Thursday, May 18, 2023

L Reuteri And Glycerol Probiotic

Diffusion Of Cargo From Dms

The Unique Probiotic Effects of L. reuteri

Initial binding of bacteria to DMs is a critical component of our formulation, however equally important is the ability to co-deliver beneficial luminal cargo needed by the adherent bacteria during transit of DMs through the gastrointestinal tract. Targeted delivery of maltose via diffusion out of the DMs directly to the probiotic bacterium over time was a desired feature of our system . However, since the method of cargo delivery would be diffusion through the porous surface of the microsphere and not its degradation, such as occurs in poly acid microspheres , the rate of diffusion is dependent upon the size of the microsphere, the mass of the solute, and the viscosity of the diluent. As proof of concept, we filled the DMs with crystal violet, a small molecular weight stain , and tested the diffusion rate of the dye out of the DMs with and without changing the viscosity of the solution in the DM lumen. As shown in Figure 4, the crystal violet diffused out of the DM lumen with a half-life of ~6 h. When the viscosity was increased by adding 40% glycerol, the half-life of release was increased to ~8 h. At 80% glycerol, the half-life of crystal violet release was further enhanced to 12 h. By 16 h > 95% of all of the crystal violet had been released under all tested conditions.

Maltose Or Sucrose Within The Lumen Of Dms Improved L Reuteri Adherence To Dms In A Gtf

Our strategy was to have probiotic bacteria adhere to a biocompatible surface to induce the formation of a biofilm . To investigate this, we differentially stained DMs with Congo Red and L. reuteri with SYTO 9, and examined binding via confocal laser scanning microscopy . As shown in Figure 2, aggregates of bacteria were associated with the surface of numerous DMs which indicated that L. reuteri was able to adhere to the DM surface within the time allotted. Since DMs are cross-linked glucan similar to the native reuteran produced by L. reuteri, we hypothesized that either an increase in GTFW or production of glucan to stimulate aggregation and biofilm formation would facilitate the adhered state of L. reuteri. To this end, we compared adherence of L. reuteri to DMs that contained luminal cargo of either sucrose or maltose . As shown in Figures 2B,C, compared to DMs that contained only water within the lumen there were greater numbers of L. reuteri adhered to DMs with either sugar as cargo.

Figure 2. L. reuteri binds to dextranomer microspheres. Confocal laser scanning microscopy of L. reuteri adhered to DMs. Water-filled DMs, sucrose-filled DMs, and maltose-filled DMs after incubation with L. reuteri for 30 min showed that L. reuteri adherence to DMs can be enhanced to incorporate biofilm-promoting cargo within the DM lumen .

Is There Lactobacillus Reuteri In Kefir

relating to Lactobacillus taxonomy, Lactobacillus species in kefir occupy a variety of subgroups , with at least one strain being present in the Lactobacillus plantarum, Lactobacillus delbrueckii, Lactobacillus casei, Lactobacillus reuteri and Lactobacillus buchneri groups, with other strains being found amongst the

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Chewable Tablets With Strawberry Flavor

For strong immunity and healthy gut flora Special density of the tablets for easier chewing Naturally pleasant taste of real strawberries Sugar free contains natural beneficial sweetener stevia Suitable for everyone, including children, pregnant women and nursing mothers

Recommended daily dose: 1-2 TABLETS A DAY CONTAIN 250-500 million alive L.Reuteri bacteria DSM 25175 = 250-500 million CFU . One packet is enough for 5-10 days. To be taken orally.

Excipients: Encapsulated citric acid, magnesium salts of fatty acids, cocoa butter, sweetener stevia, filler isomalt, natural flavouring strawberry. Excessive consumption may have a laxative effect. Net weight of one tablet 450 mg.

Microspheres Filled With Sucrose Or Maltose Improved L Reuteri Survival At Low Ph

ColiKids drops  BioGaia

Orally consumed probiotics face a significant pH challenge upon reaching the stomach, where pH values are as low as 1.5 when the stomach is empty . Enhancing the ability to deliver a maximal number of viable L. reuteri to the colon is crucial to its sustainability and effectiveness as a probiotic. We thereby hypothesized that L. reuteri bound to the surface of DMs in the form of a biofilm would increase survival upon exposure to acid, and that DMs filled with sucrose or maltose would result in even greater survival in a GTFW-dependent manner. As shown in Figure 6, less than 0.1% of WT L. reuteri without DMs survived in synthetic gastric acid after 4 h at pH 2, which resulted in a nearly 3 log loss of viable probiotic. Addition of water-filled DMs did not significantly alter the survival rate of WT L. reuteri in gastric acid however, when either DM-sucrose or DM-maltose was delivered with WT, nearly 1 log more survived the acid stress . To show that the protective effect is dependent on the microspheres and not the cargo within the DM lumen, we also incubated L. reuteri with the equivalent amount of diffusible cargo without the DMs. Acid survival in the presence of cargo only was no different than L. reuteri alone , which strongly indicated the importance of the bacterial biofilm-on-DM delivery system for the observed protective effect.

