Study on the Influence of Tea Extract on Probiotics in Skim Milk: From Probiotics Propagation to Metabolite

In this study, the influence of tea extract (TE) on the growth of probiotics in skim milk was examined. Lactobacillus plantarum ST‐III, Bifidobacterium bifidum Bb02, Lactobacillus acidophilus NCFM, and Lactobacillus rhamnosus GG were used in this study. The introduction of TE in milk significantly stimulated the propagation and acidification of L. rhamnosus GG and L. acidophilus NCFM. The antioxidant capacities and the total free amino acid contents of all fermented milk products were enhanced by the addition of TE; however, there were different antioxidant properties and free amino acid contents of fermented milk samples fermented by different bacteria. With a 9% (w/w) level, the fermentation with L. rhamnosus GG and L. acidophilus NCFM showed larger numbers of viable cells and faster acidifying rates, as well as excellent antioxidant capacity and abundant free amino acids.  The stimulative effects of TE on probiotics can be considered for industrial purposes and has practical implications for commercial applications.     

Improvement in antioxidant activity,angiotensin‐converting enzyme inhibitory activity and in vitro cellular properties of fermented pepino milk by Lactobacillus strains containing the glutamate decarboxylase gene

BACKGROUND: The purpose of this study was to evaluate the functional potential of fermented pepino extract (PE) milk by Lactobacillus strains containing the glutamate decarboxylase (GAD) gene. Three Lactobacillus strains were selected, including L. brevis BCRC 12310, L. casei BCRC 14082 and L. salivarius subsp. salivarius BCRC 14759. The contents of free amino acids, total phenolics content, total carotenoids and the associated functional and antioxidant abilities were analyzed, including angiotensin‐converting enzyme (ACE) inhibition activity, 1,1‐diphenyl‐2‐picylhydrazyl (DPPH) radical‐scavenging ability and oxygen radical absorbance capacity (ORAC). Cell proliferation of fermented PE milk was also evaluated by MTT (3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide) assay. RESULTS: Compared to the unfermented PE, fermented PE milk from Lactobacillus strains with the GAD gene showed higher levels of total phenolics, γ‐aminobutyric acid, ACE inhibitory activity, DPPH, and ORAC. The viability of human promyelocytic leukemia cells (HL‐60) determined by the MTT method decreased significantly when the cells were incubated with the PE and the fermented PE milk extracts. CONCLUSION: The consumption of fermented PE milk from Lactobacillus strains with the GAD gene is expected to benefit health. Further application as a health food is worthy of investigation. © 2012 Society of Chemical Industry     

Potential of high pressure homogenization in the control and enhancement of proteolytic and fermentative activities of some Lactobacillus species

Different species of Lactobacillus involved in dairy product fermentation and ripening were considered in order to study the effect of high pressure homogenization (HPH) on: (i) fermentation kinetics of HPH treated cells inoculated in milk; (ii) metabolic profiles; (iii) release of intracellular proteolytic enzymes; and (iv) enhance of the activity of extracellular or cellular wall located proteolytic enzymes. The HPH treatments applied were 50, 100, 150 MPa, 2 cycles at 50 and at 100 MPa. The viability loss did not exceed 1.3 log cfu/ml after the higher treatments applied. The electrophoretic profiles of α- or β-casein incubated with the different cell free filtrates shown that HPH positively affected the proteolytic activity of some strains. Moreover, HPH affected the acidification rates of the milk inoculated with the processed cells and the primary metabolism of some strains. Regarding volatile compounds, ethanol, acetoin and 2-methyl butyric acid were subjected to the major changes when the inoculum had been processed.     

Effect of Lactobacillus plantarum on the survival of acid‐tolerant non‐O157 Shiga toxin‐producing E. coli (STEC) strains in fermented goat's milk

The ability of goat's milk fermented with a Lactobacillus plantarum strain B411, and in combination with commercial starter culture, to inhibit acid‐adapted (AA) and non‐acid‐adapted (NAA) environmental non‐O157 STEC strains was investigated. Acid‐adapted and NAA non‐O157 STEC strains were not inhibited in the L. plantarum‐fermented goat's milk, while the goat's milk fermented with the combination of L. plantarum and starter culture inhibited AA more than NAA non‐O157 STEC strains. Environmental acid‐tolerant non‐O157 STEC strains were not inhibited by L. plantarum, starter culture or combination of starter culture with L. plantarum unless they were subjected to prior acid adaptation such as backslopping.     

