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Differential Reinforced Clostridial Broth Base
INTENDED USE
For cultivation of Clostridia from water.
MLSIGNUP100
Expiring in 23days 12hr7min2sec
PRODUCT SUMMARY AND EXPLANATION
Differential Reinforced Clostridial Broth Base is a differential agar medium used solely for the cultivation and counting of Clostridium species and other anaerobic bacteria. It is used extensively in food microbiology, clinical diagnostics, pharmaceutical analysis, and environmental microbiology for the isolation and counting of obligate anaerobes, especially spore-producing Clostridia. The medium facilitates the cultivation as well as differentiation of microbial populations under an anaerobic environment.
Medium also has peptones and beef extract as excellent sources of nitrogen, amino acids, and growth factors for anaerobic growth of stressed or injured anaerobic cells. Sodium acetate is added to suppress unwanted aerobic microbiota, and sodium chloride is added to contribute osmotic balance. A reducing agent like L-cysteine is usually added to give an ideal low-redox atmosphere, which is conducive to anaerobic growth. The addition of the redox indicator resazurin can give a visual indicator for anaerobic conditions by colour change relative to oxygen.
It is commonly used for sterility testing, faecal microbiota testing, and food production quality control, i.e., canned or vacuum food where Clostridium botulinum or Clostridium perfringens is a hazard. Enriched in specific carbohydrates or colouring agents, it can be made as a differential medium which can distinguish species based on the pattern of fermentation, gas production, or pH alteration.
Differential Reinforced Clostridial Broth Base is anaerobically incubated in jars, chambers, or pouches with the use of gas-generating kits to provide the necessary atmosphere. It can be utilized in combination with the Most Probable Number (MPN) technique to give measurable outcomes on the degree of microbial contamination.
Its selectivity, its very high recovery rate for anaerobes and its suitability for use with routine testing protocols render it an indispensable piece of equipment for anaerobic microbiological testing.
PRINCIPLE
The medium consists of Peptone, Beef extract, yeast extract, starch, sodium acetate which provide essential nutrients for bacterial metabolism. Glucose is the fermentable carbohydrate and serves as carbon and energy source. L-cysteine hydrochloride acts as reducing agent. Sodium sulphite and ferric citrate are added as indicators. Sulphite reducing clostridia produce sulphide from sulphite, which results in the formation of black coloured medium.
QUALITY CONTROL SPECIFICATIONS
Appearance of Powder : Cream to yellow homogeneous free flowing powder.
Appearance of prepared medium : Light yellow coloured, clear solution without any precipitate.
pH (at 25°C) : 7.2 ± 0.2
COMPOSITION
| Ingredients | Gms / Ltr |
| Peptone | 10.000 |
| Beef extract | 10.000 |
| Yeast extract | 1.500 |
| Starch | 1.000 |
| Sodium acetate, hydrated | 5.000 |
| Dextrose (Glucose) | 1.000 |
| L-Cysteine hydrochloride | 0.500 |
INSTRUCTION FOR USE
- Dissolve 29 grams in 1000 ml purified/distilled water Heat to boiling to dissolve the medium completely Sterilize by autoclaving at 15 psi pressure (121°C) for 15 minutes.
- Cool to 45-50°C. Just before use add 0.5 ml filter sterilized solution, prepared by mixing equal volumes of 4% w/v solution of sodium sulphite and 7% w/v ferric citrate, to 25 ml of single strength medium or 0.4 ml and 2 ml to 10 mland 50 ml of double strength medium respectively. Mix well.
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