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Diseases of Aquatic Organisms

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DAO 130:65-70 (2018)  -  DOI: https://doi.org/10.3354/dao03266

NOTE
Cryopreservation methods are effective for long-term storage of Labyrinthula cultures

Stacey M. Trevathan-Tackett1,2,*, Sarah Treby1,2, Frank H. Gleason3, Peter I. Macreadie1,2, Stella Loke1,4

1Deakin University, Geelong, School of Life and Environmental Sciences, Burwood, VIC 3125, Australia
2Deakin University, Geelong, Centre for Integrative Ecology, Burwood, VIC 3125, Australia
3School of Life and Environmental Sciences, University of Sydney, Sydney, NSW 2006, Australia
4Deakin University, Geelong, Centre for Cellular and Molecular Biology, Burwood, VIC 3125, Australia
*Corresponding author:

ABSTRACT: Marine heterotrophic protists of the Labyrinthulomycota are of interest for their biotechnological (e.g. thraustochytrid production of lipids) and ecological (e.g. wasting disease and rapid blight by pathogens of the genus Labyrinthula) applications; culture-based laboratory studies are a central technique of this research. However, maintaining such microorganism cultures can be labour- and cost-intensive, with a high risk of culture contamination and die-off over time. Deep-freeze storage, or cryopreservation, can be used to maintain culture back-ups, as well as to preserve the genetic and phenotypic properties of the microorganisms; however, this method has not been tested for the ubiquitous marine protists Labyrinthula spp. In this study, we trialled 12 cryopreservation protocols on 3 Labyrinthula sp. isolates of varying colony morphological traits. After 6 mo at -80°C storage, the DMSO and glycerol protocols were the most effective cryoprotectants compared to methanol (up to 90% success vs. 50% success, respectively). The addition of 30% horse serum to the cryoprotectant solution increased Labyrinthula sp. growth success by 20-30%. We expect that these protocols will provide extra security for culture-based studies, as well as opportunities for long-term research on key Labyrinthula sp. isolates.


KEY WORDS: Dimethyl sulfoxide · DMSO · Glycerol · Horse serum · Cryoprotectant · Wasting disease · Marine protist


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Cite this article as: Trevathan-Tackett SM, Treby S, Gleason FH, Macreadie PI, Loke S (2018) Cryopreservation methods are effective for long-term storage of Labyrinthula cultures. Dis Aquat Org 130:65-70. https://doi.org/10.3354/dao03266

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