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.     

牛乳低聚肽在益生菌酸奶中的应用

研究了牛乳低聚肽在益生菌酸奶中的应用。以益生菌酸乳发酵剂与传统酸乳发酵剂作为共同发酵荆．在鲜牛奶或还原奶中添：加益生菌生长促进因子一牛乳低聚肽，进行益生菌酸奶的研制。通过实验确定了最佳配方及生产工艺。     

Effect of addition of inulin and galactooligosaccharide on the survival of microencapsulated probiotics in alginate beads coated with chitosan in simulated digestive system,yogurt and fruit juice

Galactooligosaccharides (GOS) and inulin were added during microencapsulation of Lactobacillus acidophilus 5 and Lactobacillus casei 01 in alginate beads coated with chitosan at the concentrations of 0, 0.5, 1.0 and 1.5%. Addition of prebiotics significantly (p < 0.05) increased the bead size by approximately 3.8%. The presence of GOS (0.3%) in the microencapsulation provided the best protection with only 3.1 and 2.9 logs reduction for L. acidophilus 5 and L. casei 01, respectively, after incubation in simulated gastric juice (pH 1.55), followed by simulate intestinal juice containing 0.6% bile salt. The viabilities of microencapsulated probiotics containing 1.5% GOS in commercial yogurt and orange juice were also performed at refrigerated storage for 4 weeks. In yogurt, the numbers of cells with GOS were higher than those of without GOS by approximately 1.1 and 0.4 logs for L. acidophilus 5 and L. casei 01, respectively. In orange juice, the numbers of cells with GOS were higher than those of without GOS by approximately 0.5 and 0.4 logs for L. acidophilus 5 and L. casei 01, respectively. The numbers of probiotic bacteria were maintained above the recommended therapeutic minimum (10⁷ cfu g⁻¹ or mL⁻¹ of product) throughout the storage in both products.     

Microstructural studies of probiotic bacteria-loaded alginate microcapsules using standard electron microscopy techniques and anhydrous fixation

Calcium alginate microcapsules, with or without probiotic bacteria, were studied using conventional scanning electron microscopy (CSEM), cold-stage scanning electron microscopy (cryo-SEM), and transmission electron microscopy (TEM).Each type of microscopy provided unique microstructural information about the microcapsules and entrapped bacteria. Microcapsule integrity was not preserved using conventional preparation techniques with ambient temperature SEM and TEM. Only free bacteria and remnants of capsular material remained. Cryo-SEM stabilized microcapsule microstructure. It was possible to determine the size distribution of the microcapsules, to differentiate bacteria-loaded from unloaded microcapsules, and to describe characteristics of the microcapsule material. Cryo-fracturing revealed details about the microcapsule matrix, interactions of the bacteria and the microcapsule, and void spaces around the bacteria. Details of capsule microstructure and interactions with bacteria could be observed in samples prepared using an anhydrous procedure followed by TEM.What appeared to be porosity differences existed between bacteria-loaded and non-loaded microcapsules which could affect viability when exposed to gastric conditions. Such microstructural information may be important in designing microcapsules for food use as well as carriers of other substances for delivery in the body.     

Co-encapsulation of probiotics with prebiotics on alginate matrix and its effect on viability in simulated gastric environment

Two potential probiotic strains namely Staphylococcus succinus (MAbB4) and Enterococcus fecium (FIdM3) selected from previous probiotic property studies were co-encapsulated with complementary prebiotics. Two different prebiotics selected by in vitro fermentation viz. sugarbeet and chicory were separately encapsulated with both the strains in 2 g/100 mL alginate and were tested for the efficiency in improving the viability compared to free cells under in vitro acidic conditions. Results indicated significant improvement (P < 0.05) in survival of co-encapsulated cells when exposed to acidic (pH 2.0–3.0) and bile (0.3, 0.6 and 0.8 g/100 mL) conditions. The encapsulated cells showed about 98.75–88.75% of survivability in simulated gastric environment. Viability was maintained throughout the storage period and ranges from 8.1 log cfu/mL (Colony Forming Unit) to 7.9 log cfu/mL for about a period of 30 days at 4 °C. Interestingly it is the first work to use oligosaccharides rich carbohydrate source as such in encapsulation and was found to have an improved survival rate of probiotic strains along with alginate.     

