Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended use
Recommended for the selective and differential isolation of Listeria monocytogenes.
Composition
| Ingredients | Gms / Litre |
|---|---|
| HiCynth™ Peptone No.1* | 24.000 |
| HiCynth™ Peptone No.5* | 10.000 |
| Sodium pyruvate | 2.000 |
| Glucose (Dextrose) | 2.000 |
| Magnesium glycerophosphate | 1.000 |
| Magnesium sulphate | 0.500 |
| Sodium chloride | 5.000 |
| Lithium chloride | 10.000 |
| Disodium hydrogen phosphate | 2.500 |
| Chromogenic substrate | 0.050 |
| Agar | 15.000 |
| Final pH (at 25°C) | 7.2±0.2 |
**Formula adjusted, standardized to suit performance parameters
*Chemically defined peptones
Directions
Suspend 36.02 grams in 460 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. Aseptically add sterile contents of 1 vial of LP Enrichment Supplement 1 (FD214) and sterile rehydrated contents of OA Selective Supplement (FD212A). Mix well and pour into sterile Petri plates.
Principle And Interpretation
Listeria monocytogenes is a gram-positive foodborne human pathogen responsible for serious infections in pregnant women that may ultimately result in abortion, stillbirth, birth of a child with neonatal listeriosis and meningitis or primary bacteremia in adults and juveniles. The pathogenicity of Listeria ivanovii for humans is uncertain. Since L. monocytogenes and L. innocua have similar biochemical properties, they cannot be differentiated on traditional media (PALCAM, Oxford). L. mono Differential Agar Base is based on the formulation of Ottoviani and Agosti (1,2) for the selective and differential isolation of Listeria monocytogenes from food and animal feeds which is adopted by ISO Committee (3). HiCrome™ Listeria Ottaviani Agosti HiCynth™ Agar Base is prepared by completely replacing animal or vegetable based peptones with chemically defined peptones to avoid BSE/TSE risks associated with animal peptones.
HiCynth™ peptone No.1 and HiCynth™ peptone No.5 provides nitrogenous and carbonaceous substances, long chain amino acids, vitamins and essential growth nutrients. Sodium pyruvate support good growth. Glucose is the fermentable carbohydrate. Sodium chloride maintains osmotic equilibrium. Phosphate buffers the medium. Lithium chloride and added selective supplements (FD212A) inhibit accompanying microflora and allow the growth of Listeria species. Listeria species hydrolyze the chromogenic substrate which produces green coloured colonies. Differentiation of Listeria monocytogenes from other Listeria species is based on phosphatidylinositol-specific phospholipase C (PIPLC) activity. Phospholipase C enzyme hydrolyses the purified substrate (FD214) added to the medium resulting in an opaque halo around Listeria monocytogenes colonies.
Type of specimen
Food and animal feeds, environmental samples in the area of food manufacturing and handling
Specimen Collection and Handling
For food and animal feeds, environmental samples follow appropriate techniques for handling specimens as per established guidelines (3). 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
- Due to variable nutritional requirements, some strains may show poor growth on this medium.
- Further biochemical tests must be carried out to differentiate between L. monocytogenes and L.ivanovii.
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 1.5% Agar gel
Colour and Clarity of prepared medium
Light amber coloured opalescent gel forms in Petri plates
Reaction
Reaction of 7.2% w/v aqueous solution at 25°C. pH: 7.2±0.2
pH
7.00-7.40
Cultural Response
Cultural characteristics observed with added sterile LP Enrichment Supplement 1 (FD214) and OA Selective Supplement (FD212A).after an incubation at 35 - 37°C for 24 - 48 hours.
| Organism | Inoculum (CFU) | Growth | Recovery | Colour of Colony | PIPLC activity |
|---|---|---|---|---|---|
| Candida albicans ATCC 10231 (00054*) | ≥104 | inhibited | 0% | ||
| Enterococcus faecalis ATCC 29212 (00087*) | ≥104 | inhibited | 0% | ||
| Enterococcus faecalis ATCC 19433 (00009*) | ≥104 | inhibited | 0% | ||
| Escherichia coli ATCC 25922 (00013*) | ≥104 | inhibited | 0% | ||
| Escherichia coli ATCC 8739 (00012*) | ≥104 | inhibited | 0% | ||
| Pseudomonas aeruginosa ATCC 27853 (00025*) | ≥104 | inhibited | 0% | ||
| Listeria innocua ATCC 33090 (00017*) | 50-100 | luxuriant | ≥50% | greenish-blue | negative |
| Listeria grayi ATCC 19120 | 50-100 | luxuriant | ≥50% | greenish-blue | negative |
| Listeria ivanovii ATCC 19119 | 50-100 | luxuriant | ≥50% | greenish-blue | positive, opaque halo around the colony exhibiting phosphatidyl-inositol specific phospholipase activity |
| Listeria monocytogenes ATCC 35152 (00109*) | 50-100 | luxuriant | ≥50% | greenish-blue | positive, opaque halo around the colony exhibiting phosphatidylinositol specific phospholipase activity |
| Listeria monocytogenes ATCC 13932 (00021*) | 50-100 | luxuriant | ≥50% | greenish-blue | positive, opaque halo around the colony combining phosphatidylinositol specific phospholipase activity |
| Listeria monocytogenes ATCC 19112 | 50-100 | luxuriant | ≥50% | greenish-blue | positive opaque halo around the colony exhibiting phosphatidylinositol specific phospholipase activity |
| Listeria seeligeri ATCC 35967 | 50-100 | luxuriant | ≥50% | greenish-blue | negative |
| Listeria welshimeri ATCC 43549 | 50-100 | luxuriant | ≥50% | greenish-blue | negative |
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 Ottaviani Agosti HiCynth™ Agar Base |
|---|---|
| SKU | MCD1540A |
| Product Type | HiCynth™ |
| Physical Form | Powder |
| Origin | Chemically defined (HiCynth™) |
| Packaging type | HDPE |
| References | 1.Isenberg, H.D. Clinical Microbiology Procedures Handbook. 2nd Edition. 2.Jorgensen,J.H., Pfaller , M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W. (2015) Manual of Clinical Microbiology, 11th Edition. Vol. 1. |
| Customized Product Available | No |
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