Lactobacillus Fermentum ME-3 Research Articles

Lactobacillus fermentum ME-3 – a probiotic with potential for anti-atherogenic, anti-oxidative and anti-diabetic application

Lactobacillus fermentum ME-3 – a probiotic with potential for anti-atherogenic, anti-oxidative and anti-diabetic application

The Effect of Lactobacillus fermentum ME-3 Treatment on Glycation and Diabetes Complications.

Type 2 diabetes (T2D) induces organ damage associated with glycation, among other metabolic pathways. While therapeutic strategies have been tested to reduce the formation and impact of glycation products, results remain equivocal. Anti-diabetic therapies using probiotics have been proposed, but their effect upon glycation has not been reported. Here, the effects of the bacterial strain Lactobacillus fermentum ME-3 on glycation and T2D-related complications in a mouse model of T2D are investigated. Wild-type LepRdb/+ and diabetic LepRdb/db littermates receive a daily gavage of either water or the probiotic ME-3 strain (1010 CFU). Glycation markers, fructoselysine-derived furosine (FL-furosine) and carboxymethyllysine (CML), are quantified in four major organs and plasma using stable-isotope dilution LC-MS/MS. After 12 weeks of ME-3 treatment, diabetic mice gain less weight and exhibit an apparently improved glucose tolerance. The ME-3 treatment reduces median renal levels of FL-furosine in both genotypes by 12-15%, and renal and pulmonary free-CML in diabetic mice by 30% and 18%, respectively. Attenuated hepatic steatosis and an improved plasma lipid profile are also observed with treatment in both genotypes, while the gut microbiota profile is unchanged. L. fermentum ME-3 has therapeutic potential for reducing the formation/accumulation of some glycation products in kidneys and attenuating some common diabetes-related complications.

The effect of Lactobacillus fermentum ME-3 on the intestinal microbiota and urine polyamines content: A double-blind placebo-controlled pilot trial

The aim of present study was to evaluate the effects of the probiotic Lactobacillus fermentum ME-3 on composition of gut microbiota and urine polyamines level. The healthy volunteers were divided into two groups: probiotic group received kefir with L. fermentum ME-3 and control group regular kefir both for 8 weeks. The lactoflora was determined by bacteriological method, gut microbiota by real-time PCR and urine polyamines level by gas-chromatography. In probiotic group there was an increase in the richness (p = 0.025), diversity (p = 0.017) and counts (p = 0.003) of lactoflora. That was associated with the increase of the diversity of the whole gut microbiota (p = 0.003). After probiotic consumption the level of urine putrescine had negative correlation with counts of lactobacilli (r = −0.424; p = 0.0083) and bifidobacteria (r = −0.415; p = 0.00983). L. fermentum ME-3 has potential to lower the risk of gut diseases that are bound with lower richness and diversity of gut microbiota and higher putrescine level.

Regulation of plasma lipid profile by lactobacillus fermentum (probiotic strain ME-3 DSM14241) in a randomised controlled trial of clinically healthy adults

