BRILLIANT GREEN BILE BROTH WITH DURHAM TUBE

Cat. no. K09 Brilliant Green Bile Broth with Durham Tube, 20x125mm
Tube with Screw Cap, 13ml
20 tubes/box
Cat. no. K66 Brilliant Green Bile Broth with Durham Tube, 16x125mm
Tube with Screw Cap, 10ml
20 tubes/box

INTENDED USE

Hardy Diagnostics Brilliant Green Bile Broth with Durham Tube is recommended for the detection of coliforms in water and other samples.

This product is not intended to be used for the diagnosis of human disease.

SUMMARY

The formulation for Brilliant Green Bile Broth was developed by the Association of Official Analytical Communities (AOAC) and the American Public Health Association (APHA). (3-7) The basal medium is composed of peptone, which contains 2% bile, and brilliant green dye. Bile is inhibitory to gram-positive microorganisms, while brilliant green dye inhibits selected gram-negative bacilli. Lactose-fermenting organisms resistant to these inhibitors are detected by the production of gas. Gas production is noted by the appearance of bubbles in the durham tube.

FORMULA

Ingredients per liter of deionized water:*

Oxbile 20.0gm
Pancreatic Digest of Gelatin 10.0gm
Lactose 10.0gm
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

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

PRECAUTIONS

PROCEDURE

Specimen Collection: Consult listed references for information on specimen collection. (3-6) 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 medium and refrigerated until inoculation.

Method of Use: Allow medium to warm to room temperature prior to inoculation. Consult listed references for information concerning inoculation procedures. (4-6)

INTERPRETATION OF RESULTS

Production of gas within 48 +/- 2 hours of incubation is considered positive evidence of fermentation by coliform bacilli. Absence of gas production within the durham tube in 48 hours is considered a negative test for coliforms. Consult listed references for detailed results for the enumeration of coliforms using Brilliant Green Bile Broth. (3-5)

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 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
Escherichia coli
ATCC ® 25922
A 18-48hr 35°C Aerobic Growth; gas bubble in durham tube
Salmonella enterica
ATCC ® 14028
A 18-48hr 35°C Aerobic Growth; no gas bubble in durham tube
Enterococcus faecalis
ATCC ® 29212
B 18-48hr 35°C Aerobic Inhibited

User Quality Control

PHYSICAL APPEARANCE

Brilliant Green Bile Broth should appear clear and light green in color.

E. coli growing in Brilliant Green Bile Broth

Escherichia coli (ATCC ® 25922) growing in Brilliant Green Bile Broth (Cat. no. K09). The bubble in the Durham tube indicates gas production. Incubated aerobically for 24 hours at 35ºC.

S. typhimirium growing in Brilliant Green Bile Broth

Salmonella enterica (ATCC ® 14028) growing in Brilliant Green Bile Broth (Cat. no. K09). Incubated aerobically for 24 hours at 35ºC.



E. faecalis growth inhibited in Brilliant Green Bile Broth

Enterococcus faecalis (ATCC ® 29212) inhibited in Brilliant Green Bile Broth (Cat. no. K09). Incubated aerobically for 24 hours at 35ºC.



REFERENCES

1. 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.

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

3. APHA Technical Committee on Microbiological Methods for Foods. Compendium of Methods for the Microbiological Examination of Foods, APHA, Washington, D.C.

4. American Public Health Association. Standard Methods for the Examination of Dairy Products, APHA, Washington, D.C.

5. American Public Health Association. Standard Methods for the Examination of Water and Wastewater, APHA, Washington, D.C.

6. "Public Health Service Drinking Water Standards". 1962. Publication No. 956, U.S. Government Printing Office, Washington, D.C.

7. Association of Official Analytical Communities. Official Methods of Analysis . AOAC, Washington, D.C


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

040116sw