Manufacture of acid gels from skim milk using high-pressure homogenization

The effect of high-pressure homogenization (HPH) alone or in combination with a thermal treatment (TT) was investigated for the manufacture of acid gels from skim milk. Raw skim milk was subjected to HPH (0 to 350 MPa) or a TT (90°C, 5 min), or both, in the following processing combinations: 1) HPH, 2) HPH followed by TT, 3) TT followed by HPH, 4) TT, and 5) raw milk (control). After treatments, L* (lightness) values were measured, and then skim milk was acidified with 3% glucono-δ-lactone and rheological properties (G′ and gelation time), and whey holding capacity was evaluated. Treatments in which HPH and TT were combined showed greater L* values than those in which just HPH was applied. In all treatments, the L* values decreased as the pressure was increased up to 300 MPa with little change afterward. Gelation times were lower when HPH was combined with TT compared with the acid skim milk gels that were just pressure treated. The final G′ in gels obtained from skim milk subjected to the combined process (HPH and TT) was greater and pressure-dependent compared with all other gels. A maximum G′ (∼320 Pa) was observed with skim milk subjected to a combination of thermal processing before or after HPH at 350 MPa. Acid gels obtained from HPH milk at 350 MPa showed a linear decrease in whey holding capacity over time, retaining 20% more whey after centrifugation for 25 min compared with samples treated at lower pressures and all other treatments. Our results suggest that HPH in combination with TT can be used to improve the rheological properties and stability of yogurt, thus decreasing the need for additives.     

Diversity analysis and quantification of lactic acid bacteria in traditionally fermented yaks’ milk products from Tibet

Fermented yaks’ milk is a characteristic fermented dairy product with a rich microflora (especially lactic acid bacteria [LAB]), that has been produced by Tibetan people in China throughout history. Systematic analysis of the LAB composition of fermented yaks’ milk from central Tibet is essential. In the present study, 38 samples of fermented yaks’ milk were collected from three regions in central Tibet. From these, a total of 211 isolates of LAB were generated using traditional pure culture methods. To accurately analyze their biodiversity, 16S rRNA gene sequence analysis and denaturing gradient gel electrophoresis (DGGE) were used. The isolates were identified as belonging to six genera and 22 species/subspecies. Among them, Lactobacillus species predominated (117 isolates) accounting for 55.45% of all isolates. Overall, L. delbrueckii subsp. bulgaricus (60 isolates; 28.43% of the total) was the most commonly isolated species of LAB from fermented yaks’ milk from central Tibet. Seven Lactobacillus species and the Bifidobacterium genus were confirmed by quantitative PCR (q-PCR). The q-PCR results showed that the abundance of these groups in fermented yaks’ milk from central Tibet ranged from 1.12 ± 0.36 log cfu/ml (for L. sakei from Ningzhong) to 7.77 ± 0.84 log cfu/ml (for L. helvetivus from Ningzhong). Lactobacillus acidophilus, L. delbrueckii subsp. bulgaricus, L. fermentum, L. helveticus varied in concentration depending on the region. The results indicated that traditional fermented yaks’ milk from different regions of central Tibet have complex compositions of LAB species. The diversity of LAB might be related to geographical origin.     

Survival and activity of 5 probiotic lactobacilli strains in 2 types of flavored fermented milk

The viability of 5 probiotic lactobacilli strains (Lactobacillus acidophilus LA-5, Lactobacillus casei L01, Lactobacillus casei LAFTI L26, Lactobacillus paracasei Lcp37, and Lactobacillus rhamnosus HN001) was assessed in 2 types of probiotic flavored drink based on fermented milk during 21 days of refrigerated storage (5°C). Also, changes in biochemical parameters (pH, titrable acidity, and redox potential) during fermentation as well as the sensory attributes of final product were determined. Among the probiotic strains, L. casei LAFTI L26 exhibited the highest retention of viability during refrigerated storage period, while L. acidophilus LA-5 showed the highest loss of viability during this period. The decline in cell count of probiotic bacteria in strawberry fermented milk was significantly greater compared to peach fermented milk. In an overall approach, peach fermented milk containing L. casei LAFTI L26 was selected as the optimal treatment in this study in both aspects of viability and sensory accpeptibility.     

