Probiotic Functional Foods Research Articles

Enhancement of survival of alginate-encapsulatedLactobacillus caseiNCDC 298

Micro-encapsulation of hydrocolloids improves the survival of sensitive probiotic bacteria in the harsh conditions that prevail in foods and during gastrointestinal passage by segregating them from environments. Incorporation of additives in encapsulating hydrocolloids and coatings of microcapsules further improves the survival of the probiotics. In this study, the effect of incorporation of resistant-maize starch in alginate for micro-encapsulation and coating of microcapsules with poly-l-lysine, stearic acid and bees wax on the survival of encapsulated Lactobacillus casei NCDC 298 at pH 1.5, 2% high bile salt, 65 °C for 20 min and release of viable lactobacilli cells from the capsule matrix in simulated aqueous solutions of colonic pH were assessed. Addition of resistant maize starch (2%) improved the survival of encapsulated L. casei NCDC 298. Coating of microcapsules with poly-L-lysine did not further improve the protection of encapsulated cells from the harsh conditions; however, bees wax and stearic acid (2%) improved the survival under similar conditions. Incorporation of maize starch (2%) in alginate followed by coating of beads with stearic acid (2%) led to better protection and complete release of entrapped lactobacilli in simulated colonic pH solution was observed. Additional treatments improve the survival of alginate-encapsulated lactobacilli cells without hindering the release of active cells from the capsule matrix and hence, the resulting encapsulated probiotics can be exploited in the development of probiotic functional foods with better survival of sensitive probiotic organisms.

Immunobiotic lactobacilli reduce viral-associated pulmonary damage through the modulation of inflammation–coagulation interactions

The exacerbated disease due to immune- and coagulative-mediated pulmonary injury during acute respiratory viruses infection results in severe morbidity and mortality. Identifying novel approaches to modulate virus-induced inflammation-coagulation interactions could be important alternatives for treating acute respiratory viruses infections. In this study we investigated the effect of the probiotic strain Lactobacillus rhamnosus CRL1505 on lung TLR3-mediated inflammation, and its ability to modulate inflammation-coagulation interaction during respiratory viral infection. Our findings reveal for the first time that a probiotic bacterium is able to influence lung immune-coagulative reaction triggered by TLR3 activation, by modulating the production of proinflammatory and anti-inflammatory cytokines as well as expression of tissue factor and thrombomodulin in the lung. We also demonstrated that the preventive treatment with the probiotic bacteria beneficially modulates the fine tune balance between clearing respiratory viruses (respiratory syncytial virus and influenza virus) and controlling immune-coagulative responses in the lung, allowing normal lung function to be maintained in the face of a viral attack. Our data also pinpoint a crucial role for IL-10 in the immune protection induced by L. rhamnosus CRL1505 during respiratory viral infections. These observations might be helpful to propose new preventive or therapeutic approaches to better control virus-inflammatory lung damage using probiotic functional foods.

The influence of sweeteners in probiotic Petit Suisse cheese in concentrations equivalent to that of sucrose

As in the case of probiotic functional foods in recent years, demand has increased notably for light or diet foods with added sweeteners. However, little is known about the effect of different sweeteners on the microorganisms present. Thus, the objective of the current study was to establish the ideal sucrose concentration and equivalent concentrations of different sweeteners and to determine, by microbiological analyses, the influence of these compounds on the viability of the starter and probiotic cultures used in the production of strawberry-flavored Petit Suisse cheese during its shelf life. The ideal sucrose concentration was determined using the just-about-right (JAR) scale, and the equivalent concentrations of the sweeteners were subsequently determined by the magnitude estimation method. Microbiological analyses were also carried out to check the viability of the cultures during the product’s shelf life. The results showed that the compounds Neotame (NutraSweet, Chicago, IL) and stevia presented, respectively, the greatest and least sweetening power of the sweeteners tested. None of the sweeteners used in this study exerted a negative effect on the viability of the starter or probiotic cultures, and thus we were able to obtain a probiotic, functional food with reduced calorie content.