Previous studies showed that a probiotic-rich diet may improve the plasma lipid profile and lower the risk of cardiovascular disease (CVD). In a randomized controlled study we tested the possibility for regulation of plasma lipid profile using a kefir that contained the antioxidative probiotic strain, Lactobacillus fermentum ME-3 (DSM14241). The trial was performed in clinically healthy adults with borderline-high serum low-density lipoprotein-cholesterol (LDL-C) and/or high serum triglyceride (TG) levels based on guidelines from the European Cardiology Society and European Atherosclerosis Society. One hundred sixty four participants meeting the inclusion criteria were included. Participants were randomised to receive 200 ml/day kefir, either with probiotic (PG; n = 71, 58 females, mean age 49.6 ± 6.5 y) or without probiotic (CG; n = 66, 58 females, mean age 49.9 ± 6.1 y). The probiotic contained 4 × 107 cfu/ml L. fermentum ME-3 (total 8 × 109 cfu/day). At 4 weeks (n = 71 PG and n = 66 CG) and at 8 weeks (n = 43 PG and n = 33 CG), we evaluated anthropologic, blood biochemical indices, and the faecal temporal persistence of the probiotic strain was assessed by real-time PCR. After 4 weeks, the lipid profiles were mostly similar between groups: only the values of oxidised LDL (ox-LDL) and TG were significantly reduced (P < 0.001 and P = 0.005, resp.). After 8 weeks, the PG group exhibited reductions in LDL-C (5 %, P = 0.001), ox-LDL (6 %, P < 0.0002), TG (17 %, P = 0.033). Next, the ratio of LDL-C to HDL-C was decreased only in the PG (P = 0.013) while in the CG it was significantly increased (P = 0.018). After completing the trial the changes in all above listed indices were significantly (P < 0.05) different between the PG and CG groups. In faeces, the prevalence of L. fermentum ME-3 increased after 4 and 8 weeks (both P < 0.001), but the counts, determined with real-time PCR, remained constant. Eight weeks of consuming kefir with the antioxidative probiotic L. fermentum ME-3, reduced serum LDL-C, ox-LDL and TG values in clinically healthy volunteers with borderline-high lipid profile indices. Thus, L. fermentum ME-3 has potential to lower the risk of CVD that is tightly associated with maintenance of plasma lipid profile. This study was registered as current controlled trial http://www.controlled-trials.com/ISRCTN49744186 as ISRCTN49744186.

Probiotic, Antimicrobial, Antioxidant and Sensory properties of Fermented donkey milk with Lactobacillus fermentum ME-3 and Lactobacillus acidophilus (ATCC 4356)

Probiotic, Antimicrobial, Antioxidant and Sensory properties of Fermented donkey milk with Lactobacillus fermentum ME-3 and Lactobacillus acidophilus (ATCC 4356)

An antioxidant probiotic reduces postprandial lipemia and oxidative stress

Abstract Reducing postprandial oxidative stress (OxS), decreasing postprandial blood triglyceride level (TG) and improving lipoprotein status is likely to have a preventive impact on the development of cardiovascular disease (CVD). Previously we have shown that the antioxidant probiotic Lactobacillus fermentum ME-3 (DSM14241) is characterized by antiatherogenic effects. This randomized double-blind placebo-controlled study evaluated the influence of kefir enriched with an antioxidative probiotic L. fermentum ME-3 (LfKef) on postprandial OxS, blood TG response and lipoprotein status. 100 clinically healthy subjects were recruited into the study. Blood parameters of postprandial OxS, TG and lipoprotein status were determined by oxidized LDL, baseline diene conjugation in LDL (BDC-LDL), oxidized LDL complex with beta-2 glycoprotein (Beta2-GPI-oxLDL), paraoxonase (PON) activity, LDL-Chol, HDL-Chol and TG. To evaluate general body postprandial OxS-load we measured 8-isoprostanes (8-EPI) in the urine. Consumption of LfKef significantly reduced the postprandial level of oxidized LDL, BDC-LDL, Beta2-GPI-oxLDL, urinary 8-isoprostanes and postprandial TG and caused a significant increase in HDL-Chol and PON activity. This is the first evidence that kefir enriched with an antioxidant probiotic may have a positive effect on both postprandial OxS and TG response as well as on lipoprotein status.

Complete glutathione system in probiotic Lactobacillus fermentum ME-3

There is much information about glutathione (GSH) in eukaryotic cells, but relatively little is known about GSH in prokaryotes. Without GSH and glutathione redox cycle lactic acid bacteria (LAB) cannot protect themselves against reactive oxygen species. Previously we have shown the presence of GSH in Lactobacillus fermentum ME-3 (DSM14241). Results of this study show that probiotic L. fermentum ME-3 contains both glutathione peroxidase and glutathione reductase. We also present that L. fermentum ME-3 can transport GSH from environment and synthesize GSH. This means that it is characterized by a complete glutathione system: synthesis, uptake and redox turnover ability that makes L. fermentum ME-3 a perfect protector against oxidative stress. To our best knowledge studies on existence of the complete glutathione system in probiotic LAB strains are still absent and glutathione synthesis in them has not been demonstrated.

