Cetrimide Broth Base

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M862A
Recommended for cultivation of Pseudomonas aeruginosa from water samples using membrane filter technique. The composition and performance criteria of this medium are as per the specifications laid down in ISO 8360-2:1988.


Intended Use

Recommended for cultivation of Pseudomonas aeruginosa from water samples using membrane filter technique. The composition and performance criteria of this medium are as per the specifications laid down in ISO 8360-2:1988.

Composition

Ingredients g / L
Peptone 20.000
Potassium sulphate 10.000
Magnesium chloride 1.400
Cetrimide 0.500
Final pH (at 25°C) 7.2±0.2

**Formula adjusted, standardized to suit performance parameters

Directions

Suspend 31.9 grams in 1000 ml purified / distilled water. Heat, if necessary, to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 45-50°C and aseptically add 25 ml filter sterilized ethanol. Mix well before dispensing.

Principle And Interpretation

Cetrimide Broth Base is the modification of the formula designed by King, Ward and Raney (1) and is useful for the cultivation of P.aeruginosa. This medium is also selective as it contains cetrimide (Cetyl trimethyl ammonium bromide), which inhibits other bacteria except P.aeruginosa. ISO Committee recommends Cetrimide Broth Base for cultivation of P. aeruginosa using membrane filter technique (2).

Peptone provide necessary nutrients for P. aeruginosa. Cetyl trimethyl ammonium bromide (Cetrimide) is a quaternary ammonium compound, which inhibits bacteria other than Pseudomonas aeruginosa. The production of pyocyanin, a blue water-soluble pigment is stimulated by magnesium chloride and potassium sulphate (3,4).

Type of specimen

Water samples

Specimen Collection and Handling

For water samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards.(5) After use, contaminated materials must be sterilized by autoclaving before discarding.

Warning and Precautions

Read the label before opening the container. Wear protective gloves/protective clothing/eye protection/ face protection. Follow good microbiological lab practices while handling specimens and culture. Standard precautions as per established guidelines should be followed while handling specimens. Safety guidelines may be referred in individual safety data sheets.

Limitations

  1. Further isolation, biochemical and serological tests must be carried out for further identification.

Performance and Evaluation

Performance of the medium is expected when used as per the direction on the label within the expiry period when stored at recommended temperature.

Quality Control

Appearance
Cream to yellow colored homogeneous free flowing powder.

Colour and Clarity of prepared medium
Light amber coloured clear to slightly opalescent solution in tubes

Reaction
Reaction of 3.19% w/v aqueous solution at 25°C. pH: 7.2±0.2

pH
7.00-7.40

Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 24-48 hours.

Organism Inoculum (CFU) Growth
Escherichia coli ATCC 25922 (00013*) >=104 inhibited
Pseudomonas aeruginosa ATCC 27853 (00025*) 50-100 luxuriant
Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) >=104 inhibited

Key: *Corresponding WDCM numbers.

Storage and Shelf Life

Store between 10-30°C in a tightly closed container and the prepared medium at 15-30°C. Use before expiry date on the label. On opening, product should be properly stored dry, after tightly capping the bottle in order to prevent lump formation due to the hygroscopic nature of the product. Improper storage of the product may lead to lump formation. Store in dry ventilated area protected from extremes of temperature and sources of ignition. Seal the container tightly after use. Product performance is best if used within stated expiry period.

Disposal

User must ensure safe disposal by autoclaving and/or incineration of used or unusable preparations of this product. Follow established laboratory procedures in disposing of infectious materials and material that comes into contact with sample must be decontaminated and disposed of in accordance with current laboratory techniques (6,7).

More Information
Product Name Cetrimide Broth Base
SKU M862A
Product Type Regular
Physical Form Powder
Origin Animal
Packaging type HDPE
References 1. King E.O., Ward M.K. and Raney D.E., 1954, J. Lab. Clin. Med., 44(2):301. 2.USFDA Bacteriological Analytical Manual,2005, 18th Ed., AOAC, Washington, DC.3.Murray P. R., Baron J. H., Pfaller M. A., Jorgensen J. H. and Yolken R. H., (Ed.), 2003, Manual of Clinical Microbiology,8th Ed., American Society for Microbiology, Washington, D.C.4.Forbes B. A., Sahm A. S. and Weissfeld D. F., Bailey & Scotts Diagnostic Microbiology, 10th Ed., 1998, Mosby, Inc., St.Louis, Mo.5.Williams, (Ed.), 2005, Official Methods of Analysis of the Association of Official Analytical Chemists, 19th Ed., AOAC,Washington, D.C.
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