Survival and bioactivities of selected probiotic lactobacilli in yogurt fermentation and cold storage: New insights for developing a bi-functional dairy food.

In previous work, we demonstrated that two probiotic strains, namely Lactobacillus casei PRA205 and Lactobacillus rhamnosus PRA331, produce fermented milks with potent angiotensin-converting enzyme (ACE)-inhibitory and antioxidant activities. Here, we tested these strains for the survivability and the release of antihypertensive and antioxidant peptides in yogurt fermentation and cold storage. For these purposes three yogurt batches were compared: one prepared using yogurt starters alone (Lactobacillus delbrueckii subspecies bulgaricus 1932 and Streptococcus thermophilus 99), and the remaining two containing either PRA205 or PRA331 in addition to yogurt starters. Despite the lower viable counts at the fermentation end compared to PRA331, PRA205 overcame PRA331 in survivability during refrigerated storage for 28 days, leading to viable counts (>10(8) CFU/g) higher than the minimum therapeutic threshold (10(6) CFU/g). Analyses of in vitro ACE-inhibitory and antioxidant activities of peptide fractions revealed that yogurt supplemented with PRA205 displays higher amounts of antihypertensive and antioxidant peptides than that produced with PRA331 at the end of fermentation and over storage. Two ACE-inhibitory peptides, Valine-Proline-Proline (VPP) and Isoleucine-Proline-Proline (IPP), were identified and quantified. This study demonstrated that L. casei PRA205 could be used as adjunct culture for producing bi-functional yogurt enriched in bioactive peptides and in viable cells, which bring health benefits to the host as probiotics.

Effect of probiotic and storage time of thiamine and riboflavin content in the milk drinks fermented by Lactobacillus casei KNE-1.

BACKGROUND: Fermented milk drinks are unique products due to content of Lactobacillus and Bifidobacterium that are recognized as probiotics. They are a natural component of the colon microbiota as well as commonly used probiotics in functional food. OBJECTIVES: The effects of the storage time and prebiotic type (inuline or oligofructose) were studied in banana-milk drink after fermentation by Lactobacillus casei KNE-1 on the thiamine and riboflavin concentrations. MATERIAL AND METHODS: The material for the study was fermented fruit milk drinks: banana-milk prepared in laboratory conditions and fruit milk drinks purchased in a local shop, as a comparative material. The thiamine was determined by thiochrome method and the riboflavin was determined by fluorometric method. RESULTS: The storage time after the end of the fermentation process did not increase the content of thiamine and riboflavin in fermented banana-milk drink more than the output level. The addition of oligofructose significantly affected the synthesis of thiamine by Lactobacillus casei KNE-1 irrespectively of the storage time. The storage time but not the type of prebiotic affected the riboflavin concentration. Taking into account the highest content of both vitamins, the banana-milk drink fermented by Lactobacillus casei KNE-1 should be consumed immediately or 24 days after fermentation. CONCLUSIONS: This information could be used by manufacturers for the planning of technological process. The content of thiamine and riboflavin in the fermented milk drinks is the result of the type of prebiotic, the individual bacterial strain properties as well as the storage time. These factors should be investigated to optimize the content of B vitamins in fermented milk drinks in the future.

Polycyclic aromatic hydrocarbons (PAHs) in yogurt samples.

The concentrations and distributions of major polycyclic aromatic hydrocarbons (PAHs) were determined in 20 kinds of yogurt specimens collected from Italian supermarkets using reversed phase high-performance liquid chromatography equipped with fluorescence detection. The method was validated by determination of recovery percentages, precision (repeatability) and sensitivity (limits of detection) with yogurt samples fortified at 0.25, 0.5 and 1 µg/kg concentration levels. The recovery of 13 PAHs, with the exception of naphthalene and acenaphthene, ranged from 61% to 130% and from 60% to 97% at all the levels for yogurts with low (0.1%) and high (3.9%) fat content, respectively. The method is repeatable with relative standard deviation values <20% for all analytes. The results obtained demonstrate that acenaphthene, fluorantene, phenanthrene, anthracene, fluoranthene and pyrene were found in all samples with a similar distribution, but different content when yogurts with low and high fats were compared.

