Preservation of viability and antibacterial activity of Lactobacillus spp. in calcium alginate beads

The objective of the study was to produce calcium alginate beads able to deliver Lactobacillus spp. (Lactobacillus plantarum, Lactobacillus rhamnosus GG, Lactobacillus bulgaricus and Lactobacillus lactis) with preserved viability and antibacterial activity. Four types of beads, containing entrapped (E), surface and entrapped (ES), surface (S) and concentrated surface and entrapped lactobacilli (CES) were prepared and physically characterized. The antibacterial activity of lactobacilli cultures before and after immobilization, freeze-drying and throughout storage was studied in relationship to the viable number of lactobacilli. Multi-resistant clinical isolates (methicillin-resistant Staphylococcus aureus, vancomycine-resistant Enterococcus faecalis, VIM-2-metalo-β-lactamase producing Pseudomonas aeruginosa and CTX-M-15-β-lactamase producing strains: Escherichia coli and Klebsiella pneumoniae) were used as indicator strains. Alginate beads in which lactobacilli proliferated to the beads surface (ES and CES) differed significantly from the other types of beads in their physicochemical properties, showing smoother surface morphology, more spherical shape, bigger weight, lower calcium content, density and crushing force. Lactobacilli cultures, at high cell concentrations (108 cfu/ml) were active against both Gram-positive and negative multi-resistant bacteria. Beads containing both entrapped and surface lactobacilli (ES) resulted in viability and antibacterial activity most similar to non-processed lactobacilli cultures. The viability and antibacterial activity of the immobilized lactobacilli remained stable after 6 months storage.