Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Ready Prepared Media#4fa25a
Intended Use
Recommended for the selection and subculture of Clostridium sporogenes in accordance with the harmonized method of USP/EP/BP/JP/IP.
Composition**
| Ingredients | Gms / Litre |
|---|---|
| Tryptone # | 10.000 |
| HM extract ## | 5.000 |
| HM hydrolysate ### | 3.000 |
| Yeast extract | 5.000 |
| Maize starch | 1.000 |
| Sodium chloride | 5.000 |
| Agar | 15.000 |
**Formula adjusted, standardized to suit performance parameters
# Pancreatic digest of casein
## Meat peptic digest
### Heart pancreatic digest
Directions
Either streak, inoculate or surface spread the test inoculum (50-100 CFU) aseptically on the plate.
Principle And Interpretation
Columbia Blood Agar Base used as a general-purpose nutritious medium was devised by Ellner et al from Columbia University, which was further enriched by the addition of sheep blood (1). It can also be used for the isolation of organisms by addition of various supplements. Columbia Agar is prepared in accordance with the microbial limit testing harmonized methodology of USP/EP/BP/JP/IP (2-6). This medium is recommended to check the presence of Clostridium in non-sterile products like food, dietary, nutritional supplements related products. The genus Clostridium belongs to the family Clostridiaceae in the class Clostridia. The product to be examined is initially enriched in Reinforced medium for clostridia. This medium contains 0.05% Agar and cysteine, which creates anaerobic conditions, thereby allowing anaerobic organisms to grow. The enriched sample is then subcultured on Columbia Agar. Columbia Agar is used as a base for media containing blood and for selective media formulations in which different combinations of antimicrobial agents are used as additives. This medium is highly nutritious as it contains tryptone, HM extract, HM hydrolysate and yeast extract which supports rapid and luxuriant growth of fastidious as well as non-fastidious organisms. Sodium chloride maintains osmotic balance of medium. Maize starch acts as an energy source and also neutralizes toxic metabolites if produced. It is used in detection of Clostridia from pharmaceutical products. Clostridia grows under anaerobic conditions as gram positive rods giving a catalase negative test. Further confirmation is carried out by identification tests.
Type of specimen
Pharmaceutical samples
Specimen Collection and Handling
For pharmaceutical samples, follow appropriate techniques for sample collection, processing as per 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 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
- Individual strain of a microorganism may have unique growth requirements with respect to nutrients and physical conditions. Based on which the growth pattern of each varies on a medium and some even may display significant delay in development.
- Each lot of the medium has been tested for the organisms specified on the COA. It is recommended to users to validate the medium for any specific microorganism other than mentioned in the COA based on the user's unique requirement.
- It is recommended to store the plates ta 24-30°C to avoid minimum condensation.
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 Columbia Agar in 90 mm disposable plates
Colour of medium: Light amber coloured medium
Quantity of medium: 25 ml of medium in 90 mm disposable plates
pH: 7.10-7.50
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 for 48 hours. Recovery rate is considered as 100% for bacteria growth on Casein Soybean Digest Agar (Soybean Casein Digest Agar).
Growth promoting properties
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 <=100 cfu under anaerobic conditions (at 30-35°C for 48 hours).
Sterility Test
Passes release criteria
Cultural Response
| Organism | Inoculum | Growth | Observed Lot Value | Recovery | Incubation Temp. | Incubation period |
|---|---|---|---|---|---|---|
| Clostridium sporogenes ATCC 19404 (00008*) | 50-100 | luxuriant | 25-100 | >=50% | 30-35 °C | <=48 hrs |
| Clostridium sporogenes ATCC 11437 | 50-100 | good-luxuriant | 25-100 | >=50% | 30 -35 °C | <=48 hrs |
| Bacteroides vulgatus ATCC 8482 | 50-100 | luxuriant | 25-100 | >=50% | 30-35 °C | <=48 hrs |
| Bacteroides fragilis ATCC 23745 | 50-100 | luxuriant | 25-100 | >=50% | 30-35 °C | <=48 hrs |
| Streptococcus pyogenes ATCC 19615 | >=104 | inhibited | 0 | 0% | 30-35 °C | >=72 hrs |
| Neisseria meningitidis ATCC13090 | >=104 | inhibited | 0 | 0% | 30-35 °C | >=72 hrs |
| Staphylococcus epidermidis ATCC 12228 (00036*) | >=104 | inhibited | 0 | 0% | 30-35 °C | >=72 hrs |
| Clostridium perfringens ATCC 13124 (00007*) | 50-100 | luxuriant | 25-100 | >=50% | 30-35 °C | <=48 hrs |
| Staphylococcus aureus subsp. aureus ATCC 6538 (00032*) | >=104 | inhibited | 0 | 0 | 30-35 °C | >=72 hrs |
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) | >=104 | inhibited | 0 | 0% | 30-35 °C | >=72 hrs |
Key: (*) Corresponding WDCM numbers
Storage and Shelf Life
On receipt store between 20-30°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 (7,8).
| Product Name | Columbia Agar Plate |
|---|---|
| SKU | MPH144 |
| Customized Product Available | No |
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