Influence of addition of raffinose family oligosaccharides on probiotic survival in fermented milk during refrigerated storage

The influence of the addition of raffinose family oligosaccharides (RFOs) extracted from lupin seeds on the survival of Bifidobacterium lactis Bb-12 and Lactobacillus acidophilus La-5 in fermented milk during 21 days of storage in refrigerated conditions was studied. For this purpose, viability and metabolic activity (expressed as pH, lactic and acetic acid production and utilization of soluble carbohydrates) of probiotic bacteria were determined. Retention of viability of B. lactis Bb-12 and L. acidophilus La-5 was greater in fermented milk with RFOs. The pH of probiotic fermented milk at 21 days of storage was lower (4.27) compared with probiotic fermented milk with RFOs (4.37). The highest levels of lactic and acetic acid were produced in probiotic fermented milk without RFOs compared with probiotic fermented milk with RFOs during storage at 4 °C. Soluble carbohydrates were utilised in fermented milk with and without RFOs, respectively, for maintaining B. lactis Bb-12 and L. acidophilus populations during refrigerated storage. In conclusion, all these experiments provide convincing evidence that RFOs have beneficial effects on the survival of these probiotic cultures in dairy products. As a result, such stored dairy products containing both probiotics and prebiotics have synergistic actions in the promotion of health.     

Serum Cholesterol Levels in Rats Fed Skim Milk Fermented by Lactobacillus Acidophilus

The effects of reconstituted skim milk, and the same fermented by Lactobacillus acidophilus, were tested in rats. Rats were fed a stock diet and drinking water containing one of three milk treatments: (1) no supplementary milk; (2) 10% milk; or (3) 10% milk fermented by L. acidophilus. After 4 wk, rats receiving the fermented milk had lower (P < 0.05) serum cholesterol levels (65 mg/dl) than did the water-fed (78 mg/dl) or milk-fed (79 mg/dl) rats. Weight gain, feed intake, liver lipid contents and fecal lactobacilli counts were not different among treatment groups. Data indicate that factors influencing serum cholesterol levels were produced during fermentation of the milk.     

Probiotic potential of lactic acid bacteria from fermented Malaysian food or milk products

This study investigated the probiotic characteristics of lactic acid bacteria (LAB) isolated from 14 Malaysian fermented food or milk products. In total, 22.3% (121 of 542) of the LAB isolated from the local fermented products exhibited antibacterial activity against Micrococcus luteus. Twelve LAB that demonstrated better antibacterial activity against M. luteus as compared to a commercial strain Lactobacillus casei strain Shirota (LABPC) were selected for further characterisation. Based on 16S rRNA gene sequence, the LAB were identified as pediococci (seven) and lactobacilli (five). All 12 LAB showed bile tolerance, but only eight were acid tolerant at pH ≥ 3.0. The highest level of adhesion to HT‐29 cells was observed among the Lactobacillus sp. LAB 1 and 10. The LAB also showed antimicrobial activity against Escherichia coli and Staphylococcus aureus through the production of organic acids. LAB isolated from Malaysian fermented food and milk products, especially fermented tapioca, contains potential probiotic candidates.     

Development of a fermented goats' milk containing Lactobacillus rhamnosus: in vivo study of health benefits

BACKGROUND: Lactobacillus rhamnosus CRL1505, a strain of goats' milk origin, is able to stimulate mucosal immunity and protect immunocompetent mice from intestinal and respiratory infections. RESULTS: In this work we developed and characterized a fermented goats' milk containing L. rhamnosus CRL1505, and we demonstrated in a model of immunosuppression in mice that the final dairy product preserves the immunomodulatory properties of the strain. L. rhamnosus CRL1505 survived the manufacturing process of fermented milk and maintained a viability of 10⁶ cfu g^?1 during storage. The fermented goats' milk was accepted by 90.48% of the panelists and was considered as having an acid taste and pleasant aroma. We also demonstrated that the developed product, used as a supplement during the repletion of immunocompromised malnourished mice, was effective in accelerating the recovery of clinical parameters altered by malnutrition and to induce increased resistance against intestinal and respiratory infections. CONCLUSION: Goats' milk fermented with L. rhamnosus CRL1505 could be manufactured as an alternative probiotic dairy product since this new food has the ability to stimulate the common mucosal immune system and to improve defenses against respiratory and intestinal infections. Copyright © 2011 Society of Chemical Industry     

Probiotic fermented milk with high content of polyphenols: Study of viability and bioaccessibility after simulated digestion

