Evaluation of the stability of Bacillus coagulans MTCC 5856 during processing and storage of functional foods

The study was aimed to find out the stability of Bacillus coagulans MTCC 5856 in functional foods during processing and storage. B. coagulans MTCC 5856 was found to be stable during baking and storage at frozen conditions of banana muffins (92% viability) and waffles (86% viability) upto 12 months. After brewing coffee at 90 °C for 2 min, there was 87% viability and retained 66% viability even after maintaining temperature at 77 °C for 4.0 h. It showed over 95% viability in chocolate fudge frosting, hot fudge toppings, peanut butter, strawberry preserve and vegetable oil at room temperature upto 12 months. It was found to be stable in apple juice upto 6 months at refrigerated condition and concentrated glucose syrup at 4 ± 3 and 25 ± 2 °C upto 24 months. B. coagulans MTCC 5856 showed promising stability during processing and storage of functional foods and could be an excellent probiotic ingredient to use in various food applications.     

Evaluation of the in vitro cholesterol-lowering activity of the probiotic strain Bacillus coagulansMTCC 5856

Emerging scientific evidence suggests that the supplementation of probiotics may help to reduce/manage blood cholesterol levels in humans. We evaluated the in vitro cholesterol-lowering activity of the probiotic strain Bacillus coagulansMTCC 5856. This probiotic de-conjugated bile salts and liberated deoxycholic acid, confirming its bile salt hydrolase activity. Furthermore, B. coagulansMTCC 5856 also significantly (P < 0.05) reduced cholesterol levels in culture media under growing (48.42%), resting (live but suspended in buffer, 36.47%) and even heat-killed (dead, 8.5%) conditions. Bacillus coagulansMTCC 5856 significantly reduced the cholesterol levels in cholesterol-rich foods, such as egg yolk (39.79%), chicken liver (45.44%) and butter (49.51%), when incubated for 24 h in conditions mimicking the in vivo environment. Bacillus coagulansMTCC 5856 also produced significant (P < 0.05) amounts of propionic acid and butyric acid while fermenting cholesterol-rich foods. The multitudinous ways by which B. coagulansMTCC 5856 reduces cholesterol levels endorses its application in functional food formulations and as a dietary ingredient for the management of hypercholesterolemia, potentially reducing the incidence of coronary heart disease and other related disabilities.     

Performance of different microbial cultures in potentially probiotic and prebiotic yoghurts from cow and goat milks

Lactobacillus acidophilus or Bifidobacterium animalis subsp. lactis Bb‐12 and green banana pulp were used in order to obtain potentially probiotic and prebiotic yoghurts, which were compared over a 45‐day storage period. Goat milk yoghurts demonstrated probiotic effects up to 45 days of storage. Cow milk yoghurts produced with B. animalis subsp. lactis Bb‐12 showed a probiotic effect reduction during the storage period (1.74 log CFU/g). The type of milk affected the yoghurts’ chemical and physicochemical properties. Sensory acceptance was also affected, where cow milk yoghurts were better accepted than goat milk ones.     

In vitro evaluation of the fermentation of added‐value agroindustrial by‐products: cactus pear (Opuntia ficus‐indica L.) peel and pineapple (Ananas comosus) peel as functional ingredients

Agroindustrial by‐products derived from fruit processing are an important source of biocompounds that can be used as functional food ingredients. The objective of this work was to evaluate cactus pear and pineapple peel flours as an alternative carbon source during fermentation using bacteria with probiotic potential. The total fibre content of both flours was over 60%, with total soluble carbohydrate content around 20%, indicating a good carbon source for lactic acid bacteria. Kinetic parameters indicate that peel flours are a suitable carbon source because the lactic acid bacteria grow (mean growth rate constant, k, values close to glucose, 1.52 h) and acidify the culture media (maximum acidification rate, V_max, approximately 1.60 pH × 10^?3 min^?1). There was no difference in prebiotic potential or prebiotic activity score for both the peel flours. Pediococcus pentosaceus performs better during fermentation. In this respect, cactus pear and pineapple peel flours can be used as functional ingredients due to their fermentable properties.     

