Abstract
Sea cucumbers are valuable in aquaculture, but their cultivation faces challenges from diseases such as skin ulceration syndrome caused by Vibrio alginolyticus (VA). This study aimed to isolate and identify probiotics capable of combating VA and improving sea cucumber’s growth performance. Pathogenic VA was identified, through 16S rDNA sequencing, confirming its high genetic similarity (>99%) to Vibrio alginolyticus. Two Bacillus strains, Bacillus licheniformis YB-1, and Bacillus megaterium YB-2, were isolated as potential probiotics, with identification supported by 16S rDNA phylogenetic analysis and deposition in microbial culture collections. They demonstrated strong antibacterial activity against VA in vitro without exhibiting antagonism when combined. Probiotic tolerance to environmental stressors was observed, while feeding trials revealed significant growth improvements in sea cucumbers, with the highest specific growth rates observed at 1 × 106 CFU/mL for both strains. Immersion challenge tests showed that sea cucumbers treated with probiotics exhibited reduced symptoms of rotten skin syndrome and higher survival rates. The optimal combination of YB-1 and YB-2, with viable bacteria concentrations of 5 × 107 CFU/mL each, achieved a 55% survival rate after a VA challenge, demonstrating their synergistic efficacy. These findings suggest that YB-1 and YB-2 offer promising probiotic solutions for enhancing sea cucumber health and resistance to VA infections in aquaculture.