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. 2024 Jul 3;14(1):15347.
doi: 10.1038/s41598-024-64999-1.

Phenotypic characterization and genomic analysis of a Salmonella phage L223 for biocontrol of Salmonella spp. in poultry

Md Abu Sayem Khan  1 , Zahidul Islam  1 , Chayan Barua  1 , Md Murshed Hasan Sarkar  2 , Md Firoz Ahmed  3 , Sabita Rezwana Rahman  4
Affiliations

Phenotypic characterization and genomic analysis of a Salmonella phage L223 for biocontrol of Salmonella spp. in poultry

Md Abu Sayem Khan et al. Sci Rep. .

Abstract

The escalating incidence of foodborne salmonellosis poses a significant global threat to food safety and public health. As antibiotic resistance in Salmonella continues to rise, there is growing interest in bacteriophages as potential alternatives. In this study, we isolated, characterized, and evaluated the biocontrol efficacy of lytic phage L223 in chicken meat. Phage L223 demonstrated robust stability across a broad range of temperatures (20-70 °C) and pH levels (2-11) and exhibited a restricted host range targeting Salmonella spp., notably Salmonella Typhimurium and Salmonella Enteritidis. Characterization of L223 revealed a short latent period of 30 min and a substantial burst size of 515 PFU/cell. Genomic analysis classified L223 within the Caudoviricetes class, Guernseyvirinae subfamily and Jerseyvirus genus, with a dsDNA genome size of 44,321 bp and 47.9% GC content, featuring 72 coding sequences devoid of antimicrobial resistance, virulence factors, toxins, and tRNA genes. Application of L223 significantly (p < 0.005) reduced Salmonella Typhimurium ATCC 14,028 counts by 1.24, 2.17, and 1.55 log CFU/piece after 2, 4, and 6 h of incubation, respectively, in experimentally contaminated chicken breast samples. These findings highlight the potential of Salmonella phage L223 as a promising biocontrol agent for mitigating Salmonella contamination in food products, emphasizing its relevance for enhancing food safety protocols.

Keywords: Salmonella; Biocontrol; Lytic bacteriophage; Poultry; Whole-genome sequence.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
Different characteristics of Salmonella phage L223. (A) Lytic plaque on soft agar; (B) Temperature stability of L223; (C) Phage stability at different pH and (D) One step growth curve.
Figure 2
Figure 2
Bacteriolytic kinetics of Salmonella phage L223 against Salmonella Typhimurium.
Figure 3
Figure 3
Circular genome map of Salmonella phage L223.
Figure 4
Figure 4
Phylogenetic analysis of Salmonella phage L223 based on (A) genome-BLAST Distance Phylogeny (GBDP) and (B) proteomic tree based on genome wide similarities.
Figure 5
Figure 5
Phage genome network analysis using vConTACT2. (A) Salmonella phage L223 occurs in viral Cluster VC_42 (red colored), (B) A closer look on VC_42 cluster.
Figure 6
Figure 6
Comparative genomic analysis of Salmonella phage L223 and closely related phage genomes (A) VIRIDIC heatmap, (B) Whole genome comparison, and (C) PhageCloud analysis.
Figure 7
Figure 7
Pan-genome matrix of 65 Salmonella phages.
Figure 8
Figure 8
Pan-genome analysis of Salmonella phages: (A) Distributions of genes in pan-genome, (B) Changes in the number of conserved and total genes, (C) Changes in the number of new and unique genes, and (D) Average nucleotide identity between phages.
Figure 9
Figure 9
The activity of the phage L223 against Salmonella Typhimurium on chicken meat.
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