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Fermented products with probiotic qualities.

For several centuries, fermented products derived from plant or animal materials have been an acceptable and essential part of the diet in most parts of the world. Health benefits have also often been associated with them. Probiotics can be defined as fermented food containing specific live microorganisms or a live microbial food or feed supplement, which beneficially effects the human or the host animal by improving its intestinal microbial balance. Nearly all probiotics currently on the market contain Lactobacilli, Streptococci, Enterococci or Bifidobacteria. In contrast to Japan, where freeze-dried microorganisms are consumed by a substantial part of the human population, in Europe, probiotic action towards humans are only claimed for certain fermented dairy products (e.g. yoghurts). Those species that have been extensively studied so far, with several experimental trials on man, are the two yoghurt bacteria Streptocaccus thermophilus and Lactobacillus bulgaricus, L. casei and Bifidobacteria. L. acidophilus has also received important scientific interest, however, only a few human studies have been carried out. From the technological point of view a good probiotic should be stable and viable for long periods under storage, should be able to survive the low pH levels of the stomach, be able to colonise the epithelium of the gastro-intestinal tract of the host, should not be pathogenic and, last but not least, must be capable of exerting a growth promoting effect or a resistance to infectious diseases. The beneficial effects of probiotics are mainly contributed to a direct antagonistic effect against specific groups of microorganisms (Enteropathogenes), by an effect on the metabolism in the gut or by a stimulation of systemic or mucosal immunity. We will present major proven health benefits of milks fermented with those bacterial species and discuss, where possible, the impact of the specific selection and utilisation of particular strains.

Journal Article↗

Probiotics and prebiotics for gastrointestinal infections.

There is growing interest in and knowledge about the potential health-promoting benefits of both probiotics and prebiotics. Multiple mechanisms of action for the beneficial effect of probiotics and prebiotics have been postulated, including prevention of pathogenic bacteria growth, production of antimicrobial agents, stimulation of mucosal barrier function, and altering immunoregulation. Clinical trials support the efficacy of probiotics in the treatment of acute infectious diarrhea, the prevention of antibiotic associated diarrhea, and the prevention of recurrent Clostridium difficile infection. Although some data support the potential benefit of probiotic therapy in traveler's diarrhea, diverticular disease, and Helicobacter pylori, the strength of this evidence is limited. This paper will review the recent literature relevant to the mechanism of action and utility of probiotics and prebiotics in the treatment of gastrointestinal infections.

Journal Article↗

Effect of feeding diets containing an antibiotic, a probiotic, or yucca extract on growth and intestinal urease activity in broiler chicks.

A 6-wk study was conducted to determine the effect of feeding diets containing an antibiotic, a probiotic, or yucca extract on daily gain, feed conversion ratio, and urease activity and ammonia production in intestinal contents of broiler chicks. Four replicates of 10 broiler chicks (average body weight, 48 g) each were assigned to a control or diets containing 0.1% chloroxytetracycline (antibiotic), 0.1% Lactobacillus casei (probiotic), or 0.2% yucca extract. Feeding a diet containing the probiotic significantly (P < 0.05) increased average daily gain during the first 3-wk period compared to the control (30.7 vs 28.7 g). This increase was partly accounted for by increased feed intake. During the first 3 wk, feeding the diet containing probiotic significantly (P < 0.05) decreased urease activity (per gram of collected contents) in small intestinal contents but not in large intestinal contents, compared with the control. Urease activity determined at 6 wk of age was not significantly affected by diet. Our studies indicate that dietary probiotic decreases urease activity in the small intestinal contents of young chicks and thus may be beneficial for improving animal health and growth, especially during early life.

Ammonia↗

Probiotics in gastroenterology.