Effect of stress pretreatment on survival of probiotic bacteria in gastrointestinal tract simulator

The effect of stress pretreatment on survival of probiotic Lactobacillus acidophilus La-5, Lactobacillus rhamnosus GG, and Lactobacillus fermentum ME-3 cultures was investigated in the single bioreactor gastrointestinal tract simulator (GITS). The cultures were pregrown in pH-auxostat, subjected to temperature, acid, or bile stress treatment, fast frozen in liquid nitrogen (LN(2)), and tested for survival in GITS. After LN(2) freezing the colony forming ability of L. rhamnosus GG and L. fermentum ME-3 nonstressed and stressed cells was well retained (average survival of 75.4 +/- 18.3% and 88.0 +/- 7.2%, respectively). L. acidophilus La-5 strain showed good survival of auxostat nonstressed cells after fast freezing (94.2 +/- 15.0), however the survival of stress pretreated cells was considerably lower (30.8 +/- 8.5%). All LN(2) frozen auxostat cultures survived well in the acid phase of the GIT simulation (survival 81 +/- 21%); however, after the bile phase, the colony formation ability of L. acidophilus La-5, L. rhamnosus GG, and L. fermentum ME-3 decreased by approximately 1.4 +/- 0.2, 3.8 +/- 0.3, and 3.5 +/- 1.2 logarithmic units, respectively. No statistically relevant positive effect of stress pretreatments on survival of LN(2) frozen L. acidophilus La-5, L. rhamnosus GG, and L. fermentum ME-3 in GITS was observed.

Effects of a synbiotic product on blood antioxidative activity in subjects colonized withHelicobacter pylori

To evaluate the impact of the consumption of a synbiotic product on the antioxidative activity markers of blood in asymptomatic H. pylori-colonized persons. Fifty-three healthy adult volunteers without gastric symptoms participated in a randomized, double-blind placebo-controlled study. The crossover consumption of the enterocoated capsules containing antioxidative Lactobacillusfermentum ME-3, Lact. paracasei 8700:2 and Bifidobacterium longum 46 with Raftilose P95 lasted for 3 weeks and did not change the H. pylori colonization. In H. pylori-positive subjects the sera values of total antioxidative status (TAS) were significantly lower compared to H. pylori-negative subjects (0.97 vs 1.05 mmol l(-1), P = 0.008). After the consumption of the synbiotic, TAS values (0.97 vs 1.03 mmol l(-1), P = 0.004) increased, while the ratio between oxidized and reduced glutathione (0.035 vs 0.030, P = 0.016) decreased in H. pylori-positive subjects. The consumption of a synbiotic containing an antioxidative probiotic strain improved the reduced systemic antioxidative activity in H. pylori-colonized asymptomatic subjects. A synbiotic product containing an antioxidative probiotic strain may be useful in the reduction of systemic oxidative stress in H. pylori infection.

Double benefit claims for antimicrobial and antioxidative probiotic

This paper describes the in vitro assessment of the functional properties of probiotic Lactobacillus fermentum ME-3 (DSM14241) and evaluates the possibility for enhancement of the antioxidative activity of the human organism colonized or not with Helicobacter pylori (Hp) through the use of the synbiotic comprising L. fermentum ME-3, Lactobacillus paracasei 8700:2 and Bifidobacterium longum 46 with Raftilose P95 in a randomized double-blind placebo-controlled volunteer crossover trial. In vitro the probiotic strain ME-3 suppressed both clinical and reference strains of Hp and expressed the highest antioxidative activity among the probiotic bacteria. In a volunteer trial the consumption of the synbiotic suppressed the oxidative stress indices in healthy volunteers colonized or not with Hp. However, this beneficial effect was apparent without systemic antimicrobial influence of probiotic bacteria on Hp colonization. This suggests the possibility to apply the probiotic strain ME-3 with double health claims for reduction of the increased risk of cardiovascular diseases (CVD) in the composition of the synbiotic as an adjunct to Hp antimicrobial therapy. Key words: Helicobacter pylori, antioxidative activity, probiotic, lactobacilli, bifidobacteria, oxidative stress, synbiotic