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Identification Of Isolates Using A Maldi

One colony of each bacterial isolate was subcultured for 24 h and used for MALDI-TOF Biotyper analysis. The colony was acquired using a toothpick and spotted onto a polished steel MALDI target plate. One microliter of formic acid was added to the spot and dried. Subsequently, 1 µL of MALDI matrix in 50% acetonitrile/2.5% trifluoroacetic acid) was added to the spot and dried. The MALDI target plate was placed in the MALDI-TOF/Microflex LT instrument for automated measurement and data interpretation. The MALDI Biotyper output is a log between 0 and 3.0, which is calculated from a comparison of the peak list from an unknown isolate with the reference MSP in the database. A log 1.7 was indicative of a close relationship at the genus level. A log 2.0 was set as the threshold for a match at the species level. Isolates with a log 2.0 were accepted as the correct identification.

Minimal Inhibitory Volume Assay

The MIV of LRS was determined using a modified method of a previously described procedure. Pathogenic bacterial cultures generated in suitable liquid culture media as detailed above were diluted with nutrient broth to an OD600 of 0.005 or 0.05. The diluted bacterial suspensions were then treated with either MRS medium or 1×LRS, dispensed into the first well of a 96-well plate, and serially diluted into consecutive wells. Plates were incubated at 37 °C for 24 h, and the absorbance at 600 nm was measured using a microplate reader.

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Introduction Of L Reuteri

L. reuteri species are gram-positive, rod-shaped, and anaerobic microorganisms. They are a part of the natural gut microbiota of a wide range of organisms, including humans, pigs, chickens, and mice. They are also found in human breast milk. L. reuteri form chain arrangements and do not produce endospores. The species typically produces carbon dioxide, ethanol, acetate, and lactic acid from glucose fermentation. They have also been shown to produce the nutrients, folate, and vitamin B12, that are required for human health. Besides, the main feature of L. reuteri is to produce a compound called reuterin via the fermentation of glycerol. Reuterin is a potent antimicrobial that inhibits the growth of a wide array of pathogenic intestinal bacteria such as pathogenic E. coli strains. For this reason, L. reuteri has been studied as a probiotic organism for improving gastrointestinal health.

L Reuteri Survival With Dm


Overnight cultures of WT L. reuteri were aliquoted into microcentrifuge tubes, centrifuged, washed twice with sterile saline, and resuspended in either 1 ml saline or 1 ml MRS medium. Five mg of either DM-water or DM-80% glycerol were then added to the tubes and incubated at 37°C. At hourly intervals the tubes were mixed thoroughly and aliquots were taken for subsequent serial dilution and plating for viable CFU of bacteria.

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Microbial Strains And Growth Conditions

S. mutans ATCC 25175 was cultured in brainheart infusion broth and incubated in anaerobic conditions under CO2 at 37°C. P. gingivalis ATCC 33277 was cultured in BHI broth and incubated in a GasPak jar system . L. reuteri ATCC 55730 was cultured in de Man, Rogosa, Sharpe broth and incubated at 37°C under anaerobic conditions. Before exposing the HaCat cells, S. mutans and P. gingivalis were killed by heating them at 80°C for 30 min.

A Concerted Probiotic Activity To Inhibit Periodontitis

  • Paul Mathias Jansen,

    Roles Investigation, Methodology, Writing original draft

    Affiliation Division of Oral Microbiology and Immunology, Department of Operative Dentistry, Periodontology and Preventive Dentistry, Rheinisch-Westfälische Technische Hochschule Aachen University Hospital, Aachen, Germany

  • Mohamed M. H. Abdelbary,

    Roles Data curation, Methodology, Writing review & editing

    Affiliation Division of Oral Microbiology and Immunology, Department of Operative Dentistry, Periodontology and Preventive Dentistry, Rheinisch-Westfälische Technische Hochschule Aachen University Hospital, Aachen, Germany

  • * E-mail:

    Affiliation Division of Oral Microbiology and Immunology, Department of Operative Dentistry, Periodontology and Preventive Dentistry, Rheinisch-Westfälische Technische Hochschule Aachen University Hospital, Aachen, Germany

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Potential Of L Reuteri Probiotics

  • Eczema

L. reuteri supplementation, in combination with L. rhamnosus has been shown to reduce the severity of eczema in infants with atopic dermatitis and cow’s milk allergy. In a double-blind, placebo-controlled, crossover study, supplementation of L. reuteri and L. rhamnosus probiotics to 1- to 13-year-old children with atopic dermatitis for 6 weeks could improve the symptom of eczema.

