Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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MPN Presumptive Test#CC293D
Intended Use
Recommended for enumeration of coliform bacteria in water and waste water.
Composition
| Ingredients | g/L |
|---|---|
| Lactose | 20.000 |
| Dipotassium hydrogen phosphate | 1.800 |
| Sodium formate | 0.500 |
| L-Cystine | 0.040 |
| L-Aspartic acid | 0.048 |
| L-Arginine | 0.040 |
| Thiamine | 0.002 |
| Nicotinic acid (Niacin) | 0.002 |
| Pantothenic acid | 0.002 |
| Magnesium sulphate | 0.200 |
| Ferric ammonium citrate | 0.020 |
| Calcium chloride anhydrous | 0.020 |
| Bromo cresol purple | 0.020 |
| Final pH (at 25°C) | 6.7±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 22.7 grams (double strength) in 1000 ml purified/distilled water containing 12.7 grams of sodium glutamate and 5 grams of ammonium chloride. Heat if necessary to dissolve the medium completely. Dispense in tubes containing inverted Durham's tubes. Sterilize by autoclaving at A 115°C for 10 minutes.
A Correspond to 10 lbs pressure
Principle And Interpretation
Mineral Modified Glutamate Medium is used for enumeration of coliforms in water and wastewater samples. Folpmers first described a chemically defined medium based on glutamic acid for enumerating coliforms in water (1). However in the early days following its discovery, it was seen that the medium containing glucose gave many false positive results in 48 hours (2). It was then modified by Gray who incorporated formate and lactose in the medium which gave improved performance (3). This medium can also be used for detecting Escherichia coli in chlorinated waters and is better than Lauryl Tryptose Lactose Broth for the detection of small numbers of Escherichia coli.
This medium contains a variety of nutrients including salts, amino acids and vitamins. Lactose is the fermentable carbohydrate and bromo cresol purple is the pH indicator. Because of the nutrients, this medium is superior for enumerating coliforms in water and wastewater as it satisfies most of the nutritional requirements of coliforms.
Type of specimen
Water samples
Specimen Collection and Handling
For water samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards.(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
- Presumptive positive tubes must be subcultured into Lauryl Tryptose Mannitol Broth (M1070) and Brilliant Green Bile Broth (M121) and incubated at 44°C to detect indole formation at this temperature before identifying Escherichia coli.
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 green homogeneous free flowing powder
Colour and Clarity of prepared medium
Purple coloured clear solution without any precipitate
Reaction
Reaction of the medium :(mixture of 2.27% w/v of medium M643A , 1.27% w/v Sodium glutamate & 0.5% w/v ammonium chloride) is at 25°C. pH : 6.7±0.2
pH
6.50-6.90
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 18-48 hours.
| Organism | Growth | Acid | Gas |
|---|---|---|---|
| #Klebsiella aerogenes ATCC 13048 (00175*) | Luxuriant | Positive reaction | Negative reaction |
| Enterococcus faecalis ATCC 29212 (00087*) | Inhibited | ||
| Escherichia coli ATCC 25922 (00013*) | Luxuriant | Positive reaction | Positive reaction |
| Salmonella Typhi ATCC 6539 | Luxuriant | Negative reaction | Negative reaction |
| Shigella flexneri ATCC 12022 (00126*) | Luxuriant | Negative reaction | Negative reaction |
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) | Inhibited |
Key: *Corresponding WDCM numbers.
# Formerly known as Enterobacter aerogenes
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 (5,6).
| Product Name | Mineral Modified Glutamate Medium Base (Double Strength) |
|---|---|
| SKU | M643A |
| Product Type | Regular |
| Physical Form | Powder |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1.Folpmers T., 1948, Ant. V. Leeuwenhoek, J. Microbiol. Serol., 14:5 2.,,Public Health Laboratory Service, WaterCommittee, 1958, J. Hyg. Camb., 56:37 3.,,Gray R.D., 1959, J. Hyg. Camb., 57:249. |
| Customized Product Available | No |
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