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Brewer Thioglycollate HiVeg™ Medium, Modified (Thioglycollate HiVeg™ Medium, Linden)
For testing sterility of biological products and for isolation of aerobic and anaerobic organisms.
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Intended use
Recommended for testing sterility of biological products and for isolation of aerobic, anaerobic and microaerophilic organisms.
Composition**
| Ingredients | g/L |
|---|---|
| HiVeg™ hydrolysate | 17.500 |
| Soya peptone | 2.500 |
| Dextrose (Glucose) | 10.000 |
| Sodium chloride | 5.000 |
| Dipotassium hydrogen phosphate | 2.000 |
| Sodium thioglycollate | 1.000 |
| Methylene blue | 0.002 |
| Agar | 0.500 |
| Final pH (at 25°C) | 7.2±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 38.5 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. Cool to 45-50°C.
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 until the green colour disappears, and the prepared medium should be stored in the dark till use.
Principle And Interpretation
Brewer Thioglycollate HiVeg™ Medium is a modification of Linden Thioglycollate Medium (1). National Institute of Health specified the use of Brewers formula and Linden formula (2) for sterility testing, which was later referred to as Modified Brewer Thioglycollate Medium (3).
Brewer Thioglycollate HiVeg™ Medium, Modified is prepared by using vegetable peptones in place of animal based peptones which make the media free of BSE/TSE risks.
It contains highly nutritious HiVeg™ hydrolysate and soya peptone 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. The medium contains more thioglycollate and is recommended for sterility testing procedures. Organisms that ferment dextrose and lower the pH to critical levels may not survive in this medium after growth has taken place.
Growth is observed as turbidity of the medium compared to an uninoculated control. Strict aerobes tend to grow in a thin layer at the surface of the broth; obligate anaerobes will grow below the upper oxidized layer. Sometimes anaerobes can be overgrown by the more rapidly growing facultative organisms. Some anaerobes may be inhibited by acids or metabolic products produced from more rapidly growing facultative anaerobes. If the medium is to be used as a sterility testing medium incubation should be carried out for minimum 7 days under appropriate atmospheric conditions.
Type of specimen
Industrial samples for sterility testing.
Specimen Collection and Handling:
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.
Limitations :
- It is intended for the examination of clear liquid or water-soluble materials.
- 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 3.85% 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 (CFU) | Growth |
|---|---|---|
| Bacteroides melaninogenicus ATCC 25848 | 50-100 | good-luxuriant |
| Clostridium sporogenes ATCC11437 | 50-100 | good-luxuriant |
| Streptococcus mitis ATCC 9895 | 50-100 | good-luxuriant |
| Streptococcus pyogenes ATCC 19615 | 50-100 | good-luxuriant |
| Bacteroides fragilis ATCC 25285 | 50-100 | good-luxuriant |
| Staphylococcus aureus subsp. aureus ATCC ATCC 25923 (00034*) | 50-100 | good-luxuriant |
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.
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 (1,4).
| Product Name | Brewer Thioglycollate HiVeg™ Medium, Modified (Thioglycollate HiVeg™ Medium, Linden) |
|---|---|
| SKU | MV195 |
| Product Type | HiVeg™ |
| Physical Form | Powder |
| Origin | Animal Free (Veg) |
| Packaging type | HDPE |
| References | 1. Linden. 1941, National Institute of Health 2.MacFaddin J.F, 1985, Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria, Vol.1 Williams andWilkins, Baltimore. |
| Customized Product Available | No |
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