Virioplankton 'pegylation': use of PEG (polyethylene glycol) to concentrate and purify viruses in pelagic ecosystems.
We have described the use of Polyethylene glycol (PEG) for the precipitation of natural communities of aquatic viruses, and its comparison with the usual concentration method based on ultracentrifugation. Experimental samples were obtained from different freshwater ecosystems whose trophic status varied. Based on transmission electron microscope observations and counting of phage-shaped particles, our results showed that the greatest recovery efficiency for all ecosystems was obtained when we used the PEG protocol. On average, this protocol allowed the recovery of >2-fold more viruses, compared to ultracentrifugation. In addition, the diversity of virioplankton, based on genomic size profiling using pulsed field gel electrophoresis, was higher and better discriminated when we used the PEG method. We conclude that pegylation offers a valid, simple and cheaper alternative method to ultracentrifugation, for the concentration and the purification of pelagic viruses.
Colombet
J
J
Laboratoire de Biologie des Protistes, Université Blaise Pascal (Clermont-Ferrand II), UMR CNRS 6023, F-63177, Aubière Cedex, France.
Robin
A
A
Lavie
L
L
Bettarel
Y
Y
Cauchie
H M
HM
Sime-Ngando
T
T
D001435:Bacteriophages / Q000302:isolation & purification; D011232:Chemical Precipitation; D017753:Ecosystem; D004784:Environmental Monitoring / Q000379:methods; D005618:Fresh Water / Q000821:virology*; D016679:Genome, Viral / Q000235:genetics; D046529:Microscopy, Electron, Transmission; D010933:Plankton / Q000821:virology; D011092:Polyethylene Glycols / Q000737:chemistry*; D014461:Ultracentrifugation; D014776:Virus Cultivation / Q000379:methods*; D014780:Viruses / Q000235:genetics / Q000302:isolation & purification* / Q000648:ultrastructure; D014871:Water Microbiology
D011092:Polyethylene Glycols