Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended use
Recommended for the enrichment of Clostridia from pharmaceutical products in accordance with the microbial limit testing by harmonized methodology of USP, EP, BP, JP & IP.
Composition**
| Ingredients | g/L |
|---|---|
| HM Pepone B # | 10.000 |
| Peptone | 10.000 |
| Yeast extract | 3.000 |
| Soluble starch | 1.000 |
| Glucose monohydrate | 5.000 |
| Cysteine hydrochloride | 0.500 |
| Sodium chloride | 5.000 |
| Sodium acetate | 3.000 |
| Agar | 0.500 |
| Final pH (at 25°C) | 6.8±0.2 |
**Formula adjusted, standardized to suit performance parameters
# Equivalent to Beef Extract
Directions
Label the ready to use LQ130D bottle. Inoculate the sample and incubate at specified temperature and time.
Principle And Interpretation
Reinforced Medium for Clostridia was formulated by Hirsch and Grinsted (1) and is in accordance with the microbial limit testing by harmonized methodology of USP/EP/BP/JP/IP (2-6). It is recommended for sterility checking of non-sterile products, nutritional and dietary supplements. It can be used to initiate growth from small inocula and to obtain the highest viable count of Clostridia. Barnes and Ingram used the broth medium for diluting an inoculum of vegetative cells of Clostridium perfringens (7,8). It can be used in studies of spore forming anaerobes, especially Clostridium butyricum in cheese, for enumeration of Clostridia in tube dilution counts or for preparation of plates for isolation (8). Other spore forming anaerobes, Streptococci and Lactobacilli also grow in these media. These are enriched but non-selective media. Peptone, yeast extract and HM peptone B provide all the necessary nutrients for the growth of Clostridia. Glucose monohydrate is a fermentable carbohydrate in the medium while sodium chloride maintains osmotic equilibrium. Cysteine hydrochloride acts as reducing agent. Small amount of soluble starch removes toxic metabolites from the medium. Sodium acetate also acts as a good buffering agent. Small quantity of agar keeps the medium semi solid and helps in maintaining anaerobic conditions.
Type of specimen
Pharmaceutical samples
Specimen Collection and Handling
For pharmaceutical samples, follow appropriate techniques for sample collection, processing as per pharmaceutical guidelines (2-6).
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
- Some Clostridium species may show poor growth due to nutritional variations.
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 clear to slightly opalescent Reinforced Medium for Clostridia in glass bottle.
Colour
Light yellow 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 observed in an anaerobic atmosphere, after an incubation at 30-35°C for 24-48 hours.
Growth Promotion Test
Growth promotion was carried out in accordance with the harmonized method of USP/EP/BP/JP/IP, and growth was observed under anaerobic conditions after an incubation at 30-35°C <=48 hours.
Growth promoting properties
Clearly visible growth of microorganism comparable to that previously obtained with previously tested and approved lot of medium occurs at the specified temperature for not more than the shortest period of time specified inoculating not more than 100 cfu under anaerobic conditions (at 30-35°C for <=48 hours).
| Organism | Inoculum (CFU) | Growth | Incubation temperature | Incubation period |
|---|---|---|---|---|
| Growth Promoting | ||||
| Clostridium sporogenes ATCC 11437 | 50-100 | good - luxuriant | 30-35 °C | 24-48 hrs |
| Clostridium sporogenes ATCC 19404 (00008*) | 50-100 | good - luxuriant | 30-35 °C | 24-48 hrs |
| $ Phocaeicola vulgatus ATCC 8482 | 50-100 | good - luxuriant | 30-35 °C | 24-48 hrs |
Key: (*) Corresponding WDCM numbers
$ - Formerly known as Bacteroides vulgatus
Storage and Shelf Life
On receipt, 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 sample must be decontaminated and disposed of in accordance with current laboratory techniques (9,10).
| Product Name | Reinforced Medium for Clostridia |
|---|---|
| SKU | LQ130D |
| Customized Product Available | No |
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