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

Abstract

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.

Salmonellosis: An Overview of Epidemiology, Pathogenesis, and Innovative Approaches to Mitigate the Antimicrobial Resistant Infections

“…Probiotics containing multiple groups of bacteria with different mechanisms of action tend to have synergistic effects on each other and have a broad spectrum of activity [252,254]. For insistence, the MSP of Bacillus amyloliquefacrem, Enterococcus hirae, and Lysinibacillus fusiformis was able to significantly inhibit the growth and biofilm formation of Aeromonas hydrophila compared to the individual probiotics [255]. Multi-species probiotics (L. reuteri, E. faecium, B. animalis, P. acidilactici, and L. salivarius) greatly reduce S. Enteritidis infections (up to 2.7 log reduction) in poultry [256].…”

“…Probiotics are classified as mono-strain or single-strain probiotics (SSP), multi-strain probiotics (MSP), and multi-species probiotics [253,254]. Singlestrain probiotics (SSP) can provide limited health benefits to the host [255]. Probiotics containing multiple groups of bacteria with different mechanisms of action tend to have synergistic effects on each other and have a broad spectrum of activity [254,256].…”

Dynamic effects of probiotic formula ecologic®825 on human small intestinal ileostoma microbiota: a network theory approach

“… 14–16 The potential impact of multi-strain probiotic formulas, as opposed to single-strain probiotics, on the host is greater because the strains within the formula can complement each other. 17 Previous studies have shown that the oral administration of a probiotic formula, Ecologic® 825, to healthy volunteers did not result in an overall shift in fecal community composition of the microbiota. However, the LEfSe analyses identified two features belonging to a Bacteroides sp.…”

Probiotics supplementation to adult human small intestinal stoma microbiota causes dynamic increase in the community resistance to perturbations and nutrient utilization

“…Currently, probiotics are used in prevention and treatment of several disorders [13, 14]. It has been widely accepted that the impact of multi strain probiotic formulas instead of single strain probiotics is potentially greater on the host since the strains within the formula can complement each other [15]. Indeed, oral administration of a probiotic formula (Ecologic® 825) to healthy volunteers resulted in a minor shift in the fecal microbiota [16], while when administered orally to patients with irritable bowel syndrome, an increase in fecal levels of acetate and butyrate, accompanied by a reduction in fecal zonulin and symptom severity were observed [17].…”