Survival and Metabolic Activity of Microencapsulated Bifidobacterium longum in Stirred Yogurt

ABSTRACT: Live cells of Bifidobacterium longum, microencapsulated in K‐carrageenan, were added to stirred yogurt after fermentation (pH 4.6) and stored at 4.4 °C for 30 d. Cell enumeration indicated no decline of encapsulated cell number in yogurt samples, while there was significant reduction in nonencapsulated cell population (89.3% for B. longum B6 and 91.8% for B. longum ATCC 15708). Ion‐exchange high‐performance liquid chromatography showed comparable amounts of lactic and acetic acids in all samples, indicating little metabolic activity by bifidobacteria in experimental yogurts. Consumer sensory analysis of blackberry‐flavored yogurts revealed that samples containing encapsulated bifidobacteria had a grainy texture. Results suggested that microencapsulation protected bifidobacteria from the low pH of yogurt.     

Viability and Activity of Bifidobacteria During Refrigerated Storage of Yoghurt Containing Mangifera pajang Fibrous Polysaccharides

Abstract: The viability and activity of Bifidobacterium pseudocatenulatum G4, B. longum BB 536 and yoghurt cultures (Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus) were studied in yoghurt containing 0.75%Mangefira pajang fibrous polysaccharides (MPFP) and inulin. Growth of probiotic organisms, their proteolytic activities, the production of short chain fatty acids (lactic, acetic and propionic) and the pH of the yoghurt samples were determined during refrigerated storage at 4 °C for 28 d. B. pseudocatenulatum G4 and B. longum BB 536 showed better growth and activity in the presence of MPFP and inulin, which significantly increased the production of short chain fatty acids as well as the proteolytic activity of these organisms. Practical Application: This is the first study reported on produce synbiotic yoghurt as a functional food for specified health uses contains bifidobacteria and M. pajang fibrous polysaccharides. M. pajang fibrous polysaccharides can be used as a prebiotic particularly in dairy products to increase the viability and activity of bifidobacteria which can be used as probiotic to exert health benefit to the human by yoghurt that is considered common use in society; thus, the benefits of synbiotic yoghurt are readily accessible to the member of society.     

Growth, Viability and Activity of Bifidobacterium spp. in Skim Milk Containing Prebiotics

Growth, activity and mean doubling time (T_d) of five Bifidobacterium species in the presence of four types of prebiotics, and concentrations of acetic and lactic acids were determinted during fermentation and after 4 weeks of refrigerated storage. The T_d was lowest for B. animalis with raftilose and inulin. Retention of viability of bifidobacteria was greatest with hi‐amylose corn starch (hi‐maize). The average pH of skim milk at the end of 4 wks storage averaged 4.34 (for B. animalis with raftilose) to 4.07 (for B. longum with inulin). The highest levels of acetic acid and lactic acid were produced by B. pseudolongum with lactulose and B. infantis with lactulose, respectively.     

Viability of Bifidobacteria Strains in Yogurt with Added Oat Beta-Glucan and Corn Starch during Cold Storage

Abstract: Probiotics must be consumed at a level of 10⁷ CFU/mL for successful colonization of the gut. In yogurts containing beneficial cultures, the survival of probiotic strains can quickly decline below this critical concentration during cold storage. We hypothesized that beta-glucan would increase the viability of bifidobacteria strains in yogurt during cold storage. Yogurts were produced containing 0.44% beta-glucan (concentrated or freeze-dried) extracted from whole oat flour and/or 1.33% modified corn starch, and bifidobacteria (B. breve or B. longum) at a concentration of at least 10⁹ CFU/mL. All yogurts were stored at 4 °C. Bifidobacteria and yogurt cultures, Streptococcus thermophilus and Lactobacillus delbureckii subsp. bulgaricus, were enumerated from undisturbed aliquots before fermentation, after fermentation, and once a week for 5 wk. S. thermophilus and L. bulgaricus maintained a concentration of at least 10⁸ CFU/mL in yogurts containing concentrated or freeze-dried beta-glucan regardless of starch addition, and in the control with no added beta-glucan or starch. Similarly, the probiotic, Bifidobacterium breve, survived above a therapeutic level in all treatments. The addition of beta-glucan prolonged the survival of Bifidobacterium longum at a concentration of at least 10⁷ CFU/mL by up to 2 wk on average beyond the control. Further, the inclusion of concentrated beta-glucan in yogurt improved survival of B. longum above 10⁷ CFU/mL by 1 wk longer than did freeze-dried beta-glucan. Study results suggest that beta-glucan has a protective effect on bifidobacteria in yogurt when stressed by low-temperature storage. Practical Application: This study suggests that beta-glucan (oat fiber) may improve bifidobacteria survival in yogurt during refrigerated storage.     

