Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Agar Medium F (Crystal Violet, Neutral Red, Bile Agar with Glucose)
For selective cultivation of yeasts and moulds in accordance with EP/BP/IP.
Gms/Lit :50.12
8.15 Lit/500G
Gms/Lit :50.12
8.15 Lit/500G
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Intended Use
Recommended for detection and enumeration of Enterobacteriaceae in accordance with EP/BP/IP.
Composition**
| Ingredients | g/L |
|---|---|
| Gelatin peptone # | 7.000 |
| Yeast extract | 3.000 |
| Lactose monohydrate | 10.000 |
| Bile salts | 1.500 |
| Dextrose monohydrate (Glucose monohydrate) | 10.000 |
| Sodium chloride | 5.000 |
| 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
# Pancreatic digest of gelatin
(10.00 grams of Lactose monohydrate is equivalent to 9.50 grams of Lactose anhydrous)
(10.00 grams of Dextrose monohydrate is equivalent to 9.09 grams of Dextrose anhydrous)
Directions
Suspend 50.12 grams (the equivalent weight of dehydrated medium per litre) in 1000 ml purified /distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 45-50°C. Mix well and pour into sterile Petri plates.
Principle And Interpretation
It is selective medium recommended for detection of Enterobacteriaceae species as recommended by EP/BP (1,2). Mossel et al (3-5) added dextrose to the medium observing improved detection of coliforms. Incubation can be carried out at different temperatures and incubation time depending upon the group of Enterobacteriaceae to be recovered (6). Gelatin peptone and yeast extract provide nitrogenous compounds and other nutrients essential for bacterial metabolism. This media is selective due to presence of the inhibitors; bile salts and crystal violet. Crystal violet inhibits gram-positive organisms especially Staphylococci. Neutral red indicator helps to detect lactose monohydrate and dextrose monohydrate fermentation. Lactose and glucose fermenting strains grow as red or pink and may be surrounded by a zone of acid precipitated bile. 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 products, follow appropriate techniques for sample processing in case of viscous materials as mentioned under sterility (1,2). 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
- 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 performed for confirmation.
Performance and Evaluation
Performance of the medium is expected when used as per direction on the label within the expiry period when stored at the recommended temperature.
Quality Control
Appearance
Light yellow to pink 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 EP/BP and cultural characteristics are observed after an incubation at 35-37°C for 18-24 hours. Recovery rate is considered as 100% for bacteria growth on Soybean Casein Digest Agar.
Cultural response
| Organism | Inoculum (CFU) | Growth | Observed Lot value (CFU) | Recovery | Colour of colony |
|---|---|---|---|---|---|
| Escherichia coli ATCC 8739 (00012*) | 50-100 | good-luxuriant | 25-100 | >=50% | pink-red |
| ^Pseudomonas paraeruginosa ATCC 9027 (00026*) | 50-100 | good-luxuriant | 25-100 | >=50% | light pink |
| Pseudomonas aeruginosa ATCC 27853 (00025*) | 50-100 | good-luxuriant | 25-100 | >=50% | light pink |
| Staphylococcus aureus subsp. aureus ATCC 6538 (00032*) | >=103 | inhibited | 0 | 0% | |
| Escherichia coli ATCC 25922 (00013*) | 50-100 | luxuriant | 25-100 | >=50% | pink-red |
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) | >=103 | inhibited | 0 | 0% | |
| Salmonella Enteritidis ATCC 13076 (00030*) | 50-100 | luxuriant | 25-100 | >=50% | Light pink |
| #Klebsiella aerogenes ATCC 13048 (00175*) | 50-100 | luxuriant | 25-100 | >=50% | pink-red |
Key: *Corresponding WDCM numbers,
^ Formerly known as Pseudomonas aeruginosa
(#) Formerly known as Enterobacter aerogenes
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 (8,9).
| Product Name | Agar Medium F (Crystal Violet, Neutral Red, Bile Agar with Glucose) |
|---|---|
| SKU | MAP1684 |
| Customized Product Available | No |
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