This research evaluated the functional potential of a fermented milk made with Streptococcus thermophilus, a probiotic (Bifidobacterium animalis subsp. lactis) and pomegranate juice. Ferric‐reducing antioxidant potential, Trolox equivalent antioxidant capacity, scavenging effect on 1,1‐diphenyl‐2‐picrylhydrazyl free radical, oxygen radical absorbance capacity and Folin–Ciocalteu assays were used to estimate antioxidant activity and phenolic content. Probiotic survival, release and absorption of polyphenols were evaluated using in vitro gastrointestinal digestion. The concentration of polyphenols increased during digestion from 608 mg to 1417  gallic acid equivalents (GAE) /L. About 14% of polyphenols were found on the dialysate portion. The beverage presented high viability of probiotics with high survival rate after digestion (above 7 log cfu/mL).     

Enhancement of survival of probiotic and non-probiotic lactic acid bacteria by yeasts in fermented milk under non-refrigerated conditions

The effects of yeasts on the survival of probiotic and non-probiotic lactic acid bacteria (LAB) were studied in fermented milk under non-refrigerated conditions (30 °C) with a view to develop ambient-stable fermented milk with live LAB. Five yeasts tested (Saccharomyces bayanus, Williopsis saturnus var. saturnus, Yarrowia lipolytica, Candida kefyr and Kluyveromyces marxianus) enhanced the survival of Lactobacillus bulgaricus (but not Streptococcus thermophilus) in a mixed yoghurt culture in yoghurt by ~ 10² to 10⁵-fold. Seven yeasts examined (Candida krusei, Geotrichum candidum, Pichia subpelliculosa, Kloeckera apiculata, Pichia membranifaciens, Schizosaccharomyces pombe and Y. lipolytica) improved the survival of Lactobacillus rhamnosus in fermented milk by ~ 10³ to 10⁶-fold. W. saturnus var. saturnus enhanced the survival of Lactobacillus acidophilus, L. rhamnosus (probiotic) and Lactobacillus reuteri by up to 10⁶-fold, but the same yeast failed to improve the survival of Lactobacillus johnsonii (probiotic), S. thermophilus and L. bulgaricus in fermented milk. These results provide definitive evidence that yeasts possess stability-enhancing effects on LAB and that the specific effects of yeasts on LAB stability vary with yeasts as well as with LAB. However, the molecular mechanism of such interaction of yeasts with LAB remains to be found.     

Inactivation of Lactobacillus brevis in Beer Utilizing a Combination of High‐Pressure Homogenization and Lysozyme Treatment

Inactivation of Lactobacillus brevis in beer by high‐pressure homogenization (HPH) and lysozyme addition was evaluated. The minimum inhibitory concentration of lysozyme against L. brevis was determined and found to be 100 mg.L^?1. The effects of the homogenization process on lysozyme antimicrobial activity and muramidase activity, and the microbial reduction promoted by HPH, and by HPH associated with lysozyme, were evaluated. A significant reduction in lysozyme muramidase activity was observed under 250 MPa, however, no reduction in antimicrobial activity was observed after homogenization up to 300 MPa. The HPH at 100, 140 and 150 MPa promoted 1, 3 and 6 decimal reductions in L. brevis microbial counts, respectively. The HPH and lysozyme association had an additive effect on microbial inactivation, immediately after homogenization, and the lysozyme remained active during 10 days of storage, increasing the inactivation of L. brevis up to a 6 decimal reduction. Therefore, the application of lysozyme with HPH has the potential to reduce the level of pressure required for beer processing, improving the economic costs when utilizing HPH.     

Cholesterol Lowering Activity of Ropy Fermented Milk

The effect of ropy fermented milk on serum cholesterol in rats was studied. Basic diets containing slime-forming Lactococcus lactis subsp. cremoris SBT 0495, its non slime-forming variant SBT 1275, and acidified reconstituted skim milk, with 0.5% cholesterol added, were fed to F-344 rats for 7 days. Serum cholesterol level of rats fed the ropy fermented milk were the lowest among the three treatments. The serum high-density lipoprotein (HDL) cholesterol/total cholesterol ratio of rats fed ropy fermented milk was the highest. Slime materials produced by L. lactis subsp. cremoris SBT 0495 had a beneficial effect on rat cholesterol metabolism.     

