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

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022 C

030 C Store at 0

22-30 CManufacturer

Catalogue Number IVD In vitro Diagnostic

Medical Device

Hygroscopic keep container tightly closed

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

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Authorised Representative

Dehydrated Culture Media

TULIP DIAGNOSTICS (P) LTD.S-126, Phase III B, Verna Industrial Estate,Verna, Goa - 403 722, India.www.tulipgroup.com

Coagulase Mannitol Agar Base

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 14.50Final pH (at 25 C) 7.4 ± 0.2

* Formula adjusted to suit performance parameters

REF AM1028 AM5028Pack Size 100 gms 500 gms

Storage and Stability0Store the unopened container at room temperature below 30 C. 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.0 04. Sterilize by autoclaving at 118 C -121 C (12-15 lbs pressure) for

A Differential Dehydrated Culture Medium

15 minutes.05. Cool to 45-50 C.

6. 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 Response0Cultural characteristics after 18-48 hours at 35-37 C.

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 01. Inoculate and incubate at 35-37 C, 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 coagulase-positive. 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

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

3. Marvin, 1958, Am. J. Clin. Path; 30:470.

4. Esber and Faulconer, 1959, Am. J. Clin. Pathol; 32:192.

5. Data on file: Microxpress, a division of Tulip Diagnostics (P) Ltd.