Exogenous 2-keto-L-gulonic Acid Supplementation as a Novel Approach to Enhancing L-ascorbic Acid Biosynthesis in Zebrafish (Danio rerio)

SIMPLE SUMMARY: L-ascorbic acid plays a significant role in maintaining the physiological functions and survival of aquatic animals. This is the first report on elevating L-ascorbic acid accumulation in aquatic animals through exogenous 2-keto-L-gulonic acid supplementation. This study found the positive effects of exogenous 2-keto-L-gulonic acid supplementation on L-ascorbic acid metabolism and growth in zebrafish (Danio rerio), such as the enhanced L-ascorbic acid content, growth performance and feed utilization, crude protein, and crude lipid content. Our findings suggest that 2-keto-L-gulonic acid has the potential to act as a precursor of L-ascorbic acid, and supplementing with 2-keto-L-gulonic acid could be a novel strategy for enhancing L-ascorbic acid levels in aquatic animals. ABSTRACT: L-ascorbic acid (ASA) is a micronutrient that is essential for reproduction, growth, and immunity in animals. Due to the loss of enzyme L-gulono-1,4-lactone oxidase (GLO), most aquatic animals lack the capacity for ASA biosynthesis and therefore require supplementation with exogenous ASA. Recent studies have shown that 2-keto-L-gulonic acid (2KGA), a novel potential precursor of ASA, can enhance plant growth and improve stress resistance by promoting the synthesis and accumulation of ASA. Our hypothesis is that 2-keto-L-gulonic acid (2KGA) plays a similar role in aquatic animals. To investigate this, we conducted an in vivo trial to examine the effects of exogenous 2KGA supplementation on ASA metabolism and growth of zebrafish (Danio rerio). Zebrafish were categorized into groups based on their dietary intake, including a basal diet (CK group), a basal diet supplemented with 800 mg/kg ASA (ASA group), and 800 mg/kg 2KGA-Na (2KGA group) for a duration of three weeks. The results demonstrated a significant increase in ASA content in zebrafish treated with 2KGA (34.82% increase, p < 0.05) compared to the CK group, reaching a consistent level with the ASA group (39.61% increase, p < 0.05). Furthermore, the supplementation of 2KGA significantly improved growth parameters relevant to zebrafish (specific growth rate increased by 129.04%, p < 0.05) and enhanced feed utilization (feed intake increased by 15.65%, p < 0.05). Positive correlations were observed between growth parameters, feed utilization, whole-body chemical composition, and ASA content. Our findings suggest that supplementation with exogenous 2KGA can serve as a novel approach for elevating ASA synthesis in aquatic animals, and further investigation of its underlying mechanism is required.