Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Violet Red Bile Glucose Agar
For detection and enumeration of Enterobacteriaceae from pharmaceutical products in accordance with the microbial limit testing by harmonized methodology of USP/EP/BP/JP/IP.
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Intended use
Recommended for detection and enumeration of Enterobacteriaceae from pharmaceutical products in accordance with the microbial limit testing by harmonized methodology of USP/EP/BP/JP/IP .
Composition
| Ingredients | g/L |
|---|---|
| Yeast extract | 3.000 |
| Gelatin peptone # | 7.000 |
| Bile salts | 1.500 |
| Sodium chloride | 5.000 |
| Glucose monohydrate | 10.000 |
| Agar | 15.000 |
| Neutral red | 0.030 |
| Crystal violet | 0.002 |
| pH after heating (at 25°C) | 7.4±0.2 |
**Formula adjusted, standardized to suit performance parameters
# Pancreatic digest of gelatin
Directions
Suspend 40.62 grams (the equivalent weight of dehydrated medium per litre) in 1000 ml purified /distilled water. Heat to boiling to dissolve the medium completely. DO NOT HEAT IN AN AUTOCLAVE. Cool to 45 - 50°C. Mix well and pour into sterile Petri plates.
Principle And Interpretation
Violet Red Bile Glucose Agar is a selective medium recommended for detection and enumeration of Enterobacteriaceae especially the bile tolerant gram negative bacteria in accordance with the test for specified organisms by harmonized methodology of USP/EP/BP/JP/IP (1-5) from non-sterile products and pharmaceutical preparations. Gelatin peptone and yeast extract provide nitrogenous, carbonaceous compounds, long chain amino acids, vitamins and other nutrients essential for bacterial metabolism. This media is selective due to presence of the inhibitors; bile salts and crystal violet for inhibiting gram positive organisms especially Staphylococci and gram negative organisms respectively. Neutral red is an indicator that helps to detect glucose fermentation. Glucose fermenting produce red colonies with pink-red halos in the presence of neutral red. Sodium chloride maintains the osmotic equilibrium in the medium. The red colour is due to absorption of neutral red and a subsequent colour change of the dye when the pH of medium falls below 6.8.
Type of specimen
Pharmaceutical samples.
Specimen Collection and Handling
For pharmaceutical samples, follow appropriate techniques for sample collection, processing as per guidelines (1-5). 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
- 1. Though the medium is for selective isolation of Enterobacteriaceae, further biochemical and serological testing must be carried out for further confirmation.
- 2. Over incubation may result in reverting of reaction.
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 pinkish beige 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.
pH: 7.20-7.60
Growth Promotion Test
Growth Promotion is carried out in accordance with the harmonized method of USP/EP/BP/JP. Cultural response was observed after an incubation at 30-35°C for 18-24 hours. Recovery rate is considered as 100% for bacteria growth on Soyabean Casein Digest Agar.
Growth promoting properties
Growth of microorganism comparable to that previously obtained with previously tested and approved lot of medium occurs at the specified temperature for not more than the shortest period of time specified inoculating <=100 cfu (at 30-35°C for <=18 hours).
Indicative properties
Colonies are comparable in appearance and indication reaction to those previously obtained with previously tested and approved lot of medium occurs for the specified temperature for a period of time within the range specified inoculating <= 100 cfu (at 30-35°C for 18-24 hours).
Cultural Response
| Organism | Inoculum (CFU) | Growth | Observed Lot value (CFU) | Recovery | Colour of colony | Incubation temperature |
|---|---|---|---|---|---|---|
| Escherichia coli ATCC 8739 (00012*) | 50-100 | luxuriant | 25-100 | >=50% | pink-red with bile precipitate | 18-24 hrs |
| ^Pseudomonas paraeruginosa ATCC 9027 (00026*) | 50-100 | luxuriant | 25-100 | >=50% | pink to red | 18-24 hrs |
Additional Microbiological Testing
| Organism | Inoculum (CFU) | Growth | Observed Lot value (CFU) | Recovery | Colour of colony | Incubation temperature |
|---|---|---|---|---|---|---|
| Escherichia coli ATCC 25922 (00013*) | 50-100 | good-luxuriant | 25-100 | >=50% | pink-red with bile precipitate | 18-24 hrs |
| Salmonella Enteritidis ATCC 13076 (00030*) | 50-100 | good-luxuriant | 25-100 | >=50% | light pink | 18-24 hrs |
| # Klebsiella aerogenes ATCC 13048 (00175*) | 50-100 | good-luxuriant | 25-100 | >=50% | pink-red | 18-24 hrs |
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) | >=103 | inhibited | 0 | 0% | >=24 hrs | |
| Staphylococcus aureus subsp. aureus ATCC 6538 (00032*) | >=103 | inhibited | 0 | 0% | >=24 hrs |
Key :-(*) Corresponding WDCM numbers
(#) Formerly known as Enterobacter aerogenes , ^ Formerly known as Pseudomonas aeruginosa
Storage and Shelf Life
Store between 10-30°C in a tightly closed container and the prepared medium at 20-30°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 (6,7).
| Product Name | Violet Red Bile Glucose Agar |
|---|---|
| SKU | MH581 |
| Customized Product Available | No |
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