Recent evidence has suggested the potential therapeutic role for probiotics in the prevention or treatment of gastrointestinal diseases. Several studies have shown that probiotics are of benefit in gastrointestinal infections, including viral diarrhea, Clostridium difficile-associated diarrhea, traveler's diarrhea, and antibiotic-associated diarrhea. Recent data support the potential beneficial therapeutic effect in inflammatory bowel disease as well. Other possible indications for probiotic treatment include Helicobacter pylori infection, irritable bowel syndrome, and radiotherapy-associated diarrhea. It is important to select well-characterized preparations; in fact, the viability and survival of many available preparations are unproven. More precise information on the mechanisms by which probiotic strains exert their beneficial effects in vivo is needed. This may provide the scientific rationale for the selection of the best probiotic strains to use in the performance of large, double-blind, controlled clinical trials.

Journal Article↗

Probiotics against Campylobacter Pathogens.

Background: The subject matter of this study was to investigate, for the first time, the immediate-preventive effect of probiotics, composed of bacillus species, in a murine model of Campylobacter infection. Methods: An established model of Campylobacter infection in mice with a defined LD50 was utilized to assess the protective effect of probiotics. Results: The results obtained demonstrate that the level of animal protection, after a single administration of the new probiotics biosporin and subalin, reached 90-100% at LD50 and 80% at LD100. Conclusions: Such efficacy of probiotics is considered to be due to their high antagonistic activity against those pathogens registered in vitro. Antagonistic activity of other tested probiotics (bactisubtil and cereobiogen) to different cultures of Campylobacter was not manifested.

Journal Article↗

Colonization of congenitally immunodeficient mice with probiotic bacteria.

We assessed the capacity of four probiotic bacteria (Lactobacillus acidophilus, Lactobacillus reuteri, Lactobacillus casei GG, and Bifidobacterium animalis) to colonize, infect, stimulate immune responses in, and affect the growth and survival of congenitally immunodeficient gnotobiotic beige-athymic (bg/bg-nu/nu) and beige-euthymic (bg/bg-nu/+) mice. The bacteria colonized and persisted, in pure culture, in the alimentary tracts of both mouse strains for the entire study period (12 weeks). Although all adult and neonatal beige-euthymic mice survived probiotic colonization, some infant mortality occurred in beige-athymic pups born to mothers colonized with pure cultures of L. reuteri or L. casei GG. The probiotic bacteria manifested different capacities to adhere to epithelial surfaces, disseminate to internal organs, affect the body weight of adult mice and the growth of neonatal mice, and stimulate immune responses. Although the probiotic species were innocuous for adults, these results suggest that caution and further studies to assess the safety of probiotic bacteria for immunodeficient hosts, especially neonates, are required.

Animals↗

Probiotic Lactobacillus spp. diminish Helicobacter hepaticus-induced inflammatory bowel disease in interleukin-10-deficient mice.

Clinical and experimental evidence has demonstrated the potential role of probiotics in the prevention or treatment of inflammatory bowel disease. Probiotic clones with direct immunomodulatory activity may have anti-inflammatory effects in the intestine. We investigated the roles of tumor necrosis factor alpha (TNF-alpha)-inhibitory Lactobacillus clones with a pathogen-induced murine colitis model. Murine-derived probiotic lactobacilli were selected in vitro for their ability to inhibit TNF-alpha secretion by Helicobacter hepaticus-stimulated macrophages. Interleukin-10 (IL-10)-deficient mice were treated with probiotic Lactobacillus reuteri in combination with Lactobacillus paracasei and then challenged with H. hepaticus. Ten weeks postinoculation, the severity of typhlocolitis was assessed by histologic examination of the cecocolic region. Intestinal proinflammatory cytokine responses were evaluated by real-time quantitative reverse transcriptase PCR and immunoassays, and the quantities of intestinal H. hepaticus were evaluated by real-time PCR. Intestinal colonization by TNF-alpha-inhibitory lactobacilli reduced intestinal inflammation in H. hepaticus-challenged IL-10-deficient mice despite similar quantities of H. hepaticus in cocolonized animals. Proinflammatory colonic cytokine (TNF-alpha and IL-12) levels were lowered in Lactobacillus-treated animals. In this H. hepaticus-challenged IL-10-deficient murine colitis model, lactobacilli demonstrated probiotic effects by direct modulation of mucosal inflammatory responses.

