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Lactobacillus reuteri

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Lactobacillus reuteri is a species of Gram-positive, rod-shaped bacteria in the genus Lactobacillus. It is a probiotic microorganism found naturally in the gastrointestinal tracts of humans and animals.

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Lab products found in correlation

Ruminococcus gnavus, by American Type Culture Collection (3 mentions) Difco lactobacilli mrs broth, by BD (3 mentions) Lactobacillus rhamnosus, by American Type Culture Collection (2 mentions) Parabacteroides distasonis, by American Type Culture Collection (2 mentions) R gnavus, by American Type Culture Collection (2 mentions) Lactobacillus animalis, by American Type Culture Collection (1 mentions) 1 535 incubator, by Avantor (1 mentions) Lactobacillus gasseri, by American Type Culture Collection (1 mentions) Lactobacillus plantarum, by American Type Culture Collection (1 mentions) Lactobacillus casei, by American Type Culture Collection (1 mentions) Lactobacillus fermentum, by American Type Culture Collection (1 mentions) Lactobacillus helveticus, by American Type Culture Collection (1 mentions) Mrs broth, by BD (1 mentions) Lactobacillus strains, by American Type Culture Collection (1 mentions) Lactobacillus paracasei, by American Type Culture Collection (1 mentions) Lactobacillus acidophilus atcc, by American Type Culture Collection (1 mentions) Bifidobacterium breve, by American Type Culture Collection (1 mentions) Clostridium perfringens, by American Type Culture Collection (1 mentions) Bacteroides fragilis, by American Type Culture Collection (1 mentions) Lactococcus lactis, by American Type Culture Collection (1 mentions)

9 protocols using lactobacillus reuteri

1

Culturing Lactobacillus Probiotic Strains

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Lactobacillus animalis (ATCC 35046), Lactobacillus reuteri (ATCC 2837), and Lactobacillus rhamnosus (ATCC 23272) were used in this study. De Man, Rogosa and Sharpe (MRS) broth and MRS agar (BD Difco, Franklin Lakes, NJ) were used for culturing probiotics at 37°C for 48 h anaerobically (Anoxomat; Spiral Biotech, Norwood, MA; 1535 incubator; VWR International, Cornelius, OR). These strains were obtained from the American Type Culture Collection (ATCC, Manassas, VA).
Li T., Castañeda C.D., Miotto J., McDaniel C., Kiess A.S, & Zhang L. (2020). Effects of in ovo probiotic administration on the incidence of avian pathogenic Escherichia coli in broilers and an evaluation on its virulence and antimicrobial resistance properties. Poultry Science, 100(3), 100903.
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2

Probiotics Modulate Gut Microbiome in Mice

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MMb, HMb, and GF mice received orally administered Paraprevotella clara, Bacteroides uniformis, SFB, Lactobacillus reuteri (BEI HM-102), Ruminococcus gnavus (ATCC 29149), MMb cx, HMb cx, Clostridium innocuum, or Clostridium immunis (100–150 μl; ~108–109 colony-forming units). P. clara, B. uniformis, C. innocuum, and C. immunis were isolated from the feces of HMb mice; SFB was previously obtained from Y. Umesaki (Yakult; Tokyo, Japan) and propagated in SFB-monocolonized mice at Harvard Medical School. Seven days later, the mice were challenged with DSS. In some experiments, fecal samples were collected before and 7 days after probiotic administration for microbiota analysis. For Reg3γ experiments, HMb mice received orally administered (150–200 μl; ~108–109 colony-forming units) Parabacteroides distasonis (ATCC 8503; control bacteria), R. gnavus (ATCC 29149), L. reuteri (BEI HM-102), Allobaculum stercoricanis (DSM 13633), Muribaculum intestinale (DSM 28989), or Lactobacillus vaginalis (DSM 5837). Mice were sacrificed 7 days later.
Surana N.K, & Kasper D.L. (2017). Moving beyond microbiome-wide associations to causal microbe identification. Nature, 552(7684), 244-247.
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3

