An Agar-Based Method for Plating Marine Protozoan Parasites of the Genus Perkinsus
The genus Perkinsus includes protozoan parasites of mollusks responsible for losses in the aquaculture industry and hampering the recovery of natural shellfish beds worldwide, and they are a key taxon for understanding intracellular parasitism adaptations. The ability to propagate the parasite in li...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4858233/ https://www.ncbi.nlm.nih.gov/pubmed/27149378 http://dx.doi.org/10.1371/journal.pone.0155015 |
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author | Cold, Emma R. Freyria, Nastasia J. Martínez Martínez, Joaquín Fernández Robledo, José A. |
author_facet | Cold, Emma R. Freyria, Nastasia J. Martínez Martínez, Joaquín Fernández Robledo, José A. |
author_sort | Cold, Emma R. |
collection | PubMed |
description | The genus Perkinsus includes protozoan parasites of mollusks responsible for losses in the aquaculture industry and hampering the recovery of natural shellfish beds worldwide, and they are a key taxon for understanding intracellular parasitism adaptations. The ability to propagate the parasite in liquid media, in the absence of the host, has been crucial for improving understanding of its biology; however, alternative techniques to grow the parasite are needed to explore other basic aspects of the Perkinsus spp. biology. We optimized a DME: Ham’s F12–5% FBS- containing solid agar medium for plating Perkinsus marinus. This solid medium supported trophozoite propagation both by binary fission and schizogony. Colonies were visible to the naked eye 17 days after plating. We tested the suitability of this method for several applications, including the following: 1) Subcloning P. marinus isolates: single discrete P. marinus colonies were obtained from DME: Ham’s F12–5% FBS– 0.75% agar plates, which could be further propagated in liquid medium; 2) Subcloning engineered Perkinsus mediterraneus MOE[MOE]: GFP by streaking cultures on plates; 3) Chemical susceptibility: Infusing the DME: Ham’s F12–5% FBS– 0.75% agar plates with triclosan resulted in inhibition of the parasite propagation in a dose-dependent manner. Altogether, our plating method has the potential for becoming a key tool for investigating diverse aspects of Perkinsus spp. biology, developing new molecular tools, and for biotechnological applications. |
format | Online Article Text |
id | pubmed-4858233 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-48582332016-05-13 An Agar-Based Method for Plating Marine Protozoan Parasites of the Genus Perkinsus Cold, Emma R. Freyria, Nastasia J. Martínez Martínez, Joaquín Fernández Robledo, José A. PLoS One Research Article The genus Perkinsus includes protozoan parasites of mollusks responsible for losses in the aquaculture industry and hampering the recovery of natural shellfish beds worldwide, and they are a key taxon for understanding intracellular parasitism adaptations. The ability to propagate the parasite in liquid media, in the absence of the host, has been crucial for improving understanding of its biology; however, alternative techniques to grow the parasite are needed to explore other basic aspects of the Perkinsus spp. biology. We optimized a DME: Ham’s F12–5% FBS- containing solid agar medium for plating Perkinsus marinus. This solid medium supported trophozoite propagation both by binary fission and schizogony. Colonies were visible to the naked eye 17 days after plating. We tested the suitability of this method for several applications, including the following: 1) Subcloning P. marinus isolates: single discrete P. marinus colonies were obtained from DME: Ham’s F12–5% FBS– 0.75% agar plates, which could be further propagated in liquid medium; 2) Subcloning engineered Perkinsus mediterraneus MOE[MOE]: GFP by streaking cultures on plates; 3) Chemical susceptibility: Infusing the DME: Ham’s F12–5% FBS– 0.75% agar plates with triclosan resulted in inhibition of the parasite propagation in a dose-dependent manner. Altogether, our plating method has the potential for becoming a key tool for investigating diverse aspects of Perkinsus spp. biology, developing new molecular tools, and for biotechnological applications. Public Library of Science 2016-05-05 /pmc/articles/PMC4858233/ /pubmed/27149378 http://dx.doi.org/10.1371/journal.pone.0155015 Text en © 2016 Cold et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Cold, Emma R. Freyria, Nastasia J. Martínez Martínez, Joaquín Fernández Robledo, José A. An Agar-Based Method for Plating Marine Protozoan Parasites of the Genus Perkinsus |
title | An Agar-Based Method for Plating Marine Protozoan Parasites of the Genus Perkinsus |
title_full | An Agar-Based Method for Plating Marine Protozoan Parasites of the Genus Perkinsus |
title_fullStr | An Agar-Based Method for Plating Marine Protozoan Parasites of the Genus Perkinsus |
title_full_unstemmed | An Agar-Based Method for Plating Marine Protozoan Parasites of the Genus Perkinsus |
title_short | An Agar-Based Method for Plating Marine Protozoan Parasites of the Genus Perkinsus |
title_sort | agar-based method for plating marine protozoan parasites of the genus perkinsus |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4858233/ https://www.ncbi.nlm.nih.gov/pubmed/27149378 http://dx.doi.org/10.1371/journal.pone.0155015 |
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