Effect of inulin polymerization degree on various properties of synbiotic fermented milk including Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12

The effects of inulin degree of polymerization (DP) on the viabilities of Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12 and on some parameters of fermented milk, such as microbiological, rheological, biochemical, and sensory properties, were investigated during 30 d of storage. Samples were produced using L. acidophilus La-5 and B. animalis Bb-12, along with inulin having different DP as prebiotic, and the effects of high-DP (DP ≥ 23) and low-DP (DP ≤ 10) inulin on fermented milk, were determined. The viability of both strains increased when they were used with inulin having any DP. The addition of inulin increased the consistency index of all samples. During storage, we observed an increase in lactic and acetic acid contents of samples in which high-DP inulin was used, for both strains of bacteria. Of the combinations we tested, the sample produced with L. acidophilus La-5 and high-DP inulin demonstrated the highest rheological and sensory performance as well as the best viability of probiotics.     

Short communication: Incorporation of inulin and transglutaminase in fermented goat milk containing probiotic bacteria

Goat milk is a good carrier for probiotic bacteria; however, it is difficult to produce fermented goat milk with a consistency comparable to that of fermented cow milks. It can be improved by the addition of functional stabilizers, such as inulin, or treatment with transglutaminase. The aim of this study was to determine the effect of cold storage of inulin and microbial transglutaminase on the viability of Lactobacillus acidophilus La-5 and Bifidobacterium animalis ssp. lactis Bb-12 in fermented goat milk. Microbiological analysis included the determination of the probiotic bacteria cell count in fermented milk samples, whereas physico-chemical analysis included the analysis of fat content, titratable acidity, and pH of raw, pasteurized, and fermented goat milk samples. No positive influence of inulin or microbial transglutaminase on the viability of probiotics in fermented goat's milk samples was observed. Nevertheless, the population of probiotics remained above 6 log cfu/g after 8 wk of storage at 5°C.     

Impact of inulin and okara on Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12 viability in a fermented soy product and probiotic survival under in vitro simulated gastrointestinal conditions

The effect of inulin and/or okara flour on Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12 viability in a fermented soy product (FSP) and on probiotic survival under in vitro simulated gastrointestinal conditions were investigated throughout 28 days of storage at 4 °C. Employing a 2² design, four FSP trials were produced from soymilk fermented with ABT-4 culture (La-5, Bb-12, and Streptococcus thermophilus): FSP (control); FSP-I (with inulin, 3 g/100 mL of soymilk); FSP-O (with okara, 5 g/100 mL); FSP-IO (with inulin + okara, ratio 3:5 g/100 mL). Probiotic viabilities ranged from 8 to 9 log cfu/g during the 28 days of storage, and inulin and/or okara flour did not affect the viability of La-5 and Bb-12. Bb-12 resistance to the artificial gastrointestinal juices was higher than for La-5, since the Bb-12 and La-5 populations decreased approximately 0.6 log cfu/g and 3.8 log cfu/g, respectively, throughout storage period. Even though the protective effect of inulin and/or okara flour on probiotic microorganisms was not significant, when compared to a fresh culture, the FSP matrix improved Bb-12 survival on day 1 of storage and may be considered a good vehicle for Bb-12 and could play an important role in probiotic protection against gastrointestinal juices.     

Evaluation of the effect of supplementing fermented milk with quinoa flour on probiotic activity

In this work, we investigated the effect of supplementing fermented milk with quinoa flour as an option to increase probiotic activity during fermented milk production and storage. Fermented milk products were produced with increasing concentrations of quinoa flour (0, 1, 2, or 3 g/100 g) and submitted to the following analyses at 1, 14, and 28 d of refrigerated storage: postacidification, bacterial viability, resistance of probiotics to simulated gastrointestinal (GI) conditions, and adhesion of probiotics to Caco-2 cells in vitro. The kinetics of acidification were measured during the fermentation process. The time to reach maximum acidification rate, time to reach pH 5.0, and time to reach pH 4.6 (end of fermentation) were similar for all treatments. Adding quinoa flour had no effect on fermentation time; however, it did contribute to postacidification of the fermented milk during storage. Quinoa flour did not affect counts of Bifidobacterium animalis ssp. lactis BB-12 or Lactobacillus acidophilus La-5 during storage, it did not protect the probiotic strains during simulated GI transit, and it did not have a positive effect on the adhesion of probiotic bacteria to Caco-2 cells in vitro. Additionally, the adhesion of strains to Caco-2 cells decreased during refrigerated storage of fermented milk. Although the addition of up to 3% quinoa flour had a neutral effect on probiotic activity, its incorporation to fermented milk can be recommended because it is an ingredient with high nutritive value, which may increase the appeal of the product to consumers.     