In vitro effects of selected synbiotics on the human faecal microbiota composition

Synbiotics are recognized means of modulating gut microbiota composition and activities. However, whether synbiotics are superior to prebiotics and probiotics alone in moderating the gut microbiota towards a purportedly healthy composition has not been determined. Eight selected synbiotics (short-chain fructooligosaccharides or fructooligosaccharides, each combined with one of four probiotics, Lactobacillus fermentum ME-3, Lactobacillus plantarum WCFS1, Lactobacillus paracasei 8700:2 or Bifidobacterium longum 46) were added to 24-h pH-controlled anaerobic faecal batch cultures. The prebiotic and probiotic components were also tested alone to determine their respective role within the synbiotic for modulation of the faecal microbiota. Effects upon major groups of the microbiota were evaluated using FISH. Rifampicin variant probiotic strains were used to assess probiotic levels. Synbiotic and prebiotics increased bifidobacteria and the Eubacterium rectale-Clostridium coccoides group. Lower levels of Escherichia coli were retrieved with these combinations after 5 and 10 h of fermentation. Probiotics alone had little effect upon the groups, however. Multivariate analysis revealed that the effect of synbiotics differed from the prebiotics as higher levels of Lactobacillus-Enterococcus were observed when the probiotic was stimulated by the prebiotic component. Here, the synbiotic approach was more effective than prebiotic or probiotic alone to modulate the gut microbiota.

Reversible surface binding of cadmium and lead by lactic acid and bifidobacteria

Extensive cadmium and lead contamination of water has been reported to occur locally as a result of human activities. Lactic acid bacteria have been reported to remove cadmium and lead from water. The aim of this work was to clarify the mechanisms of cadmium and lead removal from water. In addition, the effect of other metals, reversibility of binding and recyclability of the biomass was assessed. Based on our earlier data, the two most promising lactic acid bacteria, Lactobacillus fermentum ME3 and Bifidobacterium longum 46, were selected for these experiments. The results showed that the presence of other cationic metals and blocking of carboxyl and phosphoryl groups reduced cadmium and lead removal. These results suggest involvement of electrostatic interactions in cadmium and lead removal, and support our earlier findings. Transmission electron micrographs showed large deposits of lead on the bacterial surface suggesting formation of metallic lead precipitates. Both cadmium and lead removal were reversible processes established by full recovery of removed metal after desorption with dilute solutions of EDTA and HNO 3. Resorption capacity of both biomasses tested was reduced after regeneration with 10 mM EDTA and 15 mM HNO 3. Taken together, the results suggest involvement of several reversible mechanisms such as ion exchange and precipitation in cadmium and lead binding by lactic acid bacteria. The results show that specific lactic acid bacteria have the potential for removal of cadmium and lead from water although reduction in resorption capacity after regeneration of the biomass may form a problem. Since the studies so far have mainly focused on removal of single metals from pure water, metal removal in conditions of natural waters should be assessed in further experiments.

Eradication of Salmonella Typhimurium infection in a murine model of typhoid fever with the combination of probiotic Lactobacillus fermentum ME-3 and ofloxacin

BackgroundThe aim of the study was to detect whether in experimental Salmonella enterica Typhimurium infection the probiotic Lactobacillus fermentum ME-3 in combination with fluoroquinolone therapy would eradicate S. Typhimurium, prevent the development of liver and spleen granulomas and improve the indices of oxidative stress in the ileum mucosa.The selected bacteriological, histological and biochemical methods were applied.ResultsCombined treatment with L. fermentum ME-3 and ofloxacin eradicated Salmonella Typhimurium from blood, ileum and liver, decreased the number of animals with liver and spleen granulomas and reduced the value of lipid peroxides in the ileum mucosa. Higher total counts of intestinal lactobacilli in all experimental groups were associated with the absence of liver granulomas.ConclusionThe antimicrobial and antioxidative probiotic L. fermentum ME-3 combined with ofloxacin enhances the eradication of experimental S. Typhimurium infection. These observations on probiotic and antimicrobial co-action may serve as basis to develop new strategies for treatment of invasive bacterial infections of the gut.