Evaluation of quality of kefir from milk obtained from goats supplemented with a diet rich in bioactive compounds.

BACKGROUND: The composition of bioactive components in dairy products depends on their content in raw milk and the processing conditions. The experimental material consisted of the milk of dairy goats supplemented with 120 g d(-1) per head of false flax cake. The aim of the study was to evaluate the quality of kefir produced from goat's milk with a higher content of bioactive components resulting from supplementation of the goats' diet with false flax cake. RESULTS: The administration of false flax cake to goats had a positive effect on the fatty acid profile of the raw milk, causing an increase in the proportion of polyunsaturated fatty acids (PUFA), including conjugated linoleic acid (CLA) and n-3 fatty acids. Their increased percentage was detected in the kefir after production as well as after storage. The processing value of the harvested milk did not differ from the qualitative characteristics of milk from goats of the control group. Increasing the proportion of bioactive components in goat's milk did not result in changes in the acidity, texture, colour, flavour, aroma or consistency of the kefir obtained. CONCLUSION: Milk and kefir obtained after the administration of false flax cake to goats contain bioactive components (PUFA including CLA, n-3 and monoenic trans fatty acids) in significant amounts. Kefir from experimental goat's milk did not differ in quality from kefir made from the milk of the control group.

Reconstituted yogurt from yogurt cultured milk powder mix has better overall characteristics than reconstituted yogurt from commercial yogurt powder.

For manufacture of commercial yogurt powder, yogurt has to go through a drying process, which substantially lowers the yogurt culture counts, so the potential health benefits of the yogurt culture bacteria are reduced. Also, upon reconstitution, commercial yogurt powder does not taste like yogurt and has an off-flavor. The objective was to study the microbial, physicochemical, and sensory characteristics of reconstituted yogurt from yogurt cultured milk powder (YCMP) mix and reconstituted yogurt from commercial yogurt powder (CYP). The CYP reconstituted yogurt was the control and YCMP mix reconstituted yogurt was the treatment. Microbial and physicochemical characteristics of the CYP reconstituted yogurt and YCMP mix reconstituted yogurt were analyzed daily for the first week and then weekly for a period of 8 wk. Sensory consumer testing of CYP reconstituted yogurt and YCMP mix reconstituted yogurt was conducted with 100 consumers. At 56 d, YCMP mix reconstituted yogurt had 5 log cfu/mL higher counts of Streptococcus thermophilus than the control (CYP reconstituted yogurt). Also, Lactobacillus bulgaricus counts of YCMP mix reconstituted yogurt were 6.55 log cfu/mL at 28 d and were 5.35 log cfu/mL at 56 d, whereas the CYP reconstituted yogurt from 28 d onwards had a count of <10 cfu/mL. The YCMP mix reconstituted yogurt also had significantly higher apparent viscosity and sensory scores for appearance, color, aroma, taste, thickness, overall liking, consumer acceptability, and purchase intent than CYP reconstituted yogurt. Overall, YCMP mix reconstituted yogurt had more desirable characteristics than CYP reconstituted yogurt.

[Biotechnological optimization of nutrient composition of fermented dairy drink].

The receipt based on the results of carried out studies is substantiated and technology of the new fermented dairy drink containing whole milk and whey with inulin (Jerusalem artichoke extract) and optimizing initial mineral composition of raw material has been developed. The starters ascertaining optimal organoleptic properties of the drink have been selected. It has been established that Jerusalem artichoke and its derivatives in the form of syrups and extracts stimulate fermentative processes of technological microflora, with maximum activity observed with Jerusalem artichoke extract. Physical-chemical and microbiological characteristics of the drink have been defined during storage. The possibility to optimize the nutrient composition of fermented dairy product by means of introducing of Jerusalem artichoke extract into milk-protein base has been demonstrated. It has been calculated that consumption of 100 g of fermented dairy drink enriched with Jerusalem artichoke extract makes it possible to satisfy the physiological needs (recommended daily allowance--RDA) for babies from 0 to 3 months in vitamins B1, B2 and B6 by 25-35% and in minerals P, K, and Ca by 20, 68, 34, 26%. For adults receiving 250 g of fermented beverage meets RDA for vitamins B1, B2 and B6 by 10-19% and in the macronutrients P, K, Ca-by 25-35%. Designed fermented dairy drink supplemented with natural plant ingredient possesses increased antioxidant activity and may be recommended for mass consumption without any limitations.

