Encapsulation of lactic acid bacteria in W₁/O/W₂ emulsions stabilized by mucilage:pectin complexes

Opuntia silvestri mucilage obtained from dried stems was explored as an emulsifier to prepare double emulsions aiming to encapsulate Lactiplantibacillus plantarum CIDCA 83114. W₁/O/W₂ emulsions were prepared using a two-step emulsification method. The aqueous phase (W₁) consisted of L. plantarum CIDCA 83114, and the oil phase (O) of sunflower oil. The second emulsion was prepared by mixing the internal W₁/O emulsion with the W₂ phase, consisting of 4 % polysaccharides, formulated with different mucilage:(citric)pectin ratios. Their stability was assessed after preparation (day 0) and during storage at 4 °C (28 days). Determinations included creaming index, color, particle size, viscosity, turbidity, and bacterial viability, along with exposure to simulated gastrointestinal conditions. Significant differences were evaluated by analysis of variance (ANOVA) and Duncan's test (P < 0.05). After 28 days storage, bacterial viability in the W₁/O/W₂ emulsions was above 6 log CFU/mL for all the pectin:mucilage ratios. Emulsions containing mucilage and pectins showed lower creaming indices after 15 days, remaining stable until the end of the storage period. Formulations including 1:1 pectin:mucilage ratio exhibited the highest bacterial viability under simulated gastrointestinal conditions and were more homogeneous in terms of droplet size distributions at day 0, hinting at a synergistic effect between mucilage components (e.g., proteins, Ca²⁺) and pectin in stabilizing the emulsions. These results showed that Opuntia silvestri mucilage enhanced the stability of emulsions during refrigerated storage, highlighting its potential for encapsulating lactic acid bacteria. This presents an economical and natural alternative to traditional encapsulating materials.