Efficacy of Single and Multi-Strain Probiotics on In Vitro Strain Compatibility, Pathogen Inhibition, Biofilm Formation Capability, and Stress Tolerance.

Compatibility of each strain in a multi-strain probiotic (MSP), along with its properties, becomes a strong base for its formulation. In this study, single-strain probiotics (SSPs) and multi-strain probiotics (MSPs) were evaluated in vitro for strain compatibility, microbial antagonism, biofilm formation capacity, and stress tolerance. Bacillus amyloliquefaciens L11, Enterococcus hirae LAB3, and Lysinibacillus fusiformis SPS11 were chosen as MSP1 candidates because they showed much stronger antagonism to Aeromonas hydrophila and Streptococcus agalactiae than a single probiotic. MSP 2 candidates were Lysinibacillus fusiformis strains SPS11, A1, and Lysinibacillus sphaericus strain NAS32 because the inhibition zone produced by MSP 2 against Vibrio harveyi and Vibrio parahaemolyticus was much higher than that produced by its constituent SSPs. MSP1 in the co-culture assay reduced (p < 0.05) A. hydrophila count from 9.89 ± 0.1 CFU mL−1 to 2.14 ± 0.2 CFU mL−1. The biofilm formation of both MSPs were significantly higher (p < 0.05) than its constituent SSPs and the pathogens. The SSPs in both MSPs generally showed resistance to high temperatures (80, 90, and 100 °C) and a wide range of pH (2 to 9). This in vitro assessment study demonstrates that MSP1 and 2 have the potential to be further explored as multi-strain probiotics on selected aquatic species.

Dietary astaxanthin augments disease resistance of Asian seabass, Lates calcarifer (Bloch, 1790), against Vibrio alginolyticus infection.

This investigation describes the impacts of dietary provisioning with astaxanthin on hemato-biochemistry, non-specific immunity, and disease resistance of the Asian seabass, Lates calcarifer, against the virulent Vibrio alginolyticus; with specific reference to dose-response associations and variations over different post-infection periods (0-, 7-, and 14-day). Triplicate groups of fish weighing 28 g, on average, were fed various diets (C, the control or astaxanthin-free; AXT50, 50 mg astaxanthin kg-1 diet; AXT100, 100 mg astaxanthin kg-1 diet; and AXT150, 150 mg astaxanthin kg-1 diet) for 90 days and subsequently challenged with V. alginolyticus at the end of the feeding period. Experimental infection unveiled that supplemented fish demonstrated significant improvements (P < 0.05) of hematological parameters (white blood cell [WBC] and red blood cell [RBC] counts, and hemoglobin and hematocrit levels) when fed diets with elevating supplemental doses of astaxanthin through distinct post-infection periods (0-, 7-, and 14-day). Furthermore, the administration of dietary astaxanthin at escalating levels markedly enhanced (P < 0.05) the serum biochemical profile (aspartate aminotransferase [AST], alanine aminotransferase [ALT], glucose, cortisol, cholesterol, and triglyceride contents) of challenged fish, resulting in better welfare. Significantly higher (P < 0.05) contents of serum total protein were observed in supplemented fish, as opposed to the control. Additionally, immunological defense mechanisms (lysozyme activity, phagocytic activity, respiratory burst activity, and total serum immunoglobulin) of challenged fish were pronouncedly elicited (P < 0.05) following the ingestion of astaxanthin. Besides, the supplementation with dietary astaxanthin significantly augmented (P < 0.05) the post-challenge survival rate of fish. Collectively, the results manifest that supplementary feeding of astaxanthin is effective in reinforcing fish immunocompetence and disease resistance against V. alginolyticus infection.