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Induced encystment improves resistance to preservation and storage of Acanthamoeba castellanii

Published online by Cambridge University Press:  21 October 2008

S. J. CAMPBELL ,
P. R. INGRAM ,
C. W. ROBERTS  and
F. L. HENRIQUEZ
S. J. CAMPBELL
Affiliation:
School of Engineering and Science, University of the West of Scotland, Paisley PA1 2BE, Scotland
P. R. INGRAM
Affiliation:
Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow G4 0NR, Scotland
C. W. ROBERTS
Affiliation:
Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow G4 0NR, Scotland
F. L. HENRIQUEZ*
Affiliation:
School of Engineering and Science, University of the West of Scotland, Paisley PA1 2BE, Scotland
*
*Corresponding author: School of Engineering and Science, University of the West of Scotland, Paisley PA1 2BE, Scotland. Tel: +01418483119. Fax: +01415484823. E-mail: [email protected]
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Summary

Several conditions that allow the preservation, storage and rapid, efficient recovery of viable Acanthamoeba castellanii organisms were investigated. The viability of trophozoites (as determined by time to confluence) significantly declined over a period of 12 months when stored at −70°C using dimethyl sulfoxide (DMSO; 5 or 10%) as cryopreservant. As A. castellanii are naturally capable of encystment, studies were undertaken to determine whether induced encystment might improve the viability of organisms under a number of storage conditions. A. castellanii cysts stored in the presence of Mg2+ at 4°C remained viable over the study period, although time to confluence was increased from approximately 8 days to approximately 24 days over the 12-month period. Storage of cysts at −70°C with DMSO (5 or 10%) or 40% glycerol, but not 80% glycerol as cryopreservants increased their viability over the 12-month study period compared with those stored at room temperature. Continued presence of Mg2+ in medium during storage had no adverse effects and generally improved recovery of viable organisms. The present study demonstrates that A. castellanii can be stored as a non-multiplicative form inexpensively, without a need for cryopreservation, for at least 12 months, but viability is increased by storage at −70°C.

Keywords

Acanthamoebapreservationstoragecysttrophozoite
Type
Original Articles
Information
Parasitology , Volume 135 , Issue 12 , October 2008 , pp. 1401 - 1405
Copyright
Copyright © 2008 Cambridge University Press

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