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Long-term passage of tachyzoites in tissue culture can attenuate virulence of Neospora caninum in vivo

Published online by Cambridge University Press:  09 June 2006

P. M. BARTLEY ,
S. WRIGHT ,
J. SALES ,
F. CHIANINI ,
D. BUXTON  and
E. A. INNES
P. M. BARTLEY
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik EH26 0PZ, Scotland, UK
S. WRIGHT
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik EH26 0PZ, Scotland, UK
J. SALES
Affiliation:
Biomathematics and Statistics Scotland, James Clerk Maxwell Building, The King's Buildings, Edinburgh EH9 3JZ, Scotland, UK
F. CHIANINI
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik EH26 0PZ, Scotland, UK
D. BUXTON
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik EH26 0PZ, Scotland, UK
E. A. INNES
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik EH26 0PZ, Scotland, UK
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Abstract

To determine whether prolonged in vitro passage would result in attenuation of virulence in vivo, Neospora caninum tachyzoites were passaged for different lengths of time in vitro and compared for their ability to cause disease in mice. Groups of Balb/c mice were inoculated intraperitoneally with 5×106 or 1×107 of low-passage or high-passage N. caninum tachyzoites. The mice were monitored for changes in their demeanour and body weight, and were culled when severe clinical symptoms of murine neosporosis were observed. Mice inoculated with the high-passage parasites survived longer (P<0·05), and showed fewer clinical symptoms of murine neosporosis, compared to the mice receiving the low-passage parasites. The parasite was detected in the brains of inoculated mice using immunohistochemistry and ITS1 PCR. Tissue cysts containing parasites were seen in mice inoculated with both low-passage and high-passage parasites. When the in vitro growth rates of the parasites were compared, the high-passage parasites initially multiplied more rapidly (P<0·001) than the low-passage parasites, suggesting that the high-passage parasites had become more adapted to tissue culture. These results would suggest that it is possible to attenuate the virulence of N. caninum tachyzoites in mice through prolonged in vitro passage.

Keywords

Neospora caninumin vitro cultureattenuation in vivo
Type
Research Article
Information
Parasitology , Volume 133 , Issue 4 , October 2006 , pp. 421 - 432
Copyright
© 2006 Cambridge University Press

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