Efficient method to isolate and purify viruses of bacteria from marine environments

Olivier Zemb (1, 2) , L. Urios (1, 2) , C. Coetsier (1, 2) , P. Lebaron (1, 2)

1 UPMC - Université Pierre et Marie Curie - Paris 6
2 OOB - Observatoire océanologique de Banyuls

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. 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. 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. Significance and Impact of the Study: This host enrichment protocol for virus detection and isolation is well-suited for aquatic viral ecology studies that require phage/host systems.

Mots clés: genetic diversity; north-sea; bacteriophage; water; hawaii

Domaines: Sciences du Vivant [q-bio]

Olivier Zemb, L. Urios, C. Coetsier, P. Lebaron. Efficient method to isolate and purify viruses of bacteria from marine environments. Letters in Applied Microbiology, 2008, 47 (1), pp.41-45. ⟨10.1111/j.1472-765X.2008.02381.x⟩. ⟨hal-02661718⟩