Effect of fermentation by pure and mixed cultures of Streptococcus thermophilus and Lactobacillus helveticus on isoflavone and B-vitamin content of a fermented soy beverage

Enhancement of nutritional or vitamin content of foods is commonly touted as a major benefit of probiotics. In this paper, we examined the ability of three probiotic bacteria either alone or in combination to enhance nutritional content. Pure and mixed cultures of Streptococcus thermophilus ST5 and Lactobacillus helveticus R0052 or Bifidobacterium longum R0175 were used to prepare fermented soy beverages. The effects of strain, acidification and mixed cultures on the deconjugation of soy isoflavones were examined. Acidification to pH 4.7 alone resulted in a 7% drop in isoflavone levels. Deconjugation levels varied between the different glucosides. Fermentation by L. helveticus R0052 resulted in the 50% reduction in total glucosides with O-malonyl glucosides being reduced 64%. Fermentation by S. thermophilus ST5 or B. longum R0175 had no significant effect on isoflavone levels. Combining a S. thermophilus strain with a L. helveticus culture reduced the effectiveness of the latter. Fermentation did not significantly modify vitamin B1 or B6 levels.     

Immunomodulatory bioactivity of soy and milk ferments on monocyte and macrophage models

Fermented products provide a functional food-based approach for probiotic delivery, containing both the bacteria and their potentially bioactive metabolites. This study investigated potential immunomodulatory effects of soy and dairy ferments prepared with Bifidobacterium longum R0175 or Lactobacillus helveticus R0052 in combination with Streptococcus thermophilus ST5 on a human monocyte model (U937). We examined the effects of both soy and dairy ferments on TGF-??1 production by U937 cells challenged with the pro-inflammatory cytokine Tumor Necrosis Factor-?? (TNF-??) at either the monocyte stage or the macrophage stage (following differentiation with all-trans retinoic acid). Effects on cell surface markers involved in macrophage activation and adhesion were also examined. Both soy and dairy ferments influenced U937 cytokine production with significant differences seen depending on the U937 differentiation stage. This suggests that potential immunomodulatory effects of fermented products can vary with respect to the target cell type and its stage of differentiation.     

High hydrostatic pressure processing on microstructure of probiotic low-fat yogurt

The effect of milk processing on the microstructure of probiotic low-fat yogurt was studied. Skim milk fortified with skim milk powder was subjected to three treatments prior to innoculation: thermal treatment at 85 °C for 30 min, high hydrostatic pressure at 676 MPa for 5 min, and combined treatments of high hydrostatic pressure (HHP) and heat. The processed milk was then fermented by using two different starter cultures containing Streptococcus thermophilus, Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus acidophilus, and Bifidobacterium longum. The microstructure of heat-treated milk yogurt had fewer interconnected chains of irregularly shaped casein micelles, forming a network that enclosed the void spaces. On the other hand, microstructure of HHP yogurt had more interconnected clusters of densely aggregated protein of reduced particle size, with an appearance more spherical in shape, exhibiting a smoother more regular surface and presenting more uniform size distribution. The combined HHP and heat milk treatments led to compact yogurt gels with increasingly larger casein micelle clusters interspaced by void spaces, and exhibited a high degree of cross-linking. The rounded micelles tended to fuse and form small irregular aggregates in association with clumps of dense amorphous material, which resulted in improved gel texture and viscosity.     

Identification and technological properties of lactic acid bacteria isolated from raw goat milk of four Algerian races

The characteristics of 725 lactic acid bacteria isolated from raw goat milk of four Algerian races were studied. They were phenotypically classified as Lactobacillus (31.6%), Lactococcus (28.4%), Leuconostoc (22.2%), Streptococcus (13.7%) and Pediococcus (4.1%). No major differences were observed as to the distribution of genus isolates in the milk of three races (Makatia, Makatia-Chamia and Kabyle). However, the number of isolates of both Leuconostoc and Lactococcus predominated in Arabia milk while the samples collected from the other races mostly contained lactobacilli isolates.The phenotypic and biochemical analyses gave a diversity of species (28 presumptive species). The most abundant species were Streptococcus thermophilus (46 isolates), Lactobacillus helveticus (45 isolates), Lactobacillus plantarum (30 isolates), Lactobacillus delbruecki subsp. bulgaricus (26 isolates), Lactobacillus brevis (25 isolates) and Lactococcus lactis subsp. lactis (25 isolates).The technological properties of the isolates were as follows: 38.6% of isolates showed a fast acidifying rate (more than 60°D during 18 h of incubation); 25.9% of isolates had a high proteolytic capacity (more than 6 ppm leucine/72 h) and 14.1% of isolates had a high diacetyl production (more than 0.8 ppm diacetyl/16 h).Nine isolates were selected to prepare yoghurt and cheese. Sensory tests performed by a trained panel revealed a pleasant yoghurt prepared with mixed culture St. thermophilus 16 TMC+L. helveticus 20TMC. Cheese prepared with isolates L. lactis subsp. lactis biovar diacetylactis 19 MCA and L. lactis subsp. lactis 10 MCM had a very good sensory quality and a pleasant taste.     

