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Survival of the probiotic, L. plantarum 299v and its effects on the faecal bacterial flora, with and without gastric acid inhibition.

INTRODUCTION: Probiotic bacteria have to survive passage through the gastrointestinal tract. In this placebo-controlled double-blind study, the effect of Lactobacillus plantarum 299v on the faecal flora was studied with and without gastric acid inhibition. METHODS: Thirty-two healthy volunteers were given pantoprazole (40 mg/day) or placebo for 3 weeks from week 2 until week 4. In addition, from week 3 until week 4, L. plantarum 299v in an oatmeal-fermented drink (10(9) CFU/ml) was given twice daily to both groups. From each healthy volunteer, faecal samples were collected at the end of week 1, 2, 4 and 8 (4 weeks after cessation of L. plantarum 299v and pantoprazole/placebo). Several aerobically and anaerobically growing bacteria were counted and short chain fatty acid concentrations were determined. RESULTS: In both the pantoprazole and the placebo group, median lactobacilli counts increased significantly in week 4 compared to week 1 (from log 4.5 to 8.0 CFU/g faeces in pantoprazole and from log 4.2 to 7.7 CFU/g faeces in placebo group) and decreased significantly in week 8 (to log 4.5 CFU/g faeces in pantoprazole and log 4.3 CFU/g faeces in placebo group). These lactobacilli were identified as L. plantarum 299v. No significant differences were observed in all other bacterial counts and short chain fatty acid concentrations. CONCLUSIONS: The comparable increase of faecal lactobacilli counts in both the pantoprazole and the placebo-treated group demonstrates that L. plantarum 299v survives passage through the gastrointestinal tract irrespective of gastric acidity. The increment of the intra-gastric pH in combination with L. plantarum 299v did not modulate bacterial composition and/or the production of short chain fatty acids.

2-Pyridinylmethylsulfinylbenzimidazoles↗

Toll-like receptors--novel targets in allergic airway disease (probiotics, friends and relatives).

Experimental and epidemiological studies enabled to hypothesize that stimulation of the immune system by selected microbial products may prevent or treat allergic diseases. According to recent advances in molecular immunology, this stimulation acts via group of conserved receptors present on antigen presenting cells, known as toll-like receptors (TLRs). These receptors play an essential role in antigen presentation and latter development of immune response into pro-allergic (Th2), cellular (Th1) or regulatory (Tr1) responses. Since toll-like receptors govern decisive points in immune regulation, an extensive research focuses on agents interfering with their immunomodulatory activities. In this report, we review information on the potential use of microbial products in allergy prevention and therapy, which are believed to target toll-like receptor network. Current toll-like receptor-based approaches, as well as potential use of lipopolysaccharide (and derivates), oligonucleotides, mycobacteria, bacterial extracts, and probiotics are discussed herein.

Animals↗

Survival of Escherichia coli O157:H7 in dry fermented sausages containing micro-encapsulated probiotic lactic acid bacteria.

Escherichia coli O157:H7 is capable of surviving the rigorous processing steps during the manufacture of dry fermented sausages. The effect of adding two probiotic organisms, Lactobacillus reuteri and Bifidobacterium longum as co-cultures with the meat starter cultures Pediococcus pentosaceus and Staphylococcus carnosus on the viability of E. coli O157:H7 in dry fermented sausages was studied. A 5 strain cocktail of E. coli O157:H7 was added at 7.4 log cfu/g to the sausage batter and challenged with either or both Lb. reuteri or B. longum before or after they were micro-encapsulated. Sausages were fermented at < or = 26 degrees C and 88% relative humidity (RH) followed by drying at 75% RH and 13 degrees C for 25 d. The pH, water activity (aw), protein, moisture, and numbers of all inoculated organisms were monitored during processing. The pH and aw decreased from 5.7 and 0.98 to 4.9 and 0.88 at the end of fermentation and drying, respectively. These processes reduced E. coli O157:H7 by 1.0 and 0.7 log cfu/g at the end of fermentation and drying, respectively. Unencapsulated Lb. reuteri with or without B. longum reduced E. coli O157:H7 by 3.0 log cfu/g and B. longum caused a 1.9 log cfu/g reduction. While micro-encapsulation increased survival of Lb. reuteri and B. longum, it reduced their inhibitory action against E. coli O157:H7.