Viability and activity of bifidobacteria in yoghurt containing fructooligosaccharide during refrigerated storage

The viability of yoghurt bacteria and two commercial strains of bifidobacteria was assessed in either yoghurt containing chicory fructooligosaccharide (FOS) or without any prebiotic, during 28 days storage at 4 °C. All the products showed a decrease in the viable count of yoghurt bacteria and bifidobacteria during storage. Numbers of Lactobacillus delbrueckii ssp. bulgaricus decreased faster than those for Streptococcus thermophilus. The viability of bifidobacteria in yoghurt was affected by the strain type and the presence of FOS. Bifidobacterium animalis exhibited better stability in the yoghurt than B. longum. The recommended level of 1 million cells was exceeded for B. animalis throughout storage. The highest viable number of bifidobacteria (3.59–2.25 × 10⁷ CFU g^?1) was obtained in the product containing B. animalis and FOS. Viability of B. longum in yoghurt containing FOS remained above 10⁶ CFU g^?1 for up to 21 days, whereas this level was maintained for only 7 days for that organism in yoghurt without any prebiotic.     

低温酸奶储存过程中氨基酸含量变化研究

目的分析低温酸奶储存过程中必需氨基酸和非必需氨基酸含量的变化。方法使用氨基酸分析仪对样品中氨基酸的组成及含量进行分析。结果搅拌型酸奶中必需氨基酸含量显著高于凝固型酸奶(P0.05),其中搅拌型酸奶-Ⅱ的必需氨基酸含量最高,达到40.93 g/100 g蛋白。5种低温酸奶中谷氨酸含量最高,显著高于其他氨基酸(P0.05),其最高值为(22.16±0.04)g/100 g蛋白。半胱氨酸含量显著低于其他氨基酸(P0.05),其最低值为(0.15±0.00)g/100 g蛋白。必需氨基酸中缬氨酸和异亮氨酸含量在储存6 d时达到最大值,亮氨酸和赖氨酸含量在储存3 d时达到最大值。非必需氨基酸中丝氨酸、脯氨酸和半胱氨酸在储存6 d时含量降到最低。结论储存过程中乳酸菌发酵对必需氨基酸和非必需氨基酸的代谢产生影响,缬氨酸、赖氨酸、异亮氨酸、亮氨酸、脯氨酸、半胱氨酸和丝氨酸含量都发生显著变化。     

Survivability of probiotics encapsulated in alginate gel microbeads using a novel impinging aerosols method

Encapsulation of probiotic bacteria in cross-linked alginate beads is of major interest for improving the survivability in harsh acid and bile environment and also in food matrices. Alginate micro beads (10-40 μm) containing the probiotics Lactobacillus rhamnosus GG and Lactobacillus acidophilus NCFM were produced by a novel technique based on dual aerosols of alginate solution and CaCl₂ cross linking solution. Extruded macro beads (approximately 2 mm diameter) produced by the conventional method and micro beads produced by novel aerosols technique offered comparable protection to L. rhamnosus in high acid and bile environment. Chitosan coating of micro beads resulted in a significant increase in survival time of L. rhamnosus from 40 to 120 min in acid condition and the reduction in cell numbers was confined to 0.94 log over this time. Alginate macro beads are more effective than micro beads in protecting L. acidophilus against high acid and bile. Chitosan coating of micro beads resulted in similar protection to L. acidophilus in macro beads in acid and extended the survival time from 90 to at least 120 min. Viability of this organism in micro beads was 3.5 log after 120 min. The continuous processing capability and scale-up potential of the dual aerosol technique offers potential for an efficient encapsulation of probiotics in very small alginate micro beads below sensorial detection limits while still being able to confer effective protection in acid and bile environment.     

Development of an Oriental-style dairy product coagulated by microcapsules containing probiotics and filtrates from fermented rice

The objectives of this study were to microencapsulate both probiotics and culture filtrates by spray drying to maintain enzyme activity and probiotic viability during storage. Thus, probiotics and culture filtrates from lao-chao were microencapsulated by spray drying with various outlet air temperatures, and the milk-clotting activity, survival of probiotics and physical properties of the microcapsules were determined. The end purpose was to create easy-to-use probiotic Kou Woan Loa cultures. In the near future, manufacturing probiotic Kou Woan Loa could be carried out by simply mixing milk with 5% microcapsules and waiting for 1 h for coagulation, which would be time saving and convenient. The present study has shown that microencapsulation of Lactobacillus acidophilus BCRC 14079, Bifidobacterium longum BCRC 14605 and culture filtrates from lao-chao by spray drying could provide a good protection for both milk-clotting enzymes and probiotics. The average of microcapsules size and density was 10 µm and 1.68 g/cm³, respectively. An increase in the microencapsulation efficiency of microcapsules and lower water activity was found when the outlet air temperature was raised. However, the survival of L. acidophilus and B. longum was reduced as the outlet air temperature increased. The numbers of probiotics were maintained above the recommended therapeutic minimum (10⁷ cfu/g) throughout storage.     