Evaluation of the prebiotic potential of five kiwifruit cultivars after simulated gastrointestinal digestion and fermentation with human faecal bacteria

We studied the in vitro prebiotic potential of five different cultivars of kiwifruit including the green‐fleshed ‘Hayward’ and ‘Zesh004’ and the gold‐fleshed ‘Hort16A’, ‘Zesy002’ and ‘Zesy003’. The kiwifruit (25 g fresh weight equivalent) were subjected to simulated gastrointestinal digestion before fermentation for 16 h with faecal microbiota from ten individual donors. Microbial metabolites including lactate were quantified while changes in microbiome composition were determined by 16S rRNA sequencing. Lactate concentrations were highest with ‘Hayward’ (P = 0.01) and correlated with the amount of the kiwifruit fibre and polyphenols, chlorogenic acid and cryptochlorogenic acid. All the kiwifruit behaved similar to inulin in increasing the relative abundance of Bifidobacterium (P < 0.001), but unlike inulin, significantly (P < 0.001) increased the abundances of Ruminococcaceae and decreased Bacteroides. In comparison with inulin, the green‐fleshed kiwifruit selectively increased Lachnospira (P = 0.008) while the gold‐fleshed kiwi fruit increased Akkermansia (P < 0.001). These data suggest that the fibre and polyphenol content of the kiwifruit play a role in modulating gut microbial metabolism. Further clinical studies with these kiwifruit cultivars are required to confirm the potential prebiotic benefits that may be achieved by normal dietary intervention.     

A new synbiotic dairy food containing lactobionic acid and Lactobacillus casei

As well as its beneficial health properties, the incorporation of the prebiotic lactobionic acid (LBA) in fermented dairy products can provide a technological advantage due to its gelling capacities. This study aimed to develop a new functional dairy product containing LBA synthesised within the process itself by the bacterium Pseudomonas taetrolens. A probiotic Lactobacillus casei strain was introduced through a sequential fermentation system. After incubation, in the case of the most effective experimental procedure, a synbiotic fermented milk with 30 g/L of prebiotic LBA was obtained, together with an active L. casei population of 10⁹ cfu/mL and <1% lactose content.     

Cream cheese as a symbiotic food carrier using Bifidobacterium animalis Bb‐12 and Lactobacillus acidophilus La‐5 and inulin

The stability of cream cheeses as a symbiotic food carrier, through supplementation with different concentrations of probiotic bacteria Bifidobacterium animalis Bb‐12 and Lactobacillus acidophilus La‐5 and the prebiotic ingredient inulin was investigated. Physicochemical parameters, pH values, total solids, fat and protein levels and the viable counts of the starter lactic culture Streptococcus thermophilus and probiotic cultures, were carried out at 1, 15, 30 and 45 days of refrigerated storage (8 ± 0.5 °C). Different physicochemical characteristics were observed in all formulations. S. thermophilus showed good viability in all the trials (6.66–9.38 log cfu/g), whereas B. animalis remained above 6 log cfu/g in all the trials during the period evaluated. However, L. acidophilus showed an accentuated decline, registering values of 3.1 log cfu/g at the end of the period studied. The results suggested that cream cheese was an adequate food matrix for supplementation with probiotic bacteria, in particular B. animalis, and the prebiotic ingredient, showing potential as a symbiotic food.     

Probiotic properties of ice creams produced with dietary fibres from by‐products of the food industry

The objective of this study was to investigate the effects of adding dietary fibre‐rich by‐products to probiotic ice creams. For this purpose, fruit (grape, apricot, apple)‐ and grain (rice, corn, sunflower, barley)‐based by‐products were added to ice cream. The viability of Lactobacillus acidophilus (ATCC 4357D‐5) and Bifidobacterium animalis subsp. lactis (ATCC 27536) was determined with microbial analyses at 1, 15, 30 and 60 days of storage. In conclusion, it was shown that dietary fibre‐rich by‐products could be used in ice cream with improved survival of the probiotic strains without any adverse effects on the physicochemical, microbiological and sensory properties of the ice cream.     

Evaluation of in vitro probiotic potential of phytase‐producing bacterial strain as a new probiotic candidate

Sixty‐three phytase‐producing bacterial strains were isolated from natural sources, and their probiotic potential was evaluated. Of these, only fifteen strains were selected on the basis of confirmatory plate assay. Among these, five phytase‐producing strains exhibiting potent probiotic properties were identified as Bacillus cereus P1, Bacillus subtilis P6, B. subtilis P7, Pseudomonas aeruginosa P12 and P. aeruginosa P15 on the basis of morphological, biochemical and molecular characteristics. Maximum phytase activity (2.74 EU mL^?1) with potential probiotic properties, that is more than 70% survivability at pH 2.0, up to 2.0% bile salt tolerance, sporulation efficiency of more than 80% and survival in anaerobic condition (94.31%), was revealed by B. subtilis P6 as compared to the well‐established commercial probiotic strains Lactobacillus sporogenes and Lactobacillus casei. Thus, phytase‐producing B. subtilis P6 with promising probiotic features can be used in food and animal feed applications for betterment of mankind after further validation.     

