Handbook of Culture Mediafor Food Microbiology, J.E.L. Corry et al. (Eds.

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9 2003 Elsevier Science B.V. All rights reserved

de Man, Rogosa and Sharpe (MRS) agar

This monograph has been assessed by members of the IUMS-ICFMH Working Party
on Culture Media and given 'Approved' status.

Description and history

MRS agar was developed by de Man, Rogosa and Sharpe (1960) primarily for the cul-
tivation of lactobacilli from various sources with the intention of producing a defined
medium as a substitute for tomato juice agar. It may be used for the cultivation of the
whole group of lactic acid bacteria. The medium shows good productivity for nearly
all lactic acid bacteria but the original version is not selective. It may be made selec-
tive for lactic acid bacteria by lowering the pH to 5.7 and the addition of 0.14% sorbic
acid (see MRS-S agar). Some strains from dairy sources may show reduced growth
rates (see Briggs agar).

Composition (grams)

Tryptic digest of casein 10.0

Beef extract 8.0
Yeast extract 4.0
Glucose 20.0
Sorbitan monooleate (Tween 80) 1.0
Di-potassium hydrogen orthophosphate 2.0
Magnesium sulphate .7H20* 0.2
Manganese (II) sulphate .4H20 § 0.05
Ammonium citrate 2.0
Sodium acetate-3H20 5.0
Agar 15.0
Distilled or deionized water 1000.0
* 20 ml of 1% solution
+ 5 ml of 1% solution

Preparation

Suspend the ingredients in the water and boil to dissolve completely. Sterilize for 15
min at 121~
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Physical properties

Appearance Light amber, clear.

pH 6.2 Adjust pH after dissolving completely at about 50~

Shelf life

Ready to use medium 14 days at 4 + 2~

Inoculation method for samples

Surface spreading over whole plate or modified Miles-Misra.

Incubation method

This depends on the particular habitat of the organisms to be cultivated. Dairy strains
should be incubated at 30~ for 2 days followed by 1 day at 22~ meat strains at 25~
for 2 days, intestinal or yoghurt strains for 2 days at either 37 or 42~ All incuba-
tions should be performed under anaerobic or microaerobic (6% 02: 10% CO 2 in N2)
conditions.

Reading of results and interpretation

MRS agar is an elective medium that gives good colony counts and a characteristic
colony size and morphology for lactobacilli and for other lactic acid bacteria. Other
microorganisms must be excluded by specific colonial appearance and by confirma-
tion tests e.g. Gram-stain and catalase test. Carnobacterium spp. either show weak
growth or fail to grow.

Quality assessment

(i) Productivity
Test strains Lactobacillus gasseri NCIMB 50040
Lactobacillus sakei ssp. sakei NCIMB 50056

Inoculation method Modified Miles-Misra, spiral plating or streaking/

ecometry.

Criteria Recovery on MRS agar should be within 1.0 logl0 of

the recovery on Briggs agar.

(ii) Characteristic appearance of colonies

Usually small greyish white colonies, flat or raised, smooth, rough or intermedi-
ate.
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Reference
de Man, J.C., Rogosa, M. and Sharpe, M.E. (1960) A medium for the cultivation of lactobacilli. J.
Appl. Bacteriol. 23, 130-135.