Short communication: Survival of the characteristic microbiota in probiotic fermented camel,cow, goat,and sheep milks during refrigerated storage

The objective of this study was to monitor the viability during storage of Lactobacillus acidophilus LA-5 (A), Bifidobacterium animalis ssp. lactis BB-12 (B), and Streptococcus thermophilus CHCC 742/2130 (T) in probiotic cultured dairy foods made from pasteurized camel, cow, goat, and sheep milks fermented by an ABT-type culture. The products manufactured were stored at 4°C for 42 d. Microbiological analyses were performed at weekly intervals. Streptococcus thermophilus CHCC 742/2130 was the most numerous culture component in all 4 products both at the beginning and at the end of storage. The viable counts of streptococci showed no significant decline in fermented camel milk throughout the entire storage period. The initial numbers of Lb. acidophilus LA-5 were over 2 orders of magnitude lower than those of Strep. thermophilus CHCC 742/2130. With the progress of time, a slow and constant decrease was observed in lactobacilli counts; however, the final viability percentages of this organism did not differ significantly in the probiotic fermented milks tested. The cultured dairy foods made from cow, sheep, and goat milks had comparable B. animalis ssp. lactis BB-12 counts on d 0, exceeding by approximately 0.5 log₁₀ cycle those in the camel milk-based product. No significant losses occurred in viability of bifidobacteria in fermented camel, cow, and sheep milks during 6 wk of refrigerated storage. In conclusion, all 4 varieties of milk proved to be suitable raw materials for the manufacture of ABT-type fermented dairy products that were microbiologically safe and beneficial for human consumption. It was suggested that milk from small ruminants be increasingly used to produce probiotic fermented dairy foods. The development of camel milk-based probiotic cultured milks appears to be even more promising because new markets could thus be conquered. It must be emphasized, however, that further microbiological and sensory studies, technology development activities, and market research are needed before such food products can be successfully commercialized.     

Tropical fruit pulps decreased probiotic survival to in vitro gastrointestinal stress in synbiotic soy yoghurt with okara during storage

The effect of mango and guava pulps on Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12 viability in a soy yoghurt (SY) and on probiotic survival under simulated gastrointestinal conditions were investigated throughout 28 days of storage at 4 °C. The impact of fruit pulps on SY sensory acceptability was also assessed. Three formulations were produced from soymilk fermented with ABT-4 culture (La-5, Bb-12, and Streptococcus thermophilus) and supplemented with inulin and okara: SYC (control), SYM (with mango pulp), and SYG (with guava pulp). All formulations showed probiotic viabilities ranging from 8 to 9 log cfu/g, and fruit pulps did not affect the probiotic viabilities. However, the fruit pulps decreased probiotic survival significantly to simulated gastrointestinal stress. Acceptability was higher for SYM and this difference was significant at 21 days. Therefore, the improved acceptability of SY through the addition of fruit pulps might lead to a reduction in probiotic functionality.     

Effects of Chlorella vulgaris and Arthrospira platensis addition on viability of probiotic bacteria in yogurt and its biochemical properties

It is a practice to add microalgae into plain and probiotic fermented milks in order to promote the functionality of these products via their direct health effects as well as the enhancing impact on viability of probiotic microorganisms in product and in gastrointestinal tract. In this study, the effects of addition of two species of microalgae including Arthrospira platensis and Chlorella vulgaris (seven yogurt treatments containing three concentrations for each microalgae—0.25, 0.50, and 1.00?%—and a control without microalgae) on pH, titrable acidity, and redox potential changes as well as on the viability of probiotic bacteria during fermentation and during a 28-day refrigerated storage period (5?°C) were investigated in yogurt. Also, the amounts of lactic and acetic acids at the end of fermentation were assessed. The culture composition of yogurt was ABY type, containing Lactobacillus acidophilus LA-5, Bifidobacterium lactis BB-12, Lactobacillus delbrueckii ssp. bulgaricus, and Stresptococcus themophilus. The addition of microalgae significantly (p?<?0.05) increased the viability of L. acidophilus and bifdobacteria at the end of fermentation and during the storage period. Treatments containing A. platensis had slower pH decline, faster acidity increase, longer incubation time, and greater final titrable acidity than those containing C. vulgaris and control. In treatments containing 0.5 or 1?% microalgae, the viability was almost higher than 10⁷ cfu/mL until the end of refrigerated storage.     

