Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended Use
Recommended for selective isolation and cultivation of Legionella.
Composition
| Ingredients | Gms / Litre |
|---|---|
| Yeast extract | 10.000 |
| Charcoal activated | 2.000 |
| ACES buffer | 10.000 |
| a-Ketoglutarate monopotassium salt | 1.000 |
| L-cysteine hydrochloride | 0.400 |
| Ferric pyrophosphate | 0.250 |
| Potassium hydroxide | 2.800 |
| Anisomycin | 0.08 |
| Colistin | 0.01 |
| Vancomycin | 0.0005 |
| Agar | 17.000 |
| Final pH (at 25°C) | 6.9±0.2 |
Formula adjusted, standardized to suit performance parameters
Directions
Either streak, inoculate or surface spread the test inoculum (50-100 CFU) aseptically on the plate. The plate should be directly inoculated with the clinical specimen or the transport medium that has been inoculated with the clinical specimen.
Principle And Interpretation
Legionella species are non-spore forming, narrow, gram-negative rods. Legionella causes pneumonia (Legionnaires disease) (1) or a milk, febrile disease (Pontiac fever). They do not oxidize or ferment carbohydrates in conventional media or grow on sheep blood agar. Amino acids are the major sources of energy for Legionella. The amino acid L-cystine holds an absolute requirement as it plays major role in growth metabolism of Legionella (5). This amino acid as well as ferric pyrophosphate helps for the growth of Legionella. Growth is much better and more rapid on Buffered Charcoal Yeast Extract Agar (3,8). Feeley et al (4) had originally formulated Charcoal Yeast Extract (CYE) Agar. This medium was a modification of the existing F-G Agar conatining starch and casein enzymic hydrolysate (3). Feely et al (3, 4) replaced these two with charcoal and yeast extract respectively, and reported better recovery of Legionella pneumophilla. Later Paseulle (9) reported that supplementation of the Charcoal Yeast Agar with ACES buffer improved the performance of the medium. Edelstein (2) further modified the medium by adding alpha-ketoglutarate. This addition helped in improving the sensitivity of the medium. BCYE Agar (Buffered Charcoal Yeast Extract Agar )is based on Edelsteins Modification.
The media contains charcoal, which acts as a detoxicant. Yeast extract acts as a rich source of vitamins, nitrogen as well as carbon. ACES Buffer maintains optimal pH for growth while L-cystine hydrochloride; ferric pyrophosphate and a-ketoglutarate stimulate growth of Legionella species. Antibiotic anisomycin, vancomycin and colistin in the medium suppress contaminating microorganisms. Anisomycin inhibits growth of fungi. Gram negative organisms are inhibited by colistin. Gram poistive organisms are inhibited by Vancomycin.
Type of Specimen
Clinical specimen: Oropharyngeal flora
Nonclinical specimen: Environmental samples, water sources such as cooling water etc.
Specimen Collection and Handling
For clinical samples follow appropriate techniques for handling specimens as per established guidelines (6,7). After use, contaminated materials must be sterilized by autoclaving before discarding.
Warning and Precautions
In Vitro diagnostic use. 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 in individual safety data sheets.
Limitations
- Few strains of Legionella would grwo well in CO2 conditions in spite of medium supplemented with L-cysteine.
- Suggestd to isolate Legionella on non selective and selctive BCYE plates in parallel.
- Biochemical tests are necessary for confirmation of Legionella.
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 BCYE Selective Agar in 90 mm disposable plates with smooth surface and absence of black particles/cracks/bubbles.
Colour
Grey-black coloured medium
Quantity of medium
25 ml of medium in 90 mm Petri plate
Reaction
6.70-7.10
Sterility Test
Passes release criteria
Cultural Response
Cultural characteristics observed in 90% humid atmosphere after an incubation at 35-37°C for 3-4 days.
| Organism | Inoculum (CFU) | Growth | Recovery | Colour of colony |
|---|---|---|---|---|
| Candida albicans ATCC 10231 (00054*) | 50-100 | inhibited | 0% | |
| Escherichia coli ATCC 25922 (00013*) | 50-100 | none-poor | <=10% | |
| Legionella dumoffii ATCC 33343 | 50-100 | luxuriant | >50% | light blue-grey |
| Legionella pneumophila ATCC 33153 | 50-100 | luxuriant | >50% | white grey to blue grey |
| Staphylococcus epidermidis ATCC 12228 (00036*) | 50-100 | none-poor | <10% |
Key: (*) Corresponding WDCM numbers.
Storage and Shelf Life
On receipt store between 2-8°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 (6,7).
| Product Name | BCYE Selective Agar Plate |
|---|---|
| SKU | MP5383 |
| Customized Product Available | No |
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