Probiotic Food Products Research Articles

Effect of cooling rate, freeze-drying, and storage on survival of free and immobilized Lactobacillus casei ATCC 393

The optimum conditions for freeze-drying of free or immobilized Lactobacillus casei ATCC 393 on apple pieces and casein were investigated. Initially, the effect of cooling rate on cell viability of free cells was assessed. Application of 5 °C/min resulted in significant higher levels (10.8 log cfu/g) after 15 days of storage at 4 °C, compared to other rates tested. In a second step, the effect of lyoprotectants (solutions of sugars, starch, polyols, nitrogenous compounds and whey) on viable cell counts of both free and immobilized cells was evaluated. Lactose and monosodium glutamate were the most efficient agents for immobilized cells on apple pieces and casein, accounting for 9.2 log cfu/g and 9.7 log cfu/g, respectively. In all cases, concentrations above 6 log cfu/g (minimum required levels for conferring a probiotic effect) were monitored after storage at 4 °C for up to 12 months, suggesting the suitability of the technology for the production of probiotic foods.

Performances of new isolates of Bifidobacterium on fermentation of soymilk.

Growth and metabolic activity of several new, human origin isolates of Bifidobacterium strains were investigated. All tested bifidobacteria strains were grown well on the native soymilk medium without any additional nutrients. The fermentation processes cultured with initial cell concentrations in 10⁵ -10⁷ cfu/ml resulted in 10⁸ cfu/ml after 8-12 h of incubation in soymilk, and were kept viable up to the end of fermentation (48 h). Volumetric productivities of B. bifidum B3.2, B. bifidum B7.1 and B. breve B9.14 were 1.6 × 10¹⁰ cfu/L.h, 4.5 × 10¹⁰ cfu/L.h and 7.6 × 10⁹ cfu/L.h, respectively, whereas these values of B. lactis Bb-12 and B. longum Bb-46 probiotic strains were 2.7 × 10⁹ cfu/L.h and 1.0 x 10¹⁰ cfu/L.h. The α-galactosidase activities were also detected in the intracellular fraction of the disrupted cells. Productions of lactic and acetic acids were in the range of 23-60 mmol/L and 2.4-5.6 mmol/L, respectively. Molar ratios of acetate to lactate in all tested strains varied from 0.05-0.1 that are very promising for further technological development of probiotic fermented soy-based food products.

Detection and Identification of Probiotic Lactobacillus plantarum Strains by Multiplex PCR Using RAPD-Derived Primers.

Lactobacillus plantarum 2035 and Lactobacillus plantarum ACA-DC 2640 are two lactic acid bacteria (LAB) strains that have been isolated from Feta cheese. Both display significant potential for the production of novel probiotic food products. The aim of the present study was the development of an accurate and efficient method for the molecular detection and identification of the above strains in a single reaction. A multiplex PCR assay was designed for each strain, based on specific primers derived from Random Amplified Polymorphic DNA (RAPD) Sequenced Characterized Amplified Region (SCAR) analysis. The specificity of the assay was tested with a total of 23 different LAB strains, for L. plantarum 2035 and L. plantarum ACA-DC 2640. The multiplex PCR assay was also successfully applied for the detection of the above cultures in yogurt samples prepared in our lab. The proposed methodology may be applied for monitoring the presence of these strains in food products, thus evaluating their probiotic character. Moreover, our strategy may be adapted for other novel LAB strains with probiotic potential, thus providing a powerful tool for molecular discrimination that could be invaluable to the food industry.

Use of anaerobic green fluorescent protein versus green fluorescent protein as reporter in lactic acid bacteria

Lactic acid bacteria (LAB) are commonly used in the production of fermented and probiotic foods. Development of molecular tools to discriminate the strains of interest from the endogenous microbiota in complex environments like food or gut is of high interest. Green fluorescent protein (GFP)-like chromophores strictly requires molecular oxygen for maturation of fluorescence, which restrict the study of microorganisms in low-oxygen environments. In this work, we have developed a noninvasive cyan-green fluorescent based reporter system for real-time tracking of LAB that is functional under anoxic conditions. The evoglow-Pp1 was cloned downstream from the promoters D-alanyl-D-alanine carboxypeptidase and elongation factor Tu of Lactobacillus reuteri CECT925 using pNZ8048 and downstream of the lactococcal P1 promoter using pT1NX. The classical gfp was also cloned in pT1NX. These recombinant expression vectors were electroporated into Lactococccus, Lactobacillus, and Enterococcus strains with biotechnological and/or probiotic interests to assess and compare their functionality under different conditions of oxygen and pH. The expression was analyzed by imaging and fluorometric methods as well as by flow cytometry. We demonstrate that reporter systems pNZ:TuR-aFP and pT1-aFP are two versatile molecular markers for monitoring LAB in food and fecal environments without the potential problems caused by oxygen and pH limitations, which could be exploited for in vivo studies. Production of the fluorescent protein did not disturb any important physiological properties of the parental strains, such as growth rate, reuterin, or bacteriocin production.

