Characterization and genome analysis of six novel Vibrio parahaemolyticus phages associated with acute hepatopancreatic necrosis disease (AHPND)

Vibrio parahaemolyticus causes acute hepatopancreatic necrosis disease (AHPND) in farmed shrimp. Due to its damage potential, which could be as high as a 100% mortality rate, bacteriophages have emerged as a promising natural control intervention other than antibiotics, yet multiple roadblocks need to be overcome. In this study, six bacteriophages isolated from seafood samples, seawater, and estuary water in Sinaloa, Mexico, demonstrated a narrow host range among Mexican AHPND-causing V. parahaemolyticus. All bacteriophages are composed of a double-stranded DNA genome with lengths ranging between 43,268 and 57,805 bp. All six phages exhibited latency periods of 10-30 min and burst sizes of 34-168 viral particles per infected cell. The optimal MOI for bacteriophage propagation was 0.01-1. No transfer RNA (tRNA), virulence, or resistance genes were found in either genome, and the life cycle of these phages was classified as virulent by the PhageAI platform. Phylogenetic and comparative genomics analyzes assigned phages M3, C2, M9, and M83 as new species not yet reported within the genus Maculvirus, Autographiviridae family. ALK and CHI phages were assigned as new members of a new genus not yet classified within the subfamily Queuovirinae. The findings highlight the potential of CHI, ALK, M3, C2, M9, and M83 as promising alternatives against AHPND-causing V. parahaemolyticus from Mexico.