Probiotic Strains as Starter Cultures Improve Angiotensin-converting Enzyme Inhibitory Activity in Soy Yogurt

Suitability of soy yogurt as a system for delivering probiotics and other bioactive compounds was assessed by fermenting soy milk using starter culture containing Lactobacillus delbrueckii ssp. bulgaricus Lb1466, Streptococcus thermophilus St1342, and probiotic organisms (Lactobacillus acidophilus LAFTI® L10, Bifidobacterium lactis LAFTI® B94, and Lactobacillus paracasei LAFTI® L26). Fermentations were terminated at different pH of 4.50, 4.55, and 4.60 and metabolic patterns of cultures (viability, proteolytic activity, organic acids production, angiotensin‐converting enzyme (ACE) inhibitory activity) were investigated during 28 d of storage at 4 °C. The presence of probiotics enhanced the growth of L. delbrueckii ssp. bulgaricus Lb1466 and S. thermophilus St134 in soy yogurt in comparison to the control produced by sole yogurt culture. In general, different termination pH had no effect (P > 0.05) on the viability of probiotic organisms that maintained good viability in soy yogurt during cold storage. Higher levels of essential growth factors in the form of peptides and amino acids in soy yogurts may have promoted the growth of L. acidophilus LAFTI® L10, B. lactis LAFTI® B94, and L. paracasei LAFTI® L26. The use of probiotic strains as a part of starter culture in soy yogurt resulted in a substantial increase in in vitro ACE inhibitory activity compared with the control produced by yogurt culture only. This improvement of ACE inhibition in soy yogurt is partly due to higher proteolytic activity of probiotics.     

Fortification of sweetened plain yogurt with insoluble dietary fiber

 Seven types of insoluble dietary fiber from five different sources (soy, rice, oat, corn and sugar beet) were used to fortify sweetened plain yogurt. Fiber addition caused an acceleration in the acidification rate of the experimental group yogurts, and most of the fortified yogurts also showed increases in their apparent viscosity. Soy I and sugar beet fibers caused a significant decrease in viscosity due to partial syneresis. In general, fiber addition led to lower overall flavor and texture scores. A grainy flavor and a gritty texture were intense in all fiber-fortified yogurts, except in those made with oat fiber. Oat II fiber gave the best results; differences with controls in terms of flavor quality scores not being statistically significant. The evolution of organic acids during the fermentation and cold storage of control and oat-II-fiber-fortified yogurts showed a similar pattern; only acetic and propionic acids were found in significantly higher amounts in the fiber- fortified product. Received: 27 June 1996/Revised version: 2 September 1996     

Probiotic viability and physico-chemical and sensory properties of plain and stirred fruit yogurts made from goat's milk