Carnobacterium divergensBacteremia in Woman

<i>Carnobacterium divergens</i>Bacteremia in Woman

Secure/Safe, Healthy, Intact and Halal of Mangosteen Small Medium Enterprise

IbM of Mangosteen UKM group is one of Agricultural Polytechnic State of Payakumbuh’s proposal program which has aim to increase the productivity value and competitiveness also to dig up the mangosteen local food potention in line with functional food paradigm development. Prebiotic and probiotic mangosteen food technology as the food quality for life safety is one of future plan effort to dig up local food potention that is Safe, Healthy, Intact and Halal (Allowed in Islamic way). By this IbM program is expected to change people’s mindset so they have willingness, be able and can to process the mangosteen to be product which about to add economical value and various functional food using technology in processing. Mangosteen (Garcinia mangostana.L) aka Queen of tropical fruit is a fruit that has many superiorities compared to another fruits like natural colour source that can be used as food coloring, also to be utilized as anti-oxidant, anti-diarrhea, anti-cancer and others. The Methods done are field survey, Focus Group Discussion (FGD), Demonstration, illumination, training that focused to the products and methods with guidance system also effective monitoring in creating functional food products and always to be preferred. In its implementation Mitra’s group in society contributing as the business doer in IbM activity. College has role giving innovation and technology transfer and evaluation and guidance to ‘Mitra’. The result expected is prebiotic and probiotic mangosteen food products like mangosteen yogurt, Mangosteen syrup, Mangosteen-dried extract and mangosteen cake, service, journal, and book.

Improving survival of probiotic bacteria using bacterial poly-γ-glutamic acid

A major hurdle in producing a useful probiotic food product is bacterial survival during storage and ingestion. The aim of this study was to test the effect of γ-PGA immobilisation on the survival of probiotic bacteria when stored in acidic fruit juice. Fruit juices provide an alternative means of probiotic delivery, especially to lactose intolerant individuals. In addition, the survival of γ-PGA-immobilised cells in simulated gastric juice was also assessed. Bifidobacteria strains (Bifidobacteria longum, Bifidobacteria breve), immobilised on 2.5% γ-PGA, survived significantly better (P<0.05) in orange and pomegranate juice for 39 and 11days respectively, compared to free cells. However, cells survived significantly better (P<0.05) when stored in orange juice compared to pomegranate juice. Moreover, both strains, when protected with 2.5% γ-PGA, survived in simulated gastric juice (pH2.0) with a marginal reduction (<0.47 log CFU/ml) or no significant reduction in viable cells after 4h, whereas free cells died within 2h. In conclusion, this research indicates that γ-PGA can be used to protect Bifidobacteria cells in fruit juice, and could also help improve the survival of cells as they pass through the harsh conditions of the gastrointestinal tract (GIT). Following our previous report on the use of γ-PGA as a cryoprotectant for probiotic bacteria, this research further suggests that γ-PGA could be used to improve probiotic survival during the various stages of preparation, storage and ingestion of probiotic cells.

Effect of electroporation on bioconversion of isoflavones and probiotic properties of parents and subsequent passages of Bifidobacterium longum.

This study aimed to evaluate the effects of electroporation on growth, bioconversion of isoflavones, and probiotic properties of parent organisms and subsequent passages of Bifidobacterium longum FTDC 8643. Electroporation with the strength of electric field at 7.5kVcm(-1) for 3.5ms was applied on B. longum FTDC 8643. The viability of B. longum FTDC 8643 increased significantly upon treatment with electroporation. Such treatment also enhanced the intracellular and extracellular β-glucosidase activity, leading to enhanced production of bioactive isoflavone aglycones in mannitol-soymilk (P < 0.05). In addition, these treated cells also exhibited better tolerance toward acidic (pH2 and pH3) and intestinal bile salt condition compared to the control (P < 0.05). The electroporated cell also possessed better adhesion ability and antimicrobial activity (P < 0.05). However, all these positive effects were only prevalent in the parent cells and were not observed in their subsequent passages of electroporated cells. Our results suggested that electroporation could enhance the bioactive and probiotic potentials of parent cells of B. longum FTDC 8643 and could be used in the production of probiotic foods with enhanced bioactivity.