Partial replacement of meat by sugar cane fibre: cooking characteristics,sensory properties of beef burgers and in vitro fermentation of sugar cane fibre

In this study, sugar cane fibre (SCF) partially replaced meat in beef burger formulations. The effects of SCF on cook yield, dimensional changes, sensory characteristics of beef burgers and in vitro gut fermentation characteristics were evaluated. Replacing beef with 1 to 5% SCF in burgers significantly increased cook yields from 13.8 ± 0.3 to 59.1 ± 0.3% due to its high water-binding capacity of 5.89 ± 0.08 g g⁻¹ and oil-binding capacity of 4.68 ± 0.03 g g⁻¹. The inclusion of SCF improved cooking properties whilst improving sensory characteristics. Burgers with 1% SCF had the highest overall acceptability. SCF was steadily fermented with a porcine faecal inoculum for up to 72 h, producing short-chain fatty acids. The characteristics of high water/oil binding and fermentability suggest that SCF has the potential to provide a range of dietary fibre benefits, and therefore deserves further study.     

Survival of Lactobacillus casei and functional characteristics of reduced sugar red beetroot yoghurt with natural sugar substitutes

In this study, probiotic gel fermentation and functional characteristics of reduced sugar yoghurt with red beetroot bioactive components and steviol glycosides from Stevia rebaudiana extracts as natural source of noncaloric sweeteners were investigated. A significant increase in Lactobacillus casei viability and fermented gel network and a decrease in syneresis were determined in yoghurt sweetened with steviol glycosides and high content of red beetroot bioactive components such as total phenolics, anthocyanins and total antioxidant capacity (DPPH and FRAP). The survival of L. casei was maintained within probiotic and therapeutic levels (>9 log₁₀ CFU/g), due to the possible prebiotic effect of steviol glycosides and fermentable fibres and phenolic compounds of red beetroot during 28 days of cold storage of the probiotic yoghurts. Colour properties (L*, a*, b*, ΔE*, C*, h*) and such sensory properties as smoothness, sweetness astringency were significantly affected. The highest organic acid content was found in red beetroot enriched yoghurt with stevia, while the most dominant organic acids were butyric, lactic, malic and citric. As a result, it was determined that fermented milk products, which are supported by phytochemical and steviol glycoside interactions, have adequate nutritional effects, high probiotic viability and acceptable sensory properties     

High-intensity ultrasound processed acerola juice containing oligosaccharides and dextran promotes Lacticaseibacillus casei NRRL B-442 growth

In the present study, a prebiotic acerola juice containing gluco-oligosaccharides and dextran was produced and processed by high-intensity ultrasound (HIUS). After a simulated in vitro digestion, the gluco-oligosaccharides and dextran maintained 90% and 80% of their initial concentration in all prebiotic’s juices. At the same time, Vitamin C and phenolic compounds concentration increased significantly by 19% and 7% (P < 0.05). After the in vitro digestion, the prebiotic juice HIUS processed by 10 min showed the highest increase in gluco-oligosaccharides and bioactive compound concentrations. The HIUS processing imparted some dextran hydrolysis and improved its fermentability by Lacticaseibacillus casei. Gluco-oligosaccharides were extensively consumed as substrate in simulated intestinal conditions, promoting the L. casei NRRL B-442 growth and production of organic acids and short-chain organic acids. The prebiotic juice HIUS processed for 6 min showed the best responses regarding the metabolism of L. casei NRRL B-442. The results showed high-intensity ultrasound processed acerola juices containing gluco-oligosaccharides and dextran as a prebiotic food.     

Inulin and probiotic concentration effects on fatty and linoleic conjugated acids in cream cheeses

The aim of this study was to evaluate the changes in the lipid profile, especially in conjugated linoleic acid (CLA), in 12 cream cheese batches with different prebiotic (inulin) and probiotic (Bifidobacterium animalis and Lactobacillus acidophilus) concentrations stored for 45 days at 8 ± 0.5 °C and analyzed every 15 days. A Central Rotational Composite Design was used to establish the prebiotic and probiotic concentrations. The effect of the pre- and probiotic concentrations on CLA contents (mg g⁻¹ of fat) was verified by plotting the response surface. The linear, quadratic, and interaction coefficients of the multiple linear regression for both prebiotic and probiotics were not significant (p > 0.05). No significant changes were observed neither in the lipid profile nor in the CLA content among the different batches, in which fatty acid composition and CLA concentrations were similar to the milk used in the preparation of the cream cheese and also to the control batch.     

