ACETAMIDE SLANT

Cat. no. L02 Acetamide Slant, 16x100mm Tube, 5.5ml Slant 20 or 100 tubes/box

INTENDED USE

Hardy Diagnostics Acetamide Agar Slant is recommended for the differentiation of Pseudomonas aeruginosa.

SUMMARY

Culture media containing acetamide as a carbon source have been used for isolation and identification of P. aeruginosa. This media has been described as a modified Simmons Citrate Agar using acetamide as the sole carbon source. A variety of modified simmons agar formulations have been used to survey the ability of various nonfermenting gram negative bacteria to deaminate acetamide. A few species were found to be capable. Acetamide deamination was most actively accomplished by Pseudomonas acidovorans, P. aeruginosa, Group III (Achromobacter xylosoxidans), and Alcaligenes odorans. The alkalization that occurs with deamination is indicated by a pronounced blue reaction in the medium. The reaction is rapid and clear cut.

FORMULA

Ingredients per liter of deionized water:*

Acetamide 10.0gm
Sodium Chloride 5.0gm
Monoammonium Phosphate 1.0gm
Dipotassium Phosphate 1.0gm
Magnesium Sulfate 0.2gm
Bromothymol Blue 0.08gm
Agar 15.0gm

Final pH 6.8 +/- 0.2 at 25ºC.

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

STORAGE AND SHELF LIFE

Storage: Upon receipt store at 2-30ºC. away from direct light. Media should not be used if there are any signs of deterioration (shrinking, cracking, or discoloration), hemolysis, contamination, or if the expiration date has passed. Product is light and temperature sensitive; protect from light, excessive heat and freezing.

PRECAUTIONS

PROCEDURE

Specimen Collection: Consult listed references for information on specimen collection.(1-4) Infectious material should be submitted directly to the laboratory without delay and protected from excessive heat and cold. If there is to be a delay in processing, the specimen should be inoculated onto an appropriate transport media and refrigerated until inoculation.

Method of Use: Performance of the Acetamide Slant is dependant upon proper inoculation of the test medium from a pure culture. To prevent delay in the initiation of growth and medium reaction, the inoculum should not be taken from a liquid or broth suspension of the organism. Allow medium to warm to room temperature. Inoculum should be taken from the center of a well isolated colony of a fresh 18-24 hour culture. Inoculate the Acetamide Slant by drawing the loop straight up the slant, or streaking the surface in a serpentine pattern. Place cap loosely on the tube to ensure adequate aeration. The test medium should be incubated at 35ºC. and checked daily for four days for signs of growth and color change.

INTERPRETATION OF RESULTS

A positive reaction is indicated by growth of the organism and a color change of the medium from green to blue.

LIMITATIONS

This product is not intended for primary isolation of patient specimens. The reactions of this medium are not sufficient for speciation.

Additional biochemical and/or serological tests are necessary for definitive identification.

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as loops, other culture media, swabs, applicator sticks, 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 24-48hr 35°C Aerobic Growth; color change from green to blue
Escherichia coli
ATCC ® 25922
B 24hr 35°C Aerobic Inhibited

User Quality Control

Physical appearance

Acetamide Slant media should appear clear, and green in color.

P. aeruginosa growing on Acetamide Slant

Pseudomonas aeruginosa (ATCC ® 27853) colonies growing on an Acetamide Slant (Cat. no. L02). Incubated aerobically for 24 hours at 35ºC.

E. coli growth inhibited on Acetamide Slant

Escherichia coli (ATCC ® 25922) inhibited on an Acetamide Slant (Cat. no. L02). Incubated aerobically for 24 hours at 35ºC.

REFERENCES

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

2. Tille, P., et al. Bailey and Scott's Diagnostic Microbiology, C.V. Mosby Company, St. Louis, MO.

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


ATCC is a registered trademark of the American Type Culture Collection.


080216gr