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 of Salmonellae other than Salmonella Typhi from faeces, food & dairy products.
Composition**
| Ingredients | g/L |
|---|---|
| Proteose peptone | 10.000 |
| Yeast extract | 3.000 |
| Lactose | 10.000 |
| Sucrose | 10.000 |
| Sodium chloride | 5.000 |
| Phenol red | 0.080 |
| Brilliant green | 0.0125 |
| Agar | 20.000 |
| Final pH (at 25°C) | 6.9±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Brilliant Green Agar is a ready to use solid media in glass bottle. The medium is pre-sterilized, hence it does not need sterilization. Medium in the bottle can be melted either by using a pre-heated water bath or any other method. Slightly loosen the cap before melting. When complete melting of medium is observed dispense the medium in tubes as butts / slants or in plates as desired and allow to solidify. If on plate, either streak, inoculate or surface spread the test inoculum (50-100 CFU) aseptically.
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. Brilliant Green Agar Base, Modified, as a primary plating medium for isolation of Salmonella species was first described by Kristensen et. al. (1) and further modified by Kauffmann (2). Brilliant Green Agar is also recommended by APHA (3,4) FDA (5) and described in EP, BP and IP (6,7,8).
This medium contains brilliant green, which inhibits growth of majority of Gram-negative and Gram-positive bacteria. Salmonella Typhi, Shigella species Escherichia coli, Pseudomonas species, Staphylococcus aureus are mostly inhibited. Clinical specimens can be directly plated on this medium. However, being highly selective, it is recommended that this medium should be used along with a less inhibitory medium to increase the chances of recovery. Often cultures enriched in Selenite or Tetrathionate Broth is plated on Brilliant Green Agar along with Bismuth Sulphite Agar, SS Agar, MacConkey Agar.
The medium contains proteose peptone 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. Sodium chloride maintains the osmotic equilibrium. Brilliant green helps to inhibit the contaminating microflora. The medium can further supplemented with sulphaacetamide (1g/l) and sodium mandelate (0.25g/l) to inhibit contaminating microorganisms when the sample is suspected to contain large number of competing organisms along with Salmonella species.
Non-lactose fermenting bacteria develop white to pinkish red colonies within 18 - 24 hours of incubation.
Type of specimen
Clinical: Faeces; Foodstuffs & dairy samples; Water samples; Pharmaceutical samples.
Specimen Collection and Handling
For clinical samples follow appropriate techniques for handling specimens as per established guidelines (9,10).
For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (3,11,12,13). For water samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards(5). After use, contaminated materials must be sterilized by autoclaving before discarding.
Warning and Precautions
In Vitro diagnostic use. For professional use only. 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
- Though this medium is selective for Salmonella other species of Enterobacteriaceae may grow.
- Salmonella Typhi and Shigella species may not grow on this medium.
- Moreover Proteus, Pseudomonas and Citrobacter species may mimic enteric pathogens by producing small red colonies.
- 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
Sterile glass bottle containing slightly opalscent Brilliant Green Agar,Modified .
Colour of medium
Greenish brown coloured medium
Quantity of medium
100 ml of medium in glass bottle.
pH
6.70-7.10
Sterility check
Passes release criteria
Cultural Response
Cultural charactistics observed after melting the medium and pouring into sterile petri plates. The plates are inoculted with following test organisms and incubation at 30-35°C for 24-48 hours. Recovery rate is considered as 100% for bacteria growth on Soyabean Casein Digest Agar.
| Organism | Inoculum (CFU) | Growth | Recovery | Colour of Colony |
|---|---|---|---|---|
| Escherichia coli ATCC 25922 (00013*) | 50-100 | none-poor | 0-10% | yellowish green |
| Escherichia coli ATCC 8739 (00012*) | 50-100 | none-poor | 0-10% | yellowish green |
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) | >=104 | inhibited | 0% | |
| Staphylococcus aureus subsp. aureus ATCC 6538 (00032*) | >=104 | inhibited | 0% | |
| Salmonella Typhi ATCC 6539 | 50-100 | fair-good | 30-40% | reddish pink |
| Salmonella Typhimurium ATCC 14028 (00031*) | 50-100 | good-luxuriant | >=50% | pinkish white |
| Salmonella Enteritidis ATCC 13076 (00030*) | 50-100 | luxuriant | >=50% | pinkish white |
| Salmonella Abony NCTC 6017 (00029*) | 50-100 | good-luxuriant | >=50% | pinkish white |
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 (9,10).
| Product Name | Brilliant Green Agar, Modified |
|---|---|
| SKU | SM016C |
| Customized Product Available | No |
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