In vitro evaluation of the probiotic and functional potential of Lactobacillus strains isolated from fermented food and human intestine

This study aims to evaluate the functional and probiotic characteristics of eight indigenous Lactobacillus strains in vitro. The selected lactobacilli include strains of Lactobacillus casei subsp. casei, Lactobacillus salivarius subsp. salicinius, Lactobacillus fermentum, Lactobacillus plantarum, Lactobacillus delbrueckii subsp. lactis, Lactobacillus delbrueckii subsp. bulgaricus, and Lactobacillus rhamnosus. All strains tolerated both pH 2 for 3 h and 1% bile salt for 24 h. The strains CICC 23174 and CGMCC 1.557 were the most adhesive strains producing the highest quantity of EPS. Although a wide variation in the ability of the eight strains to deplete cholesterol and nitrite, antagonize pathogens, scavenge free radical, and stimulate innate immune response were observed, the strains CICC 23174 and CGMCC 1.557 showed the widest range of these useful traits. Taken together, the strains CICC 23174 and CGMCC 1.557 exhibited the best probiotic properties with the potential for use in the production of probiotic fermented foods.

Lactobacillus helveticus: importance in food and health.

Lactobacillus helveticus: importance in food and health.

Toward Probiotict Food Product from Meat Through Isolation and Identification Lactic Acid Bacteria As Probiotic Culture Stater

Probiotic food products of meat can provide extensive benefits, to increase the shelf life and nutritional value also improve the taste. The use of lactic acid bacteria culture (LAB) derived from the isolation of the meat and the addition of probiotic cultures (Lactobacilli and Bifidobacteria) in fermented sausage processing is expected to produce a probiotic sausage products with nutritional value, and better shelf life and improve health. This study aimed to isolate and identify lactic acid bacteria (LAB) of meat as a starter culture fermented sausages. The parameters observed were gram test, catalase, motility, gas production, type of fermentation, growth at various temperatures and pH. The results were obtained 28 isolates, 17 isolates were able to produce acid and 8 of them are lactic acid bacteria (LAB) with the characteristics of gram-positive, catalase negative, not motile, produces gas, are hetero and homo fermentative, optimum growth temperature of 300C and a few of them are able to grow on pH 3.5. Lactic acid bacteria that able to be combined with probiotics as sausage starter culture to the probiotic food products of meat.

Comparison of rpoA and pheS Gene Sequencing to 16S rRNA Gene Sequencing in Identification and Phylogenetic Analysis of LAB from Probiotic Food Products and Supplements

An investigation of the relevance of sequenced protein-coding genes in comparison with 16S rRNA gene sequences to correctly delineate and discriminate between closely related Lactobacillus isolates is presented. These reportedly probiotic Lactobacillus strains were obtained from probiotic supplements and food products. Probiotic Lactobacillus species have been found to have several potential health benefits and technological properties which may be strain-dependent. It was deemed necessary to investigate molecular techniques for the identification and discrimination between closely related isolates. The 16S rRNA, pheS, and rpoA genes were amplified in a polymerase chain reaction using the oligonucleotide pairs 27F/1492R, pheS-21-F/pheS-23-R, and rpoA-21-F/rpoA-23-R. The PCR amplicons were separated using gel agarose electrophoresis and observed by the Bio-Rad Gel DocTM XR+ Imaging system transilluminator. Sequencing of the PCR amplicons was done in an ABI PRISMTM 3100 genetic analyzer. MEGA 5.05 software was employed in doing the phylogenetic analysis. Identification by 16S rRNA gene sequencing was confirmed by pheS and rpoA gene sequencing. Subsequent phylogenetic analysis and concatenation showed that interspecies and intraspecies degrees of diversity and similarity were better enabled by pheS and rpoA primers. Overall, findings revealed cases of inaccurate identification of Lactobacillus strains in probiotic supplements on the market, highlighting the relevance of the multigenic approach in more accurately identifying Lactobacillus strains.Supplemental materials are available for this article; see the publisher's online edition of Food Biotechnology to view the supplemental file.