In vitro evaluation of Bifidobacterium strains of human origin for potential use in probiotic functional foods

The present study investigated some in vitro properties for probiotic use of four strains of bifidobacteria isolated from faeces of healthy children (Bifidobacterium longum 51A, Bifidobacterium breve 1101A, Bifidobacterium pseudolongum 1191A and Bifidobacterium bifidum 1622A). In vitro tests were carried out to compare growth rate, aerotolerance, antagonistic activity against pathogens, antimicrobial susceptibility profile and cell wall hydrophobicity. Mean doubling time of B. longum 51A was shorter compared to the other strains. All strains were aerotolerant up to 72 h of exposure to oxygen. In vitro antagonism showed that B. longum 51A and B. pseudolongum 1191A were able to produce inhibitory diffusible compounds against all pathogenic bacteria tested, but not against Candida albicans. B. longum 51A was sensitive to all the antimicrobials tested, except neomycin. The hydrophobic property of the cell wall was highest for B. bifidum 1622A. Based on these parameters, B. longum 51A showed the best potential for probiotic use among the tested strains, presenting the greatest sensitivity to antimicrobials, the best growth rate and the highest capacity to produce antagonistic substances against various pathogenic microorganisms.

Competitive inhibition of three novel bacteria isolated from faeces of breast milk-fed infants against selected enteropathogens

Numerous in vitro and in vivo studies conducted using different probiotic micro-organisms have demonstrated their ability to interfere with the growth and virulence of a variety of enteropathogens. The reported beneficial effects of the use of probiotics to complement antibiotic therapy or prevent diarrhoea or gastrointestinal infection in infants have increased in recent years. In the present study, we demonstrated the capacity of supernatants obtained from three novel probiotics (Lactobacillus paracasei CNCM I-4034, Bifidobacterium breve CNCM I-4035 and Lactobacillus rhamnosus CNCM I-4036) isolated from the faeces of breastfed infants to inhibit the growth of enterotoxigenic and enteropathogenic (EPEC) bacteria, such as Escherichia coli, Salmonella and Shigella. To assess their potential antimicrobial activity, the 17 and 24 h cell-free supernatants broth concentrates (10×) having 1, 2 or 4 % of the three probiotics were incubated with EPEC bacteria strains. After 17 h of co-culture, the supernatants were able to inhibit the growth of E. coli, Salmonella and Shigella up to 40, 55 and 81 %, respectively. However, the inhibitory capacity of some supernatants was maintained or completely lost when the supernatants (pH 3·0) were neutralised (pH 6·5). Overall, these results demonstrated that L. paracasei CNCM I-4034, B. breve CNCM I-4035 and L. rhamnosus CNCM I-4036 produce compounds that exhibited strain-specific inhibition of enterobacteria and have the potential to be used as probiotics in functional foods.

Effect of Synbiotic Interaction of Fructooligosaccharide and Probiotics on the Acidification Profile, Textural and Rheological Characteristics of Fermented Soy Milk

Preparation of yoghurt-like product from non-dairy raw material, such as soy with probiotic and prebiotic is a novel development in the field of fermented functional foods. This research work aimed at finding the new combinations of probiotics Lactobacillus acidophilus, Lactobacillus plantarum and Lactobacillus rhamnosus, which can give good product characteristics to fermented soy milk. Fructooligosaccharide was added in an attempt to reduce the after-taste of soymilk, improve acidification rates and growth of probiotics. Acidification rate was enhanced with L. acidophilus–L. plantarum and L. acidophilus–L. plantarum–L. rhamnosus, resulting in a shorter time to reach pH 4.5. Hardness was significantly (P < 0.05) higher for soy yoghurt fermented by binary co-culture followed by mixed cultures. All the samples showed higher G′ (1,279.70–1405 Pa) and lower tan δ (0.273–0.346) values which signifies firmer and solid-like character of the gel formed by probiotic bacteria. Soy yoghurt made with L. acidophilus–L. plantarum resulted in improved product characteristics with shorter t pH 4.5 (4.28 h). Soy yoghurt fermented with L. acidophilus–L. plantarum showed more than 9 log cfu/ml count which is required for probiotic functional food.

