M1255

Tetrathionate Brilliant Green Bile Broth M1255
Intended Use:
Recommended for isolation and identification of Salmonellae.
Composition**
Ingredients Gms / Litre
Peptone 8.600
Bile 8.000
Sodium chloride 6.400
Calcium carbonate 20.000
Potassium tetrathionate 20.000
Brilliant green 0.070
Final pH ( at 25°C) 7.0±0.2
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 63.07 grams in 1000 ml purified / distilled water. Heat just to boiling. DO NOT AUTOCLAVE OR
REHEAT. Dispense into sterile tubes or flasks as desired.
Note: Due to the presence of calcium carbonate, the prepared medium forms opalescent solution with white precipitate.
Principle And Interpretation

Salmonella are gram-negative, facultatively anaerobic, non-sporulating, non-motile rods in the family Enterobacteriaceae.
They are widely distributed in animals affecting mainly the stomach and the intestines. These organisms are difficult to
differentiate biochemically from Escherichia coli. Tetrathionate Broth was originally described by Mueller (7) and later
modified by Kauffman (4,5). Tetrathionate Brilliant Green Bile Broth is used as an enrichment medium for Salmonella.
Enrichment broth is usually recommended to facilitate the recovery of small numbers of Salmonella species (4). Tetrathionate
Brilliant Green Bile Broth is also mentioned in I.P. (1) for isolation and identification of Salmonella species from foods,
water and other materials of sanitary importance.
Peptone in the medium provides nitrogenous and carbonaceous compounds, long chain amino acids, vitamins and
nutrients for growth of Salmonellae. Brilliant green and bile inhibit both gram-positive as well as some selected
gram-negative organisms. Potassium tetrathionate inhibits normal flora of faecal specimens. Sodium chloride helps in
maintaining osmotic equilibrium.
After incubation, streak the culture from Tetrathionate Brilliant Green Bile Broth (M1255) onto differential medium
for isolation and identification.
Tetrathionate Brilliant Green Bile Broth is not suitable for growth of Salmonella Typhi and Salmonella Paratyphi (6).
Type of specimen
Food samples
Specimen Collection and Handling
For food samples, follow appropriate techniques for sample collection and processing as per guidelines (9).
After use, contaminated materials must be sterilized by autoclaving before discarding.
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.
Please refer disclaimer Overleaf.

HiMedia Laboratories Technical Data
Limitations
1. For further confirmation, streak the enriched cultures after incubation, on plates of Brilliant Green Agar (M016),
MacConkey Agar (M081) and Bismuth Sulphite Agar (M027).
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.
Quality Control
Appearance
Light yellow to pale green homogeneous free flowing powder
Colour and Clarity of prepared medium
Bluish green coloured opalescent solution with white precipitate.
Reaction
Reaction of 6.3% w/v aqueous solution at 25°C. pH : 7.0±0.2
pH
6.80-7.20
Cultural Response
Cultural characteristics observed when subcultured on MacConkey Agar (M082) after an incubation at 35-37°C for 18-24
hours.
Organism Inoculum Growth Recovery Colour of
(CFU) colony
Escherichia coli ATCC 50-100 fair 20-30% pink to red with

25922 (00013*) bile precipitate
Salmonella Typhi ATCC 50-100 luxuriant >=50% colourless
6539
Salmonella Typhimurium 50-100 luxuriant >=50% colourless
ATCC 14028 (00031*)
Salmonella Enteritidis ATCC 50-100 luxuriant >=50% colourless
13076 (00030*)
Staphylococcus aureus >=104 inhibited 0%
subsp. aureus ATCC
25923 (00034*)
subsp. aureus ATCC
6538 (00032*)
Escherichia coli 50-100 fair 20-30% pink to red with

ATCC 8739 bile precipitate
Escherichia coli NCTC 9002 50-100 fair 20-30% Pink to red with
bile precipitate
NCIMB 9518
Key : *Corresponding WDCM numbers.
Storage and Shelf Life

Store between 10-30°C in a tightly closed container and the prepared medium at 15-25°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.
Please refer disclaimer Overleaf.

HiMedia Laboratories Technical Data
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 (2,3).
Reference
1. Indian Pharmacopoeia, 1996, Ministry of Health and Family Welfare, Govt. of India,
2. Isenberg, H.D. Clinical Microbiology Procedures Handbook 2nd Edition.
3. 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.
4. Kauffman F., 1930, Hyg. Abt. I. Orig., 113, 148.
5. Kauffman F., 1935, Z. Hyg. Infektionskr., 117, 26.
6. MacFaddin J. F., 1985, Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria, Vol. I, Williams
and Wilkins, Baltimore.
7. Mueller L., 1923, C. R. Soc. Biol., (Paris), 89, 434.
8. Murray P. R., Baron J. H., Pfaller M. A., Jorgensen J. H. and Yolken R. H., (Ed.). 2003, Manual of Clinical Microbiology,
8th Ed., American Society for Microbiology, Washington, D.C.
9. Salfinger Y., and Tortorello M.L., 2015, Compendium of Methods for the Microbiological Examination of Foods, 5th
Ed., American Public Health Association, Washington, D.C.
Revision : 03/ 2019
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.
HiMedia Laboratories Pvt. Ltd. Reg.office : 23, Vadhani Ind.Est., LBS Marg, Mumbai-400086, India. Customer care No.: 022-6116 9797 Corporate
office : A-516,Swastik Disha Business Park,Via Vadhani Ind. Est., LBS Marg, Mumbai-400086, India. Customer care No.: 022-6147 1919 Email:
techhelp@himedialabs.com Website: www.himedialabs.com

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Tetrathionate Brilliant Green Bile Broth M1255

Principle And Interpretation

Warning and Precautions

Please refer disclaimer Overleaf.

Performance and Evaluation

Escherichia coli ATCC 50-100 fair 20-30% pink to red with

Escherichia coli 50-100 fair 20-30% pink to red with

Key : *Corresponding WDCM numbers.

Storage and Shelf Life

Please refer disclaimer Overleaf.

Revision : 03/ 2019

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