Evaluation of the response of Lactobacillus rhamnosus VTT E-97800 to sucrose-induced osmotic stress

Environmental osmotic changes are one of the stresses live probiotics may encounter either in their natural habitats or as a result of usage in food formulations and processing. Response to osmotic stress, induced by sucrose, of the probiotic strain Lactobacillus rhamnosus VTT E-97800 (E800) was investigated. The fluorescence-based approach used, by combined staining with caboxyfluorescein (cFDA) and propidium iodide (PI) could give insights on the osmotic-induced changes of microbial esterase activity and membrane integrity; also the extrusion of intracellular accumulated carboxyfluorescein (cF) upon energizing with glucose. Comparison of the flowcytometric viability assessment with the conventional culture techniques revealed that sucrose-stressed cells had a slight loss of culturability (log N/N0∼−0.3) at 1.2 and 1.5 M sucrose concentration though they could perform an enzymatic conversion of cFDA into cF. The presence of such metabolically active bacteria in food might be critical as they may excrete toxic or food spoilage metabolites. Moreover, the perturbation of cF extrusion activities became a limiting factor for reproductive capacities. There was no change in the cell morphology. These results proved the ability of the strain of study to tolerate sucrose, even at extreme concentrations and these must be taken into consideration for its usage in the formulation/processing of sugar-based foods, e.g. jams, candies, etc.