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Cryopreservation of Paramecium bursaria Chlorella Virus-1 during an active infection cycle of its host
Best practices in laboratory culture management often include cryopreservation of microbiota, but this can be challenging with some virus particles. By preserving viral isolates researchers can mitigate genetic drift and laboratory-induced selection, thereby maintaining genetically consistent strain...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6417706/ https://www.ncbi.nlm.nih.gov/pubmed/30870463 http://dx.doi.org/10.1371/journal.pone.0211755 |
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author | Coy, Samantha R. Alsante, Alyssa N. Van Etten, James L. Wilhelm, Steven W. |
author_facet | Coy, Samantha R. Alsante, Alyssa N. Van Etten, James L. Wilhelm, Steven W. |
author_sort | Coy, Samantha R. |
collection | PubMed |
description | Best practices in laboratory culture management often include cryopreservation of microbiota, but this can be challenging with some virus particles. By preserving viral isolates researchers can mitigate genetic drift and laboratory-induced selection, thereby maintaining genetically consistent strains between experiments. To this end, we developed a method to cryopreserve the model, green-alga infecting virus, Paramecium bursaria Chlorella virus 1 (PBCV-1). We explored cryotolerance of the infectivity of this virus particle, whereby freezing without cryoprotectants was found to maintain the highest infectivity (~2.5%). We then assessed the cryopreservation potential of PBCV-1 during an active infection cycle in its Chlorella variabilis NC64A host, and found that virus survivorship was highest (69.5 ± 16.5%) when the infected host is cryopreserved during mid-late stages of infection (i.e., coinciding with virion assembly). The most optimal condition for cryopreservation was observed at 240 minutes post-infection. Overall, utilizing the cell as a vehicle for viral cryopreservation resulted in 24.9–30.1 fold increases in PBCV-1 survival based on 95% confidence intervals of frozen virus particles and virus cryopreserved at 240 minutes post-infection. Given that cryoprotectants are often naturally produced by psychrophilic organisms, we suspect that cryopreservation of infected hosts may be a reliable mechanism for virus persistence in non-growth permitting circumstances in the environment, such as ancient permafrosts. |
format | Online Article Text |
id | pubmed-6417706 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-64177062019-04-01 Cryopreservation of Paramecium bursaria Chlorella Virus-1 during an active infection cycle of its host Coy, Samantha R. Alsante, Alyssa N. Van Etten, James L. Wilhelm, Steven W. PLoS One Research Article Best practices in laboratory culture management often include cryopreservation of microbiota, but this can be challenging with some virus particles. By preserving viral isolates researchers can mitigate genetic drift and laboratory-induced selection, thereby maintaining genetically consistent strains between experiments. To this end, we developed a method to cryopreserve the model, green-alga infecting virus, Paramecium bursaria Chlorella virus 1 (PBCV-1). We explored cryotolerance of the infectivity of this virus particle, whereby freezing without cryoprotectants was found to maintain the highest infectivity (~2.5%). We then assessed the cryopreservation potential of PBCV-1 during an active infection cycle in its Chlorella variabilis NC64A host, and found that virus survivorship was highest (69.5 ± 16.5%) when the infected host is cryopreserved during mid-late stages of infection (i.e., coinciding with virion assembly). The most optimal condition for cryopreservation was observed at 240 minutes post-infection. Overall, utilizing the cell as a vehicle for viral cryopreservation resulted in 24.9–30.1 fold increases in PBCV-1 survival based on 95% confidence intervals of frozen virus particles and virus cryopreserved at 240 minutes post-infection. Given that cryoprotectants are often naturally produced by psychrophilic organisms, we suspect that cryopreservation of infected hosts may be a reliable mechanism for virus persistence in non-growth permitting circumstances in the environment, such as ancient permafrosts. Public Library of Science 2019-03-14 /pmc/articles/PMC6417706/ /pubmed/30870463 http://dx.doi.org/10.1371/journal.pone.0211755 Text en © 2019 Coy 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 Coy, Samantha R. Alsante, Alyssa N. Van Etten, James L. Wilhelm, Steven W. Cryopreservation of Paramecium bursaria Chlorella Virus-1 during an active infection cycle of its host |
title | Cryopreservation of Paramecium bursaria Chlorella Virus-1 during an active infection cycle of its host |
title_full | Cryopreservation of Paramecium bursaria Chlorella Virus-1 during an active infection cycle of its host |
title_fullStr | Cryopreservation of Paramecium bursaria Chlorella Virus-1 during an active infection cycle of its host |
title_full_unstemmed | Cryopreservation of Paramecium bursaria Chlorella Virus-1 during an active infection cycle of its host |
title_short | Cryopreservation of Paramecium bursaria Chlorella Virus-1 during an active infection cycle of its host |
title_sort | cryopreservation of paramecium bursaria chlorella virus-1 during an active infection cycle of its host |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6417706/ https://www.ncbi.nlm.nih.gov/pubmed/30870463 http://dx.doi.org/10.1371/journal.pone.0211755 |
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