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Table 1.

Variants of the preparation and storage of experimental cells using the lyophilization method.

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Fig 1.

Effect of different excipient concentrations on the viability of probiotic strains.

Viable cell counts (log CFU/mL) of five probiotic strains—Bacillus tropicus KBC8, Bacillus tequilensis KBC4, Lactobacillus salivarius KBC9, Staphylococcus hominis KBC6, and Staphylococcus gallinarum KBC10—were measured after exposure to varying concentrations (2–10%) of four excipients: (A) glucose, (B) sucrose, (C) skim milk, and (D) glycine. Each panel represents the effect of one excipient on the viability of all five strains. The experiments were conducted in triplicate, and the data are expressed as mean values. Ellipses indicate the concentration at which the maximum viability was observed for each excipient across the tested strains, representing the optimal concentration for formulation.

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Fig 2.

Long-term survival of lyophilized probiotic bacterial strains under different formulation conditions.

Survival rates (log CFU/mL) of lyophilized (A) Bacillus tropicus KBC8, (B) Bacillus tequilensis KBC4, (C) Lactobacillus salivarius KBC9, (D) Staphylococcus hominis KBC6, and (E) Staphylococcus gallinarum KBC10 were assessed over 12 months of storage. Bacterial preparations were stored under ten different formulation conditions (variants I–X) as indicated in the legends. All experiments were performed in triplicate, and the data are presented as mean.

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Table 2.

Resistance of selected probiotic isolates to gastric stress at different stages of storage under different preparation variants.

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Table 3.

Resistance of selected probiotic isolates to intestinal stress at different stages of storage under different preparation variants.

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Table 4.

Relative bacterial growth ratio (RBGR) values of the selected probiotic strains at different stages of storage under different preparation variants.

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Fig 3.

Adhesion ability of lyophilized probiotic bacterial strains during long-term storage under different formulation conditions.

Adhesion index (%) of lyophilized (A) Bacillus tropicus KBC8, (B) Bacillus tequilensis KBC4, (C) Lactobacillus salivarius KBC9, (D) Staphylococcus hominis KBC6, and (E) Staphylococcus gallinarum KBC10 measured at 0, 3, 6, 9, and 12 months of storage under ten different preparation variants (I–X). Adhesion ability was evaluated as the percentage of bacterial cells adhering to intestinal epithelial cells. Each bar represents the mean of triplicate experiments; error bars indicate standard deviations.

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