Animals↗

[Up-to-date clinical and experimental basis for the use of probiotics]

OBJECTIVE: To evaluate the potential of probiotics or biotherapeutic agents for the prevention and/or treatment of selected intestinal infections. METHODS: Medline database was searched for all relevant articles between 1990 and February 1998. Bibliographies of articles were also used. All animal experiments and placebo-controlled human studies were reviewed in order to provide information on the mechanisms of action, potential efficacy, or adverse effects of these biotherapeutic agents. RESULTS: In the first part of this review, the different mechanisms of action that are effective in the treatment of diarrhea were discussed, and they were well demonstrated in laboratory animals. The most important are: enzymatic induction of disaccharidase activity, trophic effects on the intestinal mucosa, action in blocking bacterial toxins, and also induction of the immunologic response. Therapeutic effects of probiotics in humans, mainly in the gastrointestinal tract, were reported in the second part. Placebo-controlled studies have shown that biotherapeutic agents have been used successfully in the treatment of acute diarrhea in infants, traveler s diarrhea, antibiotic-associated diarrhea, with or without Clostridium difficile-associated enterocolitis (pseudomembranous colitis), and in immunosuppression-associated diarrhea, including AIDS. Lactobacillus, Bifidobacterium and Saccharomyces boulardii were the most important biotherapeutic agents to be considered. CONCLUSIONS: Currently, there is evidence that the administration of selected microorganisms is benefic in the prevention and treatment of certain intestinal infections. According to the literature, Saccharomyces boulardii is the most important probiotic. Possible future indications were discussed, such as the probable synergic effect of many probiotics due to their different and complementary mechanisms of action. The importance of new experimental and clinical studies for the better understanding of actions and the use of probiotics in other clinical situations was emphasized.

Journal Article↗

Evaluation of cheddar cheese as a food carrier for delivery of a probiotic strain to the gastrointestinal tract.

Cheddar cheese was evaluated as a food carrier for the delivery of viable microorganisms of Enterococcus faecium (Fargo 688; Quest Int., Naarden, The Netherlands) to the gastrointestinal tract. This strain had previously been shown to possess properties required of a probiotic microorganism including the ability to relieve irritable bowel syndrome. The strain was found to survive to high numbers in Cheddar cheese during ripening at 8 degrees C for 15 mo (4 x 10(8) cfu/g) and in yogurt during storage at 4 degrees C for 21 d (4 x 10(7) cfu/g). In an in vitro model system, Cheddar cheese was found to have a greater protective effect than yogurt upon exposure of the probiotic culture to porcine gastric juice at pH 2. Subsequently, a feeding trial involving 8 pigs per group was performed in which a rifampicin-resistant variant of the probiotic strain was fed for 21 d at a mean daily intake of 1.3 x 10(10) cfu/d from Cheddar cheese or 3.7 x 10(9) cfu/d from yogurt. During the feeding period, Cheddar cheese yielded a significantly higher mean fecal probiotic count (2 x 10(6) cfu/g of feces) than did yogurt (5.2 x 10(5) cfu/g of feces). These data indicate that mature Cheddar cheese compares very favorably with fresh yogurt as a delivery system for viable probiotic microorganisms to the gastrointestinal tract.

Animal Feed↗

Probiotic-derived extracellular vesicles as food-based nanocarriers: Mechanisms, functional applications, and future perspectives in food systems.