Animals↗

Innate immune responses in rainbow trout (Oncorhynchus mykiss, Walbaum) induced by probiotics.

Carnobacterium maltaromaticum B26 and Carnobacterium divergens B33, which were isolated from the intestine of healthy rainbow trout (Oncorhynchus mykiss, Walbaum), were selected as being potentially useful as probiotics with effectiveness against Aeromonas salmonicida and Yersinia ruckeri. Thus, rainbow trout administered with feed supplemented with B26 or B33 dosed at >10(7) cells g(-1) feed conferred protection against challenge with virulent cultures of the pathogens. Moreover, both cultures persisted in the gut for up to 3 weeks after administration. The cultures enhanced the cellular and humoral immune responses. Specifically, fish fed with B26 demonstrated significantly increased phagocytic activity of the head kidney macrophages, whereas the use of B33 led to significant increases in respiratory burst and serum lysozyme activity. Also, the gut mucosal lysozyme activity for fish fed with both cultures was statistically higher than the controls.

Aeromonas↗

Transition of the probiotic bacteria, Lactobacillus casei strain Shirota, in the gastrointestinal tract of a pig.

The transition of probiotic bacteria Lactobacillus in the gastrointestinal tract was investigated in pigs that received commercially available fermented milk prepared with Lactobacillus casei strain Shirota (LCS). Three female pigs fistulated at the cecum were fed 130 ml of fermented milk that contained over 10(10) (cfu) LCS with their daily meal for 8 days. Cecal contents were sampled through a fistula every 2 h for 24 h after marker dosing. The viable cell number (log cfu/g) of LCS and the concentrations of transit markers in each sample were determined. The viable number of LCS cells ranged from 3.56 to 6.58. The number of LCS in the cecum was not stable in pigs and varied with the flow of the cecal content. The viable number of LCS cells was significantly correlated with the relative concentration of the marker. These results indicated that most LCS moved with the liquid component. The level of LCS reached the maximum (6.38) 6 h after dosing. Four doses every 6 h may be required to maintain the maximum level of LCS at the cecum.

Animals↗

Probiotic effects of Lactobacillus casei on DSS-induced ulcerative colitis in mice.

We tested the effect of Lactobacillus casei strain Shirota (LcS) on the murine model of ulcerative colitis induced by dextran sodium sulphate. The effect of LcS was tested either as a prophylactic 10 days before the onset of the disease, simultaneously with ulcerative colitis induction or continued 10 days after the disease was induced. LcS was not able to prevent the disease induction in any of the experiments. However, important clinical parameters including blood anemia indicators, body weight, and organ weight were improved in the animals receiving LcS as compared with the ulcerative colitis-induced controls. Increased colonic epithelial regeneration in the LcS treated animals was observed in the chronic stage. The results seemed better for the simultaneous short LcS treatment where some parameters remained similar to the PBS controls, including disease activity scores measured in the acute stage. We can conclude that although LcS alone cannot prevent the induction of ulcerative colitis by dextran sodium sulphate, it can improve the clinical condition of the mice. This could imply important biological consequences for the human situation. Further studies including LcS or other probiotic bacteria together with the available treatment are encouraged.

Animals↗

Optimization of medium composition for the production of a probiotic microorganism, Lactobacillus rhamnosus, using response surface methodology.

This study was undertaken to optimize yeast extract, glucose, and vitamin concentrations; and also culture pH for maximizing the growth of a probiotic bacterium, Lactobacillus rhamnosus, and to assess the effects of these factors by using response surface methodology. A central composite design was used as an experimental design for the allocation of treatment combinations. A polynomial regression model with cubic and quartic terms was used for analysis of the experimental data. It was found that the effects involving yeast extract, glucose, vitamins and pH on the growth of L. rhamnosus were significant, and the strongest effect was given by the yeast extract concentration. Estimated optimum conditions of the factors for the growth of L. rhamnosus are as follows: pH=6.9; vitamin solution=1.28% (v/v); glucose=5.01% (w/v) and yeast extract=6.0% (w/v).