The potential effect of FOS and inulin upon probiotic bacterium performance in curdled milk matrices

Inulin and fructooligosaccharides were studied for their prebiotic effect upon growth/survival of probiotic bacteria and technological potential in probiotic food processing, via characterization of glycolysis, proteolysis and lipolysis in curdled milk matrices; the ultimate goal is the manufacture of synbiotic cheeses. Prebiotic compounds did not significantly affect growth/viability of all strains studied, except Lactobacillus acidophilus La-5. Proteolysis indices revealed considerable casein degradation in probiotic and synbiotic matrices inoculated with Bifidobacterium lactis B94 and Lactobacillus casei-01; lower values were achieved in those inoculated with L. acidophilus La-5, yet a synbiotic effect was apparent in NPN values. Lipolysis was not extensive over storage, irrespective of matrix type; however, interesting differences in terms of the qualitative free fatty acids profile were observed. CLA isomers, and α-linolenic and γ−linolenic acids were detected upon 15 d of ripening of all inoculated matrices. Principal component analysis was able to discriminate the various matrices according to degree of maturation, throughout the ripening period. Microbiological and biochemical parameters unfolded a very good technological potential, especially of B. lactis B94 and L. casei-01, to produce novel types of functional dairy matrices - although extrapolation to actual cheeses should still be done with care, because e.g. syneresis was not considered.     

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.     

Characteristics of carbonated fermented milk and survival of probiotic bacteria

The carbonation of pasteurised milk was evaluated as a method for improving bacterial viability in fermented milk added with probiotic bacteria (Lactobacillus acidophilus and/or Bifidobacterium bifidum). The behaviour of microorganisms during fermentation and cold storage, and the biochemical and sensory properties of the products were assessed. In AT (Streptococcus thermophilus/L. acidophilus) and ABT (S. thermophilus/L. acidophilus/B. bifidum) products, the fermentation times to decrease the pH to 5 were significantly lowered when CO₂ or lactic acid was added to milk. The higher acidity levels of carbonated (as a result of production of carbonic acid) and lactic acidified samples enhanced growth and metabolic activity of the starter during fermentation and was the reason for this reduction in incubation time. Cell counts of S. thermophilus, L. acidophilus and B. bifidum gradually decreased through the cold storage of carbonated and non-acidified fermented milk, although the counts were always higher than 10⁶ viable cells g⁻¹. The CO₂ did not exert any influence on the viability of S. thermophilus and L. acidophilus in AT fermented milk stored at 4°C but the presence of B. bifidum and CO₂ in ABT-type products was associated with lower viability of L. acidophilus during the refrigerated storage. The higher acetate concentrations of ABT products made with non-acidified milk as compared with the carbonated products could have contributed to major survival of L. acidophilus in the former. The use of milk acidified with CO₂ had no detrimental effects on the sensory properties of ABT fermented milk. Therefore, we concluded that the carbonation of pasteurised milk prior to the starter addition could be satisfactorily used to reduce the manufacture time of fermented milk.     

A probiotic soy-based innovative product as an alternative to petit-suisse cheese

The present study aimed to develop a probiotic soy-based product similar to petit-suisse cheese and to evaluate its perspectives regarding potential for consumer health benefits, sensory acceptability, and instrumental texture during storage. Three different trials were studied: MP (milk-based petit-suisse – control); MSP (mixed product with milk cream and soy); SP (soymilk-based product). The formulations were produced with an ABT culture, containing Lactobacillus acidophilus La-5, Bifidobacterium animalis Bb-12, and the starter Streptococcus thermophilus and stored at 4 °C for up to 28 days. Bb-12 viability remained always above 8 log cfu g⁻¹ for all trials, whereas viability of La-5 was satisfactory at the end of storage for MP (7.56 log cfu g⁻¹) and MSP (6.49 log cfu g⁻¹), but only up to 21 days (6.84 log cfu g⁻¹) for SP. The pH remained stable and was lower for MSP (p < 0.05), whereas instrumental hardness and gumminess increased in soy-based products (MSP and SP) and decreased in the control (MP). SP had the highest sensory score means (6.4) on day 21, being sensorially attractive to provide consumers with a functional food without dairy ingredients and with high viability of the probiotic microorganisms.     

Innovative açaí (Euterpe oleracea,Mart., Arecaceae) functional frozen dessert exhibits high probiotic viability throughout shelf-life and supplementation with inulin improves sensory acceptance

Acceptance and the probiotic viability of an innovative non-dairy frozen açaí (Euterpe oleracea, Mart.) dessert supplemented and not supplemented with inulin during a shelf-life of 84 days at ?18°C were investigated. Desserts containing and not containing probiotic Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12 and inulin were produced, resulting in 4 mixes of frozen açaí: control (M1), pro (M2), pre (M3), and synbiotic (M4). Sensory acceptability scores and probiotic counts were obtained during 84 days of storage. Inulin, a key factor for product approval, significantly (p<0.05) increased overall product acceptance of both M3 and M4, compared with M1. La-5 viability was above 7 log CFU/g for both pro and synbiotic desserts. Bb-12 counts were above 6 log CFU/g only for M4. All frozen açaí desserts were suitable for probiotic delivery and were accepted by consumers. Addition of inulin is recommended both for increasing acceptance and for the synbiotic potential.     

