HiMedia Laboratories GmbH, Marie-Curie-Str. 3, 64683, Einhausen, Germany.
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Intended use
Recommended for isolation and selective chromogenic differentiation of Vibrio species from seafood.
Composition
| Ingredients | Gms / Litre |
|---|---|
| HiCynth™ peptone No.3 | 10.000 |
| Sodium chloride | 25.000 |
| Sodium thiosulphate | 5.000 |
| Sodium citrate | 6.000 |
| Sodium cholate | 1.000 |
| Chromogenic mixture | 5.500 |
| Agar | 15.000 |
| Final pH (at 25°C) | 8.5±0.2 |
**Formula adjusted, standardized to suit performance parameters
#- Chemically Defined peptones
Directions
Suspend 67.5 grams in 1000 ml purified / distilled water. Heat to boiling to dissolve the medium completely. DO NOT AUTOCLAVE. Cool to 45-50°C. Mix well before pouring into sterile Petri plates.
Principle And Interpretation
Vibrio's have played a significant role in human history. Outbreaks of cholera, caused by Vibrio cholerae, can be traced back in time to early recorded descriptions of enteric infections. The Vibrio's have also received the attention of marine microbiologists who observed that the readily cultured bacterial population in near-shore waters and those associated with fish and shell fish were predominantly Vibrio species (1). Vibrio species are mainly responsible for causing cholera and food poisoning in humans.
Vibrio cholerae causes cholerae due to the intake of contaminated food such as raw oysters. Vibrio parahaemolyticus is a major cause of food borne infections, causing food poisoning (2). Since Vibrio species naturally occur in sea water, worth special mention is their need for sodium chloride, although some species can grow with minimum sodium chloride concentration (1). The widely used media for Vibrio isolation media are TCBS Agar and Alkaline Peptone Water (3). However accompanying sucrose-fermenting bacteria pose a problem in the identification of Vibrio species on TCBS Agar. On HiCrome™ Vibrio Agar, the colour development by Vibrio species in not affected by the presence of colonies of other bacteria. This is because, the amount of colour developed depends on the reaction of the bacterial beta-galactosidase with the substrate contained in the media (4). HiCrome™ Vibrio HiCynth™ Agar is prepared by completely replacing animal and vegetable based peptones with chemically defined peptones to avoid BSE/TSE risks associated with animal peptones. HiCynth™ peptone No.3 provides carbonaceous, nitrogenous and essential nutrients to the organisms. High concentration of sodium chloride in addition to maintaining the osmotic equilibrium also has an inhibitory action on the accompanying microflora. Sodium thiosulphate, sodium citrate and sodium cholate are used in the formulation because they can inhibit the growth of gram positive and some gram negative bacteria, but not members of Enterobacteriaceae. The proprietary chromogenic mixture incorporated in the medium helps in the chromogenic differentiation of Vibrio cholerae and Vibrio parahaemolyticus. The high (alkaline) pH of the medium helps in selective isolation of Vibrio species.
Type of specimen
Food samples.
Specimen Collection and Handling
For food samples, follow appropriate techniques for sample collection and processing as per guidelines (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
- Being highly selective, some species may show poor growth due to nutritional variations.
- 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.
- Slight colour variation may be observed depending upon strains.
- Further biochemical tests must be carried out for confirmation.
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 light tan homogeneous free flowing powder
Gelling
Firm,comparable with 1.5% Agar gel
Colour and Clarity of prepared medium
Light yellow coloured, clear to slightly opalescent gel forms in Petri plates
Reaction
Reaction of 6.75% w/v aqueous solution at 25°C. pH: 8.5±0.2
pH
8.30-8.70
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 18-24 hours.
| Organism | Inoculum (CFU) | Growth | Recovery | Colour of colony |
|---|---|---|---|---|
| Vibrio cholerae ATCC 15748 | 50-100 | good-luxuriant | >=50% | purple |
| Vibrio parahaemolyticus ATCC 17802 (00037*) | 50-100 | good-luxuriant | >=50% | bluish green |
| Enterococcus faecalis ATCC 29212 (00087*) | >=104 | inhibited | 0% | |
| Escherichia coli ATCC 25922 (00013*) | >=104 | inhibited | 0% | |
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) | >=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 (6,7).
| Product Name | HiCrome™ Vibrio HiCynth™ Agar |
|---|---|
| SKU | MCD1682 |
| Product Type | HiCynth™, HiCrome™ |
| Physical Form | Powder |
| Origin | Chemically defined (HiCynth™) |
| Packaging type | HDPE |
| References | 1.Alcamo. E.I, 2001. Fundamentals of Microbiology, 6th ed, Jones and Bartlett Publishers, Inc. pg 254, 244.2.Clesceri, Greenberg and Eaton (ed.). 199 |
| Customized Product Available | No |
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