Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended Use
Recommended for the cultivation and enumeration of Clostridia and other anaerobes.
Composition**
| Ingredients | g/L |
|---|---|
| Tryptone | 10.000 |
| HM peptone B # | 10.000 |
| Yeast extract | 3.000 |
| Dextrose (Glucose) | 5.000 |
| Sodium chloride | 5.000 |
| Sodium acetate | 3.000 |
| Starch, soluble | 1.000 |
| L-Cysteine hydrochloride | 0.500 |
| Agar | 13.500 |
| Final pH (at 25°C) | 6.8±0.2 |
**Formula adjusted, standardized to suit performance parameters
# Equivalent to Beef extract
Directions
Reinforced Clostridial Agar is a ready to use solid media in glass bottle. The medium is pre-sterilized, hence it does not need sterilization. Medium in the bottle can be melted either by using a pre-heated water bath or any other method. Slightly loosen the cap before melting. When complete melting of medium is observed dispense the medium in tubes as butts /slants or in plates as desired and allow to solidify. If on plate, either streak, inoculate or surface spread the test inoculum (50-100 CFU) aseptically.
Principle And Interpretation
Hirsch and Grinstead formulated Semisolid Reinforced Clostridial Medium and found that clostridia from small amount of samples could be grown well on this medium with higher viable cell counts (1). This medium can also be used for growing anaerobic and facultative bacteria (2). Barnes et al used a solid (agar) version of the medium (2) to develop vegetative cells in assays of Clostridium perfringens. Reinforced Clostridial Medium was used in the enumeration of clostridia from food (3). Reinforced Clostridial Agar contains tryptone and HM peptone B as sources of carbon, nitrogen, vitamins and minerals. Yeast extract supplies B-complex vitamins which stimulate bacterial growth. Dextrose is the carbohydrate source. Sodium chloride maintains the osmotic balance. In low concentrations, soluble starch detoxifies metabolic byproducts. Cysteine hydrochloride is the reducing agent. Sodium acetate acts as a buffer. This medium can be made selective by addition of 15-20 mg polymyxin B per litre of media (3).
Type of specimen
Clinical samples- Pus samples, Food and dairy samples.
Specimen Collection and Handling
For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (4,5,6). For clinical samples follow appropriate techniques for handling specimens as per established guidelines (7,8). After use, contaminated materials must be sterilized by autoclaving before discarding.
Warning and Precautions
In Vitro diagnostic Use. For professional use only. 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 clinical specimens. Safety guidelines may be referred to in individual safety data sheets.
Limitations :
- Further serological and 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
Sterile glass bottle containing slightly opalescent Reinforced Clostridial Agar.
Colour
Amber Coloured medium
Quantity of Medium
500 ml of medium in glass bottle.
pH
6.60-7.00
Sterility Check
Passes release criteria
Cultural Response
Cultural characteristics after melting the medium and pouring into sterile petri plates. The plates are inoculated with following test organisms and incubation at 35-37°C for 48 hours under anaerobic condition.
| Organism | Inoculum (CFU) | Growth | Recovery |
|---|---|---|---|
| Bacteroides fragilis ATCC 23745 | 50-100 | good - luxuriant | >=50% |
| **Phocaeicola vulgatus** ATCC 8482 | 50-100 | good - luxuriant | >=50% |
| Clostridium butyricum ATCC 13732 | 50-100 | good - luxuriant | >=50% |
| Clostridium perfringens ATCC 13124 (00007*) | 50-100 | good - luxuriant | >=50% |
Key: (*) Corresponding WDCM numbers ** Formerly known as Bacteroides vulgatus
Storage and Shelf Life
Store between 15-30°C. 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 clinical sample must be decontaminated and disposed of in accordance with current laboratory techniques (7,8).
| Product Name | Reinforced Clostridial Agar |
|---|---|
| SKU | SM154D |
| Customized Product Available | No |
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