Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended Use
Recommended for confirmation of Pseudomonas aeruginosa in swimming pool waters.
Composition**
| Ingredients | Gms / Litre |
|---|---|
| Part A | |
| SM powder $ | 100.000 |
| Part B | |
| Peptone | 5.000 |
| Sodium chloride | 5.000 |
| HM peptone B # | 1.500 |
| Yeast extract | 1.500 |
| Agar | 15.000 |
| Final pH (at 25°C) | 7.4±0.2 |
**Formula adjusted, standardized to suit performance parameters
#Equivalent to Beef extract
$ Equivalent to Instant non-fat milk
Directions
Part A: Suspend 100 grams in 500 ml purified / distilled water. Sterilize by autoclaving at 15 lbs pressure (121°C) for 5 minutes. Cool to 55°C.
Part B: Suspend 28 grams in 500 ml purified / distilled water and sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool rapidly to 55°C. Mix Part A and Part B together and pour into sterile Petri plates.
Principle And Interpretation
Milk Agar was modified by Brown and Scott (2) for the confirmation of Pseudomonas aeruginosa in swimming pool waters. Swimming pool water is generally chlorinated potable water but it can also be from thermal springs or salt water. Microorganisms of concern are typically those from the body of the bather's including the arifices. Pseudomonas aeruginosa is one of the major supporting indicator organisms in the swimming pool. This organism is mainly responsible for ear and eye infection and is very likely to get disseminated in the swimming pool water due to constant contact of ears and eyes with the water.
SM powder, peptone, yeast extract, HM peptone B provide all the necessary nutrients mainly nitrogenous for the multiplication of Pseudomonas aeruginosa. Pseudomonas aeruginosa forms yellowish green colonies on this medium. P. aeruginosa hydrolyzes casein and produces a yellowish to green diffusible pigment on Milk Agar. For isolation, filter 200 ml or less water of the swimming pool through sterile membrane filters.
Type of specimen
Water samples- swimming pool water sample
Specimen Collection and Handling
Place each membrane filter on M-PA Agar (M1121). Incubate the plates at 41.5±0.5°C for 72 hours. Typical P. aeruginosa colonies are 0.8-2.2 mm in diameter, flat in appearance with brownish to greenish centers. For confirmation, using Brown and Scott Agar, Modified, make a single streak from an isolated colony and incubate plates at 35-37°C for 24 hours. After incubation P. aeruginosa forms pigmented colonies. 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 :
- Some strains show less growth with due to variable nutritional requirements.
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
Part A: Cream to off white homogeneous free flowing powder Part B - Cream to yellow homogeneous free flowing powder
Gelling
Firm, comparable with 1.5% Agar gel
Colour and Clarity of prepared medium
Light amber coloured opalescent gel forms in Petri plates
Reaction
Reaction of 2.8% w/v aqueous solution of Part B at 25°C. pH: 7.4±0.2
pH
7.20-7.60
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 18-24 hours.
| Organism | Inoculum (CFU) | Growth | Pigment Production |
|---|---|---|---|
| Escherichia coli ATCC 25922 (00013*) | 50-100 | good-luxuriant | no pigment |
| Pseudomonas aeruginosa ATCC 27853 (00025*) | 50-100 | good-luxuriant | yellowish green |
Key: (*) Corresponding WDCM numbers.
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 (3,4).
| Product Name | Brown and Scott Modified (Twin Pack) |
|---|---|
| SKU | M782 |
| Product Type | Regular |
| Physical Form | Powder |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1. Brown M.R.W. and Scott F. J.H., 1970, J. Clin. Pathol., 23:172. |
| Customized Product Available | No |
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