Brewer Thioglycollate Medium M019
Intended use
Brewer Thioglycollate Medium is used for testing the sterility of biological products and for isolation of aerobic and
anerobic organisms
Composition**
Ingredients Gms / Litre
Proteose peptone 10.000
HM infusion from # 500.000
Dextrose (Glucose) 5.000
Sodium chloride 5.000
Dipotassium hydrogen phosphate 2.000
Sodium thioglycollate 0.500
Methylene blue 0.002
Agar 0.500
Final pH ( at 25°C) 7.2±0.2
**Formula adjusted, standardized to suit performance parameters
# Equivalent to Beef, infusion from
Directions
Suspend 40.5 grams in 1000 ml purified/distilled water. Heat to boiling to dissolve the medium completely. Dispense in
tubes or in suitable containers as desired and sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes.
Note : If more than the upper one third layer acquires bluish-green colour (absorbs oxygen), the dissolved oxygen can be
removed by heating the medium in free flowing steam for 5-10 minutes or in a water bath untill the green colour disappears,
and the prepared medium should be stored in the dark till use.
Principle And Interpretation
Brewer thioglycollate medium is prepared as per the original formula of Brewer (1, 2).
It contains highly nutritious proteose peptone and HM infusion that provides carbon, nitrogen substances, long chain amino
acids, vitamins and minerals which support luxuriant growth of even fastidious bacteria. Sodium thioglycollate helps to
create anaerobic condition as well as neutralizes toxicity of mercurial compounds if present in the inoculum of the test
material. Sodium chloride maintains the osmotic equilibrium while dipotassium phosphate buffers the medium. Very small
amount of agar present maintains anaerobic conditions at the bottom of the broth. Methylene blue indicates oxygen content
of the medium by exhibiting bluish-green colour to the medium in presence of oxygen.
The uninoculated medium shows bluish green colour at the top indicating presence of oxygen in that part. Organisms
that ferment dextrose and lower the pH to critical levels may not survive in this medium after growth has taken place.
Type of specimen
Industrial samples for sterility testing.
Specimen Collection and Handling:
For Industrial samples, follow appropriate techniques for sample collection, processing as per guidelines (3,4) .
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. It is intended for the examination of clear liquid or water-soluble materials.
2. Further biochemical testing is required for complete identification.
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
Cream to yellow homogeneous free flowing powder
Colour and Clarity of prepared medium
Yellow coloured clear to slightly opalescent fluid with upper 10% or less medium bluish green on standing.
Reaction
Reaction of 4.05% w/v aqueous solution at 25°C. pH : 7.2±0.2
pH
7.00-7.40
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 18-48 hours. ( Clostridium and Bacteroides species
incubated anaerobically)
Organism Inoculum Growth
(CFU)
Bacteroides
50-100 good-luxuriant
melaninogenicus ATCC
25848
Clostridium sporogenes 50-100 good-luxuriant
ATCC 11437
Streptococcus mitis ATCC 50-100 good-luxuriant
9895
Streptococcus pyogenes 50-100 good-luxuriant
ATCC 19615
Bacteroides fragilis ATCC 50-100 good-luxuriant
25285
Staphylococcus aureus 50-100 good-luxuriant
subsp.aureus ATCC
25923 (00034*)
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. Use before expiry date on the label.
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 (3,4).
Reference
1. Brewer J. H., 1940, J. Bacteriol., 39:10
2. Brewer J. H., 1940, J.A.M.A., 115:598.
3. Isenberg, H.D. Clinical Microbiology Procedures Handbook. 2nd Edition.
4. Jorgensen,J.H., Pfaller , M.A., Carroll, K.C., Funke, Andry, M.L., Richter, S.S and Warnock., D.W. (2015)
Manual of Clinical Microbiology, 11th Edition. Vol. 1.
Revision : 02 / 2019
Please refer disclaimer Overleaf.
HiMedia Laboratories Technical Data
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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