Technological aspects of probiotic functional food development

Fermented food containing live cultures called probiotics are considered as fermented functional foods. Probiotics, when consumed in adequate numbers, provide a health benefit directly or indirectly by production of metabolites to the host. The development of probiotic products is a research priority and challenge for both industry and science sectors due to increasing consumer awareness of their ability to maintain good health along with the supply of essential nutrition. This review presents an overview of probiotic food development, various matrices for probiotic foods, and processing and packaging of probiotic foods.

Consumer perception of functional foods: A conjoint analysis with probiotics

The current study, performed on a sample of 600 respondents responsible of household food shopping, explored consumers evaluation of four attributes of probiotics functional foods: base product (yoghurt, orange juice and biscuits), health claim (generic, psychological and prevention), price (high, regular and low) and brand (familiar and unfamiliar). Conjoint analysis ascertained that consumers consider the base product as the most important attribute in selecting a probiotic functional food and asses prevention claim as the most valuable. In addition cluster analysis revealed that customer groups significantly differ in their evaluation of perceived healthiness of functional foods and in the importance attached to price and brand.

Isolation and Characterization of Putative Probiotic Bacterial Strain, Bacillus amyloliquefaciens, from North East Himalayan Soil Based on In Vitro and In Vivo Functional Properties.

This study is aimed to isolate some novel probiotics from the soils of North East Himalayas. Eleven Gram positive isolates were obtained in MRS with Oxgall media from soil samples. Four of the isolates withstood the in vitro gastric juice pH 3.0 and 0.45% bile salt tolerance screening. Among these, PBT 3 showed high cell surface hydrophobicity and adhered to the Caco-2 cells. 16s rDNA gene sequences of this probiotic strains were identified as Bacillus amyloliquefaciens (accession no: JF836079). In in vivo bioefficacy evaluation by DSS-induced colitis animals, B. amyloliquefaciens significantly ameliorated the loss in body weight. Further, the treatment altered the levels of myeloperoxidase, lipoperoxides, and mucous content in the colon tissues compared with normal colon. It reduced the protein and mRNA levels of pro-inflammatory cytokines such as TNF-α and IL-1β. These biochemical findings were supported by histopathological evidences. Our study reports the use of B. amyloliquefaciens isolated from Himalayan soil as probiotic and its beneficial effect on IBD for the first time and suggests that this could be used as potential probiotics in functional foods or as a curative agent.

Application of fluorescent techniques to evaluate the survival of probiotic lactobacilli to bile acid

To apply a fluorescent dye as an alternative technique to evaluate the survival of potentially probiotic lactobacilli to bile acids (BA) as first step in the design of probiotic functional foods. The use of lactic acid bacteria (LAB) in the functional food design depends on their ability to survive in the gastrointestinal tract where bile is an important natural barrier. Bile is mainly constituted by conjugated BA, which can be hydrolyzed to free BA and taurine or glycine. Changes in the transmembrane electrical potential (ΔΨ) of probiotic LAB strains due to the effect of conjugated and free BA were measured and showed that the majority of the tested LAB strains had greater sensibility to free BA than to their respective conjugated acids. Variations in the ΔΨ of the microorganism correlated well with bacterial viability determined by standard plate count method. We therefore propose the DiSC(3)-based fluorescent technique as a rapid and effective method to evaluate the resistance of probiotic lactobacilli to bile as first step for strain selection to be included in functional foods.

Impact of the Metabolic Activity of Streptococcus thermophilus on the Colon Epithelium of Gnotobiotic Rats