Effect of high pressure homogenization on lactic acid bacteria phages and probiotic bacteria phages

To investigate the effect of high pressure homogenization on virus inactivation, phages specific for Lactobacillus delbrueckii, Lactobacillus helveticus, Streptococcus thermophilus, Lactococcus lactis, Lactobacillus paracasei and Lactobacillus plantarum were studied. The influence of pressure, number of passes, suspension medium and phage concentration were studied at 25 °C. Reductions in viability were proportional to pressure and number of passes, though the inactivation extent was phage-dependent. At 100 MPa, some bacteriophages were completely inactivated (6 log₁₀ reduction) after 3 or 5 passes, while others remained infective after 8 passes. For all phages, treatment at 60 MPa was insufficient for complete inactivation, even after 8 passes. No clear influence of suspending medium was observed. Inactivation seems to depend on phage concentration; the higher the initial load, the bigger the reduction achieved. Although these results showed that several phages studied are resistant to high-pressure homogenization, this strategy could be combined with others to control their presence in raw milk.     

Identification of cultivable lactic acid bacteria isolated from Algerian raw goat's milk and evaluation of their technological properties

One hundred and fifty-eight strains of lactic acid bacteria isolated from Algerian raw goat's milk were identified and technologically characterized. Five genera were found: Lactobacillus (50.63%), Lactococcus (25.94%), Streptococcus (14.56%), Leuconostoc (7.59%) and Pediococcus (1.26%). The predominant species were Lactococcus lactis (32 strains), Streptococcus thermophilus (23 strains), Lactobacillus bulgaricus (19 strains), Lb. helveticus (16 strains) and Lb. plantarum (14 strains).Approximately 39% of the lactic acid bacteria isolated produced more than 0.6% lactic acid (w/v) after 18 h of incubation, and belonged to the Lactococcus and Lactobacillus genera. The highest proteolytic activity was approximately 3 mg tyrosine l⁻¹ for mesophilic strains and nearly 5 mg tyrosine l⁻¹ for thermophilic lactobacilli after 72 h. High aromatic activity (more than 0.8 mg diacetyl l⁻¹ after 16 h) was detected in 14% of the strains.Nine strains were used to make dairy products (a yoghurt-like product and Edam-type cheese) on a pilot scale in the laboratory. The best-liked organoleptic characteristics were noted in a yoghurt produced with a mixed culture made up of S. thermophilus (strain 16TMC+) and Lb. helveticus (strain 20TMC) and in a cheese made with a starter composed of Lc. lactis subsp. lactis (strain 10MCM) and L. lactis subsp. lactis (V.P. +) (strain 19MCM).     

Effect of Greek-style yoghurt manufacturing processes on starter and probiotic bacteria populations during storage

This study compared viable counts of Streptococcus thermophilus (starter) and Lactobacillus helveticus R0052 (probiotic), as a function of two manufacturing processes of Greek-style (GS) yoghurt: centrifugation of a curd (GS-CF process) or ultrafiltration of milk prior to fermentation (GS-UF process). Fresh GS-UF and GS-CF yoghurts had between 3 and 7 times higher counts of Lb. helveticus R0052 and S. thermophilus than the regular stirred yoghurt (Control). Strain R0052 was three times more stable in the GS-CF yoghurt than in the Control over the 44 d storage period. The highest lactose content (44 g L⁻¹) was obtained in the Control, while 60% less was obtained in the GS-UF yoghurt; the opposite trend was observed for lactic acid and galactose. Free amino acid levels were 33% higher in the GS products. GS yoghurt can be 10 times better than traditional yoghurt to deliver probiotic bacteria, as a function of the process used.     