Probiotic plain and stirred fruit yogurts were made from goat's milk using bacterial cultures comprising, Lactobacillus acidophilus LA-5, Bifidobacterium animalis subsp. lactis BB-12 and Propionibacterium jensenii 702. The products were stored at 4°C for 4weeks, during which time the viability of the yogurt starter culture and probiotic bacteria was analysed weekly. P. jensenii 702 demonstrated the highest viability (10(8)cfu/g) in all types of yogurt throughout the storage period, while the viability of the bifidobacteria (～10(7)cfu/g) also remained above the minimum therapeutic level. The viability of L. acidophilus LA-5 fell below 10(6)cfu/g in yogurts, however, the addition of fruit juice appeared to support the viability of lactobacilli, with higher microorganism numbers observed in fruit yogurts than in plain yogurt throughout the shelf life. Addition of fruit juice significantly increased the syneresis, and decreased viscosity and water holding capacity of yogurts (p<0.05), and also enhanced their sensory acceptability.     

Fortification of sweetened plain yogurt with insoluble dietary fiber

 Seven types of insoluble dietary fiber from five different sources (soy, rice, oat, corn and sugar beet) were used to fortify sweetened plain yogurt. Fiber addition caused an acceleration in the acidification rate of the experimental group yogurts, and most of the fortified yogurts also showed increases in their apparent viscosity. Soy I and sugar beet fibers caused a significant decrease in viscosity due to partial syneresis. In general, fiber addition led to lower overall flavor and texture scores. A grainy flavor and a gritty texture were intense in all fiber-fortified yogurts, except in those made with oat fiber. Oat II fiber gave the best results; differences with controls in terms of flavor quality scores not being statistically significant. The evolution of organic acids during the fermentation and cold storage of control and oat-II-fiber-fortified yogurts showed a similar pattern; only acetic and propionic acids were found in significantly higher amounts in the fiber- fortified product. Received: 27 June 1996/Revised version: 2 September 1996     

Viability of microencapsulated bifidobacteria in set yogurt during refrigerated storage

Bifidobacteria are probiotic organisms that improve the microbial balance in the human gut. They can be incorporated as live cultures in fermented dairy foods, including yogurt, for transmission to humans. Because bifidobacteria are sensitive to high acidity, their viability in yogurt is limited. The objective of the present study was to investigate the effect of microencapsulation on the viability of bifidobacteria in yogurt during refrigerated storage for 30 d. Live bifidobacterial cells were encapsulated in kappa-carrageenan. Cell enumeration, determination of titratable acidity and pH, quantitation of lactic and acetic acids, and sensory evaluation (consumer test) were carried out on the yogurt samples. Microbiological results showed a decline of 78 and 70.5% in the population of Bifidobacterium longum B6 and B. longum ATCC 15708, respectively, for the treatments containing nonencapsulated cells. No difference in bifidobacterial population was observed in the encapsulated treatments. The acetic acid content in the yogurt with nonencapsulated bifidobacteria was higher than that in the plain yogurt (control) and encapsulated treatments. The increase in lactic acid content during storage was not different among the various treatments for B. longum B6, but was greater for nonencapsulated than encapsulated B. longum 15708 and the control. Consumers judged the nonencapsulated treatment as the most sour, which was likely due to the higher acetic acid content. Consumers preferred the control and nonencapsulated treatments over the encapsulated treatment. Microencapsulation appears to increase the viability of bifidobacteria in yogurt. This technique can be used to transmit bifidobacteria via fermented products provided that sensory characteristics of the product are improved or maintained.     

Use of organic acids to inactivate Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on organic fresh apples and lettuce

