Isolation and Characterization of Novel Marine Bacteriophages from Santa Rosa Island, Florida, Using Local and Nonlocal Bacterial Strains as Hosts

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Bacteriophages, or phages, are viruses that infect bacteria and are the most prevalent biological entities in the world. While phage research has continued to develop, the investigation of phages within ecological biology remains in the early stages and requires an extensive database of environmentally derived phages, such as from marine ecosystems. The objectives of this research were to a) determine the local beach bacterial composition and select locally isolated strains as potential bacteriophage hosts, b) determine the efficacy of local bacteria as marine bacteriophage hosts (Erythrobacter citreus Pensacola AB and Microbacterium oleivorans Pensacola AB), and c) compare the host efficacy between locally isolated bacterial strains and non-local Microbacterium isolates (Microbacterium sp. Casco Bay and Microbacterium foliorum NRRL B-24224). The results suggest that of the two locally isolated bacterial strains tested, Microbacterium oleivorans was not a successful host for the scope of this research. In contrast, Erythrobacter citreus Pensacola AB was an efficient host across several trials. Of the non-local bacteria, both Microbacterium sp. Casco Bay and Microbacterium foliorum NRRL B-24224 were successful hosts at isolating phage, although not at the same consistency as Erythrobacter citreus. This research concluded that local bacterial strains could be effective for phage hunting and that the overall success of finding local marine phage was not dependent on using local bacterial strains. Four phages, two M. sp. Casco Bay phages, one M. foliorum phage, and one E. citreus phage, were selected for further purification and characterization. The four phage genomes were sequenced to characterize the molecular nature of these marine phages. KEYWORDS: Microbiome; Bacteriophage; Marine phage; Plaque Assay; Genomic sequencing; Marine ecology; Zinc chloride (ZnCl2) precipitation; PEG (polyethylene glycol) precipitation

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