CRITERION™ BRILLIANT GREEN BILE BROTH WITH MUG

Cat. no. C5280 CRITERION™ Brilliant Green Bile Broth with MUG 80gm
Cat. no. C5281 CRITERION™ Brilliant Green Bile Broth with MUG 500gm
Cat. no. C5282 CRITERION™ Brilliant Green Bile Broth with MUG 2kg
Cat. no. C5283 CRITERION™ Brilliant Green Bile Broth with MUG 10kg
Cat. no. C5284 CRITERION™ Brilliant Green Bile Broth with MUG 50kg

INTENDED USE

Hardy Diagnostics CRITERION™ Brilliant Green Bile Broth with MUG is a selective medium recommended for the detection of coliforms in water, sewage, dairy products and other samples.

The addition of 4-methylumbilliferyl-beta-D-glucuronic acid (MUG) reagent detects Escherichia coli.

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

Brilliant Green Bile Broth with MUG is a selective medium for the detection of coliforms in water, foods, dairy products and other materials.(1,4,5) Bile and brilliant green inhibit growth of most organisms other than coliforms. Coliform detection is based on lactose-fermentation with corresponding accumulation of gas within inverted fermentation vials. This medium is formulated according to APHA specifications.(4,5) The addition of MUG reagent makes this medium specific for Escherichia coli. Most strains (96-97%) of E. coli produce glucuronidase. This enzyme hydrolysis MUG reagent to 4-methylumbelliferone, a fluorescent compound, detected by (long-wavelength) UV light. Fluorescence produced in the broth is highly specific for E. coli.

FORMULA

Gram weight per liter: 40.0gm/L
Oxbile (Oxgall) 20.0gm
Pancreatic Digest of Gelatin 10.0gm
Lactose 10.0gm
MUG Reagent 50.0mg
Brilliant Green 13.3mg

Final pH 7.2 +/- 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 greenish-tan.

Store the prepared culture media at 2-8ºC.

PRECAUTIONS

METHOD OF PREPARATION FOR DEHYDRATED CULTURE MEDIA

1. Suspend 40.0gm of the dehydrated culture media in 1 liter of distilled or deionized water. Stir to mix thoroughly.

2. Heat as necessary to dissolve completely. Avoid overheating.

3. Dispense into tubes containing durham tubes.

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

PROCEDURE AND INTERPRETATION OF RESULTS

For information on procedures and interpretation of results, consult listed references.

LIMITATIONS

It may be necessary to invert the tube prior to inoculation if bubbles are trapped in the durham tube. Trapped bubbles that are not released may lead to false-positive results.

Turbidity alone is not indicative of a positive test for the presence of coliforms; turbidity with gas production is considered a positive test.

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as autoclaves, incinerators, and incubators, etc., are not provided.

QUALITY CONTROL

Test Organisms Inoculation Method* Incubation Results
Time Temperature Atmosphere
Escherichia coli
ATCC ® 25922
A 18-48hr 35°C Aerobic Growth; gas bubble in durham tube and fluorescence
Salmonella enterica
ATCC ® 14028
A 18-48hr 35°C Aerobic Growth; no gas bubble in durham tube with no fluorescence
Enterococcus faecalis
ATCC ® 29212
B 18-48hr 35°C Aerobic Inhibited

User Quality Control

PHYSICAL APPEARANCE

CRITERION™ Brilliant Green Bile Broth with MUG powder should appear homogeneous, free-flowing, and greenish-beige in color. The prepared media should appear clear, and emerald green in color.

REFERENCES

1. Eaton, Clesceri and Greenberg, (ed.). 1995. Standard Methods for the Examination of Water and Wastewater, 19th ed. APHA, Washington, D.C.

2. Association of Official Analytical Chemists. Official Methods of Analysissm, AOAC, Washington, D.C.

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

4. Vanderzant, C. and D.F. Splittstoesser, (ed.). 1992. Compendium of Methods for the Microbiological Examination of Foods, 3rd ed. APHA, Washington, D.C.

5. Marshall, Robert T. 1992. Standard Methods for the Examination of Dairy Products, 16th ed. APHA, Washington, D.C.

6. Appl. Environ. Microbiol.; 48:285. 1984.

7. Appl. Environ. Microbiol.; 43:1320. 1982.


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

051616gr