Application of Fragmented Extracellular Self-DNA (esDNA) Concept as an Alternative Prophylactic Approach against Vibrio parahaemolyticus and Vibrio harveyi Infection in Brine Shrimp Artemia

Abstract

The purpose of this study was to investigate species specific inhibitory effects of esDNA isolated from two conspecific organisms: Vibrio parahaemolyticus (VP) and Vibrio harveyi (VH), and to assess the functional role of esDNA to enhance the survival rate of Artemia sp. In an in vitro study, nine doses of Extracellular self-DNA of Vibrio parahaemolyticus (esDNAVP) and Vibrio harveyi (esDNAVH) were used as the target for the challenge test with the conspecific bacteria. In an in vivo study, the protective effect of esDNA was then tested in nauplii of the brine shrimp Artemia at various priming times and concentrations of esDNA under gnotobiotic conditions prior to challenge with VP and VH at the concentration of 5 × 105 CFU mL-1. The results from in vitro study showed that the use of esDNAVP at levels of 24.02 and 48.05 ng µl-1 and esDNAVH at concentrations of 13.33 and 26.67 ng µl-1 were able to inhibit the growth of the conspecific species when added to the culture medium at the concentration level of 5 × 105 CFU mL-1. The results from in vivo study showed that the use of 24.02; 48.05 and 72.07 ng µl-1 of esDNAVP as well as the use of 13.33; 26.67 and 40.00 ng µl-1 of esDNAVH inhibited the growth of VP and VH and enhanced the survival rate of Artemia sp compared to the control treatment (P<0.05). Taken together, we confirmed that esDNA obtained from the extraction and random fragmentation from esDNAVP and esDNAVH, produces a species-specific inhibitory effect on the same species and can serve as a potential alternative strategy for disease control to deliver the functionality of esDNA to the fish and shrimp.