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Perfringens HiVeg™ Agar Base (T.S.C. / S.F.P. HiVeg™ Agar Base)
Intended Use
Perfringens Agar Base with the addition of selective supplement and enrichment, it is used for the presumptive identification and enumeration of Clostridium perfringens.
Composition**
| Ingredients | g/L |
|---|---|
| HiVeg™ hydrolysate No. 1 | 15.000 |
| HiVeg™ extract | 5.000 |
| Soya peptone | 5.000 |
| Yeast extract | 5.000 |
| Sodium metabisulphite | 1.000 |
| Ferric ammonium citrate | 1.000 |
| Agar | 15.000 |
| Final pH (at 25°C) | 7.6±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 23.5 grams in 475 ml purified/distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°) for 15 minutes. Cool to 45-50°C. Add 25 ml of Egg Yolk Emulsion (FD045) and rehydrated contents of 1 vial of S.F.P. Selective Supplement (FD013) / T.S.C. Selective Supplement (FD014). Alternatively if fluorogenic detection is desired add rehydrated contents of CMF Selective Supplement (FD243) instead of FD013/FD014. Mix well before pouring into sterile Petri plates.
Principle And Interpretation
Tryptose Sulphite Cycloserine Agar (TSC) was originally formulated by Harmon et al (1) for the enumeration of C. perfringens from food. TSC Agar has been documented as one of the most useful media for the quantitative recovery of C. perfringens while suppressing growth of other facultative anaerobes (2). TSC Agar Base (with FD014) or SFP Agar Base (with FD013) is comparable in performance for isolation of C. perfringens (3,4) Perfringens Agar Base is also recommended by APHA (5). Perfringens Agar Base can be made selective either by addition of D-cycloserine (FD014) (1, 2) or Kanamycin and Polymyxin B (FD013) (6). Perfringens HiVeg™ Agar Base is prepared by using vegetable peptones in place of animal based peptones which make the media free of BSE/TSE risks.
HiVeg™ hydrolysate No. 1, Soya peptone, yeast extract, HiVeg™ extract provide nitrogenous compounds, carbon, sulphur, vitamin B complex and trace elements essential for clostridial growth. Sodium metabisulphite and ferric ammonium citrate act as an indicator of sulphite reduction, indicated by black coloured colonies. D-Cycloserine (FD014), Kanamycin and Polymyxin B (FD013) help in the selective isolation of C. perfringens by inhibiting accompanying flora. Egg yolk emulsion serves as a source of lecithin utilized by C. perfringens (MV837).
Type of specimen
Food samples
Specimen Collection and Handling
For food samples, follow appropriate techniques for sample collection and processing as per guidelines (5). 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
- Further biochemical and serological tests must be carried out for further identification.
- Some organism may show poor growth due to nutritional variation.
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
Light yellow to brownish yellow homogeneous free flowing powder
Gelling
Firm, comparable with 1.5% Agar gel
Colour and Clarity of prepared medium
Basal medium : Amber coloured clear to slightly opalescent gel. After Addition of Egg Yolk Emlusion (FD045) : Yellow coloured opaque gel forms in Petri plates
Reaction
Reaction of 4.7% w/v aqueous solution at 25°C. pH: 7.6±0.2
pH
7.40-7.80
Cultural Response
Cultural characteristics observed under anaerobic condition with added T.S.C. Selective Supplement (FD014)/S.F.P. Selective Supplement (FD013)/CMF Selective Supplement (FD243) and Egg Yolk Emulsion (FD045), after an incubation at 35-37°C for 18-24 hours.
| Organism | Inoculum (CFU) |
Growth | Recovery | Sulphite Reduction |
Lecithinase/ Haloes |
Fluorescence |
|---|---|---|---|---|---|---|
| Clostridium perfringens ATCC 12924 |
50-100 | luxuriant | >=50% | positive, blackening of medium |
Positive reaction, opaque zone around the colony |
Positive Reaction |
| Clostridium sordellii ATCC 9714 |
>=104 | inhibited | 0% |
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 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. 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 (4,5).
| Product Name | Perfringens HiVeg™ Agar Base (T.S.C. / S.F.P. HiVeg™ Agar Base) |
|---|---|
| SKU | MV837 |
| Product Type | HiVeg™ |
| Physical Form | Powder |
| Origin | Animal Free (Veg) |
| Packaging type | HDPE |
| References | 1. Harmon S. M., Kauttar D.A. and Peiler J. T., 1971, Appl. Microbiol., 22:688. |
| Customized Product Available | No |
