Probiotic strains Lactobacillus plantarum 299V and Lactobacillus rhamnosus GG as starter cultures for fermented sausages

Probiotic food products are a fast growing area. Although probiotic strains are currently used in dairy products, their commercial application in fermented meat products is not yet common. The aim of this study was to assess the competitiveness of two probiotic Lactobacillus strains (Lactobacillus plantarum 299V and Lactobacillus rhamnosus GG) during the manufacture of Spanish fermented sausages and their effect on the hygienic and sensory qualities of the final products. The inoculated strains were successfully monitored by Randomly Amplified Polymorphic DNA (RAPD)-PCR. Both strains prevented the growth of Enterobacteriaceae throughout the entire ripening process. L. rhamnosus GG and L. plantarum 299V at high inoculum (ca. 107 CFU/g) produced a sharp decrease of pH values and low growth of Gram-positive Catalase-positive Cocci (GCC+), leading to a negative effect on the sensory attributes evaluated. Nevertheless, L. plantarum 299V inoculated at 105 CFU/g achieved and maintained high counts until the end of ripening and storage (ca.108 CFU/g), co-dominating (60%) with the endogenous microbiota, producing functional sausages with a satisfactory overall sensory quality. No major differences in physico-chemical parameters or sensory attributes were recorded when compared to spontaneously fermented sausages, thus adding further value to this type of meat product as a probiotic vehicle.

Fermentation of vegetable juice mixture by probiotic lactic acid bacteria

Probiotics have several beneficial effects on human health. Probiotic products available on the market today are usually dairy based and cannot be consumed by individuals who suffer lactose intolerance and allergies due to milk proteins. Fruits and vegetables offer healthy alternatives for the production of probiotic foods due to their large distribution and nutritive value. While searching for alternative food matrices, the suitability of bitter gourd (Momordica charantia), bottle gourd (Lagenaria siceraria) and carrot (Daucus carota) juice as a raw material taken for the production of probiotic drinks by lactic acid bacteria (Lactobacillus acidophilus NCDC 11, Lactobacillus plantarum NCDC 414, Pediococcus pantosaceus MTCC 2819) was investigated. The vegetable juice was fermented at 30°C for up to 72 h and changes in the microbial population, pH, sugars and titrable acidity were observed during the fermentation period. The viable cell counts reached up to 8 log CFU/ml, from an initial count of 6 log cfu/ml, and pH decreased from an initial value of 5.2 to below 3.5. The viability of all strains was also determined during the storage time at 4°C within 4 weeks. The viable cell counts of the lactic cultures in the fermented vegetable juice gradually decreased during cold storage. The vegetable juice proved to be a suitable media for the production of a fermented probiotic drink and can serve as a healthy beverage for vegetarians, particularly diabetics.

Survival of alginate-prebiotic microencapsulated Lactobacillus acidophilus in mayonnaise sauce

Background: Nowadays, Microencapsulation of probiotic bacteria is the newest method for increasing the survival ability of probiotics in Food products. The Lactobacillus acidophilus as a probiotic bacterium has more beneficial effects and the low survival of this bacterium under food processing conditions and adverse environment have been studied by many researchers. OBJECTIVES: In this study, Lactobacillus acidophilus was added to mayonnaise sauce as either free cells or encapsulated and the survival of probiotic was evaluated during storage for 30 days at 4oC. METHODS: The emulsion method was performed for the microencapsulation process of Lactobacillus acidophilus. A morphological characteristic of capsules was indicated by optical microscope and scanning electron microscope. RESULTS: The microencapsulation with calcium alginate-resistant starch significantly affected (p

Isolation of Pediococcus acidilacticiKp10 with ability to secrete bacteriocin-like inhibitory substance from milk products for applications in food industry

BackgroundLactic acid bacteria (LAB) can be isolated from traditional milk products. LAB that secrete substances that inhibit pathogenic bacteria and are resistant to acid, bile, and pepsin but not vancomycin may have potential in food applications.ResultsLAB isolated from a range of traditional fermented products were screened for the production of bacteriocin-like inhibitory substances. A total of 222 LAB strains were isolated from fermented milk products in the form of fresh curds, dried curds, and ghara (a traditional flavor enhancer prepared from whey), and fermented cocoa bean. Eleven LAB isolates that produced antimicrobial substances were identified as Lactococcus lactis, Lactobacillus plantarum, and Pediococcus acidilactici strains by biochemical methods and 16S rDNA gene sequencing. Of these, the cell-free supernatant of Kp10 (P. acidilactici) most strongly inhibited Listeria monocytogenes. Further analysis identified the antimicrobial substance produced by Kp10 as proteinaceous in nature and active over a wide pH range. Kp10 (P. acidilactici) was found to be catalase-negative, able to produce β-galactosidase, resistant to bile salts (0.3%) and acidic conditions (pH 3), and susceptible to most antibiotics.ConclusionTraditionally prepared fermented milk products are good sources of LAB with characteristics suitable for industrial applications. The isolate Kp10 (P. acidilactici) shows potential for the production of probiotic and functional foods.

