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 |
|---|---|
| Peptone | 6.000 |
| Yeast extract | 2.500 |
| Bile salts mixture | 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
HiCrome™ Salmonella Agar is a ready to use solid media in glass bottle. The medium is pre-sterilized, hence it does not need sterilization. Medium in the bottle can be melted either by using a pre-heated water bath or any other method. Slightly loosen the cap before melting. When complete melting of medium is observed dispense the medium in tubes as butts /slants or in plates as desired and allow to solidify. If on plate, either streak, inoculate or surface spread the test inoculum (50-100 CFU) aseptically.
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 Agar is 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.
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 ß-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). Bile salts 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
Clinical samples: faeces, urine, etc.; Food Samples; Water samples
Specimen Collection and Handling
For clinical samples follow appropriate techniques for handling specimens as per established guidelines (4,5).
For food samples, follow appropriate techniques for sample collection and processing as per guidelines (6). For water samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards (7). After use, contaminated materials must be sterilized by autoclaving before discarding.
Warning and Precautions
In Vitro diagnostic Use. For professional use only. Read the label before opening the pack. Wear protective gloves/ protective clothing/eye protection/ face protection. Follow good microbiological lab practices while handling specimens and culture.
Limitations
- The medium is selective for Salmonella may not support the growth of other microorganisms.
- Most of the Salmonella strains show light purple colonies except few which may show colorless colonies.
- Individual organisms differ in their growth requirement and may show variable growth patterns on the medium
- 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.
- 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
Sterile glass bottle containing slightly opalescent HiCrome Salmonella Agar
Colour of medium
Light amber coloured medium
Quantity of medium
500ml of medium in glass bottle.
pH
7.50-7.90
Sterility Check
Passes release criteria
Cultural Response
Cultural charactistics observed after melting the medium and pouring into sterile petri plates. The plates are inoculated with following test organisms and incubation at 35-37°C for 24-48 hours.Recovery rate is considered as 100% for bacteria growth on Soyabean Casein Digest Agar.
| Organism | Inoculum (CFU) | Growth | Recovery | Colour of Colony |
|---|---|---|---|---|
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) | >=103 | inhibited | 0% | |
| ** Bacillus spizizenii ATCC 6633 (00003*) | >=103 | 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 w/ halo |
| Salmonella Enteritidis ATCC 13076 (00030*) | 50-100 | luxuriant | >=50% | light purple w/ halo |
| Salmonella Typhi ATCC 6539 | 50-100 | good-luxuriant | >=50% | light purple w/ halo |
Key- (*) Corresponding WDCM numbers
(**) Formerly known as Bacillus subtilis subsp. spizizenii
(##) Formerly known as Proteus vulgaris
Storage and Shelf Life
On receipt store between 2-8°C. Use before expiry date on the label. 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 clinical sample must be decontaminated and disposed of in accordance with current laboratory techniques (4,5).
| Product Name | HiCrome™ Salmonella Agar |
|---|---|
| SKU | SM1296D |
| Customized Product Available | No |
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