Microbiological quality of milk in Tanzania: from Maasai stable to African consumer table.

In Tanzania, pastoralists such as the Maasai and small urban farmers are responsible for the country's milk production, and 95% of the national milk supply is sold without regulation. This study was conducted using hygiene checklists and milk sampling to investigate milk quality and safety at various steps throughout the milk production chain. In regions of Dar es Salaam and Lake Victoria, 196 milk samples were collected: 109 samples of raw milk, 41 samples of packed or open served heat-treated products, and 46 samples of fermented products. Samples were taken from (i) the production level (pastoralists and urban farmers), (ii) the collection level (middlemen and depots), (iii) processors (dairies), and (iv) retailers (kiosks). Samples were analyzed for hygiene criteria (total bacteria, total coliforms, Escherichia coli, and coagulase-positive staphylococci) and foodborne pathogens such as Salmonella, enterohemorrhagic E. coli O157:H7, and Listeria monocytogenes. Adequate heating of milk for drinking was determined via heat labile alkaline phosphatase and lactoperoxidase analysis. Total bacterial counts indicated that only 67% (73 of 109) of raw milk samples and 46% (19 of 41) of heat-treated samples met national Tanzanian standards. Bulk milk samples taken from the traditional milking vessels of Maasai pastoralists had the lowest total bacterial counts: ≥ 1 × 10(2) CFU/ml. Foodborne pathogens such as E. coli O157:H7 and Salmonella were isolated from 10.1% (11 of 109) of raw milk samples but were not detected in heat-treated or fermented products, and 83% of heat-treated milk samples were lactoperoxidase negative, indicating overpasteurization. Coliforms were detected in 41% (17 of 41) of processed milk samples, thus indicating a high rate of recontamination. A progressive decrease in microbial quality along the milk production chain was attributed to departures from traditional methods, inadequate milk containers, long transport distances, lack of cooling, and lack of a basic understanding of hygiene.

Effects of different fermentation parameters on quality characteristics of kefir.

The main objective of the study was to determine the effects of different fermentation parameters on kefir quality. Kefir samples were produced using kefir grains or natural kefir starter culture, and fermentation was carried out under normal or modified atmosphere (10% CO(2)) conditions. The microbiological (lactobacilli, lactococci, Lactobacillus acidophilus, Bifidobacterium spp., and yeasts), chemical (pH, lactic acid, total solids, protein, ethanol, exopolysaccharide contents), rheological, and sensory properties of kefir samples were investigated during a 21-d storage period. The use of different fermentation parameters or the choice of grain versus natural kefir starter culture did not significantly affect the content of microorganisms. Lactobacilli, lactococci, and yeast contents of kefir samples varied between 9.21 and 9.28, 9.23 and 9.29, and 4.71 and 5.53 log cfu/mL, respectively, on d 1 of storage. Contents of L. acidophilus and Bifidobacterium spp. were between 5.78 and 6.43 and between 3.19 and 6.14 log cfu/mL, respectively, during 21 d of storage. During the storage period, pH, lactic acid (%), total solids (%), protein (%), acetaldehyde, and ethanol contents of kefir samples ranged from 4.29 to 4.53, from 0.81 to 0.95%, from 7.81 to 8.21%, from 3.09 to 3.48%, from 3.8 to 23.6 mg/L, and from 76.5 to 5,147 mg/L, respectively. The exopolysaccharide contents of the samples decreased during 21 d of cold storage; the samples fermented under modified atmosphere had relatively higher exopolysaccharide contents, indicating higher potential therapeutic properties. The kefir samples exhibited non-Newtonian pseudoplastic flow behavior according to the power law model. According to the sensory results, kefir produced from natural kefir starter culture under CO(2) atmosphere had the highest overall evaluation score at d 1.

