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Genomic and Biological Characterization of a Broad-Host-Range KVP40-Like Bacteriophage vB_VpM-pA3B5 for Control of

Soojin Lim^1,2, Jee Eun Han^3,4, Seon Young Park⁵

¹Department of Food Science and Biotechnology, College of BioNano Technology, Gachon University, Seongnam 13120, Republic of Korea.

Journal of Microbiology and Biotechnology

|July 15, 2026

Abstract:

Vibrio parahaemolyticus harboring the pVA1 plasmid (Vp_AHPND) causes acute hepatopancreatic necrosis disease (AHPND), a major threat to global shrimp aquaculture. Although bacteriophage (phage)-based biocontrol has emerged as a promising alternative to antimicrobial agents for controlling Vp_AHPND, most studies have focused on disease prevention in culturing shrimp rather than post-harvest applications. In this study, a KVP40-like jumbo phage, designated vB_VpM-pA3B5, was isolated and characterized, and its biocontrol potential was evaluated in both Vp_AHPND-contaminated shrimp products and shrimp bioassay. Phage vB_VpM-pA3B5 exhibited a broad host range, infecting globally distributed Vp_AHPND strains, an AHPND-causing V. campbellii strain, and three additional Vibrio species. The phage demonstrated strong lytic activity, efficient replication, and high environmental stability. Genome analysis revealed a 243,570-bp linear dsDNA genome encoding 382 predicted ORFs and 28 tRNAs, with no virulence or antimicrobial resistance genes detected. Comparative genomic analysis indicated high synteny with the Schizotequatrovirus phages KVP40 and PVA8 and identified a unique tail fiber protein (ORF115) that may contribute to its broad host range. In Vp_AHPND-contaminated shrimp products, the phage efficiently reduced bacterial loads and actively replicated during storage. In the shrimp bioassay, although cumulative mortality was not reduced, phage-treated shrimp exhibited alleviated hepatopancreatic lesions and lower pirA gene loads than the Vp_AHPND-challenged group. These findings highlight the potential of phage vB_VpM-pA3B5 as a biocontrol agent for mitigating AHPND-associated risks in shrimp aquaculture and reducing the dissemination of Vp_AHPND through shrimp products.

Insights

A novel bacteriophage, vB_VpM-pA3B5, effectively reduces Vibrio parahaemolyticus loads in shrimp products and alleviates disease symptoms. This phage shows promise for biocontrol in aquaculture and post-harvest applications against AHPND.

Area of Science:

Aquaculture
Microbiology
Biotechnology

Background:

Acute hepatopancreatic necrosis disease (AHPND), caused by Vibrio parahaemolyticus (VpAHPND), severely impacts global shrimp aquaculture.
Current control methods often rely on antimicrobials, driving the need for alternative strategies like bacteriophage biocontrol.
Existing phage research primarily focuses on disease prevention during shrimp cultivation, with limited exploration of post-harvest applications.

Purpose of the Study:

To isolate and characterize a novel bacteriophage targeting VpAHPND.
To evaluate the biocontrol efficacy of the isolated phage in shrimp products and a shrimp bioassay.
To analyze the phage's genomic features and potential for broad-spectrum activity.

Main Methods:

Isolation and characterization of a KVP40-like jumbo phage, vB_VpM-pA3B5.
Evaluation of phage host range against various Vibrio strains.
Assessment of phage lytic activity, replication, and environmental stability.
Whole-genome sequencing and comparative genomic analysis.
Testing biocontrol efficacy in VpAHPND-contaminated shrimp products and a shrimp bioassay.

Main Results:

Phage vB_VpM-pA3B5 demonstrated a broad host range, infecting multiple VpAHPND strains and other Vibrio species.
The phage exhibited strong lytic activity, efficient replication, and good environmental stability.
Genome analysis revealed a large dsDNA genome with no detected virulence or antibiotic resistance genes.
In shrimp products, the phage significantly reduced bacterial loads and replicated during storage.
In vivo, phage treatment alleviated hepatopancreatic lesions and reduced pirA gene loads, despite not reducing cumulative mortality.

Conclusions:

Phage vB_VpM-pA3B5 is a promising biocontrol agent against VpAHPND.
Its efficacy in shrimp products suggests potential for post-harvest applications to reduce disease spread.
The phage's genomic characteristics and broad host range support its development for mitigating AHPND risks in aquaculture.

Read full analysis

Keywords:

Acute hepatopancreatic necrosis disease (AHPND)Biocontrol Jumbo bacteriophage Seafood safety

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