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Buffered Tryptone Glucose Yeast Extract Broth

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M951
For cultivation and characterization of Clostridia isolated from food specimens as per APHA.

Intended Use

Recommended for cultivation and characterization of Clostridia isolated from food specimens.

Composition

Ingredients Gms / Litre
Tryptone 50.000
Peptone 5.000
Yeast extract 20.000
Dextrose (Glucose) 4.000
Disodium hydrogen phosphate 5.000
Sodium thioglycollate 1.000
Final pH (at 25°C) 7.3±0.2

Formula adjusted, standardized to suit performance parameters

Directions

Suspend 8.5 grams in 100 ml purified / distilled water. Heat, if necessary to dissolve the medium completely and dispense 15 ml into 20x150 mm test tubes or 100 ml in 170 ml bottles. Sterilize by autoclaving at 15 lbs pressure (121°C) for 8 minutes (tubes) or 15 minutes (bottles).

Principle And Interpretation

Clostridium perfringens food poisoning is one of the most common types of foodborne illness (3). A heat labile enterotoxin produced only by sporulating cells (4) induces the major symptoms of diarrhoea in perfringens poisoning (1). Although the enterotoxin is not preformed in the food, the foods in which conditions are favorable for sporulation may contain enterotoxin (2,7). Buffered Tryptone Glucose Yeast Extract Broth is prepared as recommended by APHA (8) for enrichment as well as for cultivation of Clostridia from food samples. Buffered Tryptone Glucose Yeast Extract Broth is also used to obtain pure cultures of Clostridia before proceeding for confirmation. Endospores are not usually produced in this medium(8).

The medium contains tryptone, peptone and yeast extract, which provides carbon and nitrogen, vitamins and other essential nutrients. Dextrose is the fermentable sugar. Disodium phosphate buffers the medium well. Sodium thioglycollate present in the medium acts as a reducing agent and maintains a low oxygen tension in the medium.

Type of specimen

Food samples

Specimen Collection and Handling

For enrichment 2 grams of food sample is inoculated in 15-20 ml of sterile Buffered Tryptone Glucose Yeast Extract Broth (M951). After incubation of 20-24 hours at 35-37°C the culture from the tubes shows turbidity. 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

  • Further recovery on selective media is necessary to obtain pure colonies of culture.
  • Biochemical and serological testing of pure colony 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: Light amber coloured clear solution without any precipitate

Reaction: Reaction of 8.5% w/v aqueous solution at 25°C. pH: 7.3±0.2

pH: 7.10-7.50

Cultural Response

Cultural characteristics observed after an incubation at 35-37°C for 18-48 hours.

Organism Inoculum (CFU) Growth
Clostridium botulinum ATCC 25763 50-100 good-luxuriant
Clostridium perfringens ATCC 12924 50-100 good-luxuriant
Clostridium sporogenes ATCC 11437 50-100 good-luxuriant

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 (5,6).

Reference

  1. Bartholomew et al, 1985, J. Clin. Pathol. 38:222.
  2. Craven S. E., Blankenship L.C. and McDonel J. L., 1981, Appl. Microbiol. 41: 1184.
  3. Doyle M. P., (Ed.), 1989, Foodborne Bacterial Pathogens, Marcel Dekker, New York.
  4. Duncan C. L., 1973, J. Bacteriol., 113:932.
  5. Isenberg, H.D. Clinical Microbiology Procedures Handbook 2nd Edition.
  6. 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.
  7. Naik M. S., and Duncan C. L., 1977, J. Food Safety, 1:7.
  8. 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.
More Information
Product Name Buffered Tryptone Glucose Yeast Extract Broth
SKU M951
Product Type Regular
Physical Form Powder
Origin Animal
Packaging type HDPE
References 1. Vanderzant C. and Splittstoesser D. F., (Eds.), 1992, Compendium of Methods for the Microbiological Examination ofFoods, 3rd Ed., APHA,Washington, D.C.2.Doyle M. P., (Ed.), 1989, Foodborne Bacterial Pathogens, Marcel Dekker, New York.3.Duncan C. L., 1973, J. Bacteriol., 113:932.4.Bartholomew et al, 1985, J. Clin. Pathol. 38:222.5.Craven S. E., Blankenship L.C. and McDonel J. L., 1981, Appl. Microbiol. 41 : 1184.6.Naik M. S., and Duncan C. L.,1977, J. Food Safety, 1:7.
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