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The place of probiotics in human intestinal infections.

A number of studies have been carried out on the effect of several probiotic species on treatment and prevention of intestinal infections. The most commonly used microorganisms are lactic-acid producing bacteria such as lactobacilli and bifidobacteria belonging to the human normal microflora. In vitro and animal studies have shown that probiotic microorganisms interfere with the colonisation of Helicobacter pylori and of enteropathogenic microorganisms. In humans the significance is more uncertain. Clinically significant benefits of probiotics have been demonstrated in the treatment of rotavirus induced diarrhoea and of Saccharomyces boulardii in the prevention of antibiotic-associated diarrhoea (AAD). In patients suffering from inflammatory bowel disease, several probiotic strains have been shown to be as effective as traditional medication in preventing relapses. Standardised and well performed studies are needed to elucidate further the mechanisms of action and the clinical significance of probiotics.

Digestive System↗

Probiotics: potential pharmaceutical applications.

Realisation of the importance of human gut microbiota in health restoration and maintenance has kindled an interest in probiotics. Probiotics are defined as the microbial food supplements, which beneficially affect the host by improving its intestinal microbial balance. Probiotics are the health enhancing functional food ingredients used therapeutically to prevent diarrhea, improve lactose tolerance and modulate immunity. They may also have potential to prevent cancer and lower serum cholesterol levels. Lactobacillus, Bifidobacterium and several other microbial species are perceived to exert such effects by changing the composition of the gut microbiota. However, it is important that exogenously administered bacteria reach and establish themselves in the large intestine in an intact form. The use of non-digestible oligosaccharides ('prebiotics') can fortify intestinal microflora and stimulate their growth. The present review encompasses information regarding the probiotics and their proposed uses. It addresses the concepts of prebiotics and synbiotics, the application of genetic engineering to produce newer probiotics. Finally, the list of commercially available products are reviewed with discussion of questions regarding the reliability, utility and the safety of these products.

Animals↗

Role of probiotics in the management of patients with food allergy.

OBJECTIVE: This review will concentrate on the potential use of probiotics in treating or preventing allergic disorders of the gastro-intestinal tract. In small infants, most reactions to foods are manifested in the gastro-intestinal tract. As children get older, cutaneous and even systemic or respiratory manifestations of food intolerances may be more common. In most instances, allergic disorders in the gut cause inflammation in the mucosa. DATA SOURCES: Recently, the role of gut microflora in allergic disease has gained increasing interest with reports of probiotics being used to treat food allergy. STUDY SELECTION: Pertinent scientific reports on the role of probiotics in the management of patients with food allergy will be reviewed. A recent study to determine the effect of very early probiotic colonization will be extensively reviewed as it presents an intriguing concept of a preventive strategy related to food allergy development RESULTS: Significant improvement has been noted in infants receiving Lactobacillus GG in their extensively hydrolyzed formula. CONCLUSIONS: It is likely more studies will be forthcoming with different probiotic organisms in the prevention and treatment of allergic disorders in children and adults.

Decision Making↗

Efficacy of probiotics in prevention of acute diarrhoea: a meta-analysis of masked, randomised, placebo-controlled trials.

To evaluate the evidence for the use of probiotics in the prevention of acute diarrhoea, we did a meta-analysis of the available data from 34 masked, randomised, placebo-controlled trials. Only one trial was community based and carried out in a developing country. Most of the remaining 33 studies were carried out in a developed country in a health-care setting. Evaluating the evidence by types of acute diarrhoea suggests that probiotics significantly reduced antibiotic-associated diarrhoea by 52% (95% CI 35-65%), reduced the risk of travellers' diarrhoea by 8% (-6 to 21%), and that of acute diarrhoea of diverse causes by 34% (8-53%). Probiotics reduced the associated risk of acute diarrhoea among children by 57% (35-71%), and by 26% (7-49%) among adults. The protective effect did not vary significantly among the probiotic strains Saccharomyces boulardii, Lactobacillus rhamnosus GG, Lactobacillus acidophilus, Lactobacillus bulgaricus, and other strains used alone or in combinations of two or more strains. Although there is some suggestion that probiotics may be efficacious in preventing acute diarrhoea, there is a lack of data from community-based trials and from developing countries evaluating the effect on acute diarrhoea unrelated to antibiotic usage. The effect on acute diarrhoea is dependent on the age of the host and genera of strain used.

Anti-Bacterial Agents↗

Adhesion of probiotic strains to the intestinal mucosa and interaction with pathogens.

