We investigated the three probiotic bacteria and a processed yeast (GroPro-Aqua) as the replacers of antibiotics in juvenile olive flounder. A total of seven diets were used, that is, one basal or control (CON) diet; and six other diets, of which, three diets were prepared by supplementing probiotic bacteria such as Bacillus subtilis WB60 (BSWB60) at 1 × 108 CFU/g diet, Bacillus subtilis SJ10 (BSSJ10) at 1 × 108 CFU/g diet, and Enterococcus faecium SH30 (EFSH30) at 1 × 107 CFU/g diet; one diet with processed yeast (GRO) at 0.35% diet; and two other diets were supplemented with oxytetracycline (OTC) and amoxicillin (AMO) at 4 g/kg of each. Triplicate groups of fish (average 12.1 g) were fed one of the diets for eight weeks. At the end of the feeding trial, the fish that were fed the probiotic bacteria-supplemented diets had a significantly higher final weight, weight gain, and specific growth rate compared to the CON, OTC, and AMO diets. Fish that were fed the GRO diet had significantly higher feed efficiency and protein efficiency ratios than those of the fish that were fed the CON diet. Serum glutamic pyruvic transaminase, glutamic oxaloacetic transaminase, glucose, and total protein were not affected by the diets. Lysozyme activity in fish that were fed the BSSJ10, BSWB60, and EFSH30 diets were significantly higher compared to the CON and OTC diets, whereas myeloperoxidase activity of fish fed the BSWB60 and EFSH30 diets were significantly higher than those of fish fed the CON and AMO diets. Flounder growth hormone gene expressions of fish that were fed BSWB60 and GRO diets were significantly higher compared to the CON, OTC, and AMO diets. The interleukin-1β gene expression of fish that were fed the BSSJ10, BSWB60, EFSH30, OTC, and GRO diets was significantly higher than those of fish fed the CON diet. The interleukin-10 gene expression of fish that were fed the BSSJ10, EFSH30, and GRO diets was significantly higher than those of fish fed the CON and AMO diets. Posterior intestinal histology of fish showed significantly higher villi length in fish that were fed the BSSJ10, BSWB60, EFSH30, and GRO diets compared to the CON diet. After 15 days of challenge test with pathogenic bacteria Edwardsiella tarda, the cumulative survival rate of fish that were fed the BSSJ10, BSWB60, EFSH30, and GRO diets were significantly higher than those of fish that were fed the CON diet. Overall, the results indicate that dietary supplementation of B. subtilis (108 CFU/g diet), E. faecium (107 CFU/g diet), and processed yeast (GroPro-Aqua at 0.35% diet) could replace the antibiotics in terms of improving growth, immunity, gut health, and disease resistance in juvenile olive flounder.
Keywords: Edwardsiella tarda; alternative of antibiotics; beneficial bacteria; growth; histomorphology; innate immunity; olive flounder; processed yeast.