Probiotic fermented beverages based on acid whey

Acid whey is a byproduct of cheesemaking that is difficult to use because of its low pH and less-favorable processing properties compared with rennet whey. The aim of this study was to evaluate the qualities of fermented beverages made using acid whey. In manufacturing the beverages, we used probiotic cultures Lactobacillus acidophilus LA-5 or Bifidobacterium animalis ssp. lactis BB-12. The production process included combining pasteurized acid whey with UHT milk, unsweetened condensed milk, or skim milk powder. We introduced milk to enrich casein content and obtain a product with characteristics similar to that of fermented milk drinks. The products were stored under refrigerated conditions (5 ± 1°C) for 21 d. During storage, we assessed the beverages' physicochemical properties and organoleptic characteristics. The properties of the beverages depended on their composition, microbial culture, and storage time. Beverages containing L. acidophilus had higher acidity, which increased during storage; the acidity of samples containing B. animalis was more stable. Beverages made with skim milk powder (La1 and Bb1) had higher acetaldehyde content, but this parameter decreased in all samples during storage. The hardness of the samples did not change during storage and was highest in beverage La3, made from whey, condensed milk, and L. acidophilus. Beverage La2, made from whey, milk, condensed milk, and L. acidophilus, had the best sensory properties. The whey beverages we developed provided a good medium for the probiotic bacteria; bacteria count throughout the storage period exceeded 8 log cfu/mL, distinctly higher than the minimum therapeutic dose.     

Short communication: Viability of culture organisms in honey-enriched acidophilus-bifidus-thermophilus (ABT)-type fermented camel milk

The aim of this research was to monitor the survival during refrigerated storage of Lactobacillus acidophilus LA-5 (A), Bifidobacterium animalis ssp. lactis BB-12 (B), and Streptococcus thermophilus CHCC 742/2130 (T) in cultured dairy foods made from camel and, for comparison, cow milks supplemented with black locust (Robinia pseudoacacia L.) honey and fermented by an acidophilus-bifidus-thermophilus (ABT)-type culture. Two liters of dromedary camel milk and 2 L of cow milk were heated to 90°C and held for 10 min, then cooled to 40°C. One half of both types of milk was fortified with black locust honey at the rate of 5.0% (wt/vol), whereas the other half was devoid of honey and served as a control. The camel and cow milks with and without honey were subsequently inoculated with ABT-5 culture and were fermented at 37°C until a pH value of 4.6 was reached. Thereafter, the probiotic fermented milks were cooled to 15°C in ice water and were each separated into 18 fractions that were transferred in sterile, tightly capped centrifuge tubes. After 24 h of cooling at 8°C (d 0), the samples were stored at refrigeration temperature (4°C). Three tubes of all 4 products (i.e., fermented camel and cow milks with and without honey) were taken at each sampling time (i.e., following 0, 7, 14, 21, 28, and 35 d of storage), and the counts of characteristic microorganisms and those of certain spoilage microbes (yeasts, molds, coliforms, Escherichia coli) were enumerated. The entire experimental program was repeated twice. The results showed that addition of black locust honey at 5% to heat-treated camel and cow milks did not influence the growth and survival of starter streptococci during production and subsequent refrigerated storage of fermented ABT milks. In contrast, honey improved retention of viability of B. animalis ssp. lactis BB-12 in the camel milk-based product during storage at 4°C up to 5 wk. No spoilage organisms were detected in any of the samples tested in this study. In conclusion, supplementation of cultured dairy foods, especially those made from camel milk, with honey is recommended because honey is a healthy natural sweetener with a variety of beneficial microbiological, nutritional, and sensory properties.     

