Biological Characterization of Pseudomonas fluorescens Phage Pf17397_F_PD1 and Its Application in Food Preservation

Abstract

In order to explore the application prospects of phages for controlling bacterial contamination, a lytic phage Pf17397_F_PD1 (Later abbreviated as PD1) was isolated from fish guts using Pseudomonas fluorescens ATCC 17397 as the host bacterium. The phage displayed short latency (18 min), long lysis period (212 min), and high lysis volume (1.47 × 10² PFU/each cell). It displayed wide temperature (30-70°C) and pH (4-11) tolerance. Genomic comparison revealed a maximum sequence identity of 48.65% between phage PD1 and other identified phages, indicating that PD1 was a new phage. The phage PD1 significantly inhibited the growth of P. fluorescens in milk and grass carp at 4°C and 25°C. Compared to the negative control, bacterial levels in milk stored at 25°C for 48 h were reduced by 2.71 log CFU/mL and 2.84 log CFU/mL at the multiplicity of infection (MOI) of 100 and 1,000, respectively. In contrast, when grass carp were stored at 25°C for 24 h, the bacterial load was reduced by 1.28 log CFU/g and 2.64 log CFU/g compared to the control (MOI of 100 and 1,000). When the phage was applied for preservation of grass carp blocks, total volatile salt nitrogen (TVB-N) values of phage-treated samples increased by 6.8 mg/100 g and 7.5 mg/100 g at MOI of 100 and 1,000, respectively, after 7 days of storage, which was significantly lower than that of the control group (15.83 mg/100 g). This study showed that phage PD1 was a good natural biological antimicrobial agent against P. fluorescens ATCC 17397.

Keywords: Antibacterial activity; Biological properties; Genome annotation; Phage; Pseudomonas fluorescens.