Bacteriological Techniques↗

Development of a new oat-based probiotic drink.

In the present work, a whole-grain oat substrate was fermented with lactic acid bacteria to obtain a drink, combining the health benefits of a probiotic culture with the oat prebiotic beta-glucan. The levels of several factors, such as starter culture concentration, oat flour and sucrose content, affecting the fermentation process, were established for completing a controlled fermentation for 8 h. The viable cell counts reached at the end of the process were about 7.5 x 10(10) cfu ml(-1). It was found that the addition of sweeteners aspartame, sodium cyclamate, saccharine and Huxol (12% cyclamate and 1.2% saccharine) had no effect on the dynamics of the fermentation process and on the viability of the starter culture during product storage. Beta-glucan content in the drink (0.31-0.36%) remained unchanged both throughout fermentation and storage of the drink. The shelf life of the oat drink was estimated to 21 days under refrigerated storage.

Avena↗

New therapeutic strategy for combating the increasing burden of allergic disease: Probiotics-A Nutrition, Allergy, Mucosal Immunology and Intestinal Microbiota (NAMI) Research Group report.

The dietary approach to reducing the risk of atopic diseases in infancy is evolving from passive allergen avoidance to active stimulation of the immature immune system, the aim of which is to support the establishment of tolerance. The intestinal mucosa and the mucosa-associated immune system are the primary loci of allergen contact and induction of immune responsiveness. In this review we discuss cross-talk between the intestinal microbiota and the host as it pertains to healthy immunologic maturation. Understanding these complex phenomena provides the rationale for the use of probiotics in reducing the risk and nutritional management of atopic disease.

Animals↗

D-lactic acidosis in short-bowel syndrome managed with antibiotics and probiotics.

D-lactic acidosis sometimes occurs in malabsorbed patients with short-bowel syndrome and is characterized by recurrent episodes of encephalopathy and metabolic acidosis. The characteristic neurologic abnormalities and the presence of metabolic acidosis raises a diagnostic suspicion, and the diagnosis is made when the serum level of D-lactic acid is greater than 3 mmol/L. Standard treatment consists of restricting oral carbohydrates or fasting, correction of metabolic acidosis, and a long-term suppression of pathogenic floras with antibiotics. The authors present a case of D-lactic acidosis in a 22-year-old patient with short-bowel syndrome, to whom intestinal bacterial agents (probiotics) were given in addition to oral kanamycin. Recolonization of the intestine with nonpathogenic floras should be a long-term treatment for D-lactic acidosis.

Acidosis, Lactic↗

Effects of the probiotic strain Lactobacillus johnsonii strain La1 on autonomic nerves and blood glucose in rats.

Oral administration of Lactobacillus casei reportedly reduces blood glucose concentrations in a non-insulin-dependent diabetic KK-Ay mouse model. In order to determine if other lactobacillus strains affect glucose metabolism, we evaluated the effect of the probiotic strain Lactobacillus johnsonii La1 (LJLa1) strain on glucose metabolism in rats. Oral administration of LJLa1 via drinking water for 2 weeks inhibited the hyperglycemia induced by intracranial injection of 2-deoxy-D-glucose (2DG). We found that the hyperglucagonemic response induced by 2DG was also suppressed by LJLa1. Oral administration of LJLa1 for 2 weeks also reduced the elevation of blood glucose and glucagon levels after an oral glucose load in streptozotocin-diabetic rats. In addition, we recently observed that intraduodenal injection of LJLa1 reduced renal sympathetic nerve activity and enhanced gastric vagal nerve activity, suggesting that LJLa1 might affect glucose metabolism by changing autonomic nerve activity. Therefore, we evaluated the effect of intraduodenal administration of LJLa1 on adrenal sympathetic nerve activity (ASNA) in urethane-anesthetized rats, since the autonomic nervous system, including the adrenal sympathetic nerve, may be implicated in the control of the blood glucose levels. Indeed, we found that ASNA was suppressed by intraduodenal administration of LJLa1, suggesting that LJLa1 might improve glucose tolerance by reducing glucagon secretion via alteration of autonomic nerve activities.