Probiotic-derived extracellular vesicles (PDEVs) are a promising type of postbiotic nanoparticle derived by fermentation of probiotics, and have gained growing interest as a potential application in food science and nutrition. These are lipid bilayer vesicles of nanoscale, which are naturally released by probiotic cells and contain a wide variety of bioactive molecules, such as proteins, nucleic acids, and metabolites. Moreover, PDEVs are highly stable, biocompatible, and can be easily engineered to have surfaces with high functionality, which makes them good candidates in functional engineering. In contrast to traditional live probiotics, PDEVs overcome the difficulties of preserving microbial viability during processing and storage, thus providing superior safety, stability, and predictable biological performance. This is a systematic review of the various functions of PDEVs in food systems. We conclude on the processes through which PDEVs control intestinal barrier integrity, alter gut microbiota composition, and alter host immune responses, and their potential to enhance gut health when added to functional foods. In addition to their health-promoting effects, PDEVs have shown significant potential as natural antimicrobial agents to preserve food and as effective nanocarriers of hydrophobic bioactive compounds, including fucoxanthin, to improve their stability, bioavailability, and targeted delivery. Moreover, PDEVs can be used as new regulators of microbial fermentation. However, it should be noted that a lot of the evidence that is available is still preliminary and the effectiveness of these applications in real food-processing and storage conditions has not been fully proven. Although they have potential, there are a number of challenges that still hinder the widespread use of PDEVs in the food industry. These involve the creation of scalable and cost-effective production processes, batch-to-batch consistency, vesicle stability in a variety of food matrices, and regulatory and safety considerations. Other emerging engineering approaches, such as surface functionalization and cargo loading, are also discussed in this review and could further increase the specificity, functionality, and application versatility of PDEVs in food systems. Moving forward, the incorporation of PDEVs into the next generation functional foods, novel food preservation methods, and customized nutrition plans should be prioritized in future studies. Further developments in these fields can make PDEVs useful platforms at the interface of food microbiology, nanotechnology, and human health.

Probiotics↗

Effects of multistrain probiotic supplementation on hepatic function and anthropometric parameters in patients with metabolic dysfunction-associated steatotic liver disease: a double-blind, randomized controlled trial.

BACKGROUND: Metabolic dysfunction-associated steatotic liver disease (MASLD) is increasingly prevalent on a global scale. The gut microbiota is integral to its pathogenesis, prompting extensive research into microbiota modulation as a potential adjunctive therapeutic strategy. AIM: The study aimed to evaluate the effect of multistrain probiotics supplementation on hepatic function in patients with MASLD in a double-blind, randomized, controlled trial. The primary outcomes were changes in Fibrosis-4 index (FIB-4) and fatty liver index (FLI). Secondary outcomes included changes in anthropometric parameters, selected biochemical markers, and other liver-related indices. METHODS: A total of 64 patients with MASLD were randomly assigned to two groups receiving either placebo (C) or a probiotic mixture (PRO) containing the following bacterial strains: 50% Lactococcus lactis Rosell-1058, 25% Lacticaseibacillus casei Rosell-215, 12.5% Lactobacillus helveticus Rosell-52, 12.5% Bifidobacterium bifidum Rosell-71 for 12 wk. RESULTS: Significant group &#xd7; time interactions were observed for FIB-4 (Q = 0.007), with reduction in the PRO group and increase in the C group (-0.05 vs. 0.10; P = 0.002). No significant interaction was found for FLI (Q = 0.942). Significant group &#xd7; time interactions were also observed for aspartate aminotransferase (-2.87 vs. 1.87 U/L; Q = 0.003) and APRI (-0.03 vs. 0.02; Q = 0.001), favoring the PRO group (P < 0.001 for both). No significant changes were observed in anthropometric parameters, glucose levels, or lipid profile. CONCLUSIONS: In patients with MASLD, the 12-wk probiotic supplementation had a modest but statistically significant effect on FIB-4, aspartate aminotransferase, and APRI, with no significant effect on FLI or anthropometric and metabolic parameters. These findings suggest that this probiotic formulation may have potential benefits for liver function in MASLD. However, long-term studies incorporating imaging-based and histological endpoints are required to determine the clinical significance of these findings.

Humans↗

Complete genome sequence and genomic characterization of the probiotic Limosilactobacillus reuteri PSC102.

