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Optimized conditions for the in vitro excystment of Calicophoron daubneyi metacercariae

Published online by Cambridge University Press:  14 December 2017

Kathryn M. Huson ,
Charlotte Wild ,
Caroline Fenn  and
Mark W. Robinson
Kathryn M. Huson
Affiliation:
Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, 97 Lisburn Road, Belfast, Northern Ireland
Charlotte Wild
Affiliation:
Ridgeway Research Ltd., Park Farm Buildings, Park Lane, St. Briavels, Gloucestershire, England
Caroline Fenn
Affiliation:
Ridgeway Research Ltd., Park Farm Buildings, Park Lane, St. Briavels, Gloucestershire, England
Mark W. Robinson*
Affiliation:
Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, 97 Lisburn Road, Belfast, Northern Ireland
*
Author for correspondence: Mark W. Robinson, E-mail: mark.robinson@qub.ac.uk
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Abstract

Paramphistomosis, caused by Calicophoron daubneyi, is an emerging infection of ruminants throughout Western Europe. Despite its prevalence, many questions remain regarding the basic biology of this parasite and how it interacts with its host. Consequently, there is a need to develop methods to study C. daubneyi in vitro to improve our understanding of rumen fluke biology. Towards this, we aimed to identify a suitable protocol for in vitro excystment of C. daubneyi metacercariae. Six methods that have been used to excyst metacercariae from a number of trematode species were tested with C. daubneyi metacercariae. Three of these achieved an average of >50% excystment whilst one method, which included an acid-pepsin treatment, incubation in reducing conditions and an alkaline/bile salt solution to activate the larvae, consistently gave >80% excystment. The latter protocol also showed no detrimental effect on the motility of newly excysted juvenile (NEJ) parasites when observed for up to 24 h in RPMI 1640 medium post-excystment. The successful production of C. daubneyi NEJs in vitro is a significant step forward, and will enable the discovery of infective stage-specific parasite antigens and facilitate drug screening trials, to aid the development of much needed diagnostic and therapeutic options for paramphistomosis.

Keywords

Calicophoron daubneyiExcystmentMetacercariaeParamphistome
Type
Research Article
Information
Parasitology , Volume 145 , Issue 8 , July 2018 , pp. 1015 - 1019
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Copyright
Copyright © Cambridge University Press 2017 

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