Isomaltooligosaccharide increases the Lactobacillus rhamnosus viable count in Cheddar cheese

Five batches of Cheddar cheese were manufactured containing different levels of isomaltooligosaccharide (IMO) and a probiotic strain of Lactobacillus rhamnosus to study the effect of IMO on the survival of starter lactococci and probiotic micro‐organisms, on proteolytic patterns, cheese composition and sensory properties. The cheese was exposed to conditions simulating those found in the gastrointestinal tract to evaluate the survival of Lb. rhamnosus. Results demonstrated that the addition of Lb. rhamnosus and IMO did not affect the main compositional variables of Cheddar cheese. The counts of starter culture and probiotic organisms increased in cheese which contained Isomaltooligosaccharide (Batches 3, 4 and 5) more than in the control (Batches 1 and 2) during the fermentation. The probiotic counts in fresh cheese (B‐4) was 9.23 log₁₀ cfu/g which was more than one log cycle greater than in the control (B‐2). The probiotic counts remained above 8 log₁₀ cfu/g at the end of the manufacturing process. Primary proteolysis was not affected by the addition of probiotic bacteria and IMO, but the level of secondary proteolysis was slightly higher compared with the control group. The addition of IMO improved the texture and sensory quality of the cheese and the probiotic bacterium had the same effect. Under conditions that simulated the gastrointestinal tract, the probiotic bacteria in cheese (B‐4) exhibited good survival and remained above the recommended 6–7 log₁₀ cfu/g.     

A feasibility study of Lactobacillus plantarum in fruit powders after processing and storage

The aim of this study was to develop fruit powders (apple, banana and strawberry) enriched with a probiotic strain (Lactobacillus plantarum 299v). Two methodologies were proposed: (i) drying of the fruit with probiotic culture incorporated (by convection) or (ii) drying of fruit (by convection) and addition of spray‐dried probiotic culture. In the first methodology, processing caused a notable reduction in probiotic viable counts in apple, but this reduction was lower during drying of banana and strawberry. A large reduction in viable cells was also recorded during storage. In the second methodology, the survival of L. plantarum 299v was considerably higher during spray‐drying, and fruit powders with a microbial content suitable for a probiotic food (10⁸–10⁹ cfu g^?1) were obtained. The fruit powders incorporating L. plantarum 299v can be stored at 4 °C or at room temperature, for at least 3 months. This preliminary study demonstrated that fruit powders are good carriers of probiotic cultures, but the techniques used to produce them should be carefully considered.     

The effects of ratio of cow's milk to soymilk,probiotic strain and fruit concentrate on qualitative aspects of probiotic flavoured fermented drinks

The effects of proportion of cow's milk to soymilk (100:0, 75:25, 50:50, 25:75, 0:100), probiotic bacteria (Lactobacillus acidophilus LA‐5 or Lactobacillus casei L‐01) and natural fruit concentrates (strawberry, apricot, peach and pear) on quality characteristics of soy‐based probiotic drink were investigated. The parameters were analysed at the end of fermentation and during 21 days of storage at 5 ^°C. The highest viability was observed when the equal proportion of cow's milk and soymilk and L. casei was used (50:50‐CY). During chilled storage, the flavouring apricot had the highest stimulatory effect on the survival of L. casei in 50:50 treatment. In general, the treatment 50:50‐CY was realised as the best one overall.     

Antimicrobial and antidiabetic potential of synbiotic fermented milk: A functional dairy product

The current study was based on evaluation of the therapeutic potential (antimicrobial and antidiabetic) of a synbiotic fermented milk product developed by optimising levels of the prebiotic ingredients (fructooligosaccharide and isomaltooligosaccharide). The maximum growth rate (0.27 ± 0.21a cfu/mL) of Lactobacillus acidophilus ATCC^® 4357? was observed with 2.345–2.445% of fructooligosaccharide and 2.53–2.62% of isomaltooligosaccharide. Significant antimicrobial potential of the synbiotic fermented milk was observed against Escherichia coli and Staphylococcus aureus due to the addition of prebiotics. Similarly, the final product also showed 62.9%, 71.5% and 57.0% reduction in blood glucose, urea and creatinine levels, respectively, of diabetic rabbits, when supplemented with 6% of the synbiotic fermented milk. A synbiotic fermented milk product with significant therapeutic potential could be developed using fructooligosaccharide and isomaltooligosaccharide as prebiotic ingredients.     

