Probing Mixed-Genotype Infections I: Extraction and Cloning of Infections from Hosts of the Trypanosomatid Crithidia bombi

Open access
Date
2012-11-14Type
- Journal Article
ETH Bibliography
yes
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Abstract
We here present an efficient, precise and reliable method to isolate and cultivate healthy and viable single Crithidia bombi cells from bumblebee faeces using flow cytometry. We report a precision of >99% in obtaining single trypanosomatid cells for further culture and analysis (“cloning”). In the study, we have investigated the use of different liquid media to cultivate C. bombi and present an optimal medium for obtaining viable clones from all tested, infected host donors. We show that this method can be applied to genotype a collection of clones from natural infections. Furthermore, we show how to cryo-preserve C. bombi cells to be revived to become infective clones after at least 4 years of storage. Considering the high prevalence of infections in natural populations, our method provides a powerful tool in studying the level and diversity of these infections, and thus enriches the current methodology for the studies of complex host-parasite interactions. Show more
Permanent link
https://doi.org/10.3929/ethz-b-000059580Publication status
publishedExternal links
Journal / series
PLoS ONEVolume
Pages / Article No.
Publisher
Public Library of ScienceOrganisational unit
03330 - Schmid-Hempel, Paul
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ETH Bibliography
yes
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