Abstract: This study was undertaken to investigate the antimicrobial effect of organic acids against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on whole red organic apples and lettuce. Several studies have been conducted to evaluate organic acids as sanitizers. However, no studies have compared antimicrobial effects of various organic acids on organic fresh produce, including evaluation of color changes of produce. Apples and lettuce were inoculated with a cocktail of 3 strains each of 3 foodborne pathogens provided above and treated with 1% and 2% organic acids (propionic, acetic, lactic, malic, and citric acid) for 0, 0.5, 1, 5, and 10 min. With increasing treatment time and acid concentration, organic acid treatments showed significant reduction compared to the control treatment (distilled water), and differences in antimicrobial effects between organic acids were observed. After 10 min of treatment with 1% and 2% organic acids in apples, propionic (0.92 to 2.75 log reduction), acetic (0.52 to 2.78 log reduction), lactic (1.69 to >3.42 log reduction), malic (1.48 to >3.42 log reduction), and citric acid (1.52 to >3.42 log reduction) exhibited significant (P < 0.05) antibacterial effects against 3 foodborne pathogens compared to the control treatment. In lettuce, propionic (0.93 to 1.52 log reduction), acetic (1.13 to 1.74 log reduction), lactic (1.87 to 2.54 log reduction), malic (2.32 to 2.98 log reduction), and citric acid (1.85 to 2.86 log reduction) showed significant (P < 0.05) effects compared to the control treatment. Changes in sample color subjected to organic acids treatment were not significant during storage. Practical Application: It is suggested that organic acids have a potential as sanitizers for organic fresh produce. These data may help the organic produce industry provide safe fresh produce for consumers.     

Influence of inulin and demineralised whey powder addition on the organic acid profiles of probiotic yoghurts

In this study, the effect of the different concentrations of inulin and demineralised whey (d‐whey) powder on the chemical characteristics and organic acid profiles of probiotic yoghurts were investigated. Analysis of chemical composition was performed on the first day, while pH and organic acid profiles were carried out during storage. The results indicated that different proportions of inulin and d‐whey powder significantly influenced the levels of the organic acids, produced while the duration of storage affected the orotic, pyruvic, citric, lactic and acetic acid levels. There was a direct relationship between the level of d‐whey powder and organic acid (except for butyric acid) concentration in the probiotic yoghurt.     

Survival of free and encapsulated probiotic bacteria and their effect on the sensory properties of yoghurt

The survival and effect of free and calcium-induced alginate-starch encapsulated probiotic bacteria (Lactobacillus acidophilus and Bifidobacterium lactis) on pH, exopolysaccharide production and influence on the sensory attributes of yogurt were studied over 7 weeks storage. Addition of probiotic bacteria (free or encapsulated) reduced acid development in yogurt during storage. Post-acidification in yogurt with encapsulated probiotic bacteria was slower compared to yogurt with free probiotic bacteria. More exopolysaccharides were observed in yogurts with probiotic cultures compared to those without probiotic cultures. The results showed that there was an increased survival of 2 and 1 log cell numbers of L. acidophilus and B. lactis, respectively due to protection of cells by microencapsulation. The addition of probiotic cultures either in the free or encapsulated states did not significantly affect appearance and colour, acidity, flavour and after taste of the yogurts over the storage period. There were, however, significant differences (P<0.05) in the texture (smoothness) of the yogurts. This study has shown that incorporation of free and encapsulated probiotic bacteria do not substantially alter the overall sensory characteristics of yogurts and microencapsulation helps to enhance the survival of probiotic bacteria in yogurts during storage.     

Survival,growth characteristics and bioactive potential of Lactobacillus acidophilus in a soy‐based cream cheese

BACKGROUND: Soy‐based products have received much attention lately as dairy replacers and carriers for probiotics, without the cholesterol and lactose intolerance factors. We have previously developed a soy cream cheese product and would like to evaluate its suitability as a carrier for probiotic microorganisms. Soy cream cheese is commercially uncommon, while a probiotic soy cream cheese is yet to be available in the market. RESULTS: Five strains of probiotics were screened for their α‐galactosidase activity. Lactobacillus acidophilus FTCC 0291 showed the highest α‐galactosidase‐specific activity and was incorporated into soy cream cheese for a storage study of 20 days at 25 and 4 °C. L. acidophilus FTCC 0291 in soy cream cheese at both storage temperatures maintained a viability exceeding 10⁷ CFU g^?1 over storage. Oligosaccharide and reducing sugar analyses indicated that L. acidophilus FTCC 0291 was capable of utilizing the existing reducing sugars in soymilk and concurrently hydrolyzing the oligosaccharides into simpler sugars for growth. L. acidophilus FTCC 0291 also produced organic acids, leading to decreased pH. Under low pH and high organic acid concentration, the growth of total aerobes and anaerobes was significantly (P < 0.05) suppressed compared to the control. The hydrolysis of protein in soymilk produced essential growth factors such as peptides and amino acids that may have promoted the growth of L. acidophilus FTCC 0291 and the release of bioactive peptides with in vitro angiotensin I‐converting enzyme inhibitory activity. CONCLUSION: This study showed that soy cream cheese could be used as a carrier for probiotic bacteria, with potential antihypertensive property. Copyright © 2009 Society of Chemical Industry     