Influence of gum tragacanth on the physicochemical and rheological properties of kashk.

In this study, the physicochemical properties of a low-fat dried yogurt paste (kashk) were determined, and the effects of different concentrations (0, 0·1, 0·3 and 0·5% w/w) of gum tragacanth exudates from Astragalus gossypinus on the stability and texture of the samples were investigated by measuring amount of syneresis, turbidity, particle size distribution (PSD), flow behaviour and viscoelastic properties. The flow behaviour index was not very sensitive to the concentration of gum, while a remarkable concentration dependency of the power-law consistency coefficient and Herschel-Bulkley yield stress was observed. The initial increase in the gum concentration at 0·1 and 0·3% levels led to a higher degree of syneresis, which was related to the depletion flocculation mechanism. However, the reduced amount of syneresis in samples containing 0·5% gum tragacanth was attributed to the significant increase in viscosity of the continuous phase, which is also accompanied by trapping of the aggregated casein particles. The presence of 3% salt in the samples may have led to the neutralization of charges on the surface of gum tragacanth; consequently, the non-adsorbing behaviour of high-ionic-strength polysaccharides inhibited the formation of electrostatic protein-polysaccharide complexes. Furthermore, maximum values of polydispersity, syneresis and tan δ at high frequencies were found in samples containing 0·1% gum tragacanth.

Effect of exopolysaccharide-producing strains of Streptococcus thermophilus on technological attributes of fat-free lassi.

Sixty-four exopolysaccharide-producing thermophilic lactic acid bacteria (LAB) were isolated from traditionally made Indian fermented milk products. On the basis of morphological and biochemical tests, these isolates were identified as the species of Lactobacillus, Streptococcus, and Enterococcus genera. Initial screening for technological attributes revealed that Streptococcus thermophilus IG16 was a promising isolate, and produced both capsular and ropy polysaccharides at the concentration of 211 mg/L. Exopolysaccharide produced by IG16 was a heteropolysaccharide containing rhamnose and galactose in a ratio of 5.3:1 and had a molecular weight of 3.3 x 10(4) Da. Use of IG16 as a starter culture controlled whey separation and improved viscosity, flavor, consistency, and color and appearance of lassi. Use of IG16 resulted in lassi having optimal acidity, less syneresis, high viscosity, and better scores for flavor, consistency, and color and appearance.

Effect of fat replacers on kefir quality.

The purpose of the study was to determine the effects of fat replacers on the quality of non-fat kefir. Skim milk fortified with Dairy Lo (DL) and inulin (INU) was fermented with kefir grains to manufacture kefir. The results of compositional, microbiological, rheological and sensorial analyses were compared with whole kefir (WK) and non-fat kefir (NFK) controls. Results for dry matter, pH and lactic acid ranged between 82.4 and 109.1 g kg(-1), 4.26 and 4.40, and 7.0 and 9.2 g L(-1), respectively. Acetaldehyde and ethanol contents of samples were between 2.89 and 7.28 mg L(-1), and 151.46 and 323.89 mg L(-1), respectively. In all samples, Lactobacillus spp., Streptococcus spp. and yeast counts were between 9.1 and 9.9, 9.3 and 9.9, and 5.2 and 5.6 log cfu mL(-1), respectively. Kefir samples had non-Newtonian behaviour and pseudoplastic fluid with thixotropy. At the first day, DL had the highest apparent viscosity (3.119 Pa s) while NFK had the lowest value (1.830 Pa s). In the sensory evaluation, odour and taste scores of samples were not different. Dairy Lo and inulin could be used without any adverse effect for the production of non-fat kefir.

Quality of sour cream packaged in cups with different light barrier properties measured by fluorescence spectroscopy and sensory analysis.