Administration, Oral↗

Method for direct selection of potentially probiotic Bifidobacterium strains from human feces based on their acid-adaptation ability.

A method for direct selection of acid-resistant Bifidobacterium strains was developed by prolonged exposure of human feces to homologous lethal stress conditions. The recovered strains were intrinsically resistant to acidic gastric conditions (pH 2.0) and also showed good tolerance to high concentrations of bile salts and NaCl. Bifidobacterium breve and Bifidobacterium adolescentis were, respectively, the infant- and adult-type bifidobacterial species showing the highest ability to develop an acid-tolerant phenotype. Therefore, this procedure could be applicable to the direct selection of Bifidobacterium strains with improved stability in adverse environments and, probably, contribute to expand the spectra of probiotic species of human origin currently marketed.

Adaptation, Physiological↗

Kinetic analysis of the antibacterial activity of probiotic lactobacilli towards Salmonella enterica serovar Typhimurium reveals a role for lactic acid and other inhibitory compounds.

Six Lactobacillus strains including commercial probiotic ones (L. acidophilus IBB 801, L. amylovorus DCE 471, L. casei Shirota, L. johnsonii La1, L. plantarum ACA-DC 287 and L. rhamnosus GG) were investigated, through batch fermentations under controlled conditions, for their capacity to inhibit Salmonella enterica serovar Typhimurium SL1344. All lactobacilli displayed strong antibacterial activity toward this Gram-negative pathogen and significantly inhibited invasion of the pathogen into cultured human enterocyte-like Caco-2/TC7 cells. By studying the production kinetics of antibacterial activity and applying the appropriate acid and pH control samples during a killing assay, we were able to distinguish between the effect of lactic acid and other inhibitory compounds produced. The antibacterial activity of L. acidophilus IBB 801, L. amylovorus DCE 471, L. casei Shirota and L. rhamnosus GG was solely due to the production of lactic acid. The antibacterial activity of L. johnsonii La1 and L. plantarum ACA-DC 287 was due to the production of lactic acid and (an) unknown inhibitory substance(s). The latter was (were) only active in the presence of lactic acid. In addition, the lactic acid produced was responsible for significant inhibitory activity upon invasion of Salmonella into Caco-2/TC7 cells.

Caco-2 Cells↗

Selection of potential probiotic lactobacilli from pig feces to be used as additives in pelleted feeding.

Thirty-five isolates from pig feces were identified as Lactobacillus reuteri (12 strains), Lactobacillus mucosae (7), Lactobacillus plantarum (6), Lactobacillus kitasatonis (3), Lactobacillus rossiae (2), Lactobacillus ultunensis (2), Lactobacillus crispatus (2), and Lactobacillus intestinalis (1) by partial sequence analysis of the 16S rRNA. All isolates were detected at 8-9 log CFU g(-1). Preliminarily, strains were selected based on resistance to heat treatments (ca. 70 degrees C for 10 s). The decrease in viability for some L. reuteri, L. mucosae, L. plantarum, L. kitasatonis, and L. rossiae strains was lower than 1 log cycle. Selected strains were further characterized for acid and bile salt resistance, and antibacterial activity. Except for L. kitasatonis, tolerance to simulated gastric and intestinal conditions was enhanced for all strains by addition of reconstituted skimmed milk. Antibacterial activity was found against Gram-positive and -negative potential pathogens. L. reuteri 8.1, 3S7, 6.2, and 1.2, L. mucosae 1.1R, L. plantarum 4.1, and L. rossiae 4.4 were freeze-dried and mixed (1%, w/w) into pig feed before pelleting. After pelleting, pig feed contained 10-9 log CFU kg(-1) of lactobacilli. L. plantarum 4.1, and L. reuteri 3S7 were selected based on their bile salt resistance, pH tolerance, antimicrobial activity and heat resistance. The findings in this study provide a strong basis for exploring the potential of porcine lactobacilli isolates to be used in pelleted feeding as probiotic additives.