Isolation and identification of cultivable lactic acid bacteria in traditional fermented milk of Tibet in China

One hundred and seventy‐one strains of lactic acid bacteria were isolated from 44 traditional fermented milk samples from different region of Tibet. Among these samples, the concentrations of lactic acid bacteria varied from 10 ⁵ to 10 ⁹ colony forming unit/mL. The ratio of rod bacteria (71.3%) was higher than that of cocci (28.7%), and was considered to be the major population. Lactobacillus fermentum (31%) and Lactobacillus casei (28%) were the most predominant species among all these isolates. This study systematically analysed the microbial composition of traditional fermented milk in China, which may become a valuable source for further starter selection.     

Novel probiotic-fermented milk with angiotensin I-converting enzyme inhibitory peptides produced by Bifidobacterium bifidum MF 20/5

In previous research, we have demonstrated that Bifidobacterium bifidum MF 20/5 fermented milk possessed stronger angiotensin converting enzyme (ACE) inhibitory activity than other lactic acid bacteria, including Lactobacillus helveticus DSM 13137, which produces the hypotensive casokinins Ile-Pro-Pro (IPP) and Val-Pro-Pro (VPP). The aim of this study is to investigate the ACE-inhibitory peptides released in B. bifidum MF 20/5 fermented milk. The novel ACE-inhibitory peptide LVYPFP (IC₅₀ = 132 μM) is reported here for the first time. Additionally, other bioactive peptides such as the ACE-inhibitor LPLP (IC₅₀ = 703 μM), and the antioxidant VLPVPQK were identified. Moreover, the peptide and amino acid profiles, the ACE-inhibitory activity (ACEi), pH, and degree of hydrolysis of the fermented milk were determined and compared with those obtained in milk fermented by L. helveticus DSM 13137. The sequences of the major bioactive peptides present in fermented milk of B. bifidum and L. helveticus were identified and quantified. B. bifidum released a larger amount of peptides than L. helveticus but no IPP or VPP were detected in B. bifidum fermented milk. Also the lactotripeptide concentrations and ACEi were higher in L. helveticus fermented milk when the pH was maintained at 4.6. This may represent a technical advantage for B. bifidum that reduces the pH at a slow enough rate to facilitate the peptide generation without the need for pH control. Thus these findings show the potential for the use of this probiotic strain to produce fermented milk with a wider range of health benefits including reduction of blood pressure.     

West African sorghum beer fermented with Lactobacillus delbrueckii and Saccharomyces cerevisiae: shelf‐life and consumer acceptance

The fermentation profiles, shelf‐life and consumer acceptance of traditional West African sour sorghum beer (pito) fermented with pure commercial starter cultures of lactic acid bacteria (L. delbrueckii) and yeast (Saccharomyces cerevisiae) were evaluated. The beers from this ‘pure culture’ approach were compared with the spontaneous fermentation of pito wort. Lactic acid formation, pH change and extract utilisation were monitored during fermentation. Lactic acid content was used as a measure of sourness to establish the spoilage level of over‐sourness. Further, regression models relating sourness to the time the drink was kept were used to predict the shelf‐life. Consumer acceptance of the product was evaluated using a novel nine‐point hedonic scale. The pure culture and traditionally fermented beers followed similar lactic acid and fermentation profiles but strain‐specific differences were observed. Similar levels of pH, lactic acid level and extract utilisation were achieved. An improvement in shelf‐life of two days was found over traditionally fermented pito. There was no statistical difference between the two pito products for overall liking and taste. However, there was a preference for the aroma of the pure culture pito. It is suggested that the use of pure cultures will facilitate the scale‐up of pito production. © 2019 The Institute of Brewing & Distilling     

Conjugated linoleic acid content in fermented goat milk as affected by the starter culture and the presence of free linoleic acid

The effect of yoghurt starter cultures and probiotic Lactobacillus casei on the formation of conjugated linoleic acid (CLA) and microbial populations of fermented goat milk was investigated during 35 days of cold storage. The addition of hydrolysed sunflower oil as a source of free linoleic acid was investigated. The fermentation process enhanced the content of the cis9, trans11‐CLA isomer in milk, whereas the trans10, cis12‐CLA isomer was not detected in goat milk or control fermented milks. The use of both starters generated trans10, cis12‐CLA only when hydrolysed sunflower oil was supplemented. Populations of streptococci and lactobacilli were affected by the presence of hydrolysed sunflower oil.