RODLOFF ,
M. J. ARROWOOD  and
M. NDAO
J. von OETTINGEN
Affiliation:
National Reference Centre for Parasitology, Research Institute of the McGill University Heath Centre, Montreal General Hospital Research Institute, Montreal, Canada Institute for Medical Microbiology and Epidemiology of Infectious Diseases, University of Leipzig, Leipzig, Germany
M. NATH-CHOWDHURY
Affiliation:
National Reference Centre for Parasitology, Research Institute of the McGill University Heath Centre, Montreal General Hospital Research Institute, Montreal, Canada
B. J. WARD
Affiliation:
National Reference Centre for Parasitology, Research Institute of the McGill University Heath Centre, Montreal General Hospital Research Institute, Montreal, Canada
A. C. RODLOFF
Affiliation:
Institute for Medical Microbiology and Epidemiology of Infectious Diseases, University of Leipzig, Leipzig, Germany
M. J. ARROWOOD
Affiliation:
Centers for Disease Control and Prevention, Atlanta, USA
M. NDAO*
Affiliation:
National Reference Centre for Parasitology, Research Institute of the McGill University Heath Centre, Montreal General Hospital Research Institute, Montreal, Canada
*
*Corresponding author: Research Institute of the McGill University Heath Centre, Montreal General Hospital Research Institute, Room R3-137. 1650 Cedar Ave, Montreal, Quebec, CanadaH3G 1A4. Tel: +1 514 934 8347. E-mail: momar.ndao@mail.mcgill.ca
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Summary

To date, large-scale production of Cryptosporidium parvum oocysts has only been achieved by amplification in neonatal calves and sheep. Many laboratories currently depend on supplies from external sources and store oocysts for prolonged periods which results in progressive loss of viability. Six to 8-week-old interferon γ receptor knockout (IFNγR-KO) mice on a C57BL/6 background were inoculated by gavage (2000 oocysts/animal). Fecal pellets were collected daily from 7 days post-infection (p.i.) up to 2 weeks p.i. Intestinal oocyst yield was assessed at days 11, 12 and 14 p.i. by homogenization of intestinal tissues. Ether extraction and one or more NaCl flotations were used to purify oocysts. Total recoveries averaged 2·6×106 oocysts/mouse from fecal material and 3·8×107 oocysts/mouse from intestinal tissues. Overall, 2·3×109 purified oocysts were obtained from 60 mice. Recovered oocysts were capable of sporulation and were shown to be infectious both in vitro and in vivo. Oocyst amplification was achieved in only 11–14 days with minimal expense. The simplicity of this method presents a practical alternative for the routine passage, maintenance and storage of C. parvum in biomedical laboratories.

Keywords

Cryptosporidium parvuminterferon-γ-receptor knock-out mouseoocyst amplification
Type
Original Articles
Information
Parasitology , Volume 135 , Issue 10 , September 2008 , pp. 1151 - 1156
Copyright
Copyright © 2008 Cambridge University Press

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