Arginine Decarboxylase Gene (adc) Is Essential for Vibrio anguillarum Virulence and Physiological Phenotypes

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.