Abstract
A novel virulent bacteriophage, vB_VspP_pVa5, infecting a strain of Vibrio splendidus was isolated from a sea-cage aquaculture farm in Greece, and characterized using microbiological methods and genomic analysis. Bacteriophage vB_VspP_pVa5 is a N4-like podovirus with an icosahedral head measuring 85 nm in length and a short non-contractile tail. The phage had a narrow host range infecting only the bacterial host, a latent period of 30 min and a burst size of 24 virions per infected bacterium. Its genome size was 78,145 bp and genomic analysis identified 107 densely-packed genes, 40 of which could be annotated. In addition to the very large virion encapsulated DNA-dependent RNA polymerase which is the signature of the N4-like genus, an interesting feature of the novel phage is the presence of a self-splicing group I intron in the thymidylate synthase gene. A tRNAStop interrupted by a ~2.5kb open reading frame–containing area was also identified. The absence of genes related to lysogeny along with the high efficacy observed during in vitro cell lysis trials, indicate that the vB_VspP_pVa5 is a potential candidate component in a bacteriophage cocktail suitable for the biological control of V. splendidus in aquaculture.
Highlights
Vibrio splendidus is a ubiquitous inhabitant of marine and brackish water with prodigious genotypic diversity that plays a major ecological part in the oceanic and coastal environments [1,2]
Bacterial strains were maintained in microbeads (MicroBank) at -80 ̊C and grown in sea water supplemented with 1% tryptone and 0.5% yeast extract at 25 ̊C
Following transmission electron microscope (TEM) observations, it was classified to Podoviridae family
Summary
Vibrio splendidus is a ubiquitous inhabitant of marine and brackish water with prodigious genotypic diversity that plays a major ecological part in the oceanic and coastal environments [1,2]. It has been associated with severe epizootics in many farmed aquatic animals including fishes [3,4,5], crustaceans [6], bivalves and echinoderms [7]. V. splendidus is considered to be one of the most important bacterial pathogens [8,9,10,11] responsible for severe financial losses, while in fish culture it has been reported to cause significant mortalities mostly in turbot larvae [3,12]. In fish and invertebrate larviculture where the use of antibiotics has specific drawbacks and is not applicable, control of vibriosis
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