Galacto-oligosaccharides and lactulose as protectants against desiccation of Lactobacillus delbrueckii subsp. bulcaricus.

Lactobacillus delbrueckii subsp. bulgaricus CIDCA 333 was dehydrated on desiccators containing silica gel in the presence of 20% w/w of two types of galacto-oligosaccharides (GOS Biotempo and GOS Cup Oligo H-70®) and lactulose, until no changes in water desorption were detected. After rehydration, bacterial growth was monitored at 37°C by determining: (a) the absorbance at 600 nm and (b) the near infrared spectra (NIR). Principal component analysis (PCA) was then performed on the NIR spectra of samples dehydrated in all conditions. A multiparametric flow cytometry assay was carried out using carboxyfluorescein diacetate and propidium iodide probes to determine the relative composition of damaged, viable, and dead bacteria throughout the growth kinetics. The absorbance at 600 nm and the position of the second derivative band at ∼1370 nm were plotted against the time of incubation. The efficiency of the protectants was GOS Biotempo > GOS Cup Oligo H-70® > lactulose. The better protectant capacity of GOS Biotempo was explained on the basis of the lower contribution of damaged cells immediately after rehydration (t = 0). PCA showed three groups along PC1, corresponding to the lag, exponential and stationary phases of growth, which explained 99% of the total variance. Along PC2, two groups were observed, corresponding to damaged or viable cells. The results obtained support the use of NIR to monitor the recovery of desiccated microorganisms in real time and without the need of chemical reagents. The use of GOS and lactulose as protectants in dehydration/rehydration processes was also supported.

Influence of gastrointestinal system conditions on adhesion of exopolysaccharide-producing Lactobacillus delbrueckii subsp. bulgaricus strains to caco-2 cells

This study aimed to assess the transit tolerance of potential probiotic dairy Lactobacillus strains in human uppergastrointestinal tract in vitro, and to evaluate the effect of EPS production on the viability and adhesion of these strains. Survival and adhesion of two exopolysaccharide (EPS)-producing L. delbrueckii subsp. bulgaricus strains (B3 and B2) and E. coli ATCC11229 were assessed after the exposure of different pH (gastric juice) and gastric plus pancreatic juice challenges. In the artificial gastric juice (pH 2), both the viability of the strain B3 and B2 was decreased. Artificial juice treatments significantly reduced the adhesion to caco-2 cells (P< 0.05). High EPS-producing B3 survived better in the adverse gastrointestinal conditions and showed better ability of adhesion to Caco-2 cells when assessed for competition with E. coli ATCC 11229 compared to low EPS-producing B2. This investigation showed that EPS production could be affected or be involved in the viability, adherence and competition of L. delbrueckii subsp. bulgaricus strains and support the potential of B3 strain for development of new probiotic products.

Influence of different carbon sources on exopolysaccharide production by Lactobacillus delbrueckii subsp. bulgaricus (B3, G12) and Streptococcus thermophilus (W22)

Exopolysaccharides (EPSs) production was studied by Lactobacillus delbrueckii subsp. bulgaricus (B3, G12) and Streptococcus thermophilus (W22) in the medium containing various carbon sources (glucose, fructose, sucrose or lactose). For all the strains, glucose was the most efficient carbon source and B3, G12 and W22 strains produced 211, 175 and 120 EPS mg/L respectively. Also, the influence of different concentrations of glucose (5,10,15,20,25,30 g/L) on EPS production and growth was studied. The results indicated that EPS production and growth were stimulated by the high glucose concentration (30 g/L).