Effect of ultrasound on bioconversion of isoflavones and probiotic properties of parent organisms and subsequent passages of Lactobacillus

This study evaluated the effects of ultrasound (60 W; 3 min) on growth, bioconversion of isoflavones, and probiotic properties of parent organisms and subsequent passages of Lactobacillus casei FTDC 2113. Ultrasound significantly enhanced (P < 0.05) the cell growth of the parent organisms in mannitol–soymilk fermented at 37 °C over 24 h. Such treatment also enhanced the intracellular and extracellular β-glucosidase activity, leading to increased bioconversion of isoflavones in mannitol–soymilk (P < 0.05). In addition, treated cells also exhibited better tolerance toward acidic conditions (pH 2 and pH 3) and intestinal bile salts compared to the control (P < 0.05). Ultrasound also promoted cell adhesion ability and antimicrobial activity (P < 0.05). All these positive effects were only prevalent in the parent cells without inheritance by first, second and third passage of cells. Our results suggest that ultrasound could enhance the bioactive and probiotic potentials of parent cells of L. casei FTDC 2113, and could be used in the production of probiotic foods with enhanced bioactivity.

Antioxidant properties of different milk fermented with lactic acid bacteria and yeast

SummaryThe antioxidant properties of nine different samples of freshly drawn, pasteurised and UHT milk were examined before and after fermentation with several lactobacilli, individually and in combination with the yeast Saccharomyces boulardii. The results showed no statistical differences for the content in proteins, fats and carbohydrates of the different milk. The antioxidant properties of the unfermented milk were positively affected by the thermal treatment. All samples of fermented milk had a greater antioxidant activity than unfermented but only for the freshly drawn milk that increase was significantly different. The co‐incubation of lactobacilli with the yeast determined a stronger antioxidant activity especially for the freshly drawn milk. The stability of the tested strains during storage in standard conditions was determined in UHT fermented milk to enable a possible use in probiotic food production. The results showed that the yeast incubated with lactobacilli determined an improvement of the viability of bacteria.

Encapsulation of probiotic strains in plain or cysteine‐supplemented alginate improves viability at storage below freezing temperatures

Four probiotic bacteria (Lactobacillus paracasei L26, L. casei‐01, L. acidophilus Ki, and Bifidobacterium animalis BB‐12®) were encapsulated in plain alginate or alginate supplemented with L‐cysteine·HCl, and resulting microcapsules were stored at different temperatures, namely 21, 4, −20, or −80°C for a period of up to 6 months. The results showed that the encapsulation in calcium alginate microcapsules was only effective in promoting protection at freezing temperatures, independently of the sensitivity of the strain. Storage of calcium alginate microcapsules at −80°C indicated a protective effect upon viability of all four probiotic strains and the presence of L‐cysteine·HCl in the alginate matrix improved protection upon cell viability of B. animalis BB‐12®. An increase in storage temperature of encapsulated bacteria caused an increase in rate of loss in their viability that was strain dependent. This study suggests that microencapsulation of probiotic cells in calcium alginate can be suitable for sustaining the viability of probiotics in food products that require storage below freezing temperatures, even in the absence of cryoprotectors, contributing to an increased shelf life.

Functional foods as carriers for SYNBIO®, a probiotic bacteria combination

The popularity of functional foods continues to increase as consumers desire flavorful foods that will fulfil their health needs. Among these foods, probiotics may exert positive effects on the composition of gut microbiota and overall health. However, in order to be beneficial, the bacterial cultures have to remain live and active at the time of consumption. The aim of this study was to develop new probiotic food products, such as seasoned cheeses, salami, chocolate and ice-cream with a final probiotic concentration of approximately 109CFU/daily dose of Lactobacillus rhamnosus IMC 501® and Lactobacillus paracasei IMC 502® mixed 1:1 (SYNBIO®). The survival and viability of probiotics were determined during the foods shelf-life. The values of viable probiotic bacteria of all dairy and non-dairy foods were between 107 and 109CFU/g of food at the end of the shelf-life and for some of them the values were maintained even after the expiry date. Based on the results of the current study, all the dairy (“Caciotta” cheese, “Pecorino” cheese, “Büscion” Swiss cheese and “Fiordilatte” ice-cream) and non-dairy (“Ciauscolo” salami, Larded salami, Swiss small salami, milk chocolate, dark chocolate, organic jam and chocolate mousse) food products studied would be excellent vehicles to deliver the probiotic health effects because of the high viability of probiotics during the shelf-life of foods and in some cases even after their expiry date.