Duan X et al. (2023), Isolation, characterization and application of ...

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PLoS One 2023 Jan 01;189:e0289895. doi: 10.1371/journal.pone.0289895.

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Isolation, characterization and application of a lytic phage vB_VspM_VS1 against Vibrio splendidus biofilm.

Duan X , Jiang L , Guo M , Li C .

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Vibrio splendidus is a common pathogen in the ocean that infects Apostichopus japonicus, Atlantic salmon and Crassostrea gigas, leading to a variety of diseases. In this study, a virulent phage vB_VspM_VS1, which infects V. splendidus, was isolated from aquaculture ponds in Dalian, China, and it belongs to the family Straboviridae in the order Caudoviricetes. vB_VspM_VS1 had an adsorption rate of 96% in 15 min, a latent period of 65 min, and a burst size of 140 ± 6 PFU/cell. The complete genome of phage vB_VspM_VS1 consists of a linear double-stranded DNA that is 248,270 bp in length with an average G + C content of 42.5% and 389 putative protein-coding genes; 116 genes have known functions. There are 4 tail fiber genes in the positive and negative strands of the phage vB_VspM_VS1 genome. The protein domain of the phage vB_VspM_VS1 tail fibers was obtained from the Protein Data Bank and the SMART (http://smart.embl.de) database. Bacterial challenge tests revealed that the growth of V. splendidus HS0 was apparently inhibited (OD600 < 0.01) in 12 h at an MOI of 10. In against biofilms, we also showed that the OD570 value of the vB_VspM_VS1-treated group (MOI = 1) decreased significantly to 0.04 ± 0.01 compared with that of the control group (0.48 ± 0.08) at 24 h. This study characterizes the genome of the phage vB_VspM_VS1 that infects the pathogenic bacterium V. splendidus of A. japonicus.

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References [+] :

Azam, Bacteriophage-host arm race: an update on the mechanism of phage resistance in bacteria and revenge of the phage with the perspective for phage therapy. 2019, Pubmed

	Fig 1. (a) Plaques formed by phage vB_VspM_VS1 on the host strain V. splendidus after overnight incubation at 28 °C. (b) Transmission electron micrograph showing that phage vB_VspM_VS1 belongs to the family Myoviridae and has a head of 119 × 143 ± 5 nm and a tail of 34 × 53 ± 5 nm. (c, d) Adsorption rate and population dynamics of phage vB_VspM_VS1 inoculated in V. splendidus culture. The values presented are means and standard deviations (SDs) of three independent biological repeats (n = 3).
	Fig 2. Line map of the vB_VspM_VS1 genome.In the vB_VspM_VS1 genome track, the arrows represent the ORFs and point in the direction of transcription. The colour intensity corresponds to G+C skew level.
	Fig 3. Schematic diagram domains of phage vB_VspM_VS1 tail fiber genes ORF42 (a), ORF61 (b), ORF64 (c) and ORF82 (d) obtained from the SMART (http://smart.embl.de) database.
	Fig 4. Neighbor-joining tree showing the relationship of various phages from the NCBI database to vB_VspM_VS1.The tree was drawn based on the major capsid amino acid protein of each phage. Bootstrap values were set for 500 repetitions. Phage vB_VspM_VS1 and nt-1 have very close evolutionary distance. Scale bar means distance ruler.
	Fig 5. Corresponding abundances of pfu/mL were quantified by plaque assay over a 12 h period of incubation in V. splendidus cultures in the presence of phage vB_VspM_VS2 at a multiplicity of infection (MOI) of 0.1, 1 and 10 were measured at different incubation times, respectively.
	Fig 6. Comparison of the lytic ability of phage vB_VspM_VS1 using V. splendidus as the host at various MOI from 0.1 to 10 in 2216E broth.
	Fig 7. Biofilm formed by V. splendidus at 14 h, 26 h and 48 h.Biofilm formed after phage vB_VspM_VS1 treatment was observed by optical microscope. Phage vB_VspM_VS1 inhibited the formation of V. splendidus biofilm by MOI = 1 under the same conditions at 14 h, 26 h and 48 h.
	Fig 8. Effects on V. splendidus biofilm cultured in the presence of phage vB_VspM_VS1 on MOI = 1 for 12, 24 and 48 h (OD570).