MÜLLER-KAUFFMANN BROTH (Base)

MÜLLER-KAUFFMANN BROTH (Base)
INTENDED USE
Müller-Kauffmann Broth is used as a selective enrichment medium for salmonellae in mammals and
birds, in addition to environmental samples in animal production areas. It is also adapted to the
hygiene control of pharmaceutical and cosmetic products, water, dairy products and other foodstuffs.
HISTORY
The medium was described by Müller in 1923 to favor the inhibition of coliform bacteria at the same
time as permitting the development of typhoid and paratyphoid bacilli. Kauffmann modified the
formula and obtained a greater number of positive results with this enrichment method than with the
direct method of isolating on selective media poured into plates.
PRINCIPLES
- Ox bile and brilliant green inhibit principally the development of Gram-positive bacteria.
- The production of tetrathionate resulting from the action of the iodine-iodide solution on sodium
thiosulfate, inhibits coliform bacteria and most intestinal bacteria.
- The development of Proteus can be suppressed by adding 40 mg of novobiocin per liter of
medium.
- Calcium carbonate neutralizes the sulfuric acid produced when tetrathionate is reduced. The
resultant effect is to maintain pH at a constant level.
PREPARATION
-
Suspend 88.3 g of dehydrated medium (BK135) in 1 liter of distilled or deionized water.
Slowly bring to boiling with stirring.
Continue boiling for 2 minutes.
Do not autoclave.
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INSTRUCTIONS FOR USE
-
Cool the medium to 25°C.
Dissolve 4 g of iodine in 20 mL of a solution containing 5 g of potassium iodide in a sterile flask.
Add the iodine-iodide solution to the medium
Optionally add 40 mg of a reconstituted solution of Novobiocin Selective Supplement 10 mg
(BS033) or Novobiocin 40 mg (BS056), per liter of medium.
Mix thoroughly.
Do not heat the medium once prepared.
Aseptically dispense 10 mL into sterile tubes.
To each tube, transfer 1 mL of inoculum from the preenrichment medium used : Buffered Peptone
Water (BK018, BK131, BM010, BM057, BM131, BM132) or Lactose Broth (BK082).
Verify that the pH of the medium is conform to the required specifications.
Incubate (48  4) hours at (43  1)°C.
Isolate on several selective media with a platinum loop.
Using well formed colonies, inoculate onto Nutrient agar 2% (BK184, BM118) which are the
starting points for identification.
TYPICAL COMPOSITION of the base medium, without iodine and novobiocin)
(can be adjusted to obtain optimal performance)
For 1 liter of medium :
- Tryptone .........................................................................................8.45 g
- Meat extract ..................................................................................4.23 g
- Bacteriological ox bile ...................................................................4.75 g
- Sodium chloride .............................................................................2.54 g
- Calcium carbonate .......................................................................38.04 g
- Sodium thiosulfate anhydrous .....................................................30.27 g
- Brilliant green ...............................................................................9.5 mg
QUALITY CONTROL
- Dehydrated medium : whitish powder, free-flowing and homogeneous.
- Prepared (complete) medium : bluish opaque suspension, with abundant precipitate when left
standing.
- Typical culture response after enrichment for 24 hours at 42°C, followed by subculture on XLD
Agar :
Microorganisms
Growth
Salmonella Typhimurium
+ Pseudomonas aeruginosa
+ Escherichia coli
ATCC® 14028
ATCC 27853
ATCC 25922
 10 red colonies with black center
Salmonella Enteritidis
+ Pseudomonas aeruginosa
+ Escherichia coli
CIP 82.97
ATCC 27853
ATCC 8739
 10 red colonies with black center
Enterococcus faecalis
Escherichia coli
ATCC 29212
ATCC 25922
≤ 10 colonies
inhibited, score 0
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Biokar Diagnostics – Rue des Quarante Mines – ZAC de Ther – Allonne – B.P. 10245 – F60002 Beauvais Cedex – France
Tél : + 33 (0)3 44 14 33 33 – Fax : + 33 (0)3 44 14 33 34 – www.biokar-diagnostics.com
STORAGE / SHELF LIFE
Dehydrated base medium : 2-30°C.
- The expiration date is indicated on the label.
Prepared medium (benchmark value*) :
- Base media in tubes or vials : 1 month at 2-8°C, shielded from light.
- Complete media in tubes or vials : use immediately after adding the iodine-iodide solution and
novobiocin.
Novobiocin Selective Supplements:
- Store between 2-8°C, shielded from light.
- The expiration date is indicated on the label.
Code
PACKAGING
Dehydrated base medium (without iodine nor novobiocin) :
- 500 g bottle
BK135HA
Novobiocin Selective Supplement 10 mg :
- 10 vial pack
BS03308
Novobiocin Selective Supplement 40 mg :
- 8 vial pack
BS05608
BIBLIOGRAPHY
MÜLLER, L.. 1923. Un nouveau milieu d’enrichissement pour la recherche du bacille typhique et des paratyphiques.
Comptes Rendus de la Société de Biologie, 89 : 434-437.
KAUFFMANN, F.. 1935. Weitere Erfahrungen mit dem kombinierten Anreicherungsverfahren für Salmonella bazillen.
Zeitschrift für Hygiene und Infektionskrankheit, 117 : 26-32.
JEFFRIES, L.. 1959. Novobiocin-tetrathionate broth: a medium of improved selectivity for the isolation of salmonellae in
feces. Journal of Clinical Pathology, 12 : 568-571.
NF V 59-104. Octobre 1982. Gélatine alimentaire. Recherche des Salmonella.
XP CEN ISO/TS 11133-2 (V 08-104-2). Janvier 2004. Microbiologie des aliments. Guide pour la préparation et la production
des milieux de culture. Partie 2 : Guide général pour les essais de performance des milieux de culture.
*Benchmark value refers to the expected shelf life when prepared under standard laboratory conditions following manufacturer’s instructions. It is provided as a
guide only and no warranty, implied or otherwise is associated with this information.
The information provided on the package take precedence over the formulations or instructions described in this document.
The information and specifications contained in this technical data sheet date from 2009-06-03.
They are susceptible to modification at any time, without warning.
Code document : BK135/A/2003-03 :8.
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Biokar Diagnostics – Rue des Quarante Mines – ZAC de Ther – Allonne – B.P. 10245 – F60002 Beauvais Cedex – France
Tél : + 33 (0)3 44 14 33 33 – Fax : + 33 (0)3 44 14 33 34 – www.biokar-diagnostics.com