L. reuteri species produce a broad-range antimicrobial, reuterin, which inhibits growth, binding, and invasion of harmful bacteria. They may be involved in the improvement of intestinal barrier function and contributes to suppressing the release of proinflammatory cytokines such as TNF-alpha and stimulating protective cytokines such as IL-10 and TGF-. L. reuteri probiotics have been reported to have an analgesic effect on abdominal pain. Supplementation with the probiotic L. reuteri has also been revealed to improve infantile colic without adverse events. In some studies, L. reuteri seems to reduce antibiotic-associated diarrhea and functional constipation in adults and children. Furthermore, L. reuteri has been suggested to inhibit the growth of gut pathogen Helicobacter pylori which is a leading cause of peptic ulcers.

  • High Cholesterol

For Research Use Only. Not intended for use in food manufacturing or medical procedures . Do Not Use in Humans.

Strains And Culturing Conditions

Microbiome Plus+ Probiotic Lactobacillus Reuteri NCIMB ...

Table 1. Bacterial strains, cell lines, plasmids, and oligos used in this study.

To estimate transcription from the gtfW promoter , the PgtfW-CBluc reporter plasmid was constructed by amplifying the promoter region 350 bp upstream of the gtfW start codon by PCR using oligos oSG1102-1103. The resulting DNA fragment was inserted into pJC156 using the XhoI/SalI restriction sites. The click beetle luciferase gene was amplified from the Streptococcus mutans strain ldhCBGSm using oligos oSG1067-1068 and inserted downstream of the gtfW promoter region in pJC156 using SalI/NotI restriction sites. The resulting reporter plasmid pWAR501 was transformed into L. reuteri 23272 as described above to create the reporter strain LMW501.

The E. coli gtfW overexpression strain was created by amplifying the L. reuteri gtfW open reading frame using primers oSG1120-1126. The resulting DNA fragment was inserted into pTXB1 using NheI/SapI restriction sites. The resulting plasmid, pWAR502 was then transformed into the E. coli expression strain ER2566 and selected on LB agar containing 100 g/ml ampicillin and confirmed by DNA sequencing. This strain allows the overexpression of tagless GTFW protein.

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What Are The Researched Benefits Of Taking L Reuteri

  • reduce salivary mutans streptococus
  • reduce plaque on teeth
  • reduce infant colic
  • stabilize barrier function

The cool thing about L. reuteri is that it produces an antibiotic as a product of its metabolism. It makes a molecule that we now call reuterin that slows the growth of certain harmful bacteria, viruses, yeasts, fungi, and protozoa.A It makes enough to stop the bad guys but not enough to hurt the good guys. They can eliminate gut invaders without harming other, beneficial bacteria.

Bacterial Enumeration By Fish

FISH hybridization analyses were performed as described by on fixed fecal inoculum, fermentation samples and beads, from the last 3 days and last day of each pseudo-steady-state period, respectively. Beads were dissolved in 1% EDTA solution using a Stomacher® for 5 min. Different 5 Cy3-labelled 16S rRNA oligonucleotide probes were used with hybridization conditions specific for each probe: Bif164 for Bifidobacterium spp. Bac303 for BacteroidesPrevotella cluster Lab158 for Lactobacillus and Enterococcus EC1531 for Escherichia coli Erec482 for Clostridium coccoidesEubacterium rectale group and Lbre for Lactobacillus reuteri . For total cell counts 4, 6-diamidino-2-phenylindole was added at a final concentration of 1 g mL1. To prevent fading of fluorescence, citifluor AF1 was used. Cells were counted visually using an Olympus BX 60 epifluorescence microscope on 10-well slides . To minimize the counting error due to the radial distribution of bacteria in wells, bacterial concentrations were calculated from the bacterial density corresponding to 15 annular regions as already described by . Each assay was carried out in duplicate. Results were expressed as log cells mL1 of effluent samples or g1 of gel beads or feces. The detection limit of the method was log 6.0 cells mL1 or g1 of fermentation effluents and feces, respectively, and log 7.5 cells g1 of gel beads.

