HiMedia Laboratories GmbH, Marie-Curie-Str. 3, 64683, Einhausen, Germany.
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Intended use
Recommended for selective isolation, cultivation and presumptive identification of Aeromonas hydrophila.
Composition**
| Ingredients | Gms / Litre |
|---|---|
| Yeast extract | 3.000 |
| Maltose | 3.500 |
| L-Cysteine hydrochloride | 0.300 |
| L-Lysine hydrochloride | 5.000 |
| L-Ornithine hydrochloride | 6.500 |
| Sodium thiosulphate | 6.800 |
| Ferric ammonium citrate | 0.800 |
| Synthetic detergent No. III | 1.000 |
| Sodium chloride | 5.000 |
| Bromothymol blue | 0.030 |
| Agar | 13.500 |
| Final pH (at 25°C) | 7.0±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 45.43 grams in 990 ml purified/distilled water. Heat to boiling to dissolve the medium completely. DO NOT AUTOCLAVE. Cool to 45-50°C and aseptically add rehydrated content of 1 vial of NO 5 Selective Supplement (FD096). Mix well and pour into sterile Petri plates.
Principle And Interpretation
RS Medium was formulated by Rimler and Shotts (1) based on the principle of Xylose-Lysine (XL) Agars (2,3). It is used for selective isolation and presumptive identification of Aeromonas hydrophila and other gram-negative bacteria based on their ability to decarboxylate lysine and ornithine, maltose fermentation and H2S production (4). Rimler-Shotts (RS) HiVeg™ Medium Base is prepared by completely replacing animal based peptones with vegetable peptones to avoid BSE/ TSE risks associated with animal peptones.
Yeast extract acts as a source of nutrients. Sodium thiosulphate, L-cysteine hydrochloride and ferric ammonium citrate are the indicators of H2S production. The medium contains maltose, which is mostly fermented by all Aeromonas. Maltose fermentation is indicated by bromothymol blue. Synthetic detergent No. III and novobiocin inhibit gram-positive bacteria and Vibrio species. Citrobacter freundii usually produce H2S but occasionally negative strains exist. The medium contains L-cysteine and L-ornithine, which are often decarboxylated by enteric bacteria to give alkaline products. Lysine positive and ornithine positive strains of Aeromonas may not have the typical strong yellow colour because of alkaline products produced during decarboxylation of the amino acids. Results are interpreted within 24 hours since after 26 hours slow reversion of yellow colour to a basic (green) colour occurs. Medium should be incubated at 35°C, which will eliminate possible growth of Aeromonas salmonicida, which may grow at reduced temperatures giving false-positive reaction. Test the yellow colonies with or without black centers (of H2S) for oxidase to rule out Citrobacter species. Proteus mirabilis is inhibited on this medium.
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 (5).
For water samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards (6).
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 clinical specimens.
Limitations :
- Pure isolate must be used.
- Results are interpreted within 24 hours since after 26 hours slow reversion of yellow colour to a basic (green) colour occurs.
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 green homogeneous free flowing powder
Gelling: Firm, comparable with 1.35% Agar gel.
Colour and Clarity of prepared medium: Dark green coloured clear to slightly opalescent gel forms in Petri plates
Reaction: Reaction of 4.54% w/v aqueous solution at 25°C. pH: 7.0±0.2
pH: 6.80-7.20
Cultural Response
Cultural characteristics observed with added NO 5 Selective Supplement(FD096) after an incubation at 35-37°C for 24 hours
| Organism | Inoculum (CFU) | Growth | Maltose fermentation | Lysine/ Ornithine decarboxylation | H2S |
|---|---|---|---|---|---|
| Aeromonas hydrophila ATCC 7966 (00063*) | 50-100 | good | positive reaction, yellow coloured colonies | negative reaction | negative reaction |
| Citrobacter freundii ATCC 8090 | 50-100 | good | negative reaction | variable reaction | positive, black centered colonies |
| Escherichia coli ATCC 25922 (00013*) | 50-100 | good | negative reaction | variable reaction | negative reaction |
| ## Proteus hauseri ATCC 13315 | 50-100 | good | positive reaction, yellow coloured colonies | negative reaction | positive, black centered colonies |
| Salmonella Typhi ATCC 6539 | 50-100 | good | positive reaction, yellow coloured colonies | negative reaction | negative reaction |
Key: (*) Corresponding WDCM numbers ## - Formerly known as Proteus vulgaris
Storage and Shelf Life
Store between 10-30°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 (7,8).
| Product Name | RS HiVeg™ Medium Base |
|---|---|
| SKU | MV576 |
| Product Type | HiVeg™ |
| Physical Form | Powder |
| Origin | Animal Free (Veg) |
| Packaging type | HDPE |
| References | 1.MacFaddin J. F., 1985, Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria, Vol. I, Williamsand Wilkins, Baltimore. 2.Shotts E. B. Jr. and Rimler R., 1973, Appl. Microbiol., 26(4):550. 3.Taylor W. I. and Harris B., 1965, Am. J. Clin. Pathol., 44:476. 4.Taylor W. I., 1965, Am. J. Clin. Pathol., 44:471. |
| Customized Product Available | No |
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