UNLABELLED: In this study, photooxidation of sour cream packaged in cups with different light barrier properties was investigated. The sour cream was light exposed for 36 h with a standard fluorescent light tube, simulating storage conditions in many Norwegian grocery stores. Three different cups were evaluated: a white cup, a cup with medium light barrier (LB), and a cup with high LB. The quality of the sour cream was evaluated by sensory analysis and front face fluorescence spectroscopy. The sour cream stored in white cups became very rancid during the light exposure, with a rancid flavor score of 7.8 on a scale from 1 to 9, where 9 is the highest intensity. Cups with high LB gave best protection against the light. The sensory assessors could not distinguish between sour cream stored in cups with high LB and sour cream stored in the dark. The rancid flavor intensity for the cups with medium LB was significantly different (P < 0.05) from both the white cup and the cup with high LB. The sensory flavor intensities indicated that with 36 h of light exposure, only a cup with high LB will protect the sour cream sufficiently. The fluorescence measurements corresponded well with the sensory analysis as the photosensitizers were somewhat degraded for sour cream stored in cups with medium LB and most degraded in the white cups. For the cups with high LB, the fluorescence peaks that originated from the photosensitizers were all intact. PRACTICAL APPLICATION: Dairy products such as sour cream will develop off-odor and taste faults ("sunlight taste") in a few hours if exposed to sufficient fluorescent lightning in the grocery stores. A light barrier can be incorporated in the packaging material to protect the dairy products from the light. Our experiment showed that the incorporation of a black pigment into 1 of the 3 plastic layers in the cup for packaging of sour cream protected the sour cream from degradation when exposed to light.

Fermentation characteristics and transit tolerance of probiotic Lactobacillus casei Zhang in soymilk and bovine milk during storage.

Lactobacillus casei Zhang is a novel strain that was screened out of koumiss collected in Inner Mongolia, and our previous research showed that L. casei Zhang has health benefits such as cholesterol-reducing and immunomodulating effects. The fermentation characteristics of L. casei Zhang in soymilk and bovine milk and the transit tolerance of L. casei Zhang in fermented milk products during refrigerated storage for 28 d were assessed. A faster decrease in pH and faster growth of L. casei Zhang during fermentation were observed in soymilk compared with bovine milk at various inoculation rates, probably because of the low pH buffering capacity of soymilk. The fermented bovine milk samples had much higher final titratable acidity (TA) values (between 0.80 and 0.93%) than the soymilk samples (between 0.40 and 0.46%). Dramatic increases in TA values in the fermented soymilk samples during storage were observed, and the TA values of the fermented soymilk samples changed from <0.56% to values between 0.86 and 0.98%. On the other hand, only slight increases in TA were observed in the bovine milk samples during the 28 d of storage. The survival rates of freshly prepared cultures of L. casei Zhang in simulated gastric juice at pH 2.0 and 2.5 were 31 and 69%, respectively, and the delivery of L. casei Zhang through fermented soymilk and bovine milk significantly improved the viability of L. casei Zhang in simulated gastric transit. Lactobacillus casei Zhang showed good tolerance to simulated gastric juice and intestinal juice in the fermented soymilk and bovine milk samples, and maintained high viability (>10(8) cfu/g) during storage at 4 degrees C for 28 d. Our results indicated that both soymilk and bovine milk could serve as vehicles for delivery of probiotic L. casei Zhang, and further research is needed to elucidate the mechanism of the change in pH and TA of L. casei Zhang in fermented milk samples during fermentation and storage and to understand the difference between soy- and milk-based systems.

Rheological, sensorial, and chemopreventive properties of milk fermented with exopolysaccharide-producing lactic cultures.

