AOAC MR-VP Broth (Methyl-red VOGES-PROSKAUER Broth)

BAM
Test culture medium for the methyl red test (CLARK and LUBS 1915) and the VOGES-PROSKAUER Test (VOGES and
COMPF PROSKAUER 1898), which are used for biochemical differentiation, particularly within the Coli-Aerogenes group.
MR-VP Broth (Methyl-red VOGES-PROSKAUER Broth)

ISO
SMD
SMWW

This culture medium complies with the recommendations of the potassium hydroxide solution (prepared from extra pure
ISO (1975), the DIN Norm 10160 for the examination of meat and potassium hydroxide) or 5 ml O'MEARA's reagent to the second
the DIN Norm 10181 for the examination of milk. tube. With the first two reagents a positive reaction is indicated,
if the colour of the medium changes to red within a few minutes.
Mode of Action
In the case of O'MEARA's reagent, the reaction is positive if, after
a. Some bacteria utilize glucose to form large amounts of acid frequent shaking, a pink coloration appears after approx.
with the result that the pH value of the medium falls to 20minutes beginning at the surface and becoming more intense
below 4.4. Other species produce less acid so that the fall in within 2hours.
pH is not as great. This difference can be visualized by using
methyl red which is yellow above pH 5.1 and red at pH 4.4.
b. Many microorganisms metabolize glucose to produce Colour Reaction Microorganisms
acetoin (acetylmethyl carbinol), 2,3-butanediol or diacetyl. From orange to red Escherichia coli, Citrobacter
The presence of these metabolites is established by means of and others
O'MEARA's reagent (1931) improved by LEVINE et al.
(1934), copper sulfate solution according to LEIFSON (1932), From orange to yellow Enterobacter aerogenes,
BARRIT's reagent (BARRITT 1936) or other reagents (see Enterobacter cloacae and others
references). According to HOLLNDER et al. (1982), addition
Red (positive) Enterobacter aerogenes,
of fumarate to the broth enhances this reaction.
Enterobacter cloacae and others
Details and comparative studies on the various modifica-
tions of the MR-VP test are to found in EDDY (1961), No colour change (negative) Escherichia coli, Citrobacter
SUASSUNA et al. (1961), IJUTOV (1963) and SKERMAN and others
(1969).

Typical Composition (g/litre) Literature

Suspend 17 g/litre, dispense 5 ml aliquots into tubes and auto- DIN Deutsches Institut fr Normung e.V.: Untersuchung von Fleisch und
Fleischerzeugnissen. Nachweis von Salmonellen. Referenzverfahren.
clave (15 min at 121 C). - DIN10160.
pH: 6.9 0.2 at 25 C. DIN Deutsches Institut fr Normung e.V.: Mikrobiologische Milchunter-
The broth is clear and yellowish-brown. suchung. Nachweis von Salmonellen. Referenzverfahren. DIN 10181.

Preparation of the methyl red indicator solution: Suspend 0.04 g BARRIT, M.: The intensification of the Voges-Proskauer reaction by the
addition of -naphthol. J. Path. Bact. 42; 441-454 (1936).
methyl red in 60 ml absolute ethanol, adjust the pH to a value of
approx. 5.0. The solution then becomes orange. CLARK, W., a. LUBS, H.: The differentiation of bacteria of the
Colon-Arogenes family by the use of indicators. J. Inf. Dis., 17; 160-173
Preparation of O'MEARA's reagent: Suspend 40 g potassium (1915).
hydroxide in 100 ml distilled water. Allow to cool, add 0.3g EDDY, B.P.: The Voges-Proskauer reaction and its significance: A review.
creatine (monohydrate) and dissolve. The prepared reagent - J. Appl. Bact., 24; 27-41 (1961).
solution can be stored for about 4 weeks in the refrigerator HOLLNDER, R., BHMANN, J., a. GREWING, B.: Die Verstrkung der
(+4 C). Voges-Proskauer-Reaktion durch Fumarat. Zbl. Bakt. Hyg., I Abt. Orig. A,
Preparation of copper sulfate solution acc. to LEIFSON: Suspend 252; 316-323 (1982).
1 g copper sulfate in 40 ml concentrated ammonia and add LEIFSON, E.: An improved reagent for the acetyl-methyl-carbinol test.
690ml of an approx. 10 % potassium hydroxide solution - J.Bact. 23; 353-354 (1932).
(prepared from potassium hydroxide). LEVINE, M., EPSTEIN, S.A., a. VAUGHN, R.H.: Differential reaction in the
colon group of bacteria. Publ. Hlth., 24; 505-510 (1934).
Preparation of BARRITT's reagent: Suspend 5 g naphthol(1) in
IJUTOV V.: Technique of Voges-Proskauer test. Acta path. microbiol.
100 ml absolute ethanol. scand., 58; 325-335 (1963).
Experimental Procedure and Evaluation O'MEARA, R.: A simple delicate and rapid method of detecting the formation
of acetylmethylcarbinol by bacteria fermenting carbohydrates. J. Path.
Inoculate two tubes containing MR-VP Broth with a pure culture Bact., 34; 401-406 (1931).
of the microorganisms under investigation.
SKERMAN, V.B.D.: Abstracts of microbiological methods
Incubation: up to 4 days at 35 C. (Wiley-Interscience, New York, 1969).
Carry out the following tests: SUASSUNA, J., SUASSUNA, J.R., a. EWING, W.H.: The methyl red and
Voges-Proskauer reactions of enterobacteriaceae. Publ. Hlth. Lab., 19;
Methyl red test: Add about 5 drops of the methyl red indicator 67-75 (1961).
solution to the first tube.
VOGES, O., a. PROSKAUER, B.: Beitrag zur Ernhrungsphysiologie und zur
VOGES-PROSKAUER test: Add 5 ml of copper sulfate solution Differentialdiagnose der hmorrhagischen Septicmie. Z. Hyg. Infekt., 28 ;
acc. to LEIFSON or 3 ml BARRIT's solution and 1 ml 40 % 20-32 (1898).

Merck Microbiology Manual 12th Edition 357

Ordering Information
Product Merck Cat. No. Pack size
MR-VP Broth 1.05712.0500 500 g
(Methyl-red VOGES-
PROSKAUER Broth)
Ammonia solution 25 % 1.05432.1000 1l
Copper sulfate 1.02790.0250 250 g
Creatine (monohydrate) 8.41470.0050 50 g
Ethanol absolute 1.00983.1000 1l
Methyl red 1.06076.0025 25 g
Naphthol-(1) 1.06223.0050 50 g
Potassium hydroxide 1.05033.0500 500 g
pellets

Quality control
Test strains Growth Methyl red VOGES-PROSKAUER
Escherichia coli ATCC 25922 good / very good + -
Klebsiella pneumoniae ATCC 13883 good / very good + -
Klebsiella pneumoniae ATCC 10031 good / very good + -
Enterobacter cloacae ATCC 13047 good / very good - +
Serratia marcescens ATCC 14756 good / very good +

358 Merck Microbiology Manual 12th Edition