Probiotic properties of an indigenous strain composition of Bifidobacterium bifidum ICIS-310 and Bifidobacterium longum ICIS-505 in vitro

INTRODUCTION. An active search for probiotic strains of microorganisms, particularly indigenous bifidobacteria, is crucial for the development of therapeutic and prophylactic probiotics. Evaluating the probiotic potential of candidate strains requires their assessment for bacterial viability, antibiotic sensitivity, gastrointestinal stress tolerance, lysozyme resistance, and biofilm formation capacity.

AIM. This study aimed to characterise a composition of indigenous Biffdobacterium strains, B. biffdum ICIS-310 and B. longum ICIS-505, as a potential probiotic product.

MATERIALS AND METHODS. The study tested B. biffdum ICIS-310 and B. longum ICIS-505 for resistance to antibiotics, gastric acid, and bile. Mono- and co-cultures of B. biffdum ICIS-310 and B. longum ICIS-505 were tested for the number of viable cells during culture, antilysozyme activity (lysozyme resistance), and biofilm formation capacity. Antagonistic activity was tested against test strains of bacteria and fungi, including Candida albicans ATCC 24433, Proteus mirabilis ATCC 29906, Staphylococcus aureus ATCC 29213, Escherichia coli ATCC 25922, Shigella flexneri ATCC 12022, and Klebsiella pneumoniaе ICIS-278_PBV. The content of acetic acid in the culture medium was determined by gas–liquid chromatography.

RESULTS. The indigenous strains, B. biffdum ICIS-310 and B. longum ICIS-505, were selected according to the criteria of abundance, lysozyme resistance, and biofilm formation. These strains were found to lack pathogenicity genes, exhibit resistance to a number of antimicrobials (benzylpenicillin, streptomycin, and erythromycin), and remain viable in the presence of bile for 2 hours and in the presence of gastric acid for 30 minutes. The study demonstrated the biocompatibility of Biffdobacterium cultures, with the composition of B. biffdum ICIS-310 and B. longum ICIS-505 having a higher microbial cell count after 48 hours than monocultures. When co-cultured, B. biffdum ICIS-310 and B. longum ICIS-505 demonstrated a synergistic effect, resulting in increased lysozyme resistance (up to 2.2±0.30 μg/mL×OD₄₅₀), biofilm formation (up to 0.89±0.20 OD₆₃₀ units), and acetate production (up to 33.2 mM/L). The antagonistic activity against test strains was more pronounced in the co-culture than in the monocultures, with the respective growth inhibition zones of 30–36 mm and 18–24 mm.

CONCLUSIONS. Indigenous B. biffdum ICIS-310 and B. longum ICIS-505 have demonstrated resistance to antimicrobial agents, bile salts, and gastric acid. Co-culturing the strains has revealed a synergistic effect on their lysozyme resistance, biofilm formation capacity, and antagonistic activity. The strains of bifidobacteria and the composition thereof hold promise for the development of novel probiotics. The evaluation of lysozyme resistance and biofilm formation capacity, as indicators of the adaptive potential of the microbiota, may be recommended for the selection and testing of probiotic strains.

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