Cryopreservation of Paramecium bursaria Chlorella Virus-1 during an active infection cycle of its host

Samantha R. Coy, Alyssa N. Alsante, James L. Van Etten, Steven W. Wilhelm

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Article numbere0211755
JournalPloS one
Volume14
Issue number3
DOIs
StatePublished - Mar 2019

Fingerprint

Paramecium bursaria Chlorella virus 1
Paramecium
Chlorella
Cryopreservation
Viruses
cryopreservation
virion
Virion
Infection
viruses
infection
cryoprotectants
pathogenicity
genetic drift
Genetic Drift
Chlorophyta
confidence interval
freezing
Microbiota
researchers

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

Cite this

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.

In: PloS one, Vol. 14, No. 3, e0211755, 03.2019.

Research output: Contribution to journalArticle

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. In: PloS one. 2019 ; Vol. 14, No. 3.
@article{b076cf8312e44f62bcd0926ca053a9ba,
title = "Cryopreservation of Paramecium bursaria Chlorella Virus-1 during an active infection cycle of its host",
abstract = "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.",
author = "Coy, {Samantha R.} and Alsante, {Alyssa N.} and {Van Etten}, {James L.} and Wilhelm, {Steven W.}",
year = "2019",
month = "3",
doi = "10.1371/journal.pone.0211755",
language = "English (US)",
volume = "14",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "3",

}

TY - JOUR

T1 - Cryopreservation of Paramecium bursaria Chlorella Virus-1 during an active infection cycle of its host

AU - Coy, Samantha R.

AU - Alsante, Alyssa N.

AU - Van Etten, James L.

AU - Wilhelm, Steven W.

PY - 2019/3

Y1 - 2019/3

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=85062995694&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85062995694&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0211755

DO - 10.1371/journal.pone.0211755

M3 - Article

C2 - 30870463

AN - SCOPUS:85062995694

VL - 14

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 3

M1 - e0211755

ER -