Donkey Milk for Manufacture of Novel Functional Fermented Beverages

The aim of this work was to investigate on the functional features of a donkey milk probiotic berevage as a novel food. Particularly, it was to study the decrease of lactose content and the antioxidant activity of standard yogurt (YC) and probiotic yogurt (YP; Lactobacillus acidophilus, Lactobacillus casei) from donkey milk during the storage up to 30 d at 4 ºC. The evolution of lactose content using enzymatic‐spectrophotometric kits was analyzed. Antioxidant activity of yogurt was measured using 2,2’‐azino‐bis‐3‐ethylbenzothiazoline‐6‐sulfonic acid (ABTS), ferric reducing antioxidant power (FRAP), and thiol assays. Parallel consumer sensory studies were carried out as consumer test in order to gain information about the impact of these novel fermented beverages on sensory perceptions. The statistical analysis has shown significant effect of studied factors. The results showed that the lactose content gradually decreased during storage in both yogurts, reaching values of 2.36% and 2.10% in YC and YP, respectively, at 30 d (P < 0.05). During storage of both yogurt types, the antioxidant activity increased, but YP showed a higher antioxidant activity than YC. The results suggest that the antioxidant activity of yogurt samples was affected by cultures of lactic acid bacteria (LAB). We conclude that the fermented donkey milk could be configured as health and nutraceutical food, which aims to meet nutritional requirements of certain consumers groups with lactose or cow milk protein intolerance.     

Stability and viability of synbiotic microgels incorporated into liquid,Greek and frozen yogurts

The viability of Lactobacillus acidophilus when co-encapsulated with fructooligosaccharides in alginate–gelatin microgels, for incorporation into liquid, Greek, and frozen yogurts, during storage and in vitro-simulated digestion was studied. Liquid yogurt provided the highest viability for the encapsulated probiotics during storage, followed by frozen and Greek formulations when compared to free probiotics, highlighting the influence of microencapsulation, yogurt composition, and storage conditions. Addition of up to 20% of probiotic (AG) and symbiotic (AGF) microgels did not cause significant changes in the liquid and frozen yogurts’ apparent viscosity (η_ap); however, it decreased η_ap for the Greek yogurt, indicating that microgels can alter product acceptability in this case. Both AG and AGF microparticles improved viability of cells face to gastric conditions for liquid and frozen yogurts, delivering cells in the enteric stage. Summarizing, liquid yogurt was the most appropriate for probiotic viability during storage, while frozen yogurt presented better protection along digestibility.     

Survival of Lactobacillus acidophilus and Bifidobacterium animalis ssp. lactis in stirred fruit yogurts

The effect of commercial fruit preparations (mango, mixed berry, passion fruit and strawberry) on the viability of probiotic bacteria, Lactobacillus acidophilus LAFTI^® L10 and Bifidobacterium animalis ssp. lactis LAFTI^® B94 in stirred yogurts during storage (35 days) at refrigerated temperature (4 °C) was evaluated. The results showed that addition of either 5 or 10 g/100 g fruit preparations had no significant (p>0.05) effect on the viability of the two probiotic strains except on L. acidophilus LAFTI L10 yogurt with 10 g/100 g passion fruit or mixed berry. After the addition of fruit preparation, 96% of the yogurts incorporated with fruit preparation did not exhibit a greater loss in the viability of probiotic bacteria compared to plain yogurt during the storage period. A correlation between the post-storage pH in yogurts and the survival of probiotic bacteria was observed. All the yogurts, however, contained the recommended levels of (10⁶-10⁷ cfu/g) probiotic bacteria at the end of 35-day shelf life.     