Animal Feed↗

A combinatorial construct library enables an expanded expression range of secreted therapeutic proteins by probiotic yeast.

Orally administered engineered probiotics, including Saccharomyces cerevisiae var. boulardii (Sb), are of emerging interest as protein therapeutic delivery platforms to treat gastrointestinal diseases. Tools to readily optimize protein output are required to optimize the therapeutic index of Sb-produced therapies. In this study, a 125-plex Sb secretion construct library was developed consisting of all possible combinations of five promoters, five secretion signals, and five terminators, which enabled a greater than 1800-fold range in Sb expression of a Gaussia luciferase (GLuc) reporter. Secretion signal and promoter identities had significant effects on secretion output. This library further enabled a 28-fold improvement of binding activity of Sb-secreted haPD-1, an established anti-tumor immunotherapeutic, and improved haPD-1 detection in mouse stool samples following oral gavage of Sb_haPD-1. Sb secretion trends of both GLuc and haPD-1 in vitro mirrored payload expression in vivo. This protein secretion library toolkit will serve as a valuable resource to rapidly optimize protein therapeutic output from engineered Sb.

Saccharomyces boulardii↗

Removal of the cyanobacterial toxin microcystin-LR by human probiotics.

Three human probiotics, Lactobacillus rhamnosus strains GG and LC-705, and Bifidobacterium lactis strain Bb12, were found to bind the cyanobacterial peptide toxin microcystin-LR from water solutions. The highest removal percentage was 46%, observed with heat-treated L. rhamnosus strain GG (10(10) cells/ml) and a microcystin-LR concentration of 0.5 microg/ml during an incubation of 7 h at 35 degrees C.

Bifidobacterium↗

The role of probiotics in aquaculture.

The increase of productivity in aquaculture has been accompanied by ecological impacts including emergence of a large variety of pathogens and bacterial resistance. These impacts are in part due to the indiscriminate use of chemotherapeutic agents as a result of management practices in production cycles. This review provides a summary of the use of probiotics for prevention of bacterial diseases in aquaculture, with a critical evaluation of results obtained to date.

Animals↗

Short-term consumption of probiotic-containing cheese and its effect on dental caries risk factors.

Cheese is known to contain compounds that reduce the risk of dental caries. The long-term consumption of milk containing Lactobacillus rhamnosus GG, ATCC 53103 (LGG), has been shown to reduce caries risk in children. The aim of the present study was to examine whether short-term consumption of cheese containing LGG and Lactobacillus rhamnosus LC 705 would diminish caries-associated salivary microbial counts in young adults. Altogether, 74 18-35 year-old subjects completed this double-blinded, randomised, placebo-controlled study. During the 3 week intervention, the subjects ate 5 x 15 g cheese per day. Oral examinations were made before and after the study. Stimulated salivary secretion rates, buffer capacity and counts of salivary Streptococcus mutans, yeast and lactobacilli were evaluated before and after the intervention and after a 3 week post-treatment period. The results showed no statistically significant difference between the groups in Streptococcus mutans counts after the intervention, but during the post-treatment period there was a significantly greater reduction in these counts in the intervention group compared to the control group (P=0.05). However, Streptococcus mutans counts decreased in 20% (P=0.01) and yeast counts in 27% (P=0.005) of all the subjects, regardless of the intervention group. Results from logistic regression showed a trend indicating that probiotic intervention might reduce the risk of the highest level of Streptococcus mutans (OR=0.37, 95% CI 0.08-1.75, P=0.21) and salivary yeasts (OR=0.40, 0.09-1.71, P=0.22).

Adolescent↗