Effect of cook temperature on starter and non-starter lactic acid bacteria viability,cheese composition and ripening indices of a semi-hard cheese manufactured using thermophilic cultures

Semi-hard cheeses were manufactured using Streptococcus thermophilus and Lactobacillus helveticus cultures and their ripening was characterised. During cheese manufacture, curds were cooked to a maximum temperature of 47, 50 or 53 °C, pre-pressed under whey at pH 6.15, moulded, pressed and brined. Increased cook temperature resulted in increased manufacture time, a significantly reduced growth rate of S. thermophilus during manufacture in the order 47≈50 °C>53 °C and in significantly lower mean viable cell counts of S. thermophilus up to 56 d of ripening. Increasing cook temperature had no significant effect on mean viable cell numbers of L. helveticus or non-starter lactic acid bacteria (NSLAB). Cheeses produced from curds cooked to 47 °C had significantly higher levels of moisture in non-fat substances (MNFSs), salt-in-moisture and a significantly lower pH and levels of butyrate compared with cheeses produced from curds cooked to 50 or 53 °C.     

Antihypertensive peptides from skimmed milk hydrolysate digested by cell-free extract of Lactobacillus helveticus JCM1004

In the present experiments, the proteinase, aminopeptidase and x-prolyl-dipeptidyl aminopeptidase activitives were measured at 513 ± 11.9, 376 ± 9.3 and 23.6 ± 1.6 units per gramme of cell-free extract of Lactobacillus helveticus JCM1004. ACE-inhibitory activity of skimmed milk hydrolysate produced by cell-free extract of L. helveticus JCM1004 was determined, and the optimum pH and hydrolysis time for the production of ACE-inhibition substances from skimmed milk were 6.5–7.0 and 6–10 h, respectively. Peptides of Val-Pro-Pro and Ile-Pro-Pro with potent ACE inhibitor and antihypertensive activitives were purified from the hydrolysate by three step-reverse-phase high-performance liquid chromatography. The IC₅₀ values of Val-Pro-Pro and Ile-Pro-Pro were 9.13 ± 0.21 and 5.15 ± 0.17 μM, respectively. Significant decrease (p < 0.01) in SBP of SHR was measured after a single gastric intubation of Val-Pro-Pro or Ile-Pro-Pro at 8 or 4 h, respectively.     

Development of a fermented goat's milk containing probiotic bacteria

A set-type fermented milk manufactured from goat's milk was developed. Optimal curd tension was achieved by supplementation of milk with skim milk powder and whey protein concentrate (WPC). Milk was fermented employing a commercial probiotic starter culture (ABT-2), which contained Streptococcus thermophilus ST-20Y, Lactobacillus acidophilus LA-5, and Bifidobacterium BB-12. Supplementation of milk with 3% WPC reduced fermentation time by 2 h due to the increase in viable counts of S. thermophilus and Bifidobacterium by 0.3 and 0.7 log units, respectively. Addition of WPC increased the protein content (1%) as well as potassium and magnesium content (0.3 and 0.02 g kg⁻¹, respectively). Increase of the protein content led to an increase in the apparent viscosity and gel firmness of the product, and at the same time whey syneresis was reduced. As a consequence, the product received a high score for appearance, taste, aroma, texture and overall acceptance.     

Simultaneous detection and enumeration of viable lactic acid bacteria and bifidobacteria in fermented milk by using propidium monoazide and real-time PCR

This study describes a procedure that allows specific detection and enumeration of viable bacteria in four species of lactic acid bacteria (Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus casei subsp. casei and Lactobacillus acidophilus) and of Bifidobacterium lactis, mixed in fermented milk products. The procedure is based on the combined use of propidium monoazide (PMA), able to distinguish between viable and irreversibly damaged cells, with species-specific quantitative real-time PCR (RTi-PCR). Loss of viability of the species in a fermented milk through storage at 4 °C was similarly (P < 0.05) detected by PMA–RTi-PCR and selective plate counts. Furthermore, comparison of results obtained by both methods showed a Pearson linear correlation of 0.995. The enumeration of viable bacteria by PMA–RTi-PCR could be performed in 3 h, whereas enumeration by selective plate counts required three days. The procedure developed is a fast method for the identification, enumeration and discrimination of viability of lactic acid bacteria and bifidobacteria mixed in fermented milk products.     