A Probiotic, Lactobacillus fermentum ME-3, Has Antioxidative Capacity in Soft Cheese Spreads with Different Fats

Our aim was to develop a prototype of a functional spread cheese containing both a specific probiotic and n-3 fatty acids and to analyze the viability of the probiotic and stability of n-3 fatty acids during 4 wk of shelf life. Lactobacillus fermentum ME-3 (Lf ME-3) isolated from a healthy Estonian child has been shown to have probiotic and antioxidative properties in several recent studies. In the current study this promising bacterial strain was combined with vegetable oils rich in nutritionally important α-linolenic acid and with unflavored cheese to obtain soft cheese spreads with different fat contents. Lactobacillus fermentum ME-3 survived well in all cheeses although the viable count did not increase during 4 wk of storage. The fatty acid composition of cheese triacylglycerols remained stable, whereas the profile of volatile compounds changed: hexanal and pentanal disappeared and the proportion of some alcohols increased. The changes in the profile of volatile compounds show the reductive power of Lf ME-3. A functional spread cheese containing n-3 fatty acids can be prepared with the probiotic Lactobacillus fermentum ME-3 strain leading to a reduced need for chemical anti-oxidants.

Rapid removal of lead and cadmium from water by specific lactic acid bacteria

Cadmium and lead are highly toxic metals. People are exposed to them primarily through food and water. Available conventional methods (precipitation, flocculation, ion exchange, and membrane filtration) for removal of these metals from water at low concentrations are claimed to be expensive and inefficient. Different microbes have been proposed to be an efficient and economical alternative in heavy metal removal from water. In this work, specific lactic acid bacteria (LAB) were assessed for their ability to remove cadmium and lead from water. Significant removal was observed, and it was found to be metal and bacterial strain specific. Removal was a fast, metabolism-independent surface process. It was also strongly influenced by pH, indicating that ion exchange mechanisms could be involved. The most effective metal removers were Bifidobacterium longum 46, Lactobacillus fermentum ME3 and Bifidobacterium lactis Bb12. The highest maximum cadmium and lead removal capacities of 54.7 mg metal/g and 175.7 mg/g dry biomass, respectively, were obtained with B. longum 46.

Antagonistic activity of probiotic lactobacilli and bifidobacteria against entero- and uropathogens

To develop in vitro assays for comparing the antagonistic properties and anti-oxidative activity of probiotic Lactobacillus and Bifidobacterium strains against various entero- and urinary pathogens. The antagonistic activity of five probiotic lactobacilli (Lactobacillus rhamnosus GG, Lactobacillus fermentum ME-3, Lactobacillus acidophilus La5, Lactobacillus plantarum 299v and Lactobacillus paracasei 8700:2) and two bifidobacteria (Bifidobacterium lactis Bb12, Bifidobacterium longum 46) against six target pathogens was estimated using different assays (solid and liquid media, anaerobic and microaerobic cultivation) and ranked (low, intermediate and high). Bacterial fermentation products were determined by gas chromatography, and the total anti-oxidative activity of probiotics was measured using linolenic acid test. Pyelonephritic Escherichia coli was highly suppressed by GG and both bifidobacteria strains. Lactobacilli strains 8700:2, 299v and ME-3 were the most effective against Salmonella enterica ssp. enterica in microaerobic while ME-3 and both bifidobacteria expressed high activity against Shigella sonnei in anaerobic milieu. Lact. paracasei, Lact. rhamnosus and Lact. plantarum strains showed intermediate antagonistic activity against Helicobacter pylori under microaerobic conditions on solid media. The highest anti-oxidative activity was characteristic for Lact. fermentum ME-3 (P < 0.05). No efficient antagonist against Clostridium difficile was found. The positive correlations between the pH, lactic acid production and anti-microbial activity for all tested probiotics were assessed. Developed experimental assays enable to compare the anti-microbial and -oxidative activity of Lactobacillus and/or Bifidobacterium probiotics, which have been claimed to possess the ability of suppressing the growth of various enteric and urinary pathogens. Screening Lactobacillus and Bifidobacterium sp. strains according to their activity in various environmental conditions could precede the clinical efficacy studies for adjunct treatment with probiotics in cure of different gastrointestinal and urinary tract infections.