Glucose oxidase: A potential option to decrease the oxidative stress in stirred probiotic yogurt

The effect of the addition of glucose oxidase as a technological option todecrease the oxidative stress in a stirred yogurt during 15 days of refrigerated storage was investigated. A significant increase in the viable population of Bifidobacteria longum (6.9–8.7 log CFU g^?1) was observed in all yogurts added with glucose oxidase and glucose. Additionally, the yogurts containing glucose oxidase and glucose presented and caused higher pH, proteolysis rate, and contents of organic acids and aroma compounds, and this effect was comparable to the control sample (without glucose oxidase and glucose). Overall, the combined effect of the glucose oxidase system was important to reduce the oxygen content during storage of the probiotic yogurt. These findings suggest that glucose oxidase is a potential option to decrease the oxidative stress in probiotic yogurts.     

Effects of a probiotic fermented milk beverage containing Lactobacillus casei strain Shirota on defecation frequency,intestinal microbiota,and the intestinal environment of healthy individuals with soft stools

The effects of drinking a fermented milk beverage that contains Lactobacillus casei strain Shirota (LcS) at 40 billion bacterial cells/bottle for 4 weeks (probiotics, 1 bottle/day) on defecation frequency, intestinal microbiota and the intestinal environment of healthy individuals with soft stools were evaluated. Thirty-four healthy adults who had soft stools were randomised into 2 groups, and the effects of a regular 4-week intake of probiotics were evaluated by a placebo-controlled, double-blind, parallel-group comparative design. Defecation frequency significantly decreased after the 4-week intake period compared with before the probiotic treatment. The stool quality significantly improved (hardened) compared to the placebo. Also, the water content of the stools was lower in the probiotic group than in the placebo group. Live LcS was recovered at 6.9 ± 1.3 and 7.2 ± 0.8 log₁₀ CFU per 1 g of stool after 2 and 4 weeks, respectively, of probiotic treatment. The number of bifidobacteria in the stools also increased significantly compared with the level before starting the probiotics. The organic acid levels (total, acetic acid, propionic acid, and butyric acid) significantly increased compared with the level before intake in both the probiotic and placebo groups, but they returned to the original levels after the end of the intake period. These results suggest that probiotic fermented milk beverage has an intestine-conditioning effect by improving the frequency of defecation and stool quality and increasing the intrinsic bifidobacteria in healthy individuals with soft stool.     

ORGANIC ACIDS PROFILE IN TOMATO JUICE BY HPLC WITH UV DETECTION

A simple method was developed to determine 10 organic acids simultaneously in tomato products using reversed‐phase high‐performance liquid chromatography column with the diode array detector set at 210 nm. After centrifugation and filtration, the samples were passed through an anion exchange resin and the organic acids were released using 0.1‐N HCl. The chromatographic separation was achieved with isocratic analysis in a 20‐min run. The method was reliable and sensitive, the coefficient of determination of the standard calibration curve is 0.9925 ≤ r² ≤ 0.9999 and the limit of detection ranged from 0.08 to 6.00 mg/kg for trans‐aconitic acid and acetic acid, respectively. The limit of quantification ranged from 0.19 to 15.18 mg/kg for trans‐aconitic and acetic acid, respectively. To establish the efficiency of the anion resin, the procedure was applied to a standard solution of a mixture of organic acids. The organic acids recovery ranged from 87.0% ± 1.9 for citramalic acid to 109.9% ± 5.2 for fumaric acid.