Physicochemical characterisation of eight Spanish mulberry clones: processing and fresh market aptitudes

The objective of this study was to evaluate and compare, for the first time, white and black mulberry species in terms of their main physicochemical characteristics in eight Spanish clones. The results showed significantly different characteristics between the black and white mulberry species. Fruit weight of mulberry species ranged from 2.10 to 4.15 g, and fruit juice yield, from 41% to 62%. Fructose (~61%) and glucose (~39%) were the predominant sugars in all mulberries. The MN1 clone displayed the highest total acidity (>2.6 g L^?1), and malic acid was the most abundant organic acid (6.65 g kg^?1). Cluster analysis has allowed grouping of the clones into three groups (i) MN1 and MN2; (ii) MN3 and MN4; and (iii) MA1, MA2, MA3 and MA4. Experimental results proved that Spanish mulberries have high potential for fresh consumption (attractive dark colour in Morus nigra clones, high sugars content and intense sweetness) and industrialisation (~50% juice yield, attractive juice colour, high content of crude fibre and intense sweetness). This study is also a step towards identification of this fruit as a potential healthy food, which may also be used in food industry and also have pharmaceutical interest.     

Cello-oligosaccharides production from multi-stage enzymatic hydrolysis by lignocellulosic biomass and evaluation of prebiotic potential

Cello-oligosaccharides (COS) are very recent promising water-soluble dietary fibers, which favors several application in food, chemical and pharmaceutical industries. A three-stage enzymatic hydrolysis of cellulose extracted from agroindustrial wastes (sugarcane straw and coffee husk) with the 1,4-β-endoglucanase cellulase of glycoside hydrolase family (GH12) was developed with a processing time of 48 h. Finds evidenced that removal of products (glucose and cello-oligosaccharides) at each stage provided an increase in cello-oligosaccharides enzymatic hydrolysis in yield of approximately 65%. COS promoted growth of Lactobacillus acidophilus and Lactobacillus brevis after 24 h of fermentation, although they have presented low efficiency with Bifidobacterium sp. This multi-stage enzymatic hydrolysis has potential as a successful strategy to reduction enzyme requirements for COS production with prebiotic properties.Industrial relevanceThis work proposes an innovative technology of multi-stage enzymatic hydrolysis using the enzyme adsorbed on the cellulose surface obtained from renewable sources (sugarcane straw and coffee husk). This strategy is potential to increase the COS yield production and possibly make its cost-viable. The study is also relevant due to accessing the growth promotion of a representative selection of probiotic Lactobacillus and Bifidobacterium strains in order to evidence their prebiotic potential to be used as a possible functional ingredient for the pharmaceutical and food industries.     

Innovative production of multistrain synbiotic product using Thai-pigmented rice and rice bran oil

This study found that the isolated probiotics Lactobacillus paracasei KUKPS6201, L. acidophilus KUKPS6107, L. reuteri KUKPS6103, L. rhamnosus KUKPS6007, L. salivarius KUKPS6202, Bacillus coagulans KPSTF02 and Saccharomyces boulardii KUKPS6005 had high potential for probiotic properties. All strains had antibacterial activity and high antioxidant activity of 1.654 ± 0.017 mg Trolox mL⁻¹ probiotic extract. The selected strains could survive in a simulated gastrointestinal tract under anaerobic conditions and showed no haemolytic activity. Furthermore, the probiotic strains were strongly auto-aggregated and also showed co-aggregated ability with pathogenic bacteria. The probiotic microorganisms demonstrated high ability to adhere to Thai-pigmented rice grains. The results of analysis of these probiotics showed that Riceberry rice bran oil was an excellent prebiotic. A synbiotic product containing Thai-pigmented rice grains (cultivar Riceberry, Luem Pua and Black Jasmine) and rice bran oil was produced. After 8 weeks of storage, the viability of the probiotics in terms of multistrains was 7.36 ± 0.04 log CFU g^–1 (85.78% survival rate). Microbiological safety testing indicated that the amounts of contaminants were acceptable. This study provided the first scientific report on the feasibility of applying Thai-pigment rice, rice bran oil and mixed-culture probiotics as a novel functional synbiotic product.