The thermophilic lactic acid bacterium Streptococcus thermophilus is widely and traditionally used in the dairy industry. Despite the vast level of consumption of S. thermophilus through yogurt or probiotic functional food, very few data are available about its physiology in the gastrointestinal tract (GIT). The objective of the present work was to explore both the metabolic activity and host response of S. thermophilus in vivo. Our study profiles the protein expression of S. thermophilus after its adaptation to the GIT of gnotobiotic rats and describes the impact of S. thermophilus colonization on the colonic epithelium. S. thermophilus colonized progressively the GIT of germ-free rats to reach a stable population in 30 days (10(8) cfu/g of feces). This progressive colonization suggested that S. thermophilus undergoes an adaptation process within GIT. Indeed, we showed that the main response of S. thermophilus in the rat's GIT was the massive induction of the glycolysis pathway, leading to formation of lactate in the cecum. At the level of the colonic epithelium, the abundance of monocarboxylic acid transporter mRNAs (SLC16A1 and SLC5A8) and a protein involved in the cell cycle arrest (p27(kip1)) increased in the presence of S. thermophilus compared with germ-free rats. Based on different mono-associated rats harboring two different strains of S. thermophilus (LMD-9 or LMG18311) or weak lactate-producing commensal bacteria (Bacteroides thetaiotaomicron and Ruminococcus gnavus), we propose that lactate could be a signal produced by S. thermophilus and modulating the colon epithelium.

The study to investigate the potential benefits of probiotics in yogurt, a patient-oriented, double-blind, cluster-randomised, placebo-controlled, clinical trial

Probiotic functional foods are widely advertised to consumers primarily based on probiotic supplements. Determine if consumption of yogurt containing a high dose of probiotics improves health in children ages 1-3 years attending daycare/school centers. Double-blinded, randomized, placebo-controlled, allocation concealment clinical trial. Outpatient participants in the Washington, DC area. 182 healthy children between the age of 1 and 3 years attending daycare/school at least 3 days a week. Active was a strawberry yogurt-based drink supplemented with Bifidobacterium animalis ssp. lactis (B. lactis) BB-12. The placebo was indistinguishable from the active drink, differing only in absence of the probiotic BB-12. Primary objective was to determine if consumption of a probiotic-containing yogurt-based drink decreases absences due to illnesses from daycare for children ages 1-3 years. Secondary was to determine if probiotic-containing yogurt-based drink improves overall parental satisfaction due to decreased absences from work and an overall healthier child. There were no significant differences in the days of missed school per group, with 51.9% in the active group and 47.1% in the placebo group missing at least 1 day of school throughout the study. Additionally, there were no differences in any secondary outcomes among the groups. Consumption of a yogurt-based drink delivering 10(10) CFU of Bifidobacterium animalis ssp. lactis (B. lactis) BB-12 per day did not decrease the number of days missed of school due to an illness. Additional independent research on the potential of BB-12 to reduce illness in children needs to be conducted.

Probiotic lactic acid bacteria in the gastro-intestinal tract: health benefits, safety and mode of action

Lactic acid bacteria (LAB) have received considerable attention as probiotics over the past few years. This concept has grown from traditional dairy products to a profitable market of probiotic health supplements and functional foods. Extensive research is done on novel potential probiotic strains, with specific emphasis on their health benefits and mode of action. Criteria for the selection of probiotic strains have only recently been formulated by the Food and Agriculture Organization of the United Nations and the World Health Organization (FAO/WHO). Several in vitro techniques have been developed to evaluate the probiotic properties of strains. In many cases, this is followed by in vivo tests. Safety studies are also obligatory, as a few cases of bacteremia caused by LAB have been reported. This review focuses on the health benefits and safety of LAB probiotics, the criteria used to select a probiotic, mode of action and the impact these organisms have on natural microbiota in the gastro-intestinal tract.

EVOLUTION OF PROBIOTIC CONTENT AND COLOR OF APPLES IMPREGNATED WITH LACTIC ACID BACTERIA

An important issue related with functional foods development is the stability of physiologically active compounds. For example vitamins, minerals, probiotic bacteria are very sensitive to the technological processes involved in their obtention. The objective of this paper is to evaluate the viability of a probiotic strain (Lactobacillus rhamnosus) inside a food matrix (Granny smith apple) and physical parameters such as color (L* a* b*) of vacuum impregnated apple cylinders during a refrigerated and frozen storage. The results show that the viable probiotic bacteria inside food matrix maintain the minimal value to be considered as probiotic functional food (106 CFU/g), and the color parameters which are critical in refrigerated samples during their storage.

