Pseudomonas Isolation HiCynth™ Agar Base MCD406

Pseudomonas Isolation HiCynth™ Agar Base is used for selective isolation and identification of Pseudomonas aeruginosa
from clinical and non-clinical specimens.
Composition**
Ingredients Gms / Litre
HiCynth™ Peptone No.4* 20.000
Potassium sulphate 10.000
Magnesium chloride 1.400
Triclosan (Irgasan) 0.025
Agar 13.600
Final pH ( at 25°C) 7.0±0.2
**Formula adjusted, standardized to suit performance parameters
*Chemically defined peptone

Directions
Suspend 45.03 grams in 1000 ml distilled water containing 20 ml glycerol. Heat to boiling to dissolve the medium completely.
Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 45-50°C .Mix well and pour into sterile Petri plates.

Principle And Interpretation

Pseudomonas aeruginosa is an important human pathogen commonly found in nosocomial infections. It successfully
combines adaptability to a variety of moist environments with a collection of potent virulence factors (1). Pseudomonas
infections usually occur at any site where moisture tends to accumulate e. g. tracheostomies, in-dwelling catheters, burns, the
external ear and weeping cutaneous wounds (2). Pseudomonas Isolation HiCynth™ Agar Base is a modification of
Pseudomonas Isolation Agar wherein animal peptones are substituted with chemically defined peptones to avoid BSE/TSE
risks associated with animal peptones. It is used for the selective isolation and identification of P.aeruginosa , is a
modification of Medium A, originally formulated by King, Ward and Raney (3). The medium contains pigment-enhancing
components and the selective agents, triclosan (4) which selectively inhibits non-pseudomonads. The pigment-enhancers i.e.
potassium sulphate and magnesium chloride enhance the blue or blue-green pigment production by P.aeruginosa, thus aiding
in its identification.
HiCynth™ Peptone No.4 provides nitrogenous compounds, carbon compounds, long chain amino acids, vitamins and
other essential growth nutrients. Glycerol is a source of energy and promotes pyocyanin i.e.pigment production
which is characteristic of Pseudomonas (5, 6). Potassium sulphate and magnesium chloride enhance pyocyanin production.
Triclosan (7) selectively inhibits gram-positive and gram-negative bacteria but Pseudomonas species are resistant to it. Some
pyocyanin producing strains may also produce small amounts of fluorescein, resulting in the production of a blue-green to
green pigment. Presumptive Pseudomonas should be further confirmed by performing biochemical tests, as some strains of
Pseudomonas do not produce pyocyanin (8).

Quality Control
Appearance
Cream to yellow homogeneous free flowing powder
Gelling
Firm, comparable with 1.36% Agar gel.
Colour and Clarity of prepared medium
Yellow coloured clear to slightly opalescent gel forms in Petri plates.
Reaction
Reaction of 4.5% w/v aqueous solution at 25°C. pH : 7.0±0.2
pH
6.80-7.20
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 18-48 hours.
Please refer disclaimer Overleaf.
HiMedia Laboratories Technical Data

Cultural Response
Organism Inoculum Growth Recovery Colour of
(CFU) colony
Cultural Response
Escherichia coli ATCC >=10³ inhibited 0%
25922
Proteus mirabilis ATCC >=10³ inhibited 0%
25933
Pseudomonas aeruginosa 50-100 luxuriant >=50% green
ATCC 10145
Pseudomonas aeruginosa 50-100 luxuriant >=50% blue to blue-
ATCC 27853 green
Pseudomonas aeruginosa 50-100 luxuriant >=50% blue to blue-
ATCC 9027 green

Storage and Shelf Life

Store below 30°C in tightly closed container and the prepared medium at 2 - 8°C. Use before expiry date on the label.
Reference

1. 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. ,,
2. Koneman E. W., Allen S. D., Janda W. M., Schreckenberger P. C., Winn W. C. Jr., 1992, Colour Atlas and Textbook of
Diagnostic Microbiology, 4th Ed., J. B. Lippinccott Company
3. King F. O., Ward M. K. and Raney D. E., 1954, J. Lab. Clin. Med., 44 :301.
4. Collee J. G., Fraser A. G., Marmion B. P., Simmons A., (Eds.), Mackie and McCartney, Practical Medical Microbiology,
1996, 14th Edition, Churchill Livingstone
5. Finegold S. M. and Baron E. J., 1986, Bailey and Scotts Diagnostic Microbiology, 7th Ed., The C. V. Mosby Co., St. Louis.
6. MacFaddin J. F., 1985, Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria, Vol. I, Williams
and Wilkins, Baltimore.
7. Furia T. E. and Schenkel A. G., 1968, Soap and Chemical Specialties 44:47
8. Gaby W. L. and Free E., 1958, J. Bacteriol., 76:442

Revision : 01/ 2015

Disclaimer :
User must ensure suitability of the product(s) in their application prior to use. Products conform solely to the information contained in
this and other related HiMedia™ publications. The information contained in this publication is based on our research and development
work and is to the best of our knowledge true and accurate. HiMedia™ Laboratories Pvt Ltd reserves the right to make changes to
specifications and information related to the products at any time. Products are not intended for human or animal or therapeutic use but
for laboratory,diagnostic, research or further manufacturing use only, unless otherwise specified. Statements contained herein should not
be considered as a warranty of any kind, expressed or implied, and no liability is accepted for infringement of any patents.

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