Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended Use
Recommended for promoting fast and free sporulation to distinguish between Bacillus cereus and Bacillus thuringiensis.
Composition**
| Ingredients | Gms/Litre |
|---|---|
| Peptone | 1.000 |
| Yeast extract | 0.500 |
| Phenol red | 0.025 |
| Agar | 18.000 |
| Final pH (at 25°C) | 6.8±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 19.53 grams in 900 ml purified/distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 45-50°C and aseptically add 100 ml sterile, Egg Yolk Emulsion (FD045) and sterile contents of 2 vials of reconstituted Polymyxin B Selective Supplement (FD003). Mix well and pour into sterile Petri plates.
Principle And Interpretation
Bacillus cereus, a gram-positive rod shaped and beta haemolytic bacteria, is widely distributed in nature and can be isolated from a variety of foods. B. cereus is classified as a large-celled species of Group I bacilli (species with a cell width greater than 0.9µm and whose spores do not appreciably swell the sporangium). KG Agar Base formulated by Kim and Goepfert (3) and recommended by APHA (4) is used to promote free spore formation of B.cereus, Bacillus thuringiensis within an incubation period of 20-24 hours. This feature allows a) direct confirmation of zone forming organisms as Group I bacilli by means of microscopic examination and b) immediate differentiation of B. cereus from B.thuringiensis by visualization of the endotoxin crystal in sporulated cells of the latter organism. Additionally Group 2 bacilli such as Bacillus polymyxa, which produce lecithinase, are unable to form lecithinase under the rather nutritionally poor conditions imposed by KG Agar Base. Peptone and yeast extract in the medium supports the growth of B.cereus, B.thuringiensis. Lecithinase activity is observed as an opaque zone surrounding the individual colony. B. cereus is resistant to Polymyxin B, which restricts gram-negative organisms. B.cereus and B.thuringiensis can be distinguished by means of microscopic examination of stained cells. B.thuringiensis shows endotoxin crystals in sporulated cells.
Type of specimen
Isolated Microorgansim from food samples
Specimen Collection and Handling
For food samples, follow appropriate techniques for sample collection and processing as per guidelines (4). 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 :
- Well isolated colonies must be used for differentiation on this media.
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
Light yellow to light pink homogeneous free flowing powder
Gelling
Firm, comparable with 1.8% Agar gel.
Colour and Clarity of prepared medium
Basal medium : Orange coloured clear to slightly opalescent gel After addition of Egg Yolk Emulsion : Light orange coloured opaque gel forms in Petri plates
Reaction
Reaction of 1.95% w/v aqueous solution at 25°C. pH: 6.8±0.2
pH
6.60-7.00
Cultural Response
Cultural characteristics observed with added sterile Egg Yolk Emulsion (FD045) and Polymyxin B Selective Supplement (FD003), after an incubation at 30-35°C for 24 hours.
| Organism | Inoculum (CFU) | Growth | Recovery | Lecithinase |
|---|---|---|---|---|
| Bacillus cereus ATCC 14579 | 50-100 | good-luxuriant | >=50% | positive, opaque zone around the colony |
| Bacillus thuringiensis ATCC 10792 | 50-100 | good | 40-50% | positive, opaque zone around the colony |
| Escherichia coli ATCC 25922 (00013*) | 50-100 | none-poor | <=10% | negative |
Key: (*) Corresponding WDCM numbers.
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 (1,2).
| Product Name | KG Agar Base |
|---|---|
| SKU | M658 |
| Product Type | Regular |
| Physical Form | Powder |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1.Kim H. V. and Goepfert J. M., 1971, Appl. Microbiol., 22:581. 2.Downes F. P. and Ito K., (Eds.), 2001, Compendium of Methods for the Microbiological Examination of Foods, 4th Ed.,APHA, Washington, D.C. |
| Customized Product Available | No |
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