Production of volatile aroma compounds by kefir starter cultures

Production of carbonyl compounds by single-strain cultures, kefir starter (Lactobacillus delbrueckii subsp. bulgaricus HP1+Lb. helveticus MP12+Lactococcus lactis subsp. lactis C15+Streptococcus thermophilus T15+Saccharomyces cerevisiae A13) and kefir grains during fermentation and storage of kefir was studied. The content of carbonyl compounds produced by kefir starter was greater than that produced by kefir grains. The maximum acetaldehyde concentration (18.3 μg g⁻¹) in kefir with starter culture was mainly due to the metabolic activity of Lb. delbrueckii subsp. bulgaricus HP1 isolated from kefir grains. The highest diacetyl production activity was recorded in the starter culture (1.87 μg g⁻¹) and the single-strain culture St. thermophilus T15 (1.62 μg g⁻¹), followed by Lb. helveticus MP12 (0.85 μg g⁻¹) and Lc. lactis subsp. lactis C15 (0.42 μg g⁻¹). The lactobacilli Lb. delbrueckii subsp. bulgaricus HP1 and Lb. helveticus MP12 produced acetone, which was not found in the cocci cultures. The presence of 2-butanone was related to the production ability of Lb. helveticus MP12. In comparison, Lc. lactis subsp. lactis C15 synthesized ethyl acetate more actively than the other single-strain cultures included in the starter. S. cerevisiae A13 produced ethanol and CO₂ in amounts (3975 μg g⁻¹; 1.80 g L⁻¹) that lent cultured kefir distinctive flavour and aroma characteristic of authentic kefir.     

Probiotic stability of yoghurts containing Jerusalem artichoke inulins during refrigerated storage

Experimentally prepared Jerusalem artichoke inulins (JAI) were compared with two commercial chicory root inulins for their prebiotic potentials in media broth model and growth-sustaining ability in non-fat yoghurts. Experimental yoghurts were made with 12% reconstituted skim milk (RSM) supplemented with 4% inulin powders, inoculated with mixed cultures of Lactobacillus casei LC-01, Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus (1:0.5:0.5 based on supplier’s recommendation) and incubated overnight at 37 °C. Non-supplemented yoghurt was prepared from 16% RSM and used as control. The survival and acidifying activity of lactic and probiotic cultures in all yoghurts were investigated on weekly intervals during the shelf life of 28 days at 4 °C. Incorporation of JAI resulted in improved viability of LC-01, maintaining >7.0 log CFU/g during cold storage but did not affect the viability of yoghurt bacteria in comparison with the control.     

Tea extract render probiotic Lactobacillus helveticus more resistant to oxygen exposure through lipid modification mechanism

Exposure to oxygen can cause a decrease in growth rates or a complete inhibition of growth of oxygen-sensitive probiotic bacteria. A recent study in our laboratory demonstrated that the growth of an oxygen-sensitive strain, Lactobacillus helveticus, was stimulated, under aerobic conditions, when the culture medium was enriched with green tea extracts (GTE). However, information on the mechanism by which GTE influenced the growth, in the presence of oxygen, of that strain is limited. In the present work, the effects of GTE concentrations (0 to 2000 μg/mL) and exposure to oxygen on maximal populations of L. helveticus R0052 cells and bacterial lipids were evaluated using viable counts, infrared spectroscopy and gas chromatography analyses. Supplementation of the culture medium with 0 to 500 μg/mL GTE did not have an effect on the populations reached under microaerophilic conditions and on bacterial lipid structure and composition. However, at 2000 μg/mL GTE, high population levels were reached under microaerophilic conditions concomitant with an increase in lipid order and with important changes in fatty acid composition of the bacterial lipids. Interactions between GTE components and bacterial lipids were shown by spectroscopic results. Moreover, bacterial cells have adapted to the presence of 2000 μg/mL GTE in the growth medium by changing their lipid composition. To the best of our knowledge, this work is the first to establish a relationship between the effects of GTE at 2000 μg/mL on bacterial cell's lipids and a stimulation of growth under microaerophilic conditions of an oxygen-sensitive strain.     

Effect of cryoprotectants,prebiotics and microencapsulation on survival of probiotic organisms in yoghurt and freeze-dried yoghurt

The survival of probiotic microorganisms including Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus rhamnosus and Bifidobacterium spp. was evaluated in yoghurt and freeze-dried yoghurt after processing and storage. The effectiveness of microencapsulating probiotic organisms as well as adding cryoprotectants and prebiotics in improving their viability was also investigated. The viability of Bifidobacterium infantis 17930 and L. rhamnosus GG was reduced by 0.07 log, while that of L. casei 1520 and Bifidobacterium longum 1941 was reduced by 0.28 and 0.39 log, respectively. There was a 7% improvement in the viability of L. casei 1520 when cryoprotectant ‘Unipectine™ RS 150’ was added at 2.5% (w/v). The prebiotic ‘Raftilose^®P95’ when added at 1.5% w/v to yoghurt improved the viability of the combined selected probiotic organisms by 1.42 log during four weeks of storage at 4 °C. Microencapsulation with alginate improved viability of combined selected probiotic organisms by 0.31 log in freeze-dried yoghurt stored at 21 °C.     

