Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate, Thane(W)-400604, MH, India
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Modified Buffered Charcoal HiVeg™ Agar Base
As low as
$171.60
Availability: Out of stock
Availability: In stock
Only %1 left
SKU:
MV892
Used for isolation and cultivation of Legionella species from clinical and other specimens.
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Intended Use:
Recommended for isolation and cultivation of Legionella species from various samples.
Composition
| Ingredients | Gms/Litre |
|---|---|
| HiVeg™ peptone No. 3 | 10.000 |
| Charcoal activated | 2.000 |
| ACES buffer | 10.000 |
| alpha-Ketoglutarate monopotassium salt | 1.000 |
| Agar | 17.000 |
| Final pH (at 25°C) | 6.9±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 20.0 grams in 500 ml purified/distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15lbs pressure (121°C) for 15 minutes. Cool to 45-50°C and aseptically add sterile reconstituted contents of one vial of Legi Growth Supplement w/o SS (Twin Pack) (FD041A). For selectivity of medium, add rehydrated contents of one vial of MWY Selective Supplement (MWY, FD040). Mix well and pour into sterile Petri plates with constant agitation to ensure that charcoal particles are evenly distributed. For additional selectivity, Legionella Selective Supplements (FD017, FD037 and FD038) may be added to molten medium as per choice.
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. Growth is much better and more rapid on Buffered Charcoal Yeast Extract Agar (2,3). 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 (4). This amino acid as well as ferric pyrophosphate helps for the growth of Legionella. Modified Buffered Charcoal Agar is similar to Buffered Charcoal Yeast extract Agar Base except that the yeast extract is replaced by proteose peptone. This medium is recommended for isolation and cultivation of Legionella species from clinical and environmental specimens. The medium was formulated by Feeley et al (5) and Edelstein (6) modified it further. Modified Buffered Charcoal HiVeg™ Agar Base is prepared by completely replacing animal based peptone with vegetable peptones to avoid BSE/TSE risks associate with animal peptones. The media contains charcoal, which acts as a detoxicant. HiVeg™ peptone No. 3 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. For selective isolation, antibiotic supplements can be used to suppress contaminating microorganisms. C3V Selective Supplement (FD037) containing cephalothin, colistin, vancomycin and cycloheximide (7) or MWY Selective Supplement (FD040) containing glycine, polymyxin B, anisomycin, vancomycin, bromothymol blue and bromocresol purple (8) are often used. Wear gown, mask and gloves while handling Legionella cultures. Work in a safety hood.
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 (9). 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 :
- 1. Further biochemical confirmation has to be carried out for further confirmation.
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 homogeneous free flowing powder
Gelling
Firm, comparable with 1.7% Agar gel.
Colour and Clarity of prepared medium
Grey-black coloured, opalescent gel forms in Petri plates
Reaction
Reaction of 4% w/v aqueous solution at 25°C. pH: 6.9±0.2
pH
6.70-7.10
Cultural Response
Cultural characteristics observed on addition of Legionella Supplement (FD041A and FD040) after an incubation at 35-37°C in humid atmosphere.
| Organism | Inoculum (CFU) | Growth | Recovery | Colour of colony |
|---|---|---|---|---|
| Escherichia coli ATCC 25922 (00013*) | 50-100 | none-poor | <=10% | |
| Legionella dumoffii ATCC 33343 Legionella | 50-100 | luxuriant | >=50% | light blue-grey |
| 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
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 (10,11).
| Product Name | Modified Buffered Charcoal HiVeg™ Agar Base |
|---|---|
| SKU | MV892 |
| Product Type | HiVeg™ |
| Physical Form | Powder |
| Origin | Animal Free (Veg) |
| Packaging type | HDPE |
| References | 1.Broome C. V., Fraser D. W., 1979, Epidemiol. Rev 1:1-16.2.George J. R. et al, 1980, J. Clin. Microbiol., 11:2863.Feeley J. C., Gorman G. W., Weaver R. E. et al, 1978, J. Clin. Microbiol., 8 : 320-325.4.Jones G. T., Hebert G. A., (Eds.), 1979, US Department of Health, Education and Welfare Publication No. (CDC) 79-8375,Atlanta, Centers for Disease Control.5.Feeley J. C., Gibson R. J., Gorman G. W. et al, 1979, J. Clin. Microbiol., 10:437. |
| Customized Product Available | No |
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