Viability and α‐ and β‐galactosidase activity of Bifidobacterium breve and Lactobacillus reuteri in yoghurt products supplemented with shiitake mushroom extract during refrigerated storage

This study examined the impact of shiitake mushroom extract (ME) on the viability and α/β enzyme activity of Bifidobacterium breve ATCC 15701 and Lactobacillus reuteri DSM20016 in yoghurt during refrigerated storage. The presence of ME significantly enhanced the viability of both strains. L. reuteri DSM20016 exhibited better stability in yoghurt than B. breve ATCC 15701. L. reuteri DSM20016 showed the highest viability in yoghurt samples containing 4% ME, followed by B. breve ATCC 15701, and then control samples. Results suggest that ME can be used as a natural additive to dairy products to improve the growth and viability of L. reuteri DSM20016 and B. breve ATCC 15701 strains.     

不同体细胞数原料乳对UHT贮存期间蛋白和脂肪水解的影响

不同体细胞数(21.4×104mL-1,75.8×104mL-1,118.1×104mL-1和216.2×104mL-1)原料乳生产的4组UHT乳在37℃贮存84d,对其贮存期间的蛋白水解及脂肪水解进行研究。结果表明,4组UHT乳贮存期间的蛋白水解速率无显著性差异(P>0.05),原料乳体细胞数并未对蛋白水解造成影响;4组UHT乳贮存期间的脂肪水解速率具有显著性差异(P<0.005),原料乳体细胞数与脂肪水解速率间存在极明显的正相关(R=0.9886,P<0.05)。     

不同稳定剂对凝固型牦牛酸奶在冷藏后熟过程中品质及风味的影响

为了探讨黄原胶、果胶和羧甲基纤维素钠(CMC-Na)对牦牛酸奶在后熟过程中品质及风味的变化,实验测定了添加不同稳定剂酸奶的感官、酸度、持水性、质构,并分析了不同后熟时间酸奶的挥发性风味成分。结果表明,不同稳定剂对牦牛酸奶的感官、理化性质及质构的影响不同。添加黄原胶、果胶及CMC-Na对牦牛酸奶的感官、酸度及持水性有显著影响(p<0.05),均在一定程度上提高了酸奶的硬度,但稳定剂添加量过大时会降低酸奶的粘聚性。通过正交实验确定稳定剂最佳复配组合为:果胶0.04%、黄原胶0.02%、CMC-Na 0.03%,此时牦牛酸奶的持水力为71.52%,感官评分为42。对牦牛酸奶中的挥发性风味物质检测发现,醇类是牦牛酸奶中含量最高的物质,其次为酮类、其他类(主要为芳香类),再次为醛类。随冷藏后熟时间的延长,空白组中检出的挥发性风味物质数量变化不大;在冷藏后熟2 d时,果胶组风味的稳定效果最好,其次为CMC-Na,复合稳定剂组效果最差;至冷藏后熟7 d时,与第2 d相比,复合稳定剂组风味的稳定效果最好。     

评估超高温灭菌奶安全和质量的2个重要参数

为探究初始蛋白质与脂肪含量、贮藏温度对UHT纯牛奶包装货架期的影响,以三种UHT纯牛奶为研究对象,试验测定23、30和37 ℃贮藏过程中样品褐变指数、蛋白水解度指标。将数据集整合,根据其在预测集上的表现确定具体的输入参数,开展基于BP神经网络的UHT纯牛奶包装货架期预测。结果表明,BP神经网络模型对UHT牛奶褐变指数、蛋白水解度指标的拟合度为0.9412、0.9527,相较于传统多元线性回归模型的0.8799和0.9211,经优化隐含层神经元数的BP神经网络模型对UHT纯牛奶贮藏期间的特征指标变化预测精度更高,为不同配方UHT纯牛奶货架期的快速准确预测提供技术支持。

     

UHT乳贮存过程中V_A和V_C的变化

选择市场上3种UHT乳通过25、37℃和45℃恒温进行贮存温度加速实验比较VA和Vc含量的变化。结果表明,对于VA在3个不同的贮存温度下随贮存时间延长均不断下降,下降速率最高为45℃贮存,最高下降率达到82%,且随贮存温度升高下降越快;贮存中Vc含量变化在3种产品间无差异,且随贮存时间延长略下降,不同贮存温度间变化无差异。对于贮存中UHT乳的品质变化可以选择VA作为指标之一。     

Survival and activity of selected probiotic organisms in set-type yoghurt during cold storage

The growth and metabolism of two probiotic organisms (L. acidophilus LAFTI^® L10 and Lactobacillus casei LAFTI^® L26) and a regular yoghurt culture (L. delbrueckii ssp. bulgaricus Lb1466 and Streptococcus thermophilus St1342) were studied in yoghurt containing 0.5%, 1.0%, and 1.5% (w/v) of high amylose corn starch powder (Hi-maize^®) or inulin. Viable cell counts of probiotic organisms, their metabolites and proteolytic activities, and viscosity of the yoghurts were determined during refrigerated storage for 28 d at 4 ^oC. In the presence of inulin, cultures showed better retention of viability (8.0 log cfu g⁻¹) in comparison with that of Hi-maize, which had a reduction by one log cycle. Lower concentrations of 0.5–1.0% Hi-maize improved (P<0.05) the production of propionic acid and also increased proteolytic activity of probiotic organisms substantially. A greater release of free amino acids may have sustained better growth of the organisms in yoghurts. Supplementation with either Hi-maize or inulin increased the viscosity of probiotic yoghurts significantly (P<0.05).     

