Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended Use
Recommended for identification and differentiation of Salmonella and non-Salmonella like Citrobacter from water and food samples.
Composition**
| Ingredients | Gms/Litre |
|---|---|
| HiVeg™ peptone | 10.000 |
| HiVeg™ extract | 2.000 |
| D-Cellobiose | 3.000 |
| Lactose | 10.000 |
| D-Mannitol | 1.200 |
| D-Trehalose | 1.330 |
| Chromogenic mixture | 6.600 |
| Agar | 15.000 |
| Final pH (at 25°C) | 7.6±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 49.13 grams in 1000 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. Mix well and pour into sterile Petri plates.
Principle And Interpretation
HiCrome™ MM Agar was formulated by Miller and Mallison (1) for specific isolation and detection of Salmonellae. This medium is superior to XLT4 Agar in supporting growth of Salmonella due to the presence of appropriate proportion of four sugars. Most differential and selective media are formulated with one or more sugars and pH indicators respectively. The utilization of sugars by organisms results in pH-changes. This is used as a means of distinguishing Salmonella from competing bacteria on the basis of colony colour.
Salmonella usually are unable to ferment these sugars (2) that supports growth of competing bacteria. Thus other bacteria tend to overgrow Salmonellae, masking their presence. The inclusion of sugars like mannitol, cellobiose and trehalose stimulate the better initial growth of Salmonella cells. However, the low concentrations of these sugars do not interfere with the utilization of protein and H2S production. Presence of lactose suppresses H2S production by non-salmonellae like Citrobacter freundii.
The chromogenic mixture, present in this medium helps to differentiate between lactose fermenters and nonfermenters. Lactose fermenters give bluish green coloured colonies, which would have been impossible to differentiate with an indicator based on pH change. Inclusion of tergitol 4 in the medium suppresses the presence of Proteus and Providencia colonies. HiVeg™ peptone and HiVeg™ extract provide the necessary nitrogen compounds, carbon, vitamins and also some trace ingredients necessary for the growth of bacteria.
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 (3). For water samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards. (4) 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 nutritional variations, some strains show poor growth.
- Though most of the Salmonella produce H2S certain non H2S producing Salmonella species may appear as colourless colonies.
- Certain Salmonella species which are lactose fermenters may show as bluish green coloured colonies.
- Further confirmation may be carried out on suspected colonies.
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, slightly opalescent gel forms in Petri plates
Reaction
Reaction of 4.9% w/v aqueous solution at 25°C. pH : 7.6±0.2
pH
7.40-7.80
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 18-24 hours
| Organism | Inoculum (CFU) | Growth | Recovery | Colour of Colony |
|---|---|---|---|---|
| Citrobacter freundii ATCC 8090 | 50-100 | good-luxuriant | >=50% | colourless may show bluish green colour on prolonged incubation |
| Enterococcus faecalis ATCC 29212 (00087*) | >=104 | inhibited | 0% | |
| Escherichia coli ATCC 25922 (00013*) | 50-100 | luxuriant | >=50% | light blue |
| Salmonella Typhimurium ATCC 14028 (00031*) | 50-100 | luxuriant | >=50% | black centered |
| Salmonella Enteritidis ATCC 13076 (00030*) | 50-100 | luxuriant | >=50% | black centered |
| Pseudomonas aeruginosa ATCC 27853 (00025*) | 50-100 | good-luxuriant | >=50% | colourless |
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 (5,6).
| Product Name | HiCrome™ MM HiVeg™ Agar |
|---|---|
| SKU | MV1393 |
| Product Type | HiVeg™, HiCrome™ |
| Physical Form | Powder |
| Origin | Animal Free (Veg) |
| Packaging type | HDPE |
| References | 1.Miller R.G. and Mallison E.T., 2000, J. Food Protection, 63(10), 1443-46.2.Miller R.G., Tate C.R., Mallinson E.T. and Scherrer J.A., 1991, Pault Sa 70:2429-32. |
| Customized Product Available | No |
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