The Influence of Antibacterial and Antioxidative Probiotic Lactobacilli on Gut Mucosa in a Mouse Model ofSalmonellaInfection

The purpose of the present study was to test the ability of selected probiotic Lactobacillus spp. (with high antimicrobial and antioxidative potential in in vitro tests) to compete with invasive Salmonella enterica serovar Typhimurium infection and protect the gut mucosa against excessive oxidative stress during inflammatory tissue damage in a mouse model. In total 47 mice were divided into four groups. The control group was treated with either phosphate-buffered saline (PBS, group 1) or alternatively 0.5x10 8 CFU/ml of human intestinal lactobacilli, namely Lactobacillus fermentum ME-3 (DSM 14241) and Lactobacillus acidophilus 0.5x10 8 CFU /ml (group 4), daily for 15 days. Group 2 and 3 mice were challenged with a clinical isolate of S. Typhimurium (0.5x10 5 CFU/ml). The group 3 mice were additionally orally inoculated with lactobacilli for 5 days before and 10 days after the challenge with S. Typhimurium. Counts of salmonellae and lactobacilli in blood, intestine, liver and spleen were recorded; the morphological indices of inflammation in the same organs and oxidative stress-indicative biochemical status (lipid peroxidation, total antioxidative activity, redox ratio of glutathione and iron content) of gut mucosa were assessed on the 10th day after oral inoculation. The administration of probiotic lactobacilli of human origin did not increase the total count of lactobacilli in the terminal ileum of mice; however, hyperplasia of lymph nodes was registered in group 4 mice compared with group 1 mice. In the gut the lactobacilli that were antagonistic against S. Typhimurium in vitro neither decreased the count of salmonellae nor prevented the spread of the infection. In contrast, the antioxidative potential of L. fermentum ME-3 influenced gut mucosa by the reduction of iron level (pro-oxidant), lipid peroxidation and increased total antioxidative activity, glutathione redox value. We conclude that properties of probiotic lactobacilli assessed in vitro could be expressed differentially during in vivo study. The possibility to influence the pro- and antioxidant balance by specific probiotic lactobacilli with antioxidative properties in the course of inflammatory tissue damage could be further explored. Key words: Salmonella Typhimurium, mouse model, probiotics, lactobacilli, oxidative stress, pro-oxidant, antioxidant, gut mucosa.

Antioxidative probiotic fermented goats' milk decreases oxidative stress-mediated atherogenicity in human subjects.

The increasing interest in a healthy diet is stimulating innovative development of novel scientific products in the food industry. The viable lactic acid bacteria in fermented milk products, such as yoghurt, have been associated with increased lactose tolerance, a well-balanced intestinal microflora, antimicrobial activity, stimulation of the immune system and antitumoural, anticholesterolaemic and antioxidative properties in human subjects. Recently, we have studied a human Lactobacillus spp. strain that possesses antioxidative activity. The aim of the present pilot study was to develop goats' milk fermented with the human antioxidative lactobacilli strain, Lactobacillus fermentum ME-3, and to test the effect of the fermented probiotic goats' milk on oxidative stress markers (including markers for atherosclerosis) in human blood and urine and on the gut microflora. Twenty-one healthy subjects were assigned to two treatment groups: goats' milk group and fermented goats' milk group (150 g/d) for a period of 21 d. Consumption of fermented goats' milk improved anti-atherogenicity in healthy subjects: it prolonged resistance of the lipoprotein fraction to oxidation, lowered levels of peroxidized lipoproteins, oxidized LDL, 8-isoprostanes and glutathione redox ratio, and enhanced total antioxidative activity. The consumption of fermented goats' milk also altered both the prevalence and proportion of lactic acid bacteria species in the gut microflora of the subjects. We conclude that the goats' milk fermented with our special antioxidative lactobacilli strain Lactobacillus fermentum ME-3 exhibits anti-atherogenic effects.