Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Selective Isolation#CC293D
Intended Use:
Recommended for selective cultivation of Vibrio species.
Composition**
| Ingredients | g/L |
|---|---|
| Tryptone | 4.000 |
| Proteose peptone | 3.000 |
| Yeast extract | 5.000 |
| Sucrose | 20.000 |
| Sodium thiosulphate | 6.500 |
| Sodium citrate | 10.000 |
| Sodium deoxycholate | 1.000 |
| Sodium chloride | 10.000 |
| Bile # | 5.000 |
| Sodium lauryl sulphate | 0.200 |
| China blue | 0.200 |
| Cresol red | 0.020 |
| Agar | 15.000 |
Final pH (at 25°C) 8.5±0.2
**Formula adjusted, standardized to suit performance parameters
# Equivalent to Oxgall
Directions
Suspend 79.92 grams in 1000 ml purified/distilled water. Heat to boiling to dissolve the medium completely. DO NOT AUTOCLAVE. Cool to 45-50°C. Mix well and pour into sterile Petri plates.
Principle And Interpretation
Vibrio species, like many other gram-negative bacteria, grow in the presence of relatively high levels of bile salts. They are facultatively anaerobic and grow best in alkaline conditions. Isolation is facilitated by the use of media formulated with an alkaline pH due to the tolerance of this condition by Vibrio species. Media can be made selective for Vibrios by adding appropriate selective agents. The main agents employed are bile salts, teepol, tellurite and polymyxin B and E (Colistin) (1). Vibrio Agar is a selective medium for the isolation of Vibrio cholerae, Vibrio parahaemolyticus and other Vibrios (2).
Tryptone, proteose peptone, yeast extract provide nitrogenous, carbonaceous compounds, sulphur, vitamin B complex and other essential growth nutrients. Sodium citrate, sodium deoxycholate and bile inhibit gram-positive organisms and coliforms. Sucrose is the fermentable carbohydrate. Sucrose fermentative bacteria such as V.cholerae and V.alginolyticus form blue colonies due to the indicator china blue. V.parahaemolyticus forms slightly reddish and transluscent colonies. Sodium thiosulphate in combination with ferric citrate detects H2S production. Thiosulphate also acts as a sulphur source. Alkaline pH of this medium helps in recovery of V. cholerae. China blue and cresol red are the pH indicators.
Type of specimen
Clinical samples- faeces; Food samples.
Specimen Collection and Handling
For clinical samples follow appropriate techniques for handling specimens as per established guidelines (3,4).
For food samples, follow appropriate techniques for sample collection and processing as per guidelines (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
Limitations
- Individual organisms differ in their growth requirement and may show variable growth patterns on the medium.
- Each lot of the medium has been tested for the organisms specified on the COA. It is recommended to users to validate the medium for any specific microorganism other than mentioned in the COA based on the user's unique requirement.
- 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
Light yellow to greyish yellow homogeneous free flowing powder
Gelling
Firm, comparable with 1.5% Agar gel.
Colour and Clarity of prepared medium
Reddish purple coloured clear to slightly opalescent gel forms in Petri plates.
Reaction
Reaction of 7.99% w/v aqueous solution at 25°C. pH : 8.5±0.2
pH
8.30-8.70
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 18-24 hours.
| Organism | Inoculum (CFU) | Growth | Recovery | Colour of colony |
|---|---|---|---|---|
| Enterococcus faecalis ATCC 29212 (00087*) | 50-100 | none-poor | <=10% | yellow |
| Escherichia coli ATCC 25922 (00013*) | >=104 | inhibited | 0% | - |
| Pseudomonas aeruginosa ATCC 27853(00025) | 50-100 | none-poor | <=10% | blue |
| Salmonella Typhi ATCC 6539 | >=104 | inhibited | 0% | - |
| Shigella flexneri ATCC 12022 (00126*) | >=104 | inhibited | 0% | - |
| Vibrio cholerae ATCC 15748 | 50-100 | good-luxuriant | >=50% | blue |
| Vibrio parahaemolyticus ATCC 17802 (00037*) | 50-100 | good-luxuriant | >=50% | slightly reddish |
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 clinical sample must be decontaminated and disposed of in accordance with current laboratory techniques (3,4).
| Product Name | Vibrio Agar |
|---|---|
| SKU | M820 |
| Product Type | Regular |
| Physical Form | Powder |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1. Atlas R. M. 2004, 3rd Ed., Handbook of Microbiological Media, Parks, L.C., (Ed.), CRC Press, Boca Raton. |
| Customized Product Available | No |
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