Vibrio harveyi is a common aquaculture pathogen causing diseases in a variety of aquatic animals. The extensive use of antibiotics for managing V. harveyi has led to the ban of import of fish in many countries due to the presence of antibiotic residues and the concern of the emergence of antibiotic-resistant bacteria. This study aimed to investigate the phenotypic characterization and antimicrobial susceptibility patterns of 54 V. harveyi collected from Larimichthys crocea in China. All isolates were resistant to colistin sulfate, gentamicin, tetracycline, ampicillin, co-trimoxazole, and amoxicillin. More than 92.6% were resistant to ceftiofur, streptomycin, and amikacin. The frequency of the virulence gene toxR reached up to 75.9%, while 63% strains carried at least four virulence genes. A novel V. harveyi–specific phage vB_VhaS_PcB-1G was isolated from Larimichthys crocea in the East China Sea. The sequence analysis revealed that the phage vB_VhaS_PcB-1G genome was 48,719 bp of double-stranded DNA with 80 open reading frames. The phage vB_VhaS_PcB-1G exhibited large burst sizes (210 PFU/cell) and a broad host range against V. harveyi (33/54). With the zebrafish model, vB_VhaS_PcB-1G could effectively prevent (93.33%) and control (86.67%) V. harveyi infection. Phage efficacy indicated its potential use as an antibacterial agent to control virulent V. harveyi infection in Larimichthys crocea and vibriosis in aquaculture.