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Rappaport Vassiliadis Medium
For enrichment of Salmonellae, based on its ability to multiply selectively at high osmotic pressure, low pH and at 43°C, with modest nutritional requirements.
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Intended Use
Recommended for enrichment of Salmonellae, based on its ability to multiply selectively at high osmotic pressure, low pH and at 43°C, with modest nutritional requirements.
Composition
| Ingredients | Gms/Litre |
|---|---|
| Soya peptone | 4.500 |
| Sodium chloride | 7.200 |
| Potassium dihydrogen phosphate | 1.440 |
| Magnesium chloride | 36.000 |
| Malachite green | 0.036 |
| Final pH (at 25°C) | 5.2±0.2 |
Formula adjusted, standardized to suit performance parameters
Directions
Suspend 49.17 grams in 1000 ml purified / distilled water. Heat if necessary to dissolve the medium completely. Dispense as desired into tubes and sterilize by autoclaving at 10 lbs pressure (115°C) for 15 minutes.
Principle And Interpretation
Rappaport Vassiliadis Medium is designed according to the revised formulation by Van Schothorst et al (5) and is recommended for the selective enrichment of Salmonellae from food and environmental specimens. Present medium is a modification of the Rappaport Vassiliadis Enrichment Broth described by Van Schothorst and Renauld (6). Addition of magnesium chloride to the medium was reported by Peterz et al (3). Salmonella species can be isolated from human faeces without pre-enrichment by using this medium.
Salmonella generally survive at little high osmotic pressure, grow at slightly low pH and are resistant to malachite green compared to other bacteria.
The medium contains Soya peptone which provides essential growth nutrients. Magnesium chloride raises the osmotic pressure in the medium. Malachite green is inhibitory to organisms other than Salmonella. The low pH of the medium, combined with the presence of malachite green and magnesium chloride, helps to select for the highly resistant Salmonella species. Potassium phosphate buffers the medium to maintain the constant pH. Sodium chloride maintains the osmotic balance.
Type of specimen
Food samples; Environmental samples
Specimen Collection and Handling
For foods samples, follow appropriate techniques for sample collection and processing as per guidelines (4). 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
- Due to nutritional variations some strains may show poor growth.
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 light blue homogeneous free flowing powder
Colour and Clarity of prepared medium: Bluish green coloured, clear to slightly opalescent solution with slight precipitate
Reaction: Reaction of 4.92% w/v aqueous solution at 25°C. pH: 5.2±0.2
pH: 5.00-5.40
Cultural Response: Cultural characteristics observed after an incubation at 42-43°C for 18-24 hours. After incubation, subculture on selective agar media likeMacConkey Agar (M081) or XLD Agar (M031) and incubate at 35-37°Cfor 18-24 hours.
| Organism | Inoculum (CFU) | Growth at 42±1°C | Recovery | Colour of Colony on M081 |
|---|---|---|---|---|
| Escherichia coli ATCC 25922 (00013*) | 50-100 | none-poor | <=10% | pink-red |
| Salmonella Enteritidis ATCC 13076 (00030*) | 50-100 | good-luxuriant | >=50% | colourless |
| Salmonella Typhi ATCC 6539 | 50-100 | good-luxuriant | >=50% | colourless |
| Salmonella Typhimurium ATCC 14028 (00031*) | 50-100 | good-luxuriant | >=50% | colourless |
* - Corresponding WDCM Numbers
Storage and Shelf Life
Store between 10-30°C in a tightly closed container and the prepared medium at 15-25°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 (1,2).
| Product Name | Rappaport Vassiliadis Medium |
|---|---|
| SKU | M880 |
| Product Type | Regular |
| Physical Form | Powder |
| Origin | Animal Free (Veg) |
| Packaging type | HDPE |
| References | 1. Van Schothorst M., Renauld A. and VanBeek C., 1987, Food Microbiol., 4: |
| Customized Product Available | No |
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