Probiotics and prebiotics for severe acute malnutrition (PRONUT study): a double-blind efficacy randomised controlled trial in Malawi

Severe acute malnutrition affects 13 million children worldwide and causes 1-2 million deaths every year. Our aim was to assess the clinical and nutritional efficacy of a probiotic and prebiotic functional food for the treatment of severe acute malnutrition in a HIV-prevalent setting. We recruited 795 Malawian children (age range 5 to 168 months [median 22, IQR 15 to 32]) from July 12, 2006, to March 7, 2007, into a double-blind, randomised, placebo-controlled efficacy trial. For generalisability, all admissions for severe acute malnutrition treatment were eligible for recruitment. After stabilisation with milk feeds, children were randomly assigned to ready-to-use therapeutic food either with (n=399) or without (n=396) Synbiotic2000 Forte. Average prescribed Synbiotic dose was 10(10) colony-forming units or more of lactic acid bacteria per day for the duration of treatment (median 33 days). Primary outcome was nutritional cure (weight-for-height >80% of National Center for Health Statistics median on two consecutive outpatient visits). Secondary outcomes included death, weight gain, time to cure, and prevalence of clinical symptoms (diarrhoea, fever, and respiratory problems). Analysis was on an intention-to-treat basis. This trial is registered as an International Standard Randomised Controlled Trial, number ISRCTN19364765. Nutritional cure was similar in both Synbiotic and control groups (53.9% [215 of 399] and 51.3% [203 of 396]; p=0.40). Secondary outcomes were also similar between groups. HIV seropositivity was associated with worse outcomes overall, but did not modify or confound the negative results. Subgroup analyses showed possible trends towards reduced outpatient mortality in the Synbiotic group (p=0.06). In Malawi, Synbiotic2000 Forte did not improve severe acute malnutrition outcomes. The observation of reduced outpatient mortality might be caused by bias, confounding, or chance, but is biologically plausible, has potential for public health impact, and should be explored in future studies. Department for International Development (DfID).

Probiotic properties of Lactobacillus rhamnosus and Lactobacillus paracasei isolated from human faeces

The possibility of using microbes to maintain health, and to prevent or treat disease is a topic as old as microbiology. The research of novel probiotic strains is important in order to satisfy the increasing request of the market and to obtain functional products in which the probiotic cultures are more active and with better probiotic characteristics than those already present on the market. In this study, the probiotic potential of Lactobacillus strains isolated from Italian elderly human faeces was investigated. The Lactobacillus strains were identified and examined for resistance to gastric acidity and bile toxicity, adhesion to HT-29 cells, antimicrobial activities, antibiotic susceptibility and plasmid profile. Survival of the strains through human intestine was examined in a 3 months human feeding trial. Two strains, Lactobacillus rhamnosus IMC 501 and Lactobacillus paracasei IMC 502, tolerated well low pH and bile acids. In antimicrobial activity assays, both strains showed inhibitory properties towards selected potential harmful microorganisms, particularly against Candida albicans. The two selected strains expressed high in vitro adherence to HT-29 cells increasing this characteristic when they are used in combination and they were resistant to vamcomycin, colistin sulphate, gentamicin, oxolinic acid and kanamycin. Moreover, the two strains could be recovered from stools of volunteers after the feeding trials. Lactobacillus rhamnosus IMC 501 and L. paracasei IMC 502 present favourable strain-specific properties for their utilisation as probiotics in functional foods and the high adhesion ability of the L. rhamnosus IMC 501 and L. paracasei IMC 502 used in combination, confirmed by both in vitro and in vivo study, indicate that the two bacterial strains could be used as health-promoting bacteria.

The Beneficial Use of Cereal and Cereal Components in Probiotic Foods

Cereals and cereal components can be used as fermentation substrates for probiotic organisms imparting prebiotic effects. Consumer interest in healthy functional foods has resulted in the need for food products with versatile health-benefiting properties. The conventional choice for probiotic food applications has been dairy-based products, but whole grain-based probiotic functional foods have debuted in Japan and Europe. In the US, pro- and prebiotics are mainly marketed as dietary supplements, but are moving towards inclusion in the diet as mainstream foods. Cereal constituents, such as wheat bran-based ingredients fermented with probiotics, would enhance consumer health with the benefits of probiotics, bran fiber, and healthful bioactive components.

