Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended Use
Recommended for the isolation and differentiation of Salmonella species from coliforms by chromogenic method.
Composition**
| Ingredients | g/L |
|---|---|
| HiVeg™ peptone | 6.000 |
| Yeast extract | 2.500 |
| Synthetic detergent No. I | 1.000 |
| Chromogenic mixture | 5.400 |
| Agar | 13.000 |
| Final pH (at 25°C) | 7.7±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 27.9 grams in 1000 ml purified/distilled water. Gently heat to boiling to dissolve the medium completely. DO NOT AUTOCLAVE. Cool to 45-50°C. Mix well and pour into sterile Petri plates.
Principle And Interpretation
Salmonella species have been isolated from humans and almost all animals throughout the world. They cause many types of infections from mild, self-limiting gastroenteritis to life threatening typhoid fever. Salmonella Typhi and Salmonella Paratyphi A & B cause gastroenteritis, bacteremia and enteric fever, Salmonella Choleraesuis causes gastroenteritis and enteric fever, especially in children. Salmonella Typhimurium is the most frequently isolated serotype of Salmonella. Salmonella is a cause of food poisoning (1).
HiCrome® Salmonella HiVeg® Agaris a modification of the original formulation of Rambach (2) and is used for the differentiation of Salmonella species from other enteric bacteria. Rambach formulation differentiates Salmonella based on propylene glycol utilization and presence of a chromogenic indicator. However, HiCrome® Salmonella Agar medium uses only a chromogenic mixture for identification and differentiation of Salmonella species.
HiCrome® Salmonella HiVeg® Agar is prepared by completely replacing animal peptones with vegetable peptone to avoid BSE/TSE risks.
HiVeg® peptone and yeast extract provides nitrogenous, carbonaceous compounds and other essential growth nutrients. Escherichia coli and Salmonella are easily distinguishable due to their colony characteristics. Salmonella forms light purple coloured colonies with a purple halo.
E.coli and other β-glucuronidase positive organism exhibits a characteristic blue colour, due to presence of the enzyme ß-glucuronidase. Other organisms form colourless colonies. The characteristic light purple and blue colour is due to the chromogenic mixture (3). Synthetic detergent No. I mixture inhibits gram-positive organisms.
Conventional method employees the H2S production property for Salmonella detection which is also exhibited by other non Salmonella species such as Citrobacter, Proteus, etc. Hence further biochemical confirmation is required for further identification. Salmonella species isolated from food or clinical samples exhibit light purple colour with halo due to the specific enzyme substrate reaction.
Type of specimen
Food camples; Water samples
Specimen Collection and Handling
For food samples, follow appropriate techniques for sample collection and processing as per guidelines (4).
For water samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards (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
- The medium is selective for Salmonella may not support the growth of other microorganisms.
- Most of the Salmonella strains show purple colonies except few which may show colorless colonies.
- Due to nutritional variations, some strains may show poor growth.
- Final confirmation of suspected colonies must be carried out by serological and biochemical tests.
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 1.3% Agar gel.
Colour and Clarity of prepared medium
Light amber coloured, slightly opalescent gel forms in Petri plates
Reaction
Reaction of 2.79% w/v aqueous solution at 25°C. pH: 7.7±0.2
pH
7.50-7.90
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 24-48 hours.
| Organism | Inoculum (CFU) | Growth | Recovery | Colour of Colony |
|---|---|---|---|---|
| Staphylococcus aureus subsp.aureus ATCC 25923 (00034*) | >=104 | inhibited | 0% | |
| ** Bacillus spizizenii ATCC 6633 (00003*) | >=104 | inhibited | 0% | |
| Escherichia coli ATCC 25922 (00013*) | 50-100 | luxuriant | >=50% | blue |
| ## Proteus hauseri ATCC 13315 | 50-100 | good | 40-50% | colourless |
| Salmonella Typhimurium ATCC 14028 (00031*) | 50-100 | luxuriant | >=50% | light purple |
| Salmonella Enteritidis ATCC 13076 (00030*) | 50-100 | luxuriant | >=50% | light purple |
| Salmonella Typhi ATCC 6539 | 50-100 | good-luxuriant | >=50% | light purple |
Key: *Corresponding WDCM numbers **Formerly known as Bacillus subtilis subsp. spizizenii ## Formerly known as Proteus vulgaris.
Storage and Shelf Life
Store dehydrated powder and prepared medium between 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 (6,7).
| Product Name | HiCrome™ Salmonella HiVeg™ Agar |
|---|---|
| SKU | MV1296 |
| Product Type | HiVeg™, HiCrome™ |
| Physical Form | Powder |
| Origin | Animal Free (Veg) |
| Packaging type | HDPE |
| References | 1. Baird R.B., Eaton A.D., and Rice E.W., (Eds.), 2015, Standard Methods for the Examination of Water and Wastewater,23rd ed., APHA, Washington, D.C. |
| Customized Product Available | No |
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