Probiotic lactic acid strains are live micro-organisms that, when consumed in adequate amounts as part of food, confer a health benefit on the host. The scientific basis for the use of selected probiotic strains has only recently been firmly established, and appropriate and well-conducted experimental in vitro and in vivo studies, as well as clinical studies, are now beginning to be published, especially with regard to the effectiveness of probiotic strains in antagonizing pathogens. In particular, experimental data have allowed new insights into selected probiotic strains that express strain-specific probiotic properties and into the mechanism of action of these strains. The objective of this review is to analyse the in vitro or in vivo experimental studies in which the antimicrobial activity of selected Lactobacillus and Bifidobacterium strains has been documented.

Animals↗

Probiotics and Helicobacter pylori.

There is substantial evidence that probiotics modulate Helicobacter pylori colonization of the gastric mucosa. This chapter presents the data currently available to support an interaction between probiotics and H. pylori, the importance of lactic acid production by probiotics and their capacity to release bacteriocins or secrete antibiotics. The ability of probiotics to interfere with H. pylori adhesion to epithelial cells and their capacity to attenuate H. pylori-induced gastritis in man is addressed. Finally, the potential of probiotics to modify the H. pylori eradication rate and the antibiotic-associated gastrointestinal side-effects during H. pylori eradication therapy are reviewed.

Animals↗

New therapeutic approach in the management of intestinal disease: probiotics in intestinal disease in paediatric age.

Current evidence supports the view that oral administration of probiotics may be of therapeutic usefulness in several clinical disorders by reestablishing normal flora in the gastrointestinal tract. These entities include inflammatory and infectious diseases of the gut as well as extraintestinal disorders (such as atopic eczema) in which a defective intestinal permeability plays a role. The probiotic effects are attributed to restoration to normal of increased intestinal permeability, unbalanced gut microecology, improved immunological gut barrier function, downregulation of the intestinal inflammatory responses with reduced generation of proinflammatory cytokines. Entities for which the impact of probiotic administration can be considered as proven are Rotavirus diarrhoea, Clostridium difficile diarrhoea, post-antibiotic diarrhoea, allergic diseases. On the other hand, entities for which administration of probiotics is considered under investigation are inflammatory bowel disease, necrotizing enterocolitis, cystic fibrosis, small bowel bacterial contamination, functional gastrointestinal disorders. The value of probiotics as therapy for a variety of gastrointestinal disorders in childhood still needs to be investigated in detail, through well controlled and rigorous studies, including a placebo group and strict criteria of randomisation. Much work needs to be done in this area by clearly defining indications, delivery system, costs, safety long-term effects.

Child↗

A comparative study of the preventative effects exerted by two probiotics, Lactobacillus reuteri and Lactobacillus fermentum, in the trinitrobenzenesulfonic acid model of rat colitis.

The intestinal anti-inflammatory effects of two probiotics isolated from breast milk, Lactobacillus reuteri and L. fermentum, were evaluated and compared in the trinitrobenzenesulfonic acid (TNBS) model of rat colitis. Colitis was induced in rats by intracolonic administration of 10 mg TNBS dissolved in 50% ethanol (0.25 ml). Either L. reuteri or L. fermentum was daily administered orally (5 x 10(8) colony-forming units suspended in 0.5 ml skimmed milk) to each group of rats (n 10) for 3 weeks, starting 2 weeks before colitis induction. Colonic damage was evaluated histologically and biochemically, and the colonic luminal contents were used for bacterial studies and for SCFA production. Both probiotics showed intestinal anti-inflammatory effects in this model of experimental colitis, as evidenced histologically and by a significant reduction of colonic myeloperoxidase activity (P<0.05). L. fermentum significantly counteracted the colonic glutathione depletion induced by the inflammatory process. In addition, both probiotics lowered colonic TNFalpha levels (P<0.01) and inducible NO synthase expression when compared with non-treated rats; however, the decrease in colonic cyclo-oxygenase-2 expression was only achieved with L.fermentum administration. Finally, the two probiotics induced the growth of Lactobacilli species in comparison with control colitic rats, but the production of SCFA in colonic contents was only increased when L. fermentum was given. In conclusion, L. fermentum can exert beneficial immunomodulatory properties in inflammatory bowel disease, being more effective than L. reuteri, a probiotic with reputed efficacy in promoting beneficial effects on human health.

Animals↗

Public health issues arising from microbiological and labelling quality of foods and supplements containing probiotic microorganisms.

