Mitis salivarius agar

Manufactured by Merck Group

Sourced in Germany

Mitis Salivarius Agar is a microbiological culture medium used for the isolation and enumeration of Streptococcus mutans and other oral streptococci from clinical samples. It contains selective agents that inhibit the growth of other bacterial species, allowing for the specific detection of the target organisms.

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Lab products found in correlation

Scan 500, by Interscience (1 mentions) Blood agar, by Merck Group (1 mentions) Sabouraud dextrose agar, by Thermo Fisher Scientific (1 mentions) E faecalis atcc 29212, by American Type Culture Collection (1 mentions) C albicans atcc 10231, by American Type Culture Collection (1 mentions) Irgasan, by Merck Group (1 mentions)

5 protocols using mitis salivarius agar

Quantifying Oral Bacterial Colonization

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The saliva samples were diluted by 1/10 and then they were transferred to the Mitis Salivarius Agar (MSA) and MRS-Agar (MRS-A) containing plates (Merck KGaA, Darmstadt, Germany). All plates were then incubated at 37 ºC for 24–48 h (Memmert, Hong Kong) (Table 2). Colonies of Streptococcus mutans and Lactobacillus were counted after gram staining, using an automatic colony counter (Scan® 500, Interscience, Saint Nom, France) by two experienced microbiologists who were “blinded” to the administered toothpaste type. The inter-examiner reliability was determined using the Kappa agreement coefficient (k = 0.9).Table 2

List of examined microorganisms and their culture media

Bacteria	Culture media	Manufacturer	Incubation
S. mutans^a	Mitis Salivarius Agar (MSA)	Merck KGaA, Darmstadt, Germany	24 h, in CO₂-containing incubator
L. casei^b	MRS-Agar (MRS-A)	Merck KGaA, Darmstadt, Germany	48 h

^aS. Mutans = Streptococcus mutans, ^bL. casei = Lactobacillus casei

Biria M., Rezvani Y., roodgarian R., Rabbani A, & Iranparvar P. (2022). Antibacterial effect of an herbal toothpaste containing Bamboo salt: a randomized double-blinded controlled clinical trial. BMC Oral Health, 22, 193.

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Antimicrobial Evaluation of Zataria multiflora

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Streptococcus mutans (Strain PTCC
¹1683), Enterococcus faecalis (PTCC 1394), and Candida albicans (PTCC 5027) were obtained from Iranian Research Organization for Science and Technology (IROST). In order to have fresh microbial cultures, S. mutans was seeded on Mitis Salivarius agar (Merck, Germany) containing 1% potassium tellurite, E. faecalis was cultured on blood agar (Merck, Germany) and C. albicans on Sabouraud dextrose agar (Oxoid, UK) plates. Fresh colonies of Bacterial species and Candida were inoculated in brain heart infusion (BHI) broth and incubated for 4–6 hours, to the point when growth was considered to be in the logarithmic phase. The density of the bacterial suspension was adjusted with sterile PBS to that of 0.5 McFarland standard concentration and used for the experiments.
The 0.5mg/ml Zataria multiflora extract solution (Barij Essence Pharmaceutical Co, Kashan, Iran) was used as mouthwash for evaluation of its antimicrobial effect on tested microorganisms in comparison with 0.02% CHX (Shar Daroo, Iran).

Aghili H., Jafari Nadoushan A.A, & Herandi V. (2015). Antimicrobial Effect of Zataria Multiflora Extract in Comparison with Chlorhexidine Mouthwash on Experimentally Contaminated Orthodontic Elastomeric Ligatures. Journal of Dentistry (Tehran, Iran), 12(1), 1-10.

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Orthodontic Adhesive Biofilms and CeO₂-NPs

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Twenty-five orthodontic adhesive disks with different concentrations of CeO 2 -NPs were placed in flat-bottomed 48well microtiter plates containing S. mutans suspension with a concentration of 1.5×10 8 CFU/mL. To form biofilms on the disks, the microtiter plate was incubated under the aerobic atmosphere with 5% CO 2 at 37°C for 72 hours. Afterwards, disks were rinsed in 1 mL of sterile deionized water for 1 min to remove planktonic microbial cells. Orthodontic adhesive disks were then vortexed severely in 1 mL of BHI broth for 30 s. The obtained bacterial suspensions were serially diluted, cultured in mitis salivarius agar (Merck, Darmstadt, Germany), and the microbial colony counts were determined as mentioned in the previous study. [14, 15]

Pourhajibagher M, & Bahador A. (2022). Physico-mechanical properties, antimicrobial activities, and anti-biofilm potencies of orthodontic adhesive containing cerium oxide nanoparticles against Streptococcus mutans. Folia medica, 64(2).

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Culturing E. faecalis and C. albicans

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E. faecalis ATCC 29212 and C. albicans ATCC 10231 were purchased from the American Type Culture Collection (ATCC). Microorganisms were stored at −20 °C in 5% glycerol. Unfrozen microbial cells were transferred to Brain Heart Infusion broth (BHI, Kasvi, Brazil) and incubated overnight at 37 °C. Then, one loop of the culture was transferred to Mitis Salivarius agar (Sigma-Aldrich, St. Louis, MO, USA) and Sabouraud agar (Kavsi, Brazil) for E. faecalis and C. albicans, respectively. Cultures were incubated for up to 18 h at 37 °C.

Fiallos N.D., Ribeiro Aguiar A.L., da Silva B.N., Pergentino M.L., Rocha M.F., Sidrim J.J., Maia D.C, & Cordeiro R.D. (2022). The Potential of Phenothiazines against Endodontic Pathogens: A Focus on Enterococcus-Candida Dual-Species Biofilm. Antibiotics, 11(11), 1562.

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Selective Media for Bacterial Identification

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Previously described selective media were examined for selectivity and adjusted when necessary. For all media, each selective component was examined to distinguish which components were essential for selective inhibition. For P. aeruginosa, Pseudomonas Isolation Agar (PIA; BD Diagnostics) is described which uses 25 μg/mL triclosan (Irgasan, Sigma Aldrich) as a selective agent since most bacterial species are susceptible while P. aeruginosa is resistant [30 (link),31 (link)]. For S. aureus, Mannitol Salt Agar is widely used for selection, based on a 7.5% NaCl concentration [32 (link),33 (link)]. For S. anginosus, several media were considered: NAS (nalidixic acid sulfamethazine) agar [19 (link)], Mitis Salivarius Agar (Sigma-Aldrich) and Edwards medium (Oxoid, Thermo Fisher Scientific). For A. xylosoxidans, MCXVAA medium is described [18 (link)], based on MacConkey agar supplemented with 5 mg/mL xylose, 20 μg/mL vancomycin, 20 μg/mL aztreonam and 5 μg/mL amphotericin. For R. mucilaginosa, Rothia Mucilaginosa Selective Medium (RMSM) was previously developed, containing 50 μg/mL sodium selenite and 10 μg/mL colistin [22 (link)]. Finally, for G. haemolysans, a supplemented Edwards medium with 5 μg/mL colistin sulphate and 2.5 μg/mL oxolinic acid was described [20 (link)].

Vandeplassche E., Coenye T, & Crabbé A. (2017). Developing selective media for quantification of multispecies biofilms following antibiotic treatment. PLoS ONE, 12(11), e0187540.

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