BACKGROUND: Gut microbiota are potential sources of probiotics and play an essential role in maintaining intestinal health. Limosilactobacillus reuteri PSC102 (L. reuteri PSC102), which was isolated from the feces of healthy pigs, exhibited health-beneficial properties. AIM: We aimed to conduct a whole-genome sequencing analysis of L. reuteri PSC102 to determine its molecular characteristics as a probiotic strain. METHODS: Limosilactobacillus reuteri PSC102 cells were cultured in De Man-Rogosa-Sharpe medium, followed by DNA extraction for genomic analysis using the PacBio-Illumina sequencing platform. The EzBioCloud software was used to perform gene assembly, and the genes were interpreted by the National Center for Biotechnology Information (NCBI) and the Glimmer program. Core and pan-genomic analyses were performed to assess the extent of functional conservation in the genomic sequence. Moreover, the NCBI database and the Basic Local Alignment Search Tool software were used to identify antimicrobial resistance genes and virulence factors. RESULTS: Limosilactobacillus reuteri PSC102 consists of a single circular chromosome with 2,048,626 bp, a guanine- cytosine of 38.9%, 18 rRNA genes, and 69 tRNA genes. Among the 1,846 protein-coding sequences, genes associated with probiotic characteristics were identified, including genes involved in host-microbe interactions, stress tolerance, biogenesis, and defense mechanisms. Furthermore, the genome of L. reuteri PSC102 comprises 2,446 pan-genome and 1,222 core-genome orthologous gene clusters. A total of 74 unique genes were identified in L. reuteri PSC102 genome. These genes mostly encode proteins potentially involved in the transport and metabolism of amino acids and carbohydrates. Moreover, antibacterial resistance genes and virulence factors were absent in L. reuteri PSC102. CONCLUSION: The results of the molecular insight into L. reuteri PSC102 corroborates its use as a probiotic in humans and other animals.

Limosilactobacillus reuteri↗

Sensitive and selective determination of glutathione in probiotic bacteria by capillary electrophoresis-laser induced fluorescence.

Glutathione (GSH) is a thiol with an important function in protecting tissue against the oxidative stress which has been related to carcinogenesis in the colon. For this reason the development of probiotic species producing glutathione could be of great interest. To determine the glutathione content of some probiotic bacteria of the Bifidobacterium and Lactococcus genera, a very sensitive and selective analytical method based on capillary electrophoresis coupled to laser-induced fluorescence detection has been developed. Pretreatment of cell-lysate samples is very simple--precipitation of protein with acetonitrile in 1:2 volume ratio. The fluorophore 5-iodoacetamidofluorescein (5-IAF) was chosen for glutathione derivatisation; it reacts with thiols at pH 12.5, forming a fluorescent adduct which is excited by a laser at 488 nm for detection. The reaction conditions optimised were temperature, time, and 5-IAF/GSH molar ratio. Electrophoresis was performed with a carbonate buffer (25 mmol L(-1), pH 9.8) as background electrolyte and a voltage of 30 kV; an electrophoretic run was complete in less than 7 min. There was a good linear relationship between concentration and response in the range 2.5-500 ng mL(-1) and the LOD was 0.5 ng mL(-1). The glutathione content of probiotic cells was determined by using the standard additions method to reduce matrix effects. The method was fully validated and shown to be of suitable sensitivity and selectivity for determination of GSH in probiotic cell lysates.

Acetonitriles↗

Anti-inflammatory potential of the probiotic dietary supplement Lactibiane Tolérance: in vitro and in vivo considerations.

BACKGROUND & AIMS: Probiotics are live microorganisms which when administered in adequate amounts confer a health benefit on the host. In inflammatory bowel disease (IBD), where major modifications of the intestinal microflora have been reported, there is an increasing interest in modulating the flora with probiotic products. This work addresses the anti-inflammatory potential of Lactibiane Tolérance, a probiotic dietary supplement (mixture of four strains) using in vitro and in vivo approaches. METHODS: Comparison of the four individual strains and the commercial product reconstituted from them was conducted by in vitro tests (cytokine release after 24h stimulation of human peripheral blood mononuclear cells (PBMC)). The potential immunomodulatory characteristics of Lactibiane Tolérance were determined in vivo in an acute mice model of 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis. Assessment of colitis included blinded histological and macroscopic scores. RESULTS: We showed that Lactibiane Tolérance has anti-inflammatory properties in vitro by stimulating IL-10 production and in vivo by conferring a significant protective effect in the TNBS-induced colitis model (more than 50% decrease of colitis symptoms, P<0.01). CONCLUSIONS: These results demonstrate that a probiotic dietary supplement, Lactibiane Tolérance, can significantly prevent the initial injury of TNBS and could stimulate the initiation of clinical trials in IBD.