Optimization of sour milk fermentation for the production of ACE-inhibitory peptides and purification of a novel peptide from whey protein hydrolysate

The effect of fermentation conditions on the production of angiotensin-I converting enzyme (ACE) inhibitory peptide in sour milk fermented by Lactobacillus helveticus LB10 was investigated using response-surface methodology. Optimal conditions to produce the maximum production of ACE-inhibitory peptides were found to be 4% (v/w) inoculum, 7.5 initial pH of medium and 39.0 °C. The fermented milk resulted in 75.46% inhibition in ACE activity. The cell-envelope proteinase, assisted by X-prolyldipeptidyl aminopeptidase of Lb. helveticus LB10 produced the ACE-inhibitory peptides. A novel ACE-inhibitory peptide from whey protein hydrolysate produced by crude proteinases of Lb. helveticus LB10 was purified. The separations were performed with Sephadex^® G-75 and Sephadex G-15 gel filtration chromatography and reversed-phase, high-performance liquid chromatography. The peptide with the RLSFNP sequence was isolated from β-lactoglobulin hydrolysate and its IC₅₀ while inhibiting ACE activity was 177.39 μm.     

Dietary functional benefits of Bartlett and Starkrimson pears for potential management of hyperglycemia,hypertension and ulcer bacteria Helicobacter pylori while supporting beneficial probiotic bacterial response

Phenolic-linked health benefits of Bartlett and Starkrimson pear cultivars were investigated for the potential relevance in managing type 2 diabetes and hypertension using in vitro enzyme models. Further effects of fermented (0, 24, 48, and 72-h with Lactobacillus helveticus) pear juice on inhibition of Helicobacter pylori and proliferation of probiotic Bifidobacterium longum were also evaluated. High total phenolic content along with high 2,2 diphenyl-1-picrylhydrazyl-linked free radical scavenging antioxidant activities were observed in peel extracts of both cultivars. In vitro enzyme assays with peel and pulp extracts also indicated high inhibitory activity of α-glucosidase and α-amylase used as models for anti-hyperglycemia benefits. Only the aqueous pulp extract of Bartlett pear had angiotensin I-converting enzyme inhibitory activity used as model for anti-hypertension benefits. Fermented acidic pH samples of both cultivars showed H. pylori inhibition at 48 and 72 h, while fermented sample of Starkrimson even showed inhibition at 24 h. Both cultivar extracts did not inhibit growth of probiotic B. longum.     

Influence of bacteria used as adjunct culture and sunflower oil addition on conjugated linoleic acid content in buffalo cheese

The influence of bacteria and sunflower oil addition on conjugated linoleic acid content (CLA) in buffalo cheese was determined. Fresh and short-ripened cheeses were manufactured using the same starter culture and four different adjunct strains previously selected by their CLA production rate. Lactobacillus casei, Lactobacillus rhamnosus, Bifidobacterium bifidum and Streptococcus thermophilus were individually used as adjunct culture. Sunflower oil (SO) was added to obtain a final concentration of 200 μg/ml of linoleic acid. CLA levels in cheese were higher than raw milk, especially after ripening time. SO supplementation increase CLA concentrations in fresh cheeses, except in those manufactured with S. thermophilus as adjunct culture. Both, ripening and SO supplementation showed a positive influence on CLA concentration. Similar texture, acidity and colour were determined in cheeses with or without SO supplementation. Buffalo cheeses manufactured with appropriate adjunct cultures may be a natural source of CLA for human consumption.     

Incorporation of selected nutraceuticals and probiotic bacteria into a fermented milk

Yoghurts were produced from a base milk containing three important nutraceuticals, namely ω-3-fatty acids, isoflavones and phytosterols. The cultures employed to make the yoghurts were single probiotic strains of Lactobacillus gasseri or Bifidobacterium infantis and, to achieve a short production time, a two-stage fermentation procedure was used with Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus providing the rapid acidification. Yoghurts containing counts of >1.0×10⁸ cfu mL⁻¹ of the individual probiotics and high counts of the traditional species from yoghurt were awarded overall scores for sensory acceptability >4.0 out of 5.0; the nutraceuticals appeared to have no adverse effect on flavour. Storage trials at 5 °C showed that the viability of the probiotic cultures was retained over 15 days.