Porins are crucial proteins located in the outer membrane that directly influence antimicrobial resistance mechanisms and virulence in bacteria. In this study, a porin gene (Vp-porin) was cloned in V. parahaemolyticus, and the function of Vp-Porin in biological characteristics and virulence was investigated. The results of sequence analysis showed that Vp-Porin is highly conserved in Vibrio spp., and the predicted 3D structure showed it could form a 20-strand transmembrane β-barrel domian. Membrane permeabilization provides evidence that the membrane integrity of ∆Vp-porin was damaged and the sensitivity to tetracycline, polymyxin B, rifampicin and cephalothin of ∆Vp-porin obviously increased. In addition, loss of Vp-porin damaged motility due to downregulated flagellar synthesis. In addition, ∆Vp-porin exhibited attenuated cytotoxicity to Tetrahymena. The relative survival rate of Tetrahymena infection with ∆Vp-porin was 86%, which is much higher than that with WT (49%). Taken together, the results of this study indicate that Vp-Porin in V. parahaemolyticus plays various roles in biological characteristics in membrane integrity, antimicrobial resistance and motility and contributes to virulence.

The msbB gene, encoding a lipid A phosphatease, is crucial for lipopolysaccharide (LPS) synthesis in Gram-negative bacteria and plays a critical role in their virulence. This study investigated the role of msbB in Vibrio parahaemolyticus, a significant marine pathogen causing gastroenteritis in humans and infections in aquatic animals. We constructed an msbB deletion mutant (ΔmsbB) and a complementary strain (CΔmsbB) using homologous recombination. The growth, outer membrane permeability, stress and antibiotic sensitivity, biofilm formation, swarming motility, and virulence of the wild-type (WT), ΔmsbB, and CΔmsbB strains were assessed. Additionally, the pathogenicity of ΔmsbB was evaluated using L. vannamei shrimp models. The results showed that the msbB gene was successfully deleted and complemented, and its deletion did not impair bacterial growth. However, the ΔmsbB strain exhibited an increased outer membrane permeability, reduced resistance to stresses and antibiotics, defective biofilm formation, and a reduced swarming motility. In a Tetrahymena co-culture, the ΔmsbB strain showed attenuated virulence. In shrimp infected with the ΔmsbB strain, the cumulative mortality rate was 22%, significantly lower than the 62% observed in the WT strain. Moreover, the expression levels of immune-related genes in the shrimp hepatopancreas were significantly lower in the ΔmsbB group, indicating a significant reduction in infection capability and pathogenicity. These findings indicate that the msbB gene is critical for the virulence of V. parahaemolyticus and suggest that msbB is a potential target for therapeutic interventions and vaccine development against V. parahaemolyticus infections.

Vibrio anguillarum is a major pathogenic bacterium causing vibriosis in aquatic animals, leading to substantial economic losses in the global aquaculture industry. Previous studies have indicated that L-arginine modulates the virulence of the pathogen, but the underlying molecular mechanisms remain elusive. The present study aimed to clarify the regulatory role of L-arginine metabolism in V. anguillarum virulence. We first evaluated the effects of L-arginine and its major metabolites (agmatine, putrescine, spermine) on the hemolytic activity of V. anguillarum. Results showed that L-arginine and its metabolites regulated hemolytic activity in a concentration-dependent biphasic manner, with agmatine exerting the most potent promoting effect. To identify the critical metabolic branch involved, four isogenic mutants were constructed targeting key genes in arginine metabolism (adc, astA, astD). Phenotypic analysis revealed that only the adc deletion mutant (Δadc) exhibited near-complete loss of hemolytic activity, which was dose-dependently restored by supplementation with agmatine, putrescine, or spermine. Transcriptomic analysis identified 704 significantly differentially expressed genes (DEGs) between Δadc and WT strains, with downregulated DEGs enriched in virulence-associated pathways. Key hemolysin and secretion system genes were validated to be downregulated in ∆adc by quantitative real-time PCR (qRT-PCR). Additionally, Δadc displayed attenuated anti-phagocytic ability in Tetrahymena co-culture assays, impaired biofilm formation, and increased susceptibility to multiple classes of antibiotics. Collectively, our findings demonstrate that L-arginine modulates V. anguillarum hemolysis and overall virulence through the adc-mediated agmatine biosynthesis branch. This study fills the knowledge gap in the regulatory mechanism of L-arginine on V. anguillarum virulence and provides a potential target for the control of vibriosis in aquaculture.