HiMedia Laboratories GmbH, Marie-Curie-Str. 3, 64683, Einhausen, Germany.
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Andrade Peptone Water w/ Meat Extract (Revised as Andrade Peptone Water w/HM Extract)
A basal medium for studying fermentation reactions, particularly of members of the Enterobacteriaceae.
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Intended Use
Recommended as a basal medium to which various carbohydrates can be added to study fermentation reactions, particularly of members of the Enterobacteriaceae.
Composition
| Ingredients | g/L |
|---|---|
| Peptone | 10.000 |
| HM extract | 3.000 |
| Sodium chloride | 5.000 |
| Andrade indicator | 0.100 |
| Final pH (at 25°C) | 7.1±0.2 |
**Formula adjusted, standardized to suit performance parameters
# Equivalent to Meat extract
Directions
Suspend 18.1 grams in 1000 ml purified/distilled water. Heat if necessary to dissolve the medium completely and dispense in test tubes containing inverted Durham's tubes. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to room temperature and aseptically add sterile stock solution of carbohydrate to a final concentration of 0.5% to 1.0% (w/v).
Principle And Interpretation
Bacteria differ widely in their ability to metabolize carbohydrates and related compounds. Carbohydrate fermentation reactions aids in the differentiation and identification of various bacteria. Andrade Peptone Water w/HM Extract is the most commonly used media for carbohydrate fermentation (1).
Peptone used is in the medium is free from fermentable carbohydrates (1,2) and the medium is also free from nitrates which may interfere with gas production. HM extract is an additional source of nutrients. Andrade indicator is a solution of acid fuchsin which when titrated with sodium hydroxide; changes colour from pink to yellow. The Andrade indicator changes colour from yellow to pink as the pH decreases (1). The medium is pink when hot but becomes straw coloured on cooling. Test carbohydrate solutions should be sterilized separately and aseptically added to sterile Andrade Peptone Water w/HM Extract. The biochemical identification of organisms capable of growing in this medium is made by various sugar fermentation results (1,3,4).
Use fresh cultures of organisms only which have been presumptively identified by Gram staining and colony morphology. For final identification further biochemical tests are required. Desired carbohydrate is added to the medium, which is inoculated with the test organism. If the test organism metabolizes the added carbohydrate, acids are produced, thereby lowering the pH of the medium. This causes a subsequent colour change of the indicator, from colourless to pink to red. If the added carbohydrate is not metabolized, the medium remains pale tan to straw coloured. Gas produced during fermentation is collected in the Durhams tube.
Type of specimen
Clinical samples - faeces, urine, sputum and wound exudates; Pure isolate
Specimen Collection and Handling:
For clinical samples follow appropriate techniques for handling specimens as per established guidelines (5,6). After use, contaminated materials must be sterilized by autoclaving before discarding.
Warning and Precautions
In Vitro diagnostic use only. 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 and culture. Standard precautions as per established guidelines should be followed while handling clinical specimens. Safety guidelines may be referred in individual safety data sheets.
Limitations
- Use fresh cultures of organisms only which have been presumptively identified by Gram staining and colony morphology.
- 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.
- For final identification further biochemical tests are required.
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: Cream to yellow coloured with pink tinge homogeneous free flowing powder
Colour and Clarity of prepared medium: Light pink to straw coloured clear solution without any precipitate
Reaction: Reaction of 1.81% w/v aqueous solution at 25°C. pH: 7.1±0.2
pH: 6.90-7.30
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 18-24 hours.
| Organism | Growth | Acid in absence of dextrose | Gas in absence of dextrose | Acid with added dextrose | Gas with added dextrose |
|---|---|---|---|---|---|
| Escherichia coli ATCC 25922 (00013*) | luxuriant | negative reaction | negative reaction | positive reaction, colour changes to pink-red | positive reaction |
| Klebsiella pneumoniae ATCC 13883 (00097*) | luxuriant | negative reaction | negative reaction | positive reaction, colour changes to pink-red | positive reaction |
| ## Proteus hauseri ATCC 13315 | luxuriant | negative reaction | negative reaction | positive reaction | positive reaction |
| Salmonella Typhi ATCC 6539 | luxuriant | negative reaction | negative reaction | positive reaction | negative reaction |
| Salmonella Typhimurium ATCC 14028 (00031*) | luxuriant | negative reaction | negative reaction | positive reaction | positive reaction |
| Shigella flexneri ATCC 12022 (00126*) | luxuriant | negative reaction | negative reaction | positive reaction | negative reaction |
| Shigella sonnei ATCC 25931 | luxuriant | negative reaction | negative reaction | positive reaction |
Key: (*) Corresponding WDCM numbers. ## Formerly known as Proteus vulgaris
Storage and Shelf Life
Store between 10-30°C in a tightly closed container and the prepared medium at 15-30°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 clinical sample must be decontaminated and disposed of in accordance with current laboratory techniques (5,6).
| Product Name | Andrade Peptone Water w/ Meat Extract (Revised as Andrade Peptone Water w/HM Extract) |
|---|---|
| SKU | M909 |
| Product Type | Regular |
| Physical Form | Powder |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1. MacFaddin J. F., 1985, Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria, Vol. I, Williamsand Wilkins, Baltimore. |
| Customized Product Available | No |
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