Optimization of the cultivation conditions and medium components for folate production by the highest folate producing Streptococcus thermophilus strain RD102 was carried out using a 23 central composite design and surface modelling method of response surface methodology. As folate production was observed to be growth-associated, hence the desired responses selected for the optimization were both folate production and growth. The selected factors based on preliminary investigations were incubation period, concentrations of p-amino benzoic acid (PABA) the growth precursor, and lactose as carbon source. The optimum concentrations of PABA and lactose were found to be 300 μm and 3% respectively at 72 h incubation. The optimized conditions resulted in an increase in folate production by 26% compared with control conditions (0% PABA and lactose at 37°C). Using the method of experimental factorial design and response surface analysis, it was possible to determine optimal operating conditions to obtain a higher folate production by Strep. thermophilus. Therefore, this study constitutes a step in developing strategies to modulate the folate level to a higher level.

Transit kinetics and survival rates of a bacterial species from yoghurt (i.e. Streptococcus thermophilus strain FBI3) were examined in different digestive compartments of gnotoxenic and human-microbiota-associated mice. The production of the lactose-hydrolysing enzyme (i.e. β-galactosidase) was also investigated within the digestive tract, using a chromosomal reporter system based on luciferase genes from Photorhabdus luminescens under the control of the plac promoter. In both mice models, S. thermophilus cells transited within 2h from the stomach to the caecum–colon compartment of the digestive tract where they displayed a survival rate of nearly 100%. In gnotoxenic mice, luciferase activity was found to increase in the second half of the small intestine and in the caecum–colon compartment when lactose was added to the drinking water provided to the animals. In human-microbiota-associated mice drinking lactose, luciferase activity was similarly increased in the second half of the small intestine but was drastically reduced in the caecum–colon compartment. This feature could be ascribed to the presence of the resident human microbiota.

A new method for characterization of acid production by dairy starter cultures is presented. Microplates with integrated optical pH sensors are developed. Two fluorophores, a pH-sensitive and a pH-insensitive one are immobilised at the bottom of a polystyrene 96-well microtitre plate. The pH-insensitive fluorophore serves as an internal reference and makes calibration unnecessary. The sensor measures pH accurately in optically well-defined media. Particles and fluorophores contained in the bulk medium disturbed the measurements. Despite these disturbances it was possible to clearly sense differences in inoculum type and in inoculum sizes of cultures of Lactococcus lactis and of Streptococcus thermophilus at 30 and 37°C. Besides a pH-related signal there is information about other changes during milk fermentation. The cultivation results were compared with those from the established CINAC-method. From this comparison it can be concluded that the new method can be used reliably to characterize particularly a large number of strains for screening purposes but also for quality control.

The behaviour of Streptococcus thermophilus in combination with Lactococcus lactis subsp. cremoris or subsp. lactis mesophilic starters in experimental Cheddar cheese is reported. In a standard manufacturing procedure employing a 38 °C cook temperature, even very low levels (0·007%) of Str. thermophilus combined with normal levels of the mesophilic starter (1·7%) resulted in increased rates of acid production, the formation of significant amounts of galactose (∼ 13 mmol/kg cheese), and populations nearly equivalent to those of the mesophilic lactic starter in the curd before salting. At a 41 °C cook temperature, the Str. thermophilus attained a higher maximum population (∼ log 8·2 colony forming units (cfu)/g) than the Lc. lactis subsp. cremoris (∼ log 6·8 cfu/g) and formed more galactose (∼ 28 mmol/kg). Lactobacillus rhamnosus, deliberately added to a cheese made using Str. thermophilus starter and which contained 24 mmol galactose/kg at day one, utilized all the galactose during the first 3 months of cheese ripening. Adventitious non-starter lactic acid bacteria had the potential to utilize this substrate too, and a close relationship was demonstrated between the increase in this flora and the disappearance of the galactose. Some possible consequences for cheese quality of using Str. thermophilus as a starter component are discussed.