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Violet Red Bile Glucose Agar
Intended use
Recommended for selective isolation, detection and enumeration of coli-aerogenes bacteria in water, milk other dairy food products and clinical samples.
Ingredients
| Ingredients | g/L |
|---|---|
| Peptone | 7.000 |
| Yeast extract | 3.000 |
| Sodium chloride | 5.000 |
| Bile salts mixture | 1.500 |
| Lactose | 10.000 |
| Neutral red | 0.030 |
| Crystal violet | 0.002 |
| Agar | 15.000 |
| Final pH (at 25°C) | 7.4±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Violet Red Bile 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 as desired and allow to solidify.
Principle And Interpretation
The coliform group consists of several genera of bacteria belonging to the family Enterobacteriaceae. The historical definition of this group has been based on the method used for detection i.e. lactose fermentation. This group is defined as all aerobic and facultative anaerobic, gram-negative, non-spore-forming rod shaped bacteria that ferment lactose with gas and acid formation within 48 hour at 35°C (1,2). Examination of foods, ingredients and raw materials, for the presence of marker groups such as coliforms is the one of the common tests.
Violet Red Bile Agar, a modification of MacConkey's original formulation (1) is used for the enumeration of coli- aerogenes bacterial group. It relies on the use of the selective inhibitory components crystals violet and bile salts and the indicator system lactose, and neutral red. Thus, the growth of many unwanted organisms is suppressed, while tentative identification of sought bacteria can be made. Organisms, which rapidly attack lactose, produce purple colonies surrounded by purple halos. Non-fermenters or late lactose-fermenters produce pale colonies with greenish zones (3). VRBA is recommended by APHA (4,5). Selectivity of VRBA can be increased by incubation under anaerobic conditions and/ or at elevated temperature, i.e. equal to or above 42°C (6-8). It is also recommended by ISO (9).
Peptone and yeast extract serve as sources of carbon, nitrogen, vitamins and other essential growth nutrients. Lactose is the fermentable carbohydrate, utilization of which leads to the production of acids. Neutral red indicator detects the acidity so formed. Crystal violet and bile salts mixture help to inhibit the accompanying gram-positive and unrelated flora. Sodium chloride maintains the osmotic equilibrium. Violet Red Bile Agar is not completely specific for enteric; other accompanying bacteria may give the same reaction. Further biochemical tests are necessary for positive identification (9).
Type of specimen
Clinical samples - Stool; 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 (2,4,5). For clinical samples follow appropriate techniques for handling specimens as per established guidelines (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
In Vitro diagnostic Use. For professional use only. Read the label before opening the bottle. 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
- Further biochemical tests must be carried out for 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: Sterile glass bottle containing slightly opalescent Violet Red Bile Agar.
Colour of medium: Reddish purple coloured medium
Quantity of medium: 500 ml of medium in glass bottle
pH: 7.20-7.60
Sterility Check: Passes release criteria
Cultural Response: Cultural characteristics after melting the medium and pouring into sterile Petri plates. The plates are inoculated with following test organisms and incubation at 35-37°C for 18-24 hours. Recovery rate is considered as 100% for bacteria growth on Soyabean Casein Digest Agar.
| Organism | Inoculum (CFU) | Growth | Recovery | Colour of colony |
|---|---|---|---|---|
| # Klebsiella aerogenes ATCC 13048 (00175*) | 50-100 | luxuriant | >=50% | pink to pinkish red |
| Escherichia coli ATCC 25922 (00013*) | 50-100 | luxuriant | >=50% | pinkish red with bile precipitate |
| Salmonella Enteritidis ATCC 13076 (00030*) | 50-100 | luxuriant | >=50% | Colourless to orangish yellow |
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) | >=103 | inhibited | 0% |
Key: (*) Corresponding WDCM numbers. # Formerly known as Enterobacter aerogenes
Storage and Shelf Life
Store between 15-30°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 (10,11).
| Product Name | Violet Red Bile Glucose Agar |
|---|---|
| SKU | SM049D |
| Customized Product Available | No |