Effect of probiotic cultures on the stability of anthocyanins in blueberry yoghurts

Dependent on anthocyanin pigments, colour is one of the main quality factors of berry products. In this paper we assess the influence of probiotic bacteria on the degradation rate of anthocyanins in yoghurts with a highbush blueberry preparation during storage. Four types of yoghurts were prepared: first with the yoghurt starter culture YC-X16 only (Steptococcus thermophilus and Lactobacilllus delbrueckii subsp. bulgaricus) and three more, each with the starter culture YC-X16 and one of the three probiotic cultures: Bb-12, La-5 and LCP, respectively (Bif. animalis subsp. lactis – Bb-12; Lactobacillus. acidophilus – La-5 and Lactobacillus. paracasei subsp. paracasei – LCP).Derivatives of malvidin were the predominant anthocyanins in the tested yoghurts. Degradation of pigments occurred in accordance with the first-order reaction and its half-life time depended on the bacterial cultures. Anthocyanins in the probiotic yoghurt made with LCP culture were characterized by lower stability than those made with the other bacterial cultures. The impact of lactic bacteria or their metabolic products on the stability of anthocyanins in berry yoghurts shows that an appropriate selection of culture for production of yoghurt is recommended.     

Characterization of bifidobacteria as starters in fermented milk containing raffinose family of oligosaccharides from lupin as prebiotic

The characterization of six bifidobacteria strains used as starters in the manufacture of fermented milk, containing the raffinose family of oligosaccharides (RFOs) as prebiotics, was carried out by applying in vitro methodologies. Selection criteria were the specific growth rate (μ), generation time (T_g), titratable acidity in milk, and enzymatic activities. Strain growth and acidification rate were determined in milk with and without 2% RFOs. Different growth rates, acidification rates and enzymatic patterns were observed among tested bifidobacterial strains. Bifidobacteria with higher levels of α-galactosidase activity showed growth and acidification rate enhancement when RFOs were added compared with lactose as sole carbon source. Based on the results, Bifidobacterium lactis Bb-12 was selected for further investigation as it possessed the enzymes required to enhance growth and metabolic activity in the presence of RFOs from lupins.     

Raffinose family of oligosaccharides from lupin seeds as prebiotics: application in dairy products

The raffinose family of oligosaccharides (RFOs) isolated from lupin seeds (Lupinus albus var. Multolupa) was evaluated for bifidogenic effects during the manufacture of probiotic fermented milk. A mixed starter inoculum was composed of Bifidobacterium lactis Bb-12 and Lactobacillus acidophilus (1:1). Lupins are a rich source of RFOs that can be used as functional food ingredients. The addition of RFOs to milk increased B. lactis Bb-12 and L. acidophilus populations at the final fermentation time compared with controls. Final fermentation products are positively affected by addition of RFOs, and time of fermentation was reduced from 12 to 10 h. When RFOs were added to milk, they were preferentially used as a carbon source (57.7%) compared with lactose (23.7%) at the end of fermentation. These results suggest that the eventual choice of B. lactis Bb-12 and L. acidophilus in a mixed culture at a 1:1 ratio and addition of RFOs to produce a fermented milk product would have the advantages of rapid growth and acidificationrate and would likely increase the probiotic effect of the final functional product.     

Encapsulation in an alginate–goats’ milk–inulin matrix improves survival of probiotic Bifidobacterium in simulated gastrointestinal conditions and goats’ milk yoghurt

In this work, a new encapsulating matrix, alginate–goats’ milk–inulin, was used to encapsulate Bifidobacterium animalis subsp. lactis BB‐12. The addition of inulin resulted in capsules with a compact structure, and a higher probiotic cell count under simulated gastrointestinal conditions and in probiotic goats’ milk yoghurt during refrigerated storage. Encapsulation of the probiotic bacteria led to slower post‐acidification yoghurts. The results of this study showed that the alginate–goats’ milk–inulin matrix has potential to be used as a new encapsulation material to encapsulate probiotics for use in goats’ milk‐based probiotic fermented dairy products, avoiding the cross‐contamination caused by using capsules based on cows’ milk.     

Growth Medium for Culturing Probiotic Bacteria for Applications in Vegetarian Food Products

Vegetarian probiotic foods by definition, must be free from all animal-derived ingredients. This not only includes the product ingredients but the probiotic inoculum as well. Probiotic starter cultures are traditionally grown and stored in media containing milk or meat-derived ingredients. The presence of these ingredients makes the probiotic cell concentrates unsuitable for use in vegetarian products and thus creates the need for a growth medium free from animal-derived ingredients. This study investigated the growth of two strains of Lactobacillus acidophilus (MJLA1 and La-5), L. paracasei ssp. paracasei (LCSH1 and 01) and Bifidobacterium lactis (BDBB2 and Bb-12) in five media. Growth parameters such as maximum cell concentrations, lag phase and growth rate, were calculated using the growth curves generated by DModel, Institute of Food Research, U.K. Results showed that several media were suitable, with results comparable or better than the commonly used laboratory media MRS and RCM. Media containing 25 g/L soy peptone, yeast extract and glucose monohydrate gave the most desirable results with the strains examined.