The objective of this research was to evaluate the rheological, sensorial, and chemopreventive properties of milk fermented with different exopolysaccharide (EPS)-producing lactic cultures. Reconstituted skim milk (11% wt/vol) was fermented with single strains of EPS-producing and non-EPS-producing cultures. Whey that collected on the surface of undisturbed fermented milks and after cutting was measured. All EPS-producing cultures reduced the amount of whey present on the surface of the undisturbed samples, whereas only 3 out of 5 strains reduced syneresis measured after cutting. All EPS-producing cultures except a strain of Lactobacillus delbrueckii ssp. bulgaricus reduced viscoelastic moduli in fermented milk. There was a linear correlation between ropiness and smoothness. In the chemoprevention study, 140 male Fisher rats were divided into 7 groups of 20 each. Rats in 6 groups were fed diets supplemented with fermented milks each made with a single strain of EPS-producing or non-producing cultures, whereas rats in group 7 (control) were fed a diet supplemented with milk acidified with glucono-delta-lactone (GDL). All rats were injected with azoxymethane (15 mg/kg, subcutaneous) at wk 7 and 8 of age to induce tumors and fed their respective diets ad libitum throughout the study. After 30 wk of initiation, all rats were anesthetized with ether, and their intestinal tissues were isolated and washed with cold normal saline. The number and size of tumors in the colon and small intestine were recorded. Rats fed diets supplemented with fermented milk made with 2 EPS-positive and 1 EPS-negative strains had significantly lowered incidence of colon tumor and colon tumor multiplicity. Cyclooxygenase-2 enzyme activity (the enzyme implicated in colon tumor development) was significantly lower in the colon tissue of rats fed diets containing milk fermented with 4 EPS-producing and 1 non-producing cultures than that in rats fed diets supplemented with GDL-acidified milk. Different EPS-positive cultures produced fermented milks with distinct rheological characteristics and levels of ropiness. No relationship was found between rheological properties or level of ropiness of fermented milk and its chemopreventive effect.

Strain-dependent effects of probiotic lactobacilli on EAE autoimmunity.

In this study we present new data showing strain-specific differences on the effect of commercially available probiotic drinks in an EAE rat autoimmune model for multiple sclerosis. In this particular model, we conclude that these drinks do not enhance but rather suppress the disease. We suggest that conclusions on probiotics are limited to specific strains and models and not generalised. We further suggest that physiological use (normal route, normal dose, normal growth phase, specific strain or substrain/species) is studied in all cases, so as not to overwhelm (high dose) or circumvent natural immune processing.

Numbers and strains of lactobacilli in some probiotic products.

The numbers and types of Lactobacillus strains in probiotic feed or food products for sale in Europe were compared with the information stated on the product labels. The labels of all 10 products gave information on the genus and species of Lactobacillus, and seven gave information on the numbers of a specified strain(s). Lactobacilli were recovered using a recently validated method, developed as part of the European Community Project SMT4 CT98-2235 for the official control of probiotic microorganisms used as feed additives. Polymerase chain reaction (PCR)-based methods were used to assess the accuracy of labelling with regard to genus and species, and pulsed-field gel electrophoresis (PFGE) was used to identify strains. Five products were mislabelled with respect to the numbers and three with respect to species of lactobacilli. In four cases, the specified strains were not detected. Four fermented milks sold under three trademarks contained the same strain, which was named differently on each label. As safety and functionality of probiotics are strain dependent, these results demonstrated the need to control lactobacilli present in commercially probiotic human food products, not only at the species but also at the strain level, to ensure their quality and protect the consumer. The usefulness of the official methods developed for animal feed additives to identify lactobacilli in probiotic foods for humans was demonstrated.

Effect of high pressure on some properties of lactic acid bacteria present in kefir.

The aim of the work was to determine the effect of pressurization under 700 and 1000 MPa on the following: biochemical and antibacterial activity of individual strains of streptococci and lactobacilli, isolated from kefir inoculum, and of kefir microflora. After pressurization under 1000 MPa, the studied microorganisms lost their ability to inhibit the growth of the test strains, preserving the aroma-producing ability at the same time. The pressure of 1000 MPa caused also the changes in conformation of lactic acid, produced by the bacteria in skim milk culture. The changes in metabolism of the isolated bacteria, as being the result of pressurization under 700 MPa were dependent on the strain of microorganisms.