Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended use
Recommended as a selective and differential agar medium for rapid and direct identification of Listeria species.
Composition**
| Ingredients | Gms / Litre |
|---|---|
| HiCynth™ peptone No.2 # | 23.000 |
| Sodium chloride | 5.000 |
| HiCynth™ peptone No.5 # | 6.000 |
| Lithium chloride | 5.000 |
| Rhamnose | 10.000 |
| Phenol red | 0.120 |
| Chromogenic mixture | 5.130 |
| Agar | 13.000 |
| Final pH (at 25°C) | 7.3±0.2 |
**Formula adjusted, standardized to suit performance parameters
Key: # - Chemically defined peptones
Directions
Suspend 33.62 grams in 500 ml purified / distilled water. 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. Add rehydrated contents of 1 vial of CA Selective Supplement (FD181) aseptically. Mix well and pour into sterile Petri plates.
Principle And Interpretation
HiCrome™ Listeria Agar Base, Modified is a modification of a medium first developed by Notermans et al. (1) and Mengaud et al.(2) for the detection of Listeria species from food stuffs. It allows growth of Listeria species and gives a presumptive identification of Listeria monocytogenes within 24-48 hours after pre- enrichment. This medium is based on the specific chromogenic detection of B-glucosidase activity and also rhamnose fermentation. Listeria species hydrolyse the purified chromogenic substrate in the medium giving blue coloured colonies.
HiCrome™ Listeria HiCynth™ Agar Base, Modified works on the same line wherein animal and vegetable peptones are replaced with chemically defined peptones to avoid BSE/TSE/GMO risks associated with animal peptones. Since B glucosidase activity is specific for Listeria species, other organisms cannot utilize the chromogenic substrate and therefore give white colonies. Differentiation between Listeria species is based on the property of rhamnose fermentation. The colonies of L.monocytogenes and L.innocua appear blue with a yellow halo (Rhamnose positive) while the colonies of L.ivanovii appear blue without a yellow halo (Rhamnose negative).
HiCynth™ peptone No.2 and HiCynth™ peptone No.5 provide nitrogenous, carbonaceous substances, long chain amino acids, vitamin B complex and other essential growth nutrients. Rhamnose is the fermentable carbohydrate with phenol red as an indicator. Sodium chloride maintains the osmotic equilibrium. The added lithium chloride and CA Selective Supplement (FD181) inhibit growth of most gram-positive bacteria, gram-negative bacteria, yeasts and moulds.
Type of specimen
Food samples.
Specimen Collection and Handling
For food samples, follow appropriate techniques for sample collection and processing as per guidelines (3).
After use, contaminated materials must be sterilized by autoclaving before discarding.
Warning and Precautions
For professional use only. For professional use only. 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
- Individual organisms differ in their growth requirement and may show variable growth patterns on the medium.
- Since it is an enzyme-substrate based reaction, the intensity of colour may vary with isolates.
- Further biochemical tests must be carried out for confirmation.
- Each lot of the medium has been tested for the organisms specified on the COA. It is recommended to users to validate the medium for any specific microorganism other than mentioned in the COA based on the user's unique requirement.
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 pink homogeneous free flowing powder
Gelling
Firm, comparable with 1.3% Agar gel.
Colour and Clarity of prepared medium
Red coloured, clear to slightly opalescent gel forms in Petri plates
Reaction
Reaction of 6.72% w/v aqueous solution at 25°C. pH: 7.3±0.2
pH
7.10-7.50
Cultural Response
Cultural characteristics observed w/added CA Selective Supplement (FD181), after an incubation at 35-37°C for 24-48 hours.
| Organism | Inoculum (CFU) | Growth | Recovery | Colour of colony | Rhamnose fermentation |
|---|---|---|---|---|---|
| Bacillus subtilis subsp. spizizenni ATCC 6633 (00003*) | >=104 | inhibited | 0% | ||
| Candida albicans ATCC 10231 (00054*) | >=104 | inhibited | 0% | ||
| Escherichia coli ATCC 25922 (00013*) | >=104 | inhibited | 0% | ||
| Listeria innocua ATCC 33090 (00017*) | 50-100 | luxuriant | >=50% | bluish green | positive reaction, (yellow background) |
| Listeria ivanovii ATCC 19119 (00018*) | 50-100 | luxuriant | >=50% | bluish green | negative reaction |
| Listeria monocytogenes ATCC 19118 | 50-100 | luxuriant | >=50% | bluish green | positive reaction, (yellow halo) |
| Pseudomonas aeruginosa ATCC 27853 (00025*) | >=104 | inhibited | 0% |
Key: (*) Corresponding WDCM numbers.
Storage and Shelf Life
Store between 15-25°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 sample must be decontaminated and disposed of in accordance with current laboratory techniques (4,5).
| Product Name | HiCrome™ Listeria HiCynth™ Agar Base, Modified |
|---|---|
| SKU | MCD1417 |
| Product Type | HiCynth™, HiCrome™ |
| Physical Form | Powder |
| Origin | Chemically defined (HiCynth™) |
| Packaging type | HDPE |
| References | 1.Notermans S.H. and Dufrenne J., (1991), Applied and Environmental Microbiology, 57(09): 2666-70.2.Mengaud J., Braun-Breton C. and Cossart P., (1991), Molecular Microbiology, 5(2): 367-372.P |
| Customized Product Available | No |
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