Lactobacillus Strains Preparation Protocol

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The PP mixture comprised various Lactobacillus strains supplied by the American Type Culture Collection (ATCC) (Manassas, VA, USA). The strains included Lactobacillus gasseri (ATCC 33323), Lactobacillus plantarum (ATCC BAA-793), Lactobacillus reuteri (ATCC 23272), Lactobacillus helveticus (ATCC BAA-2840), Lactobacillus fermentum (ATCC 23271), Lactobacillus rhamnosus (ATCC BAA-2836), and Lactobacillus casei (ATCC BAA-2843). Following the supplier’s protocol, we cultured the strains in MRS broth (Beckton Dickinson, Sparks, MD), and preserved them as glycerol stocks at −80°C. To initiate culture growth, we prepared 5 mL of pre-warmed MRS broth with the glycerol stocks and then incubated these starter cultures at 37°C in a 5% CO2 environment for 2 h. The bacteria were cultured overnight in 1 L of MRS broth under identical conditions until they reached the logarithmic growth phase, confirmed by measuring optical density at 600 nm (OD600). After growth, the bacterial cells were collected through several centrifugation steps at 3000×g and 4°C, followed by a wash in cold PBS. We then resuspended the bacterial pellets in 10% glycerol in PBS and promptly froze them in 1 mL aliquots for future administration to mice. The viability and concentration of the bacteria were verified by serial dilution and colony-forming unit (CFU) enumeration on agar plates.
Holcomb M., Marshall A., Flinn H., Lozano M., Soriano S., Gomez-Pinilla F., Treangen T.J, & Villapol S. (2024). Probiotic treatment causes sex-specific neuroprotection after traumatic brain injury in mice. Research Square.
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4

Culturing Diverse Anaerobic Bacteria

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Bacteria strains used are either purchased from ATCC (Lactobacillus acidophilus ATCC® 4356™, Lactobacillus paracasei ATCC® BAA-52™, Lac-tobacillus reuteri ATCC® 23272™, Lactococcus lactis ATCC® 19435™, Bifidobacterium breve ATCC® 15700™, Bifidobacterium longum subsp. longum ATCC® 15707™, Clostridium perfringens ATCC® 13124™, Ruminococcus gnavus 35913™) or obtained from the Sonnenburg lab (Bacteroides fragilis NCTC 9343, Bacteroides finegoldii DSM 17565, Bacteroides dorei DSM17855, Bacteroides ovatus ATCC 8483, Lactobacillus plantarum ATCC BAA-793, Lactobacillus rhamnosus ATCC 53103).
Lactobacilli MRS Agar plate (Hardy Diagnostic, USA) was used to culture L. acidophilus, L. paracasei, L. reuteri, L. plantarum, L. rhamnosus, L. lactis. Blood Agar plate (Hardy Diagnostic, USA) was used to culture B. breve, B. longum, C. perfringens, R. gnavus. BHI agar plate (Hardy Diagnostic, USA) was used to culture B. finegoldii, B. dorei, B. ovatus, B. fragilis.
Each bacterial strain was inoculated onto the respective culture plates under sterile conditions. The plates were then placed in an air-tight gas pouch containing one pack of BD Difco™ GasPak™ EZ Gas Generating System (ThermoFisher B260683, USA) to create an anaerobic environment. The pouches were incubated at 37 °C for 48 hrs until harvest, allowing the bacteria to grow and develop.
Zhu B., Bai Y., Yeo Y.Y., Lu X., Rovira-Clavé X., Chen H., Yeung J., Gerber G.K., Angelo M., Shalek A.K., Nolan G.P, & Jiang S. (2024). A Spatial Multi-Modal Dissection of Host-Microbiome Interactions within the Colitis Tissue Microenvironment. bioRxiv.
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5

Probiotics Modulate Gut Microbiome in Mice

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MMb, HMb, and GF mice received orally administered Paraprevotella clara, Bacteroides uniformis, SFB, Lactobacillus reuteri (BEI HM-102), Ruminococcus gnavus (ATCC 29149), MMb cx, HMb cx, Clostridium innocuum, or Clostridium immunis (100–150 μl; ~108–109 colony-forming units). P. clara, B. uniformis, C. innocuum, and C. immunis were isolated from the feces of HMb mice; SFB was previously obtained from Y. Umesaki (Yakult; Tokyo, Japan) and propagated in SFB-monocolonized mice at Harvard Medical School. Seven days later, the mice were challenged with DSS. In some experiments, fecal samples were collected before and 7 days after probiotic administration for microbiota analysis. For Reg3γ experiments, HMb mice received orally administered (150–200 μl; ~108–109 colony-forming units) Parabacteroides distasonis (ATCC 8503; control bacteria), R. gnavus (ATCC 29149), L. reuteri (BEI HM-102), Allobaculum stercoricanis (DSM 13633), Muribaculum intestinale (DSM 28989), or Lactobacillus vaginalis (DSM 5837). Mice were sacrificed 7 days later.
Surana N.K, & Kasper D.L. (2017). Moving beyond microbiome-wide associations to causal microbe identification. Nature, 552(7684), 244-247.
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6