Animals↗

Probiotic and other functional microbes: from markets to mechanisms.

Insight into the diversity and function of the human intestinal microbiota has been stimulated by clinical studies with bacteria that exhibit specific functions and which are marketed as probiotics to positively affect our health. Initial efforts concentrated on establishing sound scientific support for the efficacy of these probiotic bacteria, which mainly include Lactobacillus and Bifidobacterium species. Following these evidence-based functional approaches, considerable research is now focused on the mechanisms of action of probiotic bacteria. The mechanisms identified to date mainly relate to the stimulation of host defence systems, immune modulation and the competitive exclusion of pathogens. Recent efficacy, molecular and genomics-based studies have also been reported for some probiotic strains that have found their position in the market place.

Animals↗

Immune modulation and expression of cytokine genes in rainbow trout Oncorhynchus mykiss upon probiotic feeding.

This study elucidates the immune modulation including the expression of cytokine genes following dietary administration of three selected probiotic bacteria--Lactobacillus rhamnosus, Enterococcus faecium and Bacillus subtilis to fish, rainbow trout Oncorhynchus mykiss. They were fed for 45 days on either a basal control diet or one of the three probiotic diets containing the specific bacteria in freeze-dried form at a density of 10(9)CFUgfeed-1. The non-specific immune parameters examined--superoxide anion production by the head kidney leukocytes and the alternate complement activity of serum was improved by probiotic feeding. Besides this, the relative gene expressions of interleukin-1beta1, tumor necrosis factor 1 and 2 and transforming growth factor-beta were up regulated in the spleen and the head kidney. The comparatively better performance of E. faecium could possibly be linked to their suitable ambient temperature conditions. Thus, probiotic bacteria delivered in feed exerts its influence on the immune system of fish, both at cellular and molecular levels.

Animal Feed↗

Modulation of the microbial ecology of the human colon by probiotics, prebiotics and synbiotics to enhance human health: an overview of enabling science and potential applications.

The application of probiotics and prebiotics to the manipulation of the microbial ecology of the human colon has recently seen many scientific advances. The sequencing of probiotic genomes is providing a wealth of new information on the biology of these microorganisms. In addition, we are learning more about the interactions of probiotics with human cells and with pathogenic bacteria. An alternative means of modulating the colonic microbial community is by the use of prebiotic oligosaccharides. Increasing knowledge of the metabolism of prebiotics by probiotics is allowing us to consider specifically targeting such dietary intervention tools at specific population groups and specific disease states.

Colon↗

The use of bacterial spore formers as probiotics.

The field of probiosis has emerged as a new science with applications in farming and aqaculture as alternatives to antibiotics as well as prophylactics in humans. Probiotics are being developed commercially for both human use, primarily as novel foods or dietary supplements, and in animal feeds for the prevention of gastrointestinal infections, with extensive use in the poultry and aquaculture industries. The impending ban of antibiotics in animal feed, the current concern over the spread of antibiotic resistance genes, the failure to identify new antibiotics and the inherent problems with developing new vaccines make a compelling case for developing alternative prophylactics. Among the large number of probiotic products in use today are bacterial spore formers, mostly of the genus Bacillus. Used primarily in their spore form, these products have been shown to prevent gastrointestinal disorders and the diversity of species used and their applications are astonishing. Understanding the nature of this probiotic effect is complicated, not only because of the complexities of understanding the microbial interactions that occur within the gastrointestinal tract (GIT), but also because Bacillus species are considered allochthonous microorganisms. This review summarizes the commercial applications of Bacillus probiotics. A case will be made that many Bacillus species should not be considered allochthonous microorganisms but, instead, ones that have a bimodal life cycle of growth and sporulation in the environment as well as within the GIT. Specific mechanisms for how Bacillus species can inhibit gastrointestinal infections will be covered, including immunomodulation and the synthesis of antimicrobials. Finally, the safety and licensing issues that affect the use of Bacillus species for commercial development will be summarized, together with evidence showing the growing need to evaluate the safety of individual Bacillus strains as well as species on a case by case by basis.

Animals↗