OBJECTIVE: To assess the accuracy and helpfulness of labelling on products containing probiotic bacteria. DESIGN AND SETTING: 52 such products - 44 from the UK (21 supplements, 15 fermented functional foods, eight 'health-care' products) and eight from continental Europe - have been tested for microbiological content, and results compared to the information available on their labels. Products were stored in the dark at 4 degrees C and analysed before their expiry or sell-by date. Careful note was taken of wording on labels, package inserts, packaging, promotional literature and catalogue descriptions, as applicable. Products were cultured on appropriate bacteriological media, and organisms grown were counted and identified. RESULTS: Bioyoghurts gave no indication of numbers, and only five accurately described their bacterial content; results of culture were usually satisfactory. 'Healthcare' products (mostly intended for the bowel) usually indicated the presence of bacteria, but the numerical content was hard to ascertain, and cultural results fell short of label claims. Supplements were sometimes incorrectly labelled in bacteriological terms, and often contained markedly reduced numbers and/or had extraneous strains and/or strains specified on the label were missing. Products from continental Europe (that were sold for specific medical indications) seemed of a higher microbiological standard. The potential pathogen Enterococcus faecium was found in nine products. The most successful of the new functional foods in Britain now contain probiotics, and probiotic preparations are prominent among the expanding range of nutritional supplements presently available to consumers. CONCLUSIONS: Our findings have public health implications, and suggest that improvements are needed in labelling and quality assurance procedures for products containing probiotic organisms. The presence of the potential pathogen Enterococcus faecium (intentionally or as a contaminant) in some products calls for a review of the value of this species as a probiotic.

Dietary Supplements↗

Screening of intestinal microflora for effective probiotic bacteria.

Increasing consumer awareness of health-promoting intestinal bacteria has fueled the addition of viable probiotic bacteria as functional ingredients in certain foods. However, to effectively market the enhanced attributes of these foods, the added probiotic bacteria need to have scientific credibility. The scientific rationale for using many of the strains of probiotic bacteria currently on the market is weak. Furthering the current understanding of what features a bacterium needs to have for effective probiotic functionality will enable the selection of strains with a more credible scientific rationale. To screen for effective strains, one must understand the microbial diversity in the intestines of healthy individuals. The advent of molecular tools has greatly enhanced our ability to accomplish this. These tools comprise genetic fingerprinting, specific probes, molecular speciation, and techniques for the in situ analysis of specific microbial groups in the intestine. This review will detail these scientific approaches and how their impact will improve criteria for selection of probiotic bacteria.

Bifidobacterium↗

Engineered probiotic overcomes pathogen defences using signal interference and antibiotic production to treat infection in mice.

Probiotic supplements are suggested to promote human health by preventing pathogen colonization. However, the mechanistic bases for their efficacy in vivo are largely uncharacterized. Here using metabolomics and bacterial genetics, we show that the human oral probiotic Streptococcus salivarius K12 (SAL) produces salivabactin, an antibiotic that effectively inhibits pathogenic Streptococcus pyogenes (GAS) in vitro and in mice. However, prophylactic dosing with SAL enhanced GAS colonization in mice and ex vivo in human saliva. We showed that, on co-colonization, GAS responds to a SAL intercellular peptide signal that controls SAL salivabactin production. GAS produces a secreted protease, SpeB, that targets SAL-derived salivaricins and enhances GAS survival. Using this knowledge, we re-engineered probiotic SAL to prevent signal eavesdropping by GAS and potentiate SAL antimicrobials. This engineered probiotic demonstrated superior efficacy in preventing GAS colonization in vivo. Our findings show that knowledge of interspecies interactions can identify antibiotic- and probiotic-based strategies to combat infection.

Animals↗

Whole-genome sequencing reveals hidden antimicrobial resistance genes in phenotypically susceptible probiotic candidate lactic acid bacteria.

Phenotypic assays commonly used to evaluate probiotic safety may fail to detect clinically relevant antimicrobial resistance (AMR), potentially allowing genetically concerning strains to appear acceptable based on MIC testing alone. To explore this issue, we applied whole-genome sequencing (WGS) to three lactic acid bacteria (LAB) isolates previously identified as probiotic candidates based on acid and bile tolerance, antagonism against enteric pathogens, and biofilm formation in vitro: Lactiplantibacillus plantarum L25F and L22F (from pigs) and Ligilactobacillus salivarius AF2319 (from a chicken). Genome annotation identified extensive repertoires of probiotic-associated genes (46-47 per strain) linked to stress tolerance, adhesion, immunomodulation, and quorum sensing, supporting functional potential. The two L. plantarum strains exhibited broader predicted metabolic capacities than L. salivarius AF2319. However, genomic analysis revealed acquired AMR genes with complex genotype-phenotype relationships not fully apparent from phenotypic testing. The L. plantarum strains harbored lnu(A) (99.79% identity) on extrachromosomal DNA, conferring the L-phenotype (lincomycin resistance, clindamycin susceptibility); clindamycin MICs (1&#xa0;mg/L) were concordant with this genotype, though lincomycin MICs were not determined. L. salivarius AF2319 carried tet(M), tet(L), and erm(C) (99.48%, 99.49%, and 99.45% identity by ResFinder, respectively) on extrachromosomal DNA; notably, the erythromycin MIC (1&#xa0;mg/L) was precisely at the EFSA breakpoint (&#x2264;&#x2009;1&#xa0;mg/L), representing borderline genotype-phenotype discordance potentially due to silent gene expression. Under current EFSA QPS criteria, these acquired ARGs would preclude all three strains from approval as probiotic feed additives despite favorable functional profiles, underscoring the indispensable role of WGS-based AMR gene detection in modern probiotic safety evaluation.

