HiMedia Laboratories GmbH, Marie-Curie-Str. 3, 64683, Einhausen, Germany.
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Intended Use
Recommended for detection and enumeration of Enterobacteriaceae in raw food samples.
Composition**
| Ingredients | Gms / Litre |
|---|---|
| HiCynth™ Peptone No.2* | 7.000 |
| HiCynth™ Peptone No.5* | 3.000 |
| Sodium chloride | 5.000 |
| Synthetic detergent | 1.500 |
| Dextrose (Glucose) | 10.000 |
| Neutral red | 0.030 |
| Crystal violet | 0.002 |
| Agar | 12.000 |
| Final pH (at 25°C) | 7.4±0.2 |
**Formula adjusted, standardized to suit performance parameters
*Chemically defined peptones
Directions
Suspend 38.53 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 and pour into sterile Petri plates.
Principle And Interpretation
Violet Red Bile Agar, a modification of MacConkey original formulation (1) is used for the enumeration of coli-aerogenes bacterial group.
Violet Red Bile Glucose Agar w/o Lactose, a modification of VRBA (M049), was designed for the enumeration of Enterobacteriaceae (2). Violet Red Bile Glucose HiCynth™ Agar w/o Lactose is a modification of Violet Red Bile Glucose Agar wherein animal or vegetable peptones are replaced with chemically defined peptone to avoid BSE/TSE/GMO risks associated with animal peptones. It employs the selective inhibitory components crystals violet and bile salts and the indicator system glucose and neutral red. Sought bacteria will dissimilate glucose and produce purple zones around the colonies (3). ISO committee has also recommended this medium (4). Selectivity of VRBGA can be increased by incubation under anaerobic conditions and/ or at elevated temperature, i.e. equal to or above 42°C (5-7).
HiCynth™ Peptone No.2 and HiCynth™ Peptone No.5 serve as sources of carbon, nitrogen, vitamins and other essential growth nutrients. Glucose is the fermentable carbohydrate, utilization of which leads to the production of acids. Neutral red indicator detects the acidity so formed. Crystal violet and bile salts mixture help to inhibit the accompanying gram-positive and unrelated flora. Sodium chloride maintains the osmotic equilibrium. Further biochemical tests are necessary for positive identification (8).
Type of specimen
Food and dairy samples; Water samples
Specimen Collection and Handling
For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (9-11). For water samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards (12).
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 shall be followed while handling specimens. Safety guidelines may be referred in individual safety data sheets.
Limitations
- Over incubation may result in reverting of reaction.
- Further biochemical tests must be carried out for confirmation.
- Individual organisms differ in their growth requirement and may show variable growth patterns on the medium.
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 pinkish beige homogeneous free flowing powder
Gelling
Firm, comparable with 1.2% Agar gel.
Colour and Clarity of prepared medium
Reddish purple coloured clear to slightly opalescent gel forms in Petri plates.
Reaction
Reaction of 3.85% w/v of aqueous solution at 25°C. pH: 7.4±0.2
pH
7.20-7.60
Cultural Response
Cultural characteristics was observed after an incubation at 35-37°C for 18-24 hours. Recovery rate is considered as 100%for bacteria growth on Tryptone Soya HiCynth™ Agar.
| Organism | Inoculum (CFU) | Growth | Recovery | Colour of colony |
|---|---|---|---|---|
| Escherichia coli ATCC 25922 (00013*) | 50-100 | good-luxuriant | >=50% | pink-red |
| Escherichia coli ATCC 8739 (00012*) | 50-100 | luxuriant | >=50% | pink-red with bile precipitate |
| Escherichia coli NCTC 9002 | 50-100 | luxuriant | >=50% | pink-red with bile precipitate |
| Pseudomonas aeruginosa ATCC 9027 (00026*) | 50-100 | good-luxuriant | >=50% | pink to red |
| Salmonella Enteritidis ATCC 13076 (