Administration of Lactobacillus casei and Lactobacillus plantarum affects the diversity of murine intestinal lactobacilli, but not the overall bacterial community structure

Lactobacilli are normal inhabitants of the gastrointestinal tract (GIT) of many mammalian hosts. Their administration as probiotics in functional foods is currently a frequent practice, mainly because of their benefits to host health. It is therefore of interest to study the impact of administration of exogenous strains of Lactobacillus normally used as probiotics upon endogenous microbial populations. For this purpose, fecal and intestinal tissue samples were analyzed in a mouse model fed with a mixture of Lactobacillus plantarum and Lactobacillus casei isolated from commercially available dairy products. The murine intestinal microbiota was studied by means of cultivation-independent 16S rRNA gene-targeted techniques, namely denaturing gradient gel electrophoresis (DGGE), terminal restriction fragment length polymorphism (T-RFLP) and sequence analysis of clone libraries. Multivariate statistical analysis was used to integrate datasets obtained from the different techniques applied. Whereas no differences were detected in the composition of the overall fecal bacterial community, changes were observed for intestinal tissue samples. Moreover, an increase in the diversity of gut lactobacilli was observed in fecal as well as intestinal tissue samples when mice received the mixture of L. casei and L. plantarum.

Divergent Patterns of Colonization and Immune Response Elicited from Two IntestinalLactobacillusStrains That Display Similar Properties In Vitro

Lactobacilli derived from the endogenous flora of normal donors are being increasingly used as probiotics in functional foods and as vaccine carriers. However, a variety of studies done with distinct strains of lactobacilli has suggested heterogeneous and strain-specific effects. To dissect this heterogeneity at the immunological level, we selected two strains of lactobacilli that displayed similar properties in vitro and studied their impact on mucosal and systemic B-cell responses in monoxenic mice. Germfree mice were colonized with Lactobacillus johnsonii (NCC 533) or Lactobacillus paracasei (NCC 2461). Bacterial loads were monitored for 30 days in intestinal tissues, and mucosal and systemic B-cell responses were measured. Although both Lactobacillus strains displayed similar growth, survival, and adherence properties in vitro, they colonized the intestinal lumen and translocated into mucosal lymphoid organs at different densities. L. johnsonii colonized the intestine very efficiently at high levels, whereas the number of L. paracasei decreased rapidly and it colonized at low levels. We determined whether this difference in colonization correlated with an induction of different types of immune responses. We observed that colonization with either strain induced similar germinal center formation and immunoglobulin A-bearing lymphocytes in the mucosa, suggesting that both strains were able to activate mucosal B-cell responses. However, clear differences in patterns of immunoglobulins were observed between the two strains in the mucosa and in the periphery. Therefore, despite similar in vitro probiotic properties, distinct Lactobacillus strains may colonize the gut differently and generate divergent immune responses.

Inventing probiotic functional foods for patients with allergic disease

The primary objective of this review is to discuss the rationale for and applicability of probiotics as part of the nutritional approach to the prevention and management of allergic disease. This review represents a synthesis of recent literature published in the fields of probiotics, nutrition, and allergic disease. The expert opinions of the authors were used to select the relevant data for the review. The hygiene hypothesis suggests that allergic disease may arise from a lack of counterbalancing microbial exposure at an early age. Thus, the initial compositional development of gut microbiota is considered a key determinant in the development of both the immune responder phenotype and the normal gut barrier functions. A wide range of data documents the specific actions of selected probiotics. The regulatory role of probiotics in allergic disease has been demonstrated as improving the clinical course or preventing the development of atopic eczema in infants given a probiotic-supplemented diet. In addition, certain types of fatty acids and antioxidants have been proposed for protective effects against the development of allergy and to diminish the inflammatory response in allergic disease. The increase in the prevalence of allergic disease during the past decades is likely to be explained by changes in the environment, including reduced microbial exposure and altered food consumption. Scientifically composed functional foods containing probiotics and other functional components offer a nutritional strategy for both the prevention and the management of allergic disease. Further research is needed to characterize the gut microbiota and to clarify the mechanisms of action that control specific physiologic processes not only in the evolution of allergic disease in at-risk populations but also in the management of allergic diseases.