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Technical Data
Brilliant Green Bile Broth M121I
Intended Use:
Recommended for isolation and cultivation of coliform organisms from cream, yogurt and raw milk. The composition and
performance criteria of this medium are as per the specifications laid down in ISO 4831:2006, ISO 11133:2014 &
Amd.2 :2020 (E).
Composition**
ISO Specifications : BGBLB Brilliant Green Bile Broth M121I
Ingredients g/L Ingredients g/L
Enzymatic digest of casein 10.000 Tryptone$ 10.000
Lactose 10.000 Lactose monohydrate 10.000
Dehydrated Ox bile 20.000 Dehydrated bile 20.000
Brilliant green 0.0133 Brilliant green 0.0133
Final pH ( at 25°C) 7.2±0.2 Final pH ( at 25°C) 7.2±0.2
**Formula adjusted, standardized to suit performance parameters
$ Equivalent to Enzymatic digest of casein
Directions
Suspend 39.51 gram (the equivalent weight of dehydrated medium per liter) in 1000 ml purified / distilled water. Heat if
necessary to dissolve the medium completely. Dispense the medium in quantities of 10ml in test tubes of approximately
16mm x 160mm containing Durham tubes. Sterilize in an autoclave set at 121°C for 15 minutes. Cool to 45-50°C.
Note: The Durham tube shall not contain air bubbles after sterilization.
Principle And Interpretation
Brilliant Green Bile Broth is formulated as per ISO for confirmation of coliform bacteria (1,2) present in food samples or
environmental samples in the area of food handling or food sampling.
Brilliant green and dehydrated bile present in the medium inhibit gram-positive bacteria including lactose fermenting
Clostridia (3). Production of gas from lactose fermentation is detected by incorporating inverted Durham’s tube, indicates a
positive evidence of faecal coliforms since nonfaecal coliforms growing in this medium do not produce gas.
During examination of samples, growth from presumptive positive tubes showing gas in Lauryl Tryptose Broth
(M080) is inoculated in Brilliant Green Bile Broth wherein gas formation within 48 ± 2 hours confirms the presumptive test
(1). Gram-positive spore-formers may produce gas if the bile or brilliant green inhibition is weakened by food material.
Type of specimen
Food samples

Specimen Collection and Handling:

ISO 4831:2006 (1,2)
Depending on the limit of detection that is required, x ml of the test sample if liquid, or x ml of the initial suspension in the
case of other products, is transferred to a tube containing 10 ml of double-strength selective enrichment medium. Incubate at
30°C or 37°C (as agreed) for 24 h ± 2 h, continue incubation for another 24 h ± 2 h for gas formation. Gas formation is
considered as positive.
Warning and Precautions :
Read the label before opening the container. Wear protective gloves/protective clothing/eye protection/ face protection.
Follow good microbiological lab practices while handling specimens and culture. Standard precautions as per established
guidelines should be followed while handling specimens. Safety guidelines may be referred in individual safety data sheets.

Limitations :
1. Individual organisms differ in their growth requirement and may show variable growth patterns on the medium.
2. Further biochemical & serological identification is necessary for confirmation.
Performance and Evaluation
Performance of the medium is expected when used as per the direction on the label within the expiry period when stored at
recommended temperature.
Please refer disclaimer Overleaf.
HiMedia Laboratories Technical Data

Quality Control
Appearance
Cream to pale green homogeneous free flowing powder
Colour and Clarity of prepared medium
Emerald green coloured, clear solution without any precipitate.
Reaction
Reaction of 3.95% w/v aqueous solution at 25°C. pH : 7.2±0.2
pH
7.00-7.40
Cultural Response
Productivity : Cultural characteristics observed after an incubation at 30±1°C for 24±2h to 48±2h.
Selectivity : Cultural characteristics observed after an incubation at 30±1°C for 24±2h to 48±2h.
Organism Inoculum Growth Gas
(CFU)
Productivity
Escherichia coli ATCC 50-100 good-luxuriant positive
25922 (00013*) reaction
Escherichia coli ATCC 50-100 good-luxuriant positive
8739 (00012*) reaction
Citrobacter freundii
50-100 good-luxuriant positive
ATCC 43864 (00006*)
reaction
Selectivity
Enterococcus faecalis ATCC50-100 none-poor negative
29212 (00087*) reaction
Enterococcus faecalis 50-100 none-poor negative
ATCC 19433 (00009*) reaction
Key : * - Corresponding WDCM numbers
Storage and Shelf Life
Store between 10-30°C in a tightly closed container and the prepared medium at 15-30°C. Use before expiry date on the
label. On opening, product should be properly stored dry, after tightly capping the bottle in order to prevent lump formation
due to the hygroscopic nature of the product. Improper storage of the product may lead to lump formation. Store in
dry ventilated area protected from extremes of temperature and sources of ignition. Seal the container tightly after use.
Product performance is best if used within stated expiry period.
Disposal
User must ensure safe disposal by autoclaving and/or incineration of used or unusable preparations of this product. Follow
established laboratory procedures in disposing of infectious materials and material that comes into contact with sample must
be decontaminated and disposed of in accordance with current laboratory techniques (4,5).
Reference
1. International Standard, ISO 4831:2006 (E). Microbiology of food and animal feeding stuffs — Horizontal method for
the detection and enumeration of coliforms — Most probable number technique.
2. Microbiology of food, animal feeding stuffs and water- Preparation, production,storage and performance testing of culture
media, EN ISO 11133:2014(E) /Amd.: 2020 .
3. McCrady and Langerin, 1932, J. Dairy Science, 15:321.
4. Isenberg, H.D. Clinical Microbiology Procedures Handbook 2nd Edition.
5. Jorgensen, J.H., Pfaller, M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W. (2015) Manual
of Clinical Microbiology, 11th Edition. Vol. 1.

Revision:04/2024

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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