Short- and Long-term Probiotic Effects of Enterococcus hirae Isolated from Fermented Vegetable Wastes on the Growth, Immune Responses, and Disease Resistance of Hybrid Catfish (Clarias gariepinus × Clarias macrocephalus).

This study evaluated the short- and long-term effects of dietary supplementation with Enterococcus hirae strain UPM02 on the growth performance, immunity, and disease resistance of hybrid catfish (Clarias gariepinus × Clarias macrocephalus) against Aeromonas hydrophila infection. In the long-term trial, fingerling fish were fed diets containing 0 (control), 2×105, or 2×107 CFU/g E. hirae UPM02 for 120 days. Administration of E. hirae UPM02 had significant effects on the specific growth rate (SGR), feed utilization efficiency, body indices (P<0.05), and gut villus physiology of the catfish. E. hirae UPM02 application also significantly increased the complete blood cell counts, phagocytic activity, respiratory burst, lysozyme activity, and alternative complement pathway hemolytic (ACH50) activity in tested catfish throughout the experimental periods (P<0.05). Dietary E. hirae UPM02 at both concentrations significantly increased the expression levels of the alpha-2-macroglobulin (α2M), CC chemokines, CXC chemokines, lysozyme c (LYZC), myeloperoxidase (MYE), NF-kappa-B1 p105 subunit (NF-K), and bactericidal permeability-increasing protein (BPIP) genes in the head kidney, liver, and spleen (P<0.05) at days 80, 100 and 120 after application. However, heat shock protein 70 (HSP70) gene expression was slightly downregulated in these organs. Interestingly, fish fed the diets containing 2×105 and 2×107 CFU/g E. hirae UPM02 exhibited a significantly lower (P<0.05) postchallenge mortality rates (32% and 30%, respectively) after 14 days of A. hydrophila challenge than the control fish (58%). In short-term (28 days) application to juvenile catfish, the two concentrations of E. hirae did not affect all growth parameters. Nevertheless, these concentrations markedly elevated all tested immune parameters, similarly to long-term application. Immune-related gene expression was significantly upregulated at day 28 in the head kidney, at day 14 in the liver, and at day 7 in the spleen in fish treated with the two concentrations of the probiotics (P<0.05). Mortality at 14 days after challenge with A. hydrophila in the groups receiving the two concentrations of the probiotic was significantly lower than that in the control group, at 28, 24, and 48%, respectively (P<0.05). These results collectively suggest that dietary supplementation with E. hirae UPM02 at 2×105 and 2×107 CFU/g effectively influenced immune responses, enhanced disease protection, and stimulated immunity-related gene expression in hybrid catfish under both short- and long-term application. However, growth enhancement was significantly evidenced with long-term application only.