Andrade Peptone Water w/ HiVeg™ Extract No. 1

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
HiVeg™™ peptone	10.000
HiVeg™ extract No. 1	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

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). Andrade Peptone Water w/ HiVeg™™ Extract No. 1 is prepared by using vegetable peptones in place of animal based peptones which make the media free of BSE/TSE risks.

HiVeg™™ 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. HiVeg™™ extract No. 1 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 (2,3,4). 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

Pure isolate

Specimen Collection and Handling:

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. Use fresh cultures of organisms only which have been presumptively identified by Gram staining and colony morphology.
2. 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	Inoculum (CFU)	Growth	Acid in absence of dextrose	Gas in absence of dextrose	Acid with added dextrose	Gas with added dextrose
Escherichia coli ATCC 25922 (00013*)	50-100	luxuriant	negative reaction	negative reaction	positive reaction, colour changes to pink-red	positive reaction
Klebsiella pneumoniae ATCC 13883 (00097*)	50-100	luxuriant	negative reaction	negative reaction	positive reaction, colour changes to pink-red	positive reaction
## Proteus hauseri ATCC 13315	50-100	luxuriant	negative reaction	negative reaction	positive reaction	positive reaction
Salmonella Typhi ATCC 6539	50-100	luxuriant	negative reaction	negative reaction	positive reaction	negative reaction
Salmonella Typhimurium ATCC 14028 (00031*)	50-100	luxuriant	negative reaction	negative reaction	positive reaction	positive reaction
Shigella flexneri ATCC 12022 (00126*)	50-100	luxuriant	negative reaction	negative reaction	positive reaction	negative reaction
Shigella sonnei ATCC 25931	50-100	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 15-25°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	Andrade Peptone Water w/ HiVeg™ Extract No. 1
SKU	MV909
Product Type	HiVeg™
Physical Form	Powder
Origin	Animal Free (Veg)
Packaging type	HDPE
References	1. MacFaddin J. F., 1985, Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria, Vol. I, Williamsand Wilkins, Baltimore.2.Cowan S. T. and Steel K. J., 1974, Manual of Identification of Medical Bacteria, 2nd Ed., Cambridge United Press.3.Finegold S. M. and Baron E. J., 1986, Bailey and Scotts Diagnostic Microbiology, 7th Ed., The C.V. Mosby Co., St. Louis.4.Murray P. R., Baron E. J., Jorgensen J. H., Pfaller M. A., Yolken R.H., (Eds.), 8th Ed., 2003, Manual of Clinical Microbiology,ASM, Washington, D.C.
Customized Product Available	No

Andrade Peptone Water w/ HiVeg™ Extract No. 1