High doses of sodium ascorbate act as a prooxidant and protect gnotobiotic brine shrimp larvae (Artemia franciscana) against Vibrio harveyi infection coinciding with heat shock protein 70 activation

Ascorbate is an essential nutrient commonly regarded as an antioxidant. In this study, using axenic brine shrimp and pathogenic strain Vibrio harveyi as the host-pathogen model, we confirmed that pretreatment of sodium ascorbate (NaAs), at an optimum concentration, was a prooxidant by generation of hydrogen peroxide, inducing protective effects in the brine shrimp against V. harveyi infection. Such a protective effect could be neutralized by the addition of an antioxidant enzyme catalase. We further showed that generation of oxygen radicals is linked to the induction of heat shock protein 70 (Hsp70), which is involved in eliciting the antioxidant protection system including superoxidase dismutase (SOD) and possibly many other immune responses. Furthermore, using RNA interference technique, we found that the pretreatment of sodium ascorbate increased the survival significantly in the control knockdown groups (using green fluorescent protein, GFP) but not in Hsp70 knockdown groups and the result directly suggested that the up-regulated Hsp70 induced by sodium ascorbate pretreatment induced the protective effect. These results provide a mechanistic rationale for exploring the further use of ascorbate for antimicrobial therapy in aquaculture.