Man rogosa sharpe agar

Manufactured by Thermo Fisher Scientific

Sourced in United Kingdom

Man-Rogosa-Sharpe agar is a culture medium used for the selective isolation and enumeration of lactobacilli from food, dairy, and other samples. It provides the necessary nutrients and growth factors for the cultivation of Lactobacillus species.

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Lab products found in correlation

Columbia blood agar, by BD (1 mentions) Protein assay solution, by Bio-Rad (1 mentions) 0.1 mm glass beads, by Biospec (1 mentions) Wilkins chalgren agar, by Thermo Fisher Scientific (1 mentions) Mannitol salt phenol red agar, by Thermo Fisher Scientific (1 mentions) Anaerocult a, by Merck Group (1 mentions) Violet red bile dextrose agar, by Merck Group (1 mentions) Rose bengal chloramphenicol agar, by Merck Group (1 mentions) Macconkey medium, by Thermo Fisher Scientific (1 mentions)

4 protocols using man rogosa sharpe agar

Characterization of Enterococcus and Lactobacillus Strains

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We used a collection of 6 E. faecium and 4 S. gallolyticus well-characterized strains causing bacteraemia/endocarditis and gut colonization (CEIC-106/09) (Table 1).
The Enterococcus faecalis ATCC 29212 (www.atc.org) and the Lactobacillus reuteri (Spanish Type Culture Collection, CECT 925 T) reference strains were used as controls. All strains were grown on Columbia blood agar (Becton, Dickinson, MI, USA) for 24–48 h at 37°C except for L. reuteri which was cultured under anaerobic conditions on Man-Rogosa-Sharpe agar (MRS, Oxoid, Basingstoke, Hampshire, UK) supplemented with L-cysteine (0.5 g/L) for 48 h at 37°C.

Sánchez-Díaz A.M., Romero-Hernández B., Conde-Moreno E., Kwak Y.K., Zamora J., Colque-Navarro P., Möllby R., Ruiz-Garbajosa P., Cantón R., García-Bermejo L, & del Campo R. (2016). New Insights into the Enterococcus faecium and Streptococcus gallolyticus subsp. gallolyticus Host Interaction Mechanisms. PLoS ONE, 11(7), e0159159.

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Cultivation and Lysis of Probiotic Bacteria

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Wilkins-Chalgren agar (Oxoid, UK) was used to culture B. adolescentis DSM 20083 (ATCC 15703) and DSM 20086 (ATCC 15705) and B. longum DSM 20088 (ATCC 15697). Man-Rogosa-Sharpe agar (Oxoid, UK) was used to culture L. rhamnosus GG. Wilkins-Chalgren agar plates were incubated in an anaerobic cabinet (Concept, UK; with gas mixture of 5% CO₂, 5% H₂, and 90% N₂) and Man-Rogosa-Sharpe agar in a microaerobic environment (Joan, France) with a gas mixture of 10% CO₂ for 48 h.
Bacterial cells were collected, suspended in phosphate-buffered saline solution (PBS, pH = 7.4), and washed 3 times with the same buffer. Subsequently, cells were disrupted with 0.1 mm glass beads (Biospec Products, USA) in PBS in the presence of complete protease inhibitors (Boehringen Mannheim-Roche, Switzerland) on ice. The total protein concentration in lysates was determined using the Protein Assay solution (Bio-Rad, USA) using bovine serum albumin as a standard and kept in aliquots at −20°C until used.

Talja I., Kubo A.L., Veijola R., Knip M., Simell O., Ilonen J., Vähä-Mäkilä M., Sepp E., Mikelsaar M., Utt M, & Uibo R. (2014). Antibodies to Lactobacilli and Bifidobacteria in Young Children with Different Propensity to Develop Islet Autoimmunity. Journal of Immunology Research, 2014, 325938.

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Enumeration of Microbial Populations in Food Samples

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Twenty-five grams of each sample was placed into sterile stomacher bags, and 225 mL of sterile physiological saline (0.85% NaCl) was added. After homogenizing in a stomacher (Lab Stomacher Blender 400-BA 7021, Seward, West Sussex, UK), appropriate dilutions were spread on selective agar plates.
de Man–Rogosa–Sharpe agar (Oxoid, Basingstoke, England) was used for the enumeration of lactic acid bacteria, and the plates were incubated under anaerobic conditions (Anaerocult A, Merck, Darmstadt, Germany) for 48 h at 30°C. Micrococcus/Staphylococcus was enumerated on mannitol salt phenol-red agar (Oxoid, Basingstoke, England) for 48 h at 30°C; Enterobacteriaceae was enumerated on Violet Red Bile Dextrose agar (Merck, Darmstadt, Germany) for 48 h at 30°C under anaerobic conditions (Anaerocult A, Merck, Darmstadt, Germany); molds–yeasts were enumerated on Rose Bengal chloramphenicol agar (Merck, Darmstadt, Germany) for 5 D at 25°C.

Kaban G., Kızılkaya P., Börekçi B.S., Hazar F.Y., Kabil E, & Kaya M. (2020). Microbiological properties and volatile compounds of salted-dried goose. Poultry Science, 99(4), 2293-2299.

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Microbiological Enumeration of Gut Digesta

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The microbiological counts in the large intestinal digesta were determined by the plate-count technique (Shen et al. 2009) . One gram of digesta was homogenized in salt solution (0.9% NaCl) and serially diluted in sterlile saline ranging from 10 -1 to 10 -10 . Then each sample was inoculated on the following agar plates, using standard protocols: for total bacterial count, Bile Esculine Agar (BEA, Oxoid, Milan, Italy) for Streptococci spp count, MacConkey medium (Oxoid, Milan, Italy) for Enterobacteriaceae count, and Man-Rogosa-Sharpe Agar (MRS Oxoid, Milan, Italy) for Lactobacillus spp count. The plates were incubated in an aerobic atmosphere at 20°C for 24 h (for TSA and MacConkey medium), and at 20°C for 48 -72 h under 5% CO2 atmosphere (for BEA and MRS medium). The microbial populations were log transformed before statistical analysis and data are expressed as log10 colony-forming units (CFU) / g of digesta.

Di Giancamillo A., Rossi R., Martino P.A., Aidos L., Maghin F., Domeneghini C, & Corino C. (2018). Copper sulphate forms in piglet diets: Microbiota, intestinal morphology and enteric nervous system glial cells. Animal science journal = Nihon chikusan Gakkaiho, 89(3).

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