Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate, Thane(W)-400604, MH, India
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Legionella Agar Base, Granulated
Availability: Out of stock
Availability: Out of stock
Availability: Out of stock
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SKU:
GM809A
With addition of supplements it is used for cultivation of Legionella species.
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Intended Use
With addition of supplements it is used for cultivation of Legionella species.
Composition**
| Ingredients | g/L |
|---|---|
| Charcoal activated | 2.000 |
| Yeast extract | 10.000 |
| Agar | 13.000 |
| Final pH (at 25°C) | 6.9±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 12.5 grams in 440 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 contents of 1 vial of BCYE Growth Supplement (FD142). In case of non incorporation of GVPC Selective Supplement (FD143) or GVPA Selective Supplement, Modified (FD160), add aseptically 10 ml sterile distilled water to bring the total volume to 500 ml of medium. The final pH of the medium will be 6.9 ± 0.2. Mix well and pour into sterile petri plates. Stir the medium while dispensing to prevent the settling of charcoal particles. If desired, the medium can be made selective by aseptically adding rehydrated contents of 1 vial of either BMPA Selective Supplement (FD144) or GVPC Selective Supplement, (FD143) or GVPA Selective Supplement, Modified (FD160), along with 1 vial of BCYE Growth Supplement (FD142) to 440 ml sterile molten, cooled Legionella Agar Base. Simultaneously, a medium without L-Cysteine may be prepared by adding aseptically contents of 1 vial of Legi Growth Supplement w/o L-Cysteine (FD206).
Principle And Interpretation
Legionella Agar initially called as F-G agar was modified by Feely et al (1) by replacing Starch with charcoal and casein hydrolysate with yeast extract which resulted in better recovery of Legionella pneumophila (2). Pasculle et al (3) reported that the addition of ACES (N-2-acetamido-2-amino ethane sulphonic acid) buffer improved the nutritive value of medium. Edelstein (4) suggested addition of α-Ketoglutarate to increase the sensitivity of this medium. The medium contains yeast extract to provide the necessary nitrogenous nutrients for Legionella growth. Activated charcoal nullifies toxic compounds that either accumulate in the medium during growth or develop during sterilization of medium. Addition of ACES buffer helps in maintaining proper pH of the medium for the optimal growth of Legionella. Antibiotics in the supplement inhibits the growth of various contaminating bacteria and fungi (4,5). Legionella species have an absolute nutritional requirement for L-Cysteine. Presumptive Legionella species colonies can be subcultured onto both Legionella Agar Base with FD142 and with FD206 (Medium without L-Cysteine). All plates are incubated at 35°C. Colonies which grow on Legionella Agar Base with FD142, with L-Cysteine, but not on Legionella Agar Base with FD206 without L-Cysteine, can be regarded as presumptive Legionella species.
Type of specimen
Water samples
Specimen Collection and Handling
For water samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards (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
- Some strains may show poor growth due to variable nutritional requirements.
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
Grey to black coloured granular medium
Gelling
Firm, comparable with 1.3% Agar gel.
Colour and Clarity of prepared medium
Black coloured opaque gel forms in petri plates.
Reaction
Reaction of 2.5% w/v aqueous solution on addition of BCYE Growth Supplement (FD142) at 25°C. pH: 6.9±0.2
pH
6.70-7.10
Cultural Response
Cultural characteristics observed with added Sterile BCYE Growth Supplement (FD142) or Legi Growth Supplement w/o L-Cysteine (FD206) and GVPC Selective Supplement (FD143) or GVPA Selective Supplement, Modified (FD160), after an incubation at 35-37°C for 48-72 hours.
| Organism | Growth (with FD142 & FD143) | Growth (With FD206) |
|---|---|---|
| Escherichia coli ATCC 25922 (00013*) | inhibited | good |
| Legionella dumoffii ATCC 33343 | good-luxuriant | inhibited |
| Legionella pneumophila ATCC 33153 | good-luxuriant | inhibited |
Key: (*) corresponding WDCM numbers
Storage and Shelf Life
Store between 10-30°C in a tightly closed container and the prepared medium at 2-8°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 | Legionella Agar Base, Granulated |
|---|---|
| SKU | GM809A |
| Product Type | Granulated |
| Physical Form | Granular |
| Origin | Animal Free (Microbial) |
| Packaging type | HDPE |
| References | 1. Feely J. C., et al, 1978, J. Clin. Microbiol., 8(3):320. 2. Feely, Gibson, Gorman, et al, 1979, J. Clin. Microbiol., 10(4):437. 3. Pasculle, Feely, Gibson et al, 1980, J. Infect. Dis., 141:727. 4. Edelstein, 1981, J. Clin. Microbiol., 14:298. 5. Dennis et al, 1984, Proceeding of the 2nd International Symposium, Washington D.C. Am. Soc. Microbiol. PP 294-296 |
| Customized Product Available | No |
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