Ozone nanobubble treatments improve survivability of Nile tilapia (Oreochromis niloticus) challenged with a pathogenic multi-drug-resistant Aeromonas hydrophila

A rapid increase in multi-drug-resistant (MDR) bacteria in aquaculture highlights the risk of production losses due to diseases and potential public health concerns. Previously, we reported that ozone nanobubbles (NB-O-3) were effective at reducing concentrations of pathogenic bacteria in water and modulating fish immunity against pathogens; however, multiple treatments with direct NB-O-3 exposures caused alterations to the gills of exposed fish. Here, we set up a modified recirculation system (MRS) assembled with an NB-O-3 device (MRS-NB-O-3) to investigate whether MRS-NB-O-3 (a) were safe for tilapia (Oreochromis niloticus), (b) were effective at reducing bacterial load in rearing water and (c) improved survivability of Nile tilapia following an immersion challenge with a lethal dose of MDR Aeromonas hydrophila. The results showed no behavioural abnormalities or mortality of Nile tilapia during the 14-day study using the MRS-NB-O-3 system. In the immersion challenge, although high bacterial concentration (similar to 2 x 10(7) CFU/ml) was used, multiple NB-O-3 treatments in the first two days reduced the bacteria between 15.9% and 35.6% of bacterial load in water, while bacterial concentration increased from 13.1% to 27.9% in the untreated control. There was slight up-regulation of non-specific immune-related genes in the gills of the fish receiving NB-O-3 treatments. Most importantly, this treatment significantly improved survivability of Nile tilapia with relative percentage survival (RPS) of 64.7%-66.7% in treated fish and surviving fish developed specific antibody against MDR A. hydrophila. In summary, the result suggests that NB-O-3 is a promising non-antibiotic approach to control bacterial diseases, including MDR bacteria, and has high potential for application in recirculation aquaculture system (RAS).