Viability of microencapsulated Bifidobacterium animalis ssp. lactis BB12 in kefir during refrigerated storage

The viability of free and immobilized cells of Bifidobacterium animalis ssp. lactis BB12 incorporated into kefir was studied for 28 days during refrigerated storage. The immobilized bifidobacteria were added directly to previously prepared kefir. Titratable acidity, pH, ethanol, fat, protein and lactose were evaluated in the kefir with bifidobacteria after the storage. The survival of the free and microencapsulated bifidobacteria was evaluated during the storage period and in simulated gastric juice. The pH of kefirs ranged from 4.3 to 4.6. Encapsulation improved significantly the survival of bifidobacteria during exposure to nisin, during the storage period and in simulated gastric juice.     

The effect of adding sour cherry pulp into yoghurt on the physicochemical properties,phenolic content and antioxidant activity during storage

This study investigated the effect of adding sour cherry pulp into yoghurt on its physicochemical properties, phenolic content, antioxidant activity and sensory characteristics. Sour cherry pulp was added at 0%, 8%, 12% and 16% into the yoghurt, and measures were checked through 14 days cold storage. The increasing sour cherry pulp concentration in yoghurt resulted in increasing pH and whey separation, whereas the values of the other parameters, total solid, fat, protein, ash, titratable acidity and viscosity decreased. During storage, total phenolic contents and antioxidant activity in yoghurts ranged from 20 to 81 μg gallic acid equivalent per mg of sample and from 48% to 86%, respectively.     

Protein lactosylation in UHT milk during storage measured by Liquid Chromatography–Mass Spectrometry and quantification of furosine

The initial stage of the Maillard reaction, protein lactosylation, occurs during heat treatment of milk and continues during subsequent storage. We compared the initial lactosylation as well as the rate of lactosylation of milk proteins during storage in UHT milk subjected to direct or indirect heat treatment using liquid chromatography (LC) coupled with electrospray injection mass spectrometry (ESI‐MS). Furosine content was used as an overall marker to allow for a quantitative correlation of lactosylation measured by LC‐ESI‐MS in the UHT milks. Protein lactosylation increased during the storage period of 6 months at 20 °C. Both the initial extent and the rate of lactosylation positively correlated with the number of lysine residues in the different proteins. An exponential or linear correlation with furosine concentration could be established for major and minor lactosylated proteins, respectively.     

Viability of Lactobacillus acidophilus in rice bran‐enriched stirred yoghurt and the physicochemical and sensory characteristics of product during refrigerated storage

This study was aimed at investigating the fortification of probiotic yoghurt with rice bran to increase nutritional properties of the product. The different levels of rice bran (0.3%, 0.6%, 0.9% and 1.2%) were incorporated into milk. The yoghurt samples were produced after pasteurisation, addition of starter culture and 1% Lactobacillus acidophilus suspension (6 × 10⁸ CFU mL^?1) and incubation. During sample storage in refrigerator, the viability of L. acidophilus, viscosity and physicochemical and sensory properties of product were investigated. Rice bran significantly increased the viability of L. acidophilus (P < 0.05). In addition, all probiotic yoghurts incorporating rice bran indicated higher viscosity and acidity and lower pH and syneresis compared to plain yoghurts. Furthermore, increments in rice bran incorporation levels resulted in a reduction in consumers' sample preferences. In general, the addition of rice bran at a suitable level could increase L. acidophilus viability and improve quality attributes of yoghurt.     

Release kinetics of antioxidant compounds from Hibiscus sabdariffa L. encapsulated in gelatin beads and coated with sodium alginate

Gelatin beads containing a concentrated extract of Roselle (Hibiscus sabdariffa L.) calyx rich in polyphenolic compounds were coated with sodium alginate and ionotropically gelled using CaCl₂. Single‐coated beads and double‐coated beads were obtained by this technique, and the release pattern of the loaded extract was evaluated. As a result, release pattern of these compounds fits properly to a first–order Weibull distribution equation. The release rate constant decreased linearly with the number of alginate coats and with the increase in immersion time in CaCl₂ and the Lag period increased significantly with the number of alginate coats. The release of H. sabdariffa's polyphenols can be well controlled manipulating the number of alginate coats and the immersion time in a CaCl₂ solution, allowing not only to control the gastrointestinal segment where they could be released but also to control the release rate with the certainty that the initial concentration will be completely released showing a highly significant antioxidant activity as well.