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Strain Isolation And Identification

Novel Lactobacillus reuteri strains were isolated from seven infants aged 37 years and from the small intestines of 13 6-month-old female pigs in the Republic of Korea. The pigs were fed a mixed diet. The intestinal contents from each child or pig were resuspended and serially diluted in sterile 0.85% NaCl. Aliquots were cultured anaerobically on MRS agar in an atmosphere of CO2:H2:N2 atmosphere. After 23 days, single colonies were subcultured on fresh MRS agar. All colonies were selected irrespective of their shape and size. Genomic DNA was extracted and purified from cells grown on MRS agar as described previously. The gDNA was used for 16S rRNA gene amplification and sequencing, and whole genome sequencing. The complete 16S rRNA gene sequence was amplified using universal primers: 27F and 1492R . The amplified genes were sequenced and compared with sequences obtained from the EzBioCloud and GenBank/EMBL/DDBJ databases.

Growth Inhibition Of The Periodontal Pathogens By Probiotic Strains And Vice Versa

Top 5 Reasons To Take L REUTERI for HAIR LOSS

A qRT-PCR was performed to measure the genome number of both, the pathogenic and the therapeutic probiotic strains, before and after inhibition assays. The DNA of the stock suspensions was serially diluted in tenfold step with nuclease-free water to create a standard curve. The qRT-PCR was performed by the aid of a QuantStudio 3 and in 96 well plate block formats . Except for the reciprocal inhibition experiments , every pathogen/probiotic combination and controls were run in biological triplicates and the DNA extracted from each well was measured in technical triplicates. The PowerUp SYBR Green Master Mix was used to create a reaction mix. Each well contained 20 l of the reaction mix with the following components: PowerUp SYBR Green Master Mix , Forward Primer , Reverse Primer , nuclease-free water , Template . The concentrations of all primers were 100 M. DNA of all pathogens andfor inverse experimentstwo probiotics was amplified and quantified with strain specific primers . As a negative control, nuclease-free water was added instead of the template.

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Bacterial Enumeration By Plate Counts

The concentration of L. reuteri ATCC 55730 in pure culture was determined by plate counts on MRS agar. Lactobacillus reuteri ATCC 55730 was recently shown to be resistant to tetracycline . The LAMVAB agar, selective for isolating lactobacilli in feces , and Lactobacillus Anaerobic MRS with Vancomycin and Bromocresol green agar supplemented with tetracycline were used to enumerate total lactobacilli and L. reuteri ATCC 55730, respectively, in fecal and intestinal fermentation samples. In a first attempt to selectively enumerate L. reuteri ATCC 55730, a concentration of 200 g mL1 tetracycline was added to LAMVAB medium. This tetracycline concentration was chosen after preliminary tests with pure cultures of L. reuteri ATCC 55730 and other Lactobacillus spp. commonly encountered in human feces. Spiked fecal samples with the same lactobacilli cultures were also tested . Agar plates were incubated in anaerobic jars at 37 °C for up to 5 days. Results were expressed as log CFU mL1 of effluent samples. The detection limit of the method was determined to be log 4.

Key Takeaways For Lactobacillus Reuteri

  • Lactobacillus reuteri is officially known as Limosilactobacillus reuteri it contains well studied strains within the species shown to be particularly helpful in womens and childrens health.
  • The benefits of L. reuteri include producing antimicrobial substances along with folate, vitamin B12 and reuterin.
  • Two of the most well studied strains of L. reuteri are Lactobacillus reuteri RC-14® and Lactobacillus reuteri Protectis®.

For more insights and professional updates on probiotics, please visit the Probiotic Professionals pages.

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The Ultimate Probiotic For Testosterone: Lactobacillus Reuteri

As some of you know, my second favorite health topic after testosterone is gut health. Im happy to finally tackle some of the effects that probiotics have on hormonal health, and especially the interaction between Lactobacillus reuteri and testosterone.

For many years, I have spoken and written about beneficial gut bacteria and the importance of intestinal health. Yet, I was still surprised when I saw some of the preliminary studies on probiotics and testosterone.

Really, it shouldnt maybe have been that big of a surprise, because we already know how important the gut microbiome is to almost all aspects of our health.

The digestive tract is really quite an astonishing machine. We can stuff our mouths with anything that even remotely resembles food, and our guts are able to deal with it.

The food is first broken down in the stomach and any unwanted microbes and pathogens are neutralized in the acidic environment.

At the same time, many organs around the digestive tract have kicked into high gear. The pancreas and stomach produce digestive enzymes that further break down carbs, protein, and fats. The liver produces bile, which the gallbladder then releases to help with the digestion of fats.

In the 20-feet-long small intestine, the food matter is further broken down before the nutrients are absorbed into the bloodstream.

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