Efficient method to isolate and purify viruses of bacteria from marine environments.
OBJECTIVE
To isolate viruses of specific heterotrophic bacterial strains from marine environments using a host addition/virus amplification protocol (HAVAP) for use in phage/host systems.


RESULTS
Bacteria-free seawater samples containing natural viruses assemblages were inoculated with a single laboratory grown bacterial host of interest in a nutrient-enriched [peptone, Fe(III) and yeast extract] seawater suspension. These conditions enhanced the replication of only those virus(s) capable of infecting the host bacterium. After incubation, free viruses were recovered at concentrations ranging 10(5)-10(10) infectious virus particles per ml of seawater. Using this approach, 15 viruses were isolated and represented 12 unique phage/host systems. Two of the hosts tested were infected by more than one virus.


CONCLUSIONS
Isolation of high concentrations of specific viruses is possible even if their initial concentrations in native waters are low. This approach allows the recovery of phage/host systems that may not be numerically dominant.


CONCLUSIONS
This host enrichment protocol for virus detection and isolation is well-suited for aquatic viral ecology studies that require phage/host systems.
Zemb
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UPMC Univ Paris, LOBB, Observatoire Océanologique, Banyuls/mer, France.
Urios
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Coetsier
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Lebaron
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D001419:Bacteria / Q000254:growth & development / Q000821:virology*; D001435:Bacteriophages / Q000145:classification / Q000254:growth & development / Q000302:isolation & purification* / Q000502:physiology; D004777:Environment; D012623:Seawater / Q000821:virology*; D014871:Water Microbiology