Abstract

Foodborne salmonellosis remains a major public health concern, with poultry products serving as key reservoirs for Salmonella enterica. The increasing prevalence of S. enterica serovar Agona in broiler production highlights the need for safe and sustainable serovar-specific control strategies. In this study, Salmonella phage WP85 was comprehensively characterized to assess its potential as a biocontrol agent against S. Agona in chicken meat. Broad lytic activity was observed against 57% of tested S. Agona isolates, and efficient propagation was achieved on susceptible hosts. Infection kinetics showed rapid adsorption (80% within 25 min), a short latent period (20 min), and a high burst size (134 particles per infected cell), indicating strong lytic capacity. Transmission electron microscopy (TEM) demonstrated that phage WP85 exhibits siphovirus-like morphology. Whole genome sequencing (WGS) identified a 109,049-bp linear double-stranded DNA genome comprising of 201 open reading frames and 21 tRNAs. Phylogenetic and comparative genomic analyses classified phage WP85 within the Tequintavirus genus and suggested that it represents a novel species. Structural modeling and docking of the receptor-binding protein supported a plausible host-recognition mechanism. High environmental robustness was demonstrated, with phage viability retained between 4 and 60 °C, pH 5–11, and up to 15% NaCl. Strong multiplicity-of-infection (MOI)-dependent growth suppression was observed in vitro, with sustained inhibition of bacterial turbidity at high MOIs. When applied to spiked chicken meat during refrigerated storage, phage WP85 significantly reduced S. Agona populations by up to 3.2 log CFU/mL (99.9%). Importantly, no significant changes were detected in proximate composition, lipid oxidation indices, or overall meat quality. Collectively, the biological efficiency, genomic characterization, environmental stability, and practical decontamination efficacy demonstrated, established phage WP85 as a safe, stable, and effective candidate for postharvest poultry meat biocontrol and supports phage application as a sustainable, residue-free intervention to improve food safety within the poultry value chain.