Probiotics↗

Dose-response study of probiotic bacteria Bifidobacterium animalis subsp lactis BB-12 and Lactobacillus paracasei subsp paracasei CRL-341 in healthy young adults.

OBJECTIVE: This study was performed to investigate the dose-response effects of supplementation with Bifidobacterium animalis subsp lactis (BB-12) and Lactobacillus paracasei subsp paracasei (CRL-431) on blood lipids, recovery from feces and bowel habits. Changes of the fecal microflora was analyzed in the 10(10) CFU/day probiotic and placebo group. DESIGN: The study was designed as a randomized, placebo-controlled, double-blinded, parallel dose-response study. SUBJECTS: Healthy young adults (18-40 years) were recruited by advertising in local newspapers. Of the 75 persons enrolled, 71 (46 women, 25 men, mean age 25.6 years (range 18-40 years)) completed the study. INTERVENTION: The volunteers were randomly assigned into five groups receiving either placebo or a mixture of the two probiotics in the concentration of 10(8), 10(9), 10(10) or 10(11) CFU/day in 2 weeks run-in period, 3 weeks intervention and 2 weeks wash-out. Diary reporting bowel habits and well being (abdominal bloating, flatulence and headache) was kept for all 7 weeks and blood lipids, fecal recovery of BB-12 and CRL-431, as well as fecal microflora was tested before, immediately and 2 weeks after intervention. RESULTS: The fecal recovery of BB-12 increased significantly (P < 0.001) with increasing dose. In the group receiving 10(11) CFU/day BB-12 was recovered from 13 out of 15 volunteers. CRL-431 was not recovered in any of the fecal samples. Supplementation with probiotics did not change the fecal bacterial composition. A significant linear increase in fecal consistency (looser stool) with increasing probiotic dose (P = 0.018) was observed. No overall dose-response effect was found on the blood lipids. High doses of probiotics were well tolerated. CONCLUSION: A dose-related recovery of BB-12 from feces was observed.

Adolescent↗

Use of complementary and alternative therapies and probiotic agents by children attending gastroenterology outpatient clinics.

OBJECTIVES: Complementary and alternative medicines (CAM) and probiotic therapies appear to be increasingly accepted and used. This questionnaire-based study aimed to ascertain the frequency of use and the acceptance of these therapies by children attending outpatient gastroenterology clinics. METHODS: Parents accompanying children to their appointments were asked to complete a questionnaire in order to determine usage of probiotic and alternative therapies. Questions also ascertained relevant background information and parental acceptance of alternative therapies. RESULTS: Ninety-two questionnaires were completed. The ages of the children varied from 6 months to 16 years (mean +/- SD; 6.5 +/- 4.3 years) and they had been prescribed an average of 1.7 +/- 1.3 (range 0-6) conventional medications. Thirty-three children (35.9%) were taking CAM and 98.6% of parents answered that they would be prepared to administer CAM to their child. Symptomatic improvements were attributed to CAM by the parents of 24 out of 33 children given these therapies. In addition, probiotic therapies were utilized by 23.8% of children, and 93.0% of parents would administer probiotic agents if recommended for their child's condition. CONCLUSION: Complementary and alternative medicines and probiotic therapies are used frequently by children attending gastroenterology clinics and are accepted widely by their parents.

Adolescent↗

Adhesion of inactivated probiotic strains to intestinal mucus.

It has been suggested that probiotics should be viable in order to elicit beneficial health effects. Inactivation of probiotics has been suggested to interfere with the binding to the mucosa and thereby with the immune modulating activity of probiotics. The effect of different inactivation methods on the mucus adhesion of nine probiotic strains was studied. Inactivation by heat or gamma-irradiation generally decreased the adhesive abilities. However, heat treatment increased the adhesion of Propionibacterium freudenreichii and gamma-irradiation enhanced the adhesion of Lactobacillus casei Shirota. Inactivation by u.v. was not observed to modulate the adhesion of the tested strains and it was concluded to be the most appropriate method for studying non-viable probiotics and preparing control products.

