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Efficacy of Single and Multi-Strain Probiotics on In Vitro Strain Compatibility, Pathogen Inhibition, Biofilm
Puvaneswari Puvanasundram1,2, Chou Min Chong1,2, Suriana Sabri3
1Laboratory of Aquatic Animal Health and Therapeutics, Institute of Biosciences, University Putra Malaysia, Serdang 43400, Selangor, Malaysia.
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.
Insights
This study evaluated multi-strain probiotics (MSPs) for compatibility and efficacy. Promising results suggest MSPs can enhance microbial antagonism and biofilm formation for potential use in aquatic species.
Area of Science:
- Microbiology
- Aquaculture
- Probiotics
Background:
- Strain compatibility and properties are crucial for multi-strain probiotic (MSP) formulation.
- In vitro evaluation of single-strain probiotics (SSPs) and MSPs is essential for assessing their potential.
Purpose of the Study:
- To evaluate the in vitro compatibility, microbial antagonism, biofilm formation, and stress tolerance of selected SSPs and MSPs.
- To identify potential MSP candidates for application in aquatic species.
Main Methods:
- In vitro assays were conducted to assess strain compatibility, microbial antagonism against specific pathogens (Aeromonas hydrophila, Streptococcus agalactiae, Vibrio harveyi, Vibrio parahaemolyticus), biofilm formation capacity, and stress tolerance (high temperature and wide pH range).
- Co-culture assays were used to quantify pathogen reduction by MSP1.
- Biofilm formation was compared between MSPs, SSPs, and pathogens.
Main Results:
- MSP1 demonstrated significantly stronger antagonism against Aeromonas hydrophila and Streptococcus agalactiae compared to individual SSPs, reducing A. hydrophila counts from 9.89 ± 0.1 CFU mL−1 to 2.14 ± 0.2 CFU mL−1.
- MSP2 exhibited higher inhibition zones against Vibrio harveyi and Vibrio parahaemolyticus than its constituent SSPs.
- Both MSPs showed significantly higher biofilm formation capacity than their SSPs and tested pathogens.
- The SSPs within both MSPs generally exhibited resistance to high temperatures (80-100 °C) and a broad pH range (2-9).
Conclusions:
- The evaluated MSPs (MSP1 and MSP2) show significant potential as effective probiotics.
- Further in vitro and in vivo studies are warranted to explore the application of these MSPs in selected aquatic species for improved health and disease resistance.
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