Isolation, characterization, and application of bacteriophage on Vibrio parahaemolyticus biofilm to control seafood contamination

•Lytic bacteriophage CAU_VPP01 might be a new species in the family Siphoviridae of the order Caudovirales.•The bacteriophage was stable at wide range of pH(5–9), and temperature(0–60 degree).•At MOI 10, phage CAU_VPP01 effectively reduced biofilm proliferation and biomass volume, thickness, and roughness at 4 °C.•CAU_VPP01 could be an eco-friendly alternative to control V. parahaemolyticus biofilm. This study intended to isolate a Vibrio-particular phage from the natural environment, analyse its characteristics and genome sequence, and investigate its reduction effect on V. parahaemolyticus biofilm as a biocontrol agent in squid and mackerel. Among 21 phages, phage CAU_VPP01, isolated from beach mud, was chosen for further experiments based on host range and EOP tests. When examining the reduction effect of phage CAU_VPP01 against Vibrio parahaemolyticus biofilms on surfaces (stainless steel [SS] and polyethylene terephthalate [PET]) and food surfaces (squid and mackerel). The phage showed the most excellent reduction effect at a multiplicity-of-infection (MOI) 10. Three-dimensional images acquired with confocal laser scanning microscopy (CLSM) analysis were quantified using COMSTAT, which showed that biomass, average thickness, and roughness coefficient decreased when treated with the phage. Colour and texture analysis confirmed that the quality of squid and mackerel was maintained after the phage treatment. Finally, a comparison of gene expression levels determined by qRT-PCR analysis showed that the phage treatment induced a decrease in the gene expression of flaA, vp0962, andluxS, as examples. This study indicated that Vibrio-specific phage CAU_VPP01 effectively controlled V. parahaemolyticus biofilms under various conditions and confirmed that the isolated phage could possibly be used as an effective biocontrol weapon in the seafood manufacturing industry. [Display omitted]