Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Brilliant Green HiVeg™ Agar Base w/ Phosphates
For selective isolation and identification of Salmonellae from mixed flora while inhibiting Escherichia coli , Proteus and Pseudomonas species.
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Animal Free Culture Media (HiVeg™)#4fa25a
Intended Use
Recommended for selective isolation and identification of Salmonellae from mixed flora while inhibiting Escherichia coli, Proteus and Pseudomonas species.
Composition**
| Ingredients | g/L |
|---|---|
| HiVeg™ peptone | 10.000 |
| HiVeg™ extract | 5.000 |
| Yeast extract | 3.000 |
| Lactose | 10.000 |
| Saccharose (Sucrose) | 10.000 |
| Disodium hydrogen phosphate | 1.000 |
| Sodium dihydrogen phosphate | 0.600 |
| Phenol red | 0.090 |
| Brilliant green | 0.0047 |
| Agar | 12.000 |
| Final pH (at 25°C) | 6.9±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 25.84 grams in 500 ml purified/distilled water. Heat with occasional agitation and bring just to the boil to dissolve the medium completely. DO NOT AUTOCLAVE. For more selectivity and maximum recovery aseptically add the rehydrated contents of 1 vial of S Selective Supplement (FD068). Mix well before pouring into sterile Petri plates.
Principle And Interpretation
Salmonella species cause many types of infections, from mild self-limiting gastroenteritis to life threatening typhoid fever. The most common form of Salmonella disease is self-limiting gastroenteritis with fever lasting less than 2 days and diarrhoea lasting less than 7 days (1).
Brilliant Green Agar Base w/phosphates is formulated as per the recommendation of Rijks Institute Voorde Volksgezondheld (National Institute for Public Health), Utrecht (2,3). It is also recommended by the ISO Committee (4,5,6), because of its improved performance with respect to recovery of smaller numbers of Salmonella species, inhibition of Escherichia coli, Proteus species and Pseudomonas species (7). Brilliant Green HiVeg™ Agar Base w/ Phosphates is prepared by completely replacing animal based peptone with vegetable peptones to avoid BSE/TSE risks associate with animal peptones. The medium contains HiVeg™ peptone, HiVeg™ extract and yeast extract as sources of carbon, nitrogen, vitamins, amino acids and essential nutrients. The two sugars namely lactose and sucrose serve as energy sources. Fermentation of lactose and / or sucrose in the medium results in the formation of acidic pH which is detected by phenol red indicator. Phosphates (M971) buffer the medium. Brilliant green helps to inhibit the contaminating microflora. The medium can further supplemented with sulphacetamide (1g/l) and sodium mandelate (0.25g/1) to inhibit contaminating microorganisms when the sample is suspected to contain large number of competing organisms along with Salmonella species (8). Brilliant Green Agar w/Phosphates being highly selective is recommended to be used along with a less inhibitory medium to improve the chances of recovery. Often cultures are enriched in (Selenite Cystine HiVeg™ Broth (MV025) or Tetrathionate HiVeg™ Broth Base w/o Iodine & BG (MV032). These enriched cultures are then isolated simultaneously on Brilliant Green HiVeg™ Agar Base, Modified (MV016/MV971), SS HiVeg™ Agar (MV108), Bismuth Sulphite HiVegTM Agar (MV027) and MacConkey HiVeg™ Agar (MV081).
Type of specimen
Food and dairy samples; Water samples
Specimen Collection and Handling
For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (9,10,11). For water samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards (12). 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
- Though this medium is selective for Salmonella other species of Enterobacteriaceae may grow.
- Further confirmation has to be carried out on presumptive Salmonella isolates.
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 pink homogeneous free flowing powder
Gelling
Firm, comparable with 1.2% Agar gel.
Colour and Clarity of prepared medium
Greenish brown coloured clear to slightly opalescent gel forms in Petri plates.
Reaction
Reaction of 5.2% w/v aqueous solution at 25°C. pH : 6.9±0.2
pH
6.70-7.10
Cultural Response
Cultural characteristics observed after an incubation at 35 - 37°C for 18 - 24 hours.
| Organism | Inoculum (CFU) | Growth | Recovery | Colour of colony |
|---|---|---|---|---|
| Escherichia coli ATCC 25922 (00013*) |
>=104 | inhibited | 0% | |
| ^Proteus hauseri ATCC 13315 |
50-100 | none-poor | <=10% | red |
| Pseudomonas aeruginosa ATCC 10145 (00024*) |
50-100 | none-poor | <=10% | red |
| Salmonella Enteritidis ATCC 13076 (00030*) |
50-100 | luxuriant | >=50% | bright red |
| Salmonella Typhimurium ATCC 14028 (00031*) |
50-100 | luxuriant | >=50% | bright red |
Key: *Corresponding WDCM numbers ^- Formerly known as Proteus vulgaris.
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 (13,14)
| Product Name | Brilliant Green HiVeg™ Agar Base w/ Phosphates |
|---|---|
| SKU | MV971 |
| Product Type | HiVeg™ |
| Physical Form | Powder |
| Origin | Animal Free (Veg) |
| Packaging type | HDPE |
| References | 1. Murray P. R., Baron J. H., Pfaller M. A., Jorgensen J. H. and Yolken R. H., (Ed.), 2003, Manual of Clinical Microbiology,8th Ed., American Society for Microbiology, Washington, D.C. |
| Customized Product Available | No |
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