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Columbia Agar, Granulated
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$200.20
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Availability: In stock
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SKU:
GMH144
For detection of Clostridium sporogenes from pharmaceutical products in accordance with the microbial limit testing by harmonized methodology of USP/EP/BP/IP/JP.
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Granulated™ Media#4fa25a
Intended use
Recommended for detection of Clostridium sporogenes from pharmaceutical products in accordance with the microbial limit testing by harmonized methodology of USP/EP/BP/JP/IP (Medium 15).
Composition**
| Ingredients | g/L |
|---|---|
| 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 |
If necessary adjust the pH so that after sterilization it is 7.3±0.2
* pH can also be measured after sterilization at 25°C
**Formula adjusted, standardized to suit performance parameters
# Pancreatic digest of casein
## Meat peptic digest
### Heart pancreatic digest
Directions
Suspend 44.0 grams in 1000 ml purified/distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes or as per validated cycle. Cool to 45-50°C, if required add the rehydrated contents of 1 vial of Gen Selective Supplement (FD252). Mix well and pour into sterile Petri plates.
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 and HM hydrolysate and yeast extract provides carbonaceous and nitrogenous substances, long chain amino acids, vitamins of B complex group and other essential nutrients for the 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. Gen Selective Supplement (FD252) inhibits a number of contaminating gram-negative organisms and Staphylococcus species. 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 (2,3,4,5,6). 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 to individual safety data sheets.
Limitations
- Individual organisms differ in their growth requirement and may show variable growth patterns on the medium.
- 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.
- Further biochemical tests must be carried out 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 coloured granular medium
Gelling
Firm, comparable with 1.5% Agar gel
Colour and Clarity of prepared medium
Light amber coloured clear to slightly opalescent gel forms in Petri 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, and growth was observed under anaerobic conditions after an incubation at 30-35°C for 24-48 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).
Cultural Response
| Organism | Inoculum (CFU) |
Growth | Observed Lot value (CFU) |
Recovery | Incubation temperature |
Incubation period |
|---|---|---|---|---|---|---|
| Growth Promoting | ||||||
| Clostridium sporogenes ATCC 19404 (00008*) | 50-100 | luxuriant | 25-100 | >=50% | 30-35 °C | <=48 hrs |
| Clostridium sporogenes ATCC 11437 | 50-100 | luxuriant | 25-100 | >=50% | 30-35 °C | <=48 hrs |
| ** Phocaeicola vulgatus ATCC 8482 | 50-100 | luxuriant | 25-100 | >=50% | 30-35 °C | <=48 hrs |
| Additional Microbiological testing | ||||||
| Clostridium perfringens ATCC 13124 (00007*) | 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 |
Key: (*) Corresponding WDCM numbers **Formerly known as Bacteroides vulgatus
Storage and Shelf Life
Store between 10-30°C in a tightly closed container and the prepared medium at 20-30°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 (7,8).
| Product Name | Columbia Agar, Granulated |
|---|---|
| SKU | GMH144 |
| Product Type | Granulated |
| Physical Form | Granular |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1. Ellner P. P., Stoessel C. J., Drakeford E. and Vasi F., 1966, Am. J. Clin. Pathol., 45:502. 2.Fildes P., 1920, Br. J. Exp. Pathol., 1:129. 3.Fildes P., 1921, Br. J. Exp. Pathol., 2:16. 4.Chapin K. C. and Doern G. V., 1983, J. Clin. Microbiol., 17:1163. 5.Bailey R. K., Voss J. L. and Smith R. F., 1979, J. Clin. Microbiol., 9 ; 65-71 6.Salfinger Y., and Tortorello M.L. Fifth (Ed.), 2015, Compendium of Methods for the Microbiological Examination ofFoods, 5th Ed., American Public Health Association, Washington, D.C. 7.Baird R.B., Eaton A.D., and Rice E.W., (Eds.), 2015, Standard Methods for the Examination of Water andWastewater, 23rd ed., APHA, Washington, D.C. 8.Wehr H. M. and Frank J. H., 2004, Standard Methods for the Microbiological Examination of Dairy Products, 17th Ed.,APHA Inc., Washington, D.C. 10.Jorgensen,J.H., Pfaller , M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W. (2015) Manualof Clinical Microbiology, 11th Edition. Vol. 1. |
| Customized Product Available | No |
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