Oral Administration of L. reuteri to Mice for SEB Mitigation

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Bacterial strain Lactobacillus reuteri was purchased from American Type Culture Collection (ATCC, Manassas, VA) under deposited name Lactobacillus fermentum Beijerinck (ATCC23272). Based on the vendor recommendation, the frozen bacterial cells were cultured aerobically in Difco™ Lactobacilli MRS Broth (BD, San Jose, CA, USA) at 37 °C for 24 h. Cultured L. reuteri bacterial cells were harvested at the mid-log phase of growth, spun down at 2000 g at 25 °C for 5 min, washed with sterile PBS twice, suspended in sterile PBS and used to transfer into mice exposed to SEB. At the same time, the supernatant was saved, purified using 0.2 μm filter, and stored at −80 °C for further use as raw bacterial metabolites (named as the total culture metabolite, TCM). Mice received the oral dose of 2 × 1010 bacterial cells/mouse per day for 5 consecutive days prior to SEB administration.
Alghetaa H., Mohammed A., Zhou J., Singh N., Nagarkatti M, & Nagarkatti P. (2021). Resveratrol-mediated attenuation of superantigen-driven acute respiratory distress syndrome is mediated by microbiota in the lungs and gut. Pharmacological research, 167, 105548.
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7

Culturing Lactobacillus reuteri for Research

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Lactobacillus reuteri (ATCC 53608) was purchased from American Type Culture Collection (ATCC) and were cultured in De Man, Rogosa and Sharpe (MRS) broth medium (M264-02) at 37°C and under anaerobic conditions. The bacterial suspension was washed with sterile PBS, centrifuged (3,200g, 5 min) and resuspended in PBS until OD600 = 1, which was approximately 1 × 108 colony forming units (CFUs)/ml (Jiménez-Flores et al., 2010 (link)).
Zhao B., Zhou B., Dong C., Zhang R., Xie D., Tian Y, & Yang L. (2021). Lactobacillus reuteri Alleviates Gastrointestinal Toxicity of Rituximab by Regulating the Proinflammatory T Cells in vivo. Frontiers in Microbiology, 12, 645500.
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8

Oral Delivery of L. reuteri Probiotic

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Lactobacillus reuteri was purchased from the American Type Culture Collection (ATCC 23272) and grown in Difco Lactobacilli MRS Broth (Becton Dickinson) for approximately 24 hr at 37°C with 5% CO2. The probiotic was washed with PBS, adjusted to 1×109 CFU/ml, and 100 μl was administered to mice via oral gavage (i.e., 1×108 CFU). Mice were given the probiotic after each exposure to the SDR stressor. Vehicle-treated mice were dosed orally with PBS as a control for the daily handling and repeated gavages.
Mackos A.R., Galley J.D., Eubank T.D., Easterling R.S., Parry N.M., Fox J.G., Lyte M, & Bailey M.T. (2015). Social stress-enhanced severity of Citrobacter rodentium induced colitis is CCL2-dependent and attenuated by probiotic Lactobacillus reuteri. Mucosal immunology, 9(2), 515-526.
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9

Oral Delivery of L. reuteri Probiotic

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Lactobacillus reuteri was purchased from the American Type Culture Collection (ATCC 23272) and grown in Difco Lactobacilli MRS Broth (Becton Dickinson) for approximately 24 hr at 37°C with 5% CO2. The probiotic was washed with PBS, adjusted to 1×109 CFU/ml, and 100 μl was administered to mice via oral gavage (i.e., 1×108 CFU). Mice were given the probiotic after each exposure to the SDR stressor. Vehicle-treated mice were dosed orally with PBS as a control for the daily handling and repeated gavages.
Mackos A.R., Galley J.D., Eubank T.D., Easterling R.S., Parry N.M., Fox J.G., Lyte M, & Bailey M.T. (2015). Social stress-enhanced severity of Citrobacter rodentium induced colitis is CCL2-dependent and attenuated by probiotic Lactobacillus reuteri. Mucosal immunology, 9(2), 515-526.
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