HiMedia Laboratories GmbH, Marie-Curie-Str. 3, 64683, Einhausen, Germany.
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Intended use
Recommended for the selective isolation of Pseudomonas species.
Composition**
| Ingredients | g/L |
|---|---|
| HiVeg® hydrolysate | 10.000 |
| HiVeg® peptone No.2 | 16.000 |
| Potassium sulphate | 10.000 |
| Magnesium chloride, anhydrous | 1.400 |
| Agar | 11.000 |
| Final pH (at 25°C) | 7.1±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 24.2 grams in 500 ml purified/ distilled water containing 5 ml glycerol. Heat to boiling 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 sterile rehydrated contents of either CetriNix Supplement (FD029) or CFC Supplement (FD036) as desired. Mix well and pour into sterile Petri plates.
Note : Do not keep the molten agar for longer than 4 hours.
Principle And Interpretation
Pseudomonas Agar Base is a modification of Kings A medium (1) which contains magnesium chloride and potassium sulphate to enhance pigment production. Goto and Enomoto (2) formulated CetriNix supplement for the selective isolation of Pseudomonas aeruginosa from clinical specimens. Lowbury and Collins (3) studied cetrimide as a selective agent. CetriNix supplement suppresses Klebsiella, Proteus and Providencia species. Pseudomonas HiVeg® Agar Base is prepared by completely replacing animal based peptones with vegetable peptones to avoid BSE/TSE risks associated with animal peptones.
HiVeg® hydrolysate and HiVeg® peptone No.2 supplies nitrogenous and carbonaceous compounds, long chain amino acids, and other essential growth nutrients.
C-F-C Supplement was formulated by Mead and Adams (4) making the medium specific for isolation of Pseudomonas from chilled foods and processing plants, environmental samples and water. This medium is recommended for enumeration of Pseudomonas species from meat and meat products. It can also be used for clinical samples.
Examine inoculated plates after 24 hours and 48 hours using both white and UV light. The presence of blue-green or brown pigmentation may be considered as presumptive evidence of Pseudomonas aeruginosa. Alteromonas species may form brown or pink colonies on the medium.
Type of specimen
Food samples; Water samples.
Specimen Collection and Handling:
For food samples, follow appropriate techniques for sample collection and processing as per guidelines (5).
For water samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards.(6)
After use, contaminated materials must be sterilized by autoclaving before discarding.
Warning and Precautions
Read the label before opening the container. The media should be handled by trained personnel only. 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
- Due to nutritional variation, some strains may show poor growth.
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
Gelling
Firm, comparable with 1.1% Agar gel.
Colour and Clarity of prepared medium
Yellow coloured clear to slightly opalescent gel forms in Petri plates
Reaction
Reaction of 4.84% w/v aqueous solution containing 1% v/v glycerol at 25°C. pH : 7.1±0.2
pH
6.90-7.30
Cultural Response
Cultural characteristics observed after an incubation for 40-48 hours. Recovery rate is considered as 100% for growth on Soyabean Casein Digest Agar.
| Organism | Inoculum (CFU) | Growth (at 34-38°C with FD029) | Recovery (at 34-38°C with FD029) | Growth (at 24-26°C with FD036) | Recovery (at 24-26°C with FD036) | Colour/ Fluorescence under uv |
|---|---|---|---|---|---|---|
| $Proteus hauseri ATCC 13315 | >=104 | inhibited | 0% | - | - | - |
| Pseudomonas aeruginosa ATCC 27853 (00025*) | 50-100 | good-luxuriant | >=50% | - | - | blue-green /positive |
| ^Pseudomonas paraeruginosa ATCC 9027 (00026*) | 50-100 | good-luxuriant | >=50% | - | - | blue-green /positive |
| Pseudomonas aeruginosa ATCC 10145(00024*) | 50-100 | good-luxuriant | >=50% | - | - | blue-green /positive |
| Pseudomonas cepacia ATCC 10661 | 50-100 | - | - | good-luxuriant | >=50% | - |
| Pseudomonas fluorescens ATCC 13525 (00115*) | 50-100 | - | - | good-luxuriant | >=50% | - |
| Pseudomonas fragi ATCC 4973 (00116*) | 50-100 | - | - | good-luxuriant | >=50% | - |
| Enterococcus faecalis ATCC 29212 (00087*) | >=104 | inhibited | 0% | - | - | - |
| Enterococcus faecalis ATCC 19433 (00009*) | >=104 | inhibited | 0% | - | - | - |
| Escherichia coli ATCC 25922 (00013*) | >=104 | inhibited | 0% | inhibited | 0% | - |
| Escherichia coli ATCC 8739 (00012*) | >=104 | inhibited | 0% | inhibited | 0% | - |
Cultural characteristics observed after an incubation for 40-48 hours. Recovery rate is considered by comparing with previously approved lot of the same medium.
Key: * - Corresponding WDCM numbers
($) Formerly known as Proteus vulgaris
(^) Formerly known as Pseudomonas aeruginosa
Storage and Shelf Life
Store between 10-30°C in a tightly closed container and the prepared medium at 2-8°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 (7,8).
| Product Name | Pseudomonas HiVeg™ Agar Base |
|---|---|
| SKU | MV085 |
| Customized Product Available | No |
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