Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Salt Polymyxin Broth Base
Recommended as a selective enrichment medium for Vibrio species for its further detection and enumeration from food and animal feed products. The composition and performance criteria of this medium are as per the specifications laid down in ISO 8914:1990.
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Enrichment#CC293D
Intended use
Recommended for detection and enumeration of Vibrio species. The composition and performance criteria of this medium are as per the specifications laid down in ISO 8914:1990.
Composition
| Ingredients | g/L |
|---|---|
| Peptone # | 9.000 |
| Yeast extract | 2.700 |
| Sodium chloride | 18.000 |
| Final pH (at 25°C) | 7.4±0.2 |
**Formula adjusted, standardized to suit performance parameters
# Peptic digest of animal tissue
Directions
Suspend 14.85 grams in 500 ml purified / distilled water. Heat if necessary to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 45 - 50°C and aseptically add rehydrated contents of 1 vial of PolyB Selective Supplement (FD003). Mix well and dispense into sterile tubes or flasks as desired.
Principle And Interpretation
Vibrios are fairly easy to isolate from both clinical and environmental material, though some species may require growth factors and/or vitamins. Vibrio parahaemolyticus is the leading cause of bacterial diarrhoea associated with the consumption of contaminated food products. Salt Polymyxin Broth (M821) is formulated as per the recommendation of APHA (1). ISO Committee (2) has recommended a modification (M8211) of this medium for detection and enumeration of salt tolerant Vibrio parahaemolyticus
Peptone and yeast extract provide nitrogenous compounds, carbon, sulphur, trace elements, long chain amino acids and vitamin B complex, essential for the growth. Polymyxin B sulphate inhibits gram-positive organisms.
Weigh 50 grams of sample into a blender. Add 450 ml phosphate buffer saline dilution water and blend for 1 minute at 8000 rpm. This constitutes 1:10 dilution. Prepare 1:100, 1:1000, 1:10000 dilutions or higher, if necessary, in PCB. Inoculate 3 x 10 ml portion of the 1: 10 dilutions into 3 tubes containing 10 ml of enrichment broth i. e. Salt Polymyxin Broth Base-2x concentration. This represents the 1 gram portion. Similarly inoculate 3 x 1 ml of dilutions into 10 ml of single strength Salt Polymyxin Broth Base. Incubate tubes at 35 ± 2°C for 24 hours.
After incubation a loopful is subcultured on solid medium such as TCBS Agar (M189) for further studies. V. parahaemolyticus appears as round, green or bluish colonies, 2-3 mm in diameter while V. cholerae forms yellow coloured colonies.
Type of specimen
Food samples and animal feeding stuffs
Specimen Collection and Handling
For food samples, follow appropriate techniques for sample collection and processing as per guidelines (1,2).
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.
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 free flowing powder
Colour and Clarity of prepared medium: Light yellow coloured clear solution without any precipitate
Reaction: Reaction of 2.97% w/v aqueous solution 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 24-48 hours with added PolyB Selective Supplement (FD003).
| Organism | Inoculum (CFU) | Growth |
|---|---|---|
| Vibrio cholerae ATCC 14035 | 50-100 | luxuriant |
| Vibrio parahaemolyticus ATCC 17802 | 50-100 | luxuriant |
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 sample must be decontaminated and disposed of in accordance with current laboratory techniques (3,4).
| Product Name | Salt Polymyxin Broth Base |
|---|---|
| SKU | M821I |
| Product Type | Regular |
| Physical Form | Powder |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1. International Organization for Standardization (ISO), 1990, Draft ISO/DIS 8914 |
| Customized Product Available | No |
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