Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Azide Dextrose Broth, Granulated
A selective medium for detection of Streptococci in water, sewage, food and other materials suspected of sewage contamination.
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Intended use
Azide Dextrose Broth, Granulated is used for detection and enumeration of Streptococci in water, sewage, food and other materials suspected of sewage contamination.
Composition
| Ingredients | Gms / Litre |
|---|---|
| Peptone, special | 15.000 |
| HM peptone B # | 4.500 |
| Dextrose (Glucose) | 7.500 |
| Sodium chloride | 7.500 |
| Sodium azide | 0.200 |
| Final pH (at 25°C) | 7.2±0.2 |
**Formula adjusted, standardized to suit performance parameters
# Equivalent to Beef extract
Directions
Suspend 34.7 grams in 1000 ml distilled water for preparing single strength broth or use 69.4 grams in 1000 ml distilled water for double strength broth. Heat, if necessary, to ensure complete solution. Dispense in test tubes and sterilize by autoclaving at 118°C for 15 minutes.
Principle And Interpretation
Enterococci are more resistant to chlorine in water, hence are better indicators of sewage pollution than Escherichia coli .
Until 1984, members of the genus Enterococcus were classified as Group D Streptococci. Upon genomic DNA analysis, a seperate genus status was provided to them. (7). Azide Dextrose Broth is recommended by APHA for enumeration of faecal Streptococci by MPN technique. Azide Dextrose Broth was initially formulated by Rothe, Mullmann and Seligmann (2,3) for quantitative determination of Enterococci in water, sewage, foods and other materials suspected of contamination with sewage. When large volumes of water samples are to be examined, double strength medium is used. Turbidity in tubes indicates presence of Enterococci, however, it should be further confirmed by inoculation in Ethyl Violet Azide Broth (M426).
Azide Dextrose Broth is a highly nutritious medium due to the presence of nutrient rich peptone special, HM peptone B and dextrose. Sodium azide inhibits growth of gram-negative bacteria, allowing Enterococci to grow (1,4,5). Streptococci detected by the above media should be further identified using chemicals (6).
Type of specimen
Food samples; Water and sewage samples
Specimen Collection and Handling
For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (8,9,10).
For water samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards.(11)
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 guidleines should be followed while handling specimens. Saftey guidelines may be referred in
individual safety data sheets
Limitations
Further biochemical tests must be carried out for confirmation.
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 homogeneous granular media
Colour and Clarity of prepared medium: Amber coloured clear solution without any precipitate.
Reaction: Reaction of 3.47% w/v aqueous solution at 25°C. pH : 7.2±0.2
pH: 7.00-7.40
Cultural Response
M345: Cultural characteristics observed after an incubation at 35-37°C for 18-24 hours
| Organism | Inoculum (CFU) | Growth |
|---|---|---|
| Escherichia coli ATCC 25922 (00013*) | >=103 | inhibited |
| Enterococcus faecalis ATCC 50-100 29212 (00087*) | good-luxuriant |
Key: *Corresponding WDCM numbers.
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 inorder 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. Use before expiry date on the label. 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 (12,13).
| Product Name | Azide Dextrose Broth, Granulated |
|---|---|
| SKU | GM345 |
| Product Type | Granulated |
| Physical Form | Granular |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1. Eaton A.D.,Clesceri L.S., and Greenberg A.E.,(Eds), 1998, Standard Methods for the Examination of Water and Wastewater,20th Ed., APHA, Washington, D.C. 2.Mallmann and Seligmann , 1950, Am. J. Publ. Health, 40:286. 3.Rothe, 1948, Illinois State Health Department. 4.Edwards S.J., 1933, J. Comp. Path. Therap., 46:21 5.Hartman G., 1937, Milchw. Forsch, 18:166. 6.MacFaddin J.F.,1985, Media for Isolation-Cultivation-Identification-Maintenance of Medical bacteria, Vol.1. Williams&Wilkins, Baltimore, Md. 7.Schleider K.H., Kilpper Bolz R., 1984, Int.J.Sys.Bacteriol., 34:31 9.Salfinger Y., and Tortorello M.L. Fifth (Ed.), 2015, Compendium of Methods for the Microbiological Examination ofFoods, 5th Ed., American Public Health Association, Washington, D.C.10.Wehr H. M. and Frank J. H., 2004, Standard Methods for the Microbiological Examination of Dairy Products, 17th Ed.,APHA Inc., Washington, D.C. 11.Baird R.B., Eaton A.D., and Rice E.W., (Eds.), 2015, Standard Methods for the Examination of Water and Wastewater,23rd ed., APHA, Washington, D.C. 12.Isenberg, H.D. Clinical Microbiology Procedures Handb0ook. 2nd Edition. 13.Jorgensen,J.H., Pfaller , M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W. (2015)Manual of Clinical Microbiology, 11th Edition. Vol. 1. |
| Customized Product Available | No |
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