CRITERION™ PSEUDOMONAS ISOLATION AGAR BASE

Cat. no. C6680 CRITERION™ Pseudomonas Isolation Agar Base 90gm
Cat. no. C6681 CRITERION™ Pseudomonas Isolation Agar Base 500gm
Cat. no. C6682 CRITERION™ Pseudomonas Isolation Agar Base 2kg
Cat. no. C6683 CRITERION™ Pseudomonas Isolation Agar Base 10kg
Cat. no. C6684 CRITERION™ Pseudomonas Isolation Agar Base 50kg

INTENDED USE

Hardy Diagnostics CRITERION™ Pseudomonas Isolation Agar Base is recommended for the selective isolation and differentiation of Pseudomonas aeruginosa .

This dehydrated culture medium is a raw material intended to be used in the making of prepared media products, which will require further processing, additional ingredients, or supplements.

SUMMARY

Non-fermentative gram-negative bacteria (NFB) can exploit the wet environments in hospitals and are often associated with nosocomial infections. For instance, non-fermentative bacteria have been implicated in the contamination of faucet aerators, respiratory therapy equipment, as well as sterile solutions and medications. Among the NFB, Pseudomonas aeruginosa is the most clinically significant bacteria.(8)

Hardy Diagnostics CRITERION™ Pseudomonas Isolation Agar Base is a modification of the formula developed by King, et al.(1) The medium is designed to enhance pyocyanin production, thereby improving the differentiation of pseudomonads. The modified formula contains a low phosphorous content, added glycerol, magnesium chloride and potassium sulfate, all of which promote pyocyanin production.(1) Pyocyanin production is unique to Pseudomonas aeruginosa and is noted as a blue-green water soluble pigment that imparts a greenish color into the media.(8)

The addition of glycerol (Cat. no. 228210) to Hardy Diagnostics CRITERION™ Pseudomonas Isolation Agar Base provides an energy source. Peptone provides nutrients necessary for bacterial growth. Irgasan® is incorporated into the medium to inhibit the growth of many gram-positive and gram-negative microorganisms other than Pseudomonas spp.(2)

FORMULA

Gram weight per liter: 45.0gm/L
Gelatin Peptone 20.0gm
Potassium Sulfate 10.0gm
Magnesium Chloride 1.4gm
Irgasan® 25.0mg
Agar 13.6gm

Final pH 7.0 +/- 0.2 at 25ºC.

* Adjusted and/or supplemented as required to meet performance criteria.

STORAGE AND SHELF LIFE

Store the sealed bottle(s) containing dehydrated culture medium at 2-30ºC. Dehydrated culture medium is very hygroscopic. Keep lid tightly sealed. Protect dehydrated culture media from moisture and light. The dehydrated culture media should be discarded if it is not free-flowing or if the color has changed from its original light beige.

Store the prepared culture media at 2-8ºC.

PRECAUTIONS

METHOD OF PREPARATION FOR DEHYDRATED CULTURE MEDIA

1. Suspend 45.0gm of the dehydrated culture media in 980ml of deionized water. Stir to mix thoroughly.

2. Add 20ml glycerol (Cat. no. 228210).

3. Boil to dissolve completely. DO NOT OVERHEAT.

4. Sterilize in the autoclave at 121ºC. for 15 minutes.

5. Cool to 45-50ºC.

6. Dispense into sterile petri dishes.

PROCEDURE AND INTERPRETATION OF RESULTS

For information on procedures and interpretation of results, consult listed references or refer to the prepared media Instructions for Use (IFU) for Cat. No. G219.

LIMITATIONS

Some pseudomonads do not grow well at 35ºC. If no growth appears after 24 hours of incubation at 35ºC., the plates should be reincubated at 25ºC. for 24 hours.

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as loops, slides, staining supplies, other culture media, glycerol (Cat. no. 228210), microscopes, incinerators, and incubators, etc., as well as serological and biochemical reagents, are not provided.

QUALITY CONTROL

Test Organisms Inoculation Method* Incubation Results
Time Temperature Atmosphere
Pseudomonas aeruginosa
ATCC ® 27853
A 18-48hr 35°C Aerobic Growth; blue-green colonies
Pseudomonas aeruginosa
ATCC ® 9027
A 18-48hr 35°C Aerobic Growth; blue-green colonies
Burkholderia ( Pseudomonas ) cepacia
ATCC ® 25416
A 18-48hr 35°C Aerobic Growth; no pigment
Escherichia coli
ATCC ® 25922
B 18-48hr 35°C Aerobic Inhibited
Staphylococcus aureus
ATCC ® 25923
B 18-48hr 35°C Aerobic Inhibited

User Quality Control

Physical Appearance

CRITERION™ Pseudomonas Isolation Agar Base powder should appear homogeneous, free-flowing, and light beige in color. The prepared media should appear slightly opalescent, and light amber in color.

REFERENCES

1. King, E.O., et al. 1954. J. Lab. Clin. Med.; 44:301.

2. Soap and Chemical Specialties, January 1968.

3. Anderson, N.L., et al. Cumitech 3B; Quality Systems in the Clinical Microbiology Laboratory, Coordinating ed., A.S. Weissfeld. American Society for Microbiology, Washington, D.C.

4. Jorgensen., et al. Manual of Clinical Microbiology, American Society for Microbiology, Washington, D.C.

5. Isenberg, H.D. Clinical Microbiology Procedures Handbook, Vol. I, II & III. American Society for Microbiology, Washington, D.C.

6. MacFaddin, J.F. 1985. Media for Isolation, Cultivation, Identification, Maintenance of Bacteria, Vol. I. Williams & Wilkins, Baltimore, MD.

7. Quality Assurance for Commercially Prepared Microbiological Culture Media, M22. Clinical and Laboratory Standards Institute (CLSI - formerly NCCLS), Wayne, PA.

8. Bodey, G.D., et al. 1989. "Infections caused by P. aeruginosa". Rev. Infect. Dis.; 5:279-313.


ATCC is a registered trademark of the American Type Culture Collection.
Irgasan is a registered trademark of Geigy Chemical Corp.

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