Accumix

TM

Coagulase Mannitol Agar Base

used and by chance contaminated must be safely disinfected or

sterilized. The environment in which microbiological cultures are
handled must also be taken into account.
Bibliography
1. Chapman.1946. J. Bact. Vol. 51:409.

2.
3.
4.
5.

Zebovit, Evans and Nivens 1955. J. Bact; 70: 686.

Marvin, 1958, Am. J. Clin. Path; 30:470.
Esber and Faulconer, 1959, Am. J. Clin. Pathol; 32:192.
Data on file: Microxpress, a division of Tulip Diagnostics (P) Ltd.

Hygroscopic keep
container tightly closed

A Differential Dehydrated Culture Medium

REF
Pack Size

AM1028
100 gms

AM5028
500 gms

Storage and Stability

Store the unopened container at room temperature below 300C. Under
optimal conditions, the medium has a shelf life of 2-5 years. When the
container is opened for the first time, note the time and date on the
label space provided on the container. After the desired amount of
medium has been taken out replace the cap tightly to protect from
hydration.
Directions
1. Suspend 47.02 gms of the powder in 1000 ml distilled water.
2. Mix thoroughly.
3. Boil with frequent agitation to dissolve the powder completely.
4. Sterilize by autoclaving at 1180C -1210C (12-15 lbs pressure) for

15 minutes.
Cool to 45-500C.
Add 7-15% v/v sterile, pre-tested, rabbit plasma to the basal
medium.
7. Mix well and pour into sterile petri plates.
Quality Control
Dehydrated Appearance
Light grey coloured, homogeneous, free flowing powder.
Prepared Appearance
Purple coloured, slightly opalescent gel.
Cultural Response
Cultural characteristics after 18-48 hours at 35-370C.
Organisms (ATCC) Growth
Mannitol
Coagulase
Fermentation
production
Staphylococcus
Luxuriant
+ (yellow)
+ (opaque
aureus (25923)
zone)
Staphylococcus
Luxuriant
- (purple)
epidermidis (12228)
The performance standards of the above medium conform to the
NCCLS standards of quality assurance for commercially prepared
microbiological culture media.
Procedure
1. Inoculate and incubate at 35-370C, and examine for growth after
18-24 hours.
2. Avoid prolonged incubation because it may cause the opaque
zones surrounding coagulase-positive organisms to become
clear.
Interpretation of Results
1. Coagulase-positive organisms will produce opaque zones;
coagulase-negative organisms will produce no opacity.
2. Organisms that utilize mannitol produce yellow zones. S.aureus
may be presumptively identified as those colonies with opaque,
yellow zones around them.
Precautions
1. Some old or mutant strains of S.aureus may be weak coagulase
producers or exhibit negative coagulase reaction and should be
subcultured and retested if in doubt.
2.
E.coli also uses mannitol and may be weakly coagulasepositive. Colonial morphology and a gram stain should readily
allow for differentiation from S.aureus.
Use and Disposal of Dehydrated Culture Media
Inoculation of culture media with bacteria, deliberately or accidentally,
leads to a very great number of organisms being produced. High
concentrations of any organisms are potentially hazardous and must
be disposed off safely. Therefore, after use, prepared plates, samples,
sample containers or other contaminated material must be sterilized
or incinerated before discarding. All autoclaved biohazards should be
disposed off in accordance with state and local environmental
regulations.
Only qualified personnel who have been trained in microbiological
procedures should handle all infected specimens and inoculated
culture media. User should ensure that any machinery or apparatus
5.
6.

Dehydrated Culture Media, Bases, Supplements

300C

22 C

REF

ACC CMAB/0906/AE/VER-1

Use
Coagulase Mannitol Agar Base with added plasma is used for isolation
and differentiation of staphylococci from clinical specimens or for
classifying pure cultures.
Summary
Coagulase-positive and coagulase-negative Staphylococcus species
have major medical significance. Coagulase producing staphylococci
(S.aureus) may be differentiated and presumptively identified on this
medium based on production of coagulase and mannitol utilization.
Chapman (16) introduced the first selective medium for isolating and
differentiating staphylococcal species. Several years later, Zebovit et
al (124) and Marwin (79) introduced tellurite-glycine media designed
to selectively isolate coagulase positive staphylococcal species. The
present formulation is based on Esber and Faulconer (26) formulation.
Principle
Coagulase production is dependent on the presence of mannitol and a
protein factor in the brain heart infusion and blood serum (plasma).
During utilization of mannitol, the pH of the medium drops, causing the
bromocresol purple indicator to change from purple to yellow,
producing yellow zones around these colonies. An opaque area of
coagulated plasma forms around the colonies of organisms that also
produce coagulase. In contrast some coagulase-negative species
that do not utilize mannitol, such as Staphylococcus epidermidis, do
not change the colour of the medium and it remains clear. Other
coagulase-negative species may utilize mannitol and produce a
yellow zone around the colony, but an opaque zone will not be
produced.
Formula*
Ingredients in grams per liter
Tryptone
10.5
Mannitol
10.0
Brain Heart Infusion
5.0
Sodium Chloride
3.5
Soya Peptone
3.5
Bromocresol Purple
0.02
Agar
14.5
Final pH (at 250C) 7.4 0.2
* Formula adjusted to suit performance parameters

Store at
0
22-30 C

Manufacturer

Catalogue
Number

Consult
Instructions
for use

LOT

Batch
Number

IVD

Use by

In vitro Diagnostic
Medical Device

DCM

RO

Dehydrated Culture Media

Received on

EC REP

OO

Authorised Representative

Opened on

TULIP DIAGNOSTICS (P) LTD.

S-126, Phase III B, Verna Industrial Estate,
Verna, Goa - 403 722, India.
www.tulipgroup.com

EC

REP

k
Quicable y
Rei l biolog
ro
Mic

Qarad b.v.b.a. Volmolenheide 13, B-2400 Mol, Belgium

Dehydrated Culture Media, Bases, Supplements, Ready to use Media, Indicators & Stains, Test Kits