AIM: 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. METHODS AND 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. 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.
AIM: 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. METHODS AND 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. 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.
Authors: Alex Echeverría-Vega; Pablo Morales-Vicencio; Camila Saez-Saavedra; Felipe Gordillo-Fuenzalida; Rubén Araya Journal: MethodsX Date: 2019-11-07