Bacterial Adhesion↗

Binding of extracellular matrix molecules by probiotic bacteria.

AIMS: The aim of this study was to investigate extracellular matrix (ECM) and mucin binding of selected bacterial isolates with probiotic features in comparison with commercially used probiotic bacteria. METHODS AND RESULTS: ECM molecules were immobilized in microtitre plates (mucin and fetuin) or on the surface of latex beads. Porcine mucin was bound by all 13 probiotic strains tested with important inter-strain differences; however, fetuin binding was similar (weak) for all 14 strains tested. Strongly positive (three) binding of bovine fibrinogen was expressed by strains from fermented food (Lactobacillus rhamnosus GG, L. casei Shirota and L. johnsonii La1) as well as by L. casei L.c., Lactobacillus sp. 2I3 and by L. plantarum LP. The other strains expressed moderate (2) or weakly positive (1) binding of bovine fibrinogen. Strongly positive (3) binding of porcine fibronectin was observed only with two strains; however, all other strains also bound this molecule. Bovine lactoferrin was bound to a higher extent than transferrins. SIGNIFICANCE AND IMPACT OF THE STUDY: Some animal strains (at least L. casei L.c. and Lactobacillus sp. 2I3) are comparable with the commercially used strains with respect to their ECM binding ability. As this feature is important for probiotic bacteria to be able to colonize intestine, these strains should be considered for their wider use in fermented feed (or probiotic preparations) for animals.

Animals↗

The effect of probiotic LSP 122 on the control of post-weaning diarrhoea syndrome of piglets.

Post-weaning diarrhoea syndrome (PWDS) of piglets is caused mainly by enterotoxigenic Escherichia coli (ETEC) strains. A new in-feed probiotic, LSP 122 (Alpharma), containing viable spores of Bacillus licheniformis was tested for its efficacy to control PWDS in piglets in a low health-status farm, using four groups with a total of 256 weaned piglets for a 28-day period. One group (negative control) was offered antimicrobial-free and probiotics-free fed, one group was offered feed supplemented with 10(6)viable spores of Bacillus toyoi (Toyocerin(R)) per gram of feed and two groups were offered feed supplemented with 10(6)and 10(7)viable spores of B. licheniformis per gram of feed, respectively, and were compared with regard to the appearance of clinical signs, mortality, weight gain and feed conversion. The results showed that all groups supplemented with probiotics exhibited a reduced incidence and severity of diarrhoea. Mortality in all probiotic supplemented pigs was significantly lower compared with the negative control group (P<0.05). The evaluation of the weight gain data, as well as feed conversion ratio, indicated that the three treated groups performed remarkably better than the negative control group (P<0.05) and the group receiving the high inclusion of LSP 122 performed better than the two other groups receiving probiotics (P<0.05). No ETEC strains were detected on day 22 in the high inclusion of LSP 122 and Toyocerin groups as compared with the untreated control. It was concluded that the high dosage schedule of LSP 122, providing 10(7)viable spores of B. licheniformis per g of feed, is a very useful agent for the control of PWDS due to ETEC.

Animals↗

Probiotics during pregnancy and breast-feeding might confer immunomodulatory protection against atopic disease in the infant.

The prevalence of atopic diseases is increasing throughout the Western world, and means of primary prevention are needed to reverse this trend. The role of breast-feeding, the best source of infant nutrition, in protection against atopic disease remains elusive. In this double-blinded, placebo-controlled study of 62 mother-infant pairs, it is shown that administering probiotics to the pregnant and lactating mother increased the immunoprotective potential of breast milk, as assessed by the amount of anti-inflammatory transforming growth factor beta2 (TGF-beta2) in the milk (2885 pg/mL [95% CI, 1624-4146] in mothers receiving probiotics vs 1340 pg/mL [95% CI, 978-1702] in mothers receiving placebo; P =.018). The risk of developing atopic eczema during the first 2 years of life in infants whose mothers received probiotics was significantly reduced in comparison with that in infants whose mothers received placebo (15% and 47%, respectively; relative risk, 0.32 [95% CI, 0.12-0.85]; P =.0098). Maternal atopy was a clear risk factor for atopic eczema in the infant. The infants most likely to benefit from maternal probiotic supplementation were those with an elevated cord blood IgE concentration. Administering probiotics during pregnancy and breast-feeding thus offers a safe and effective mode of promoting the immunoprotective potential of breast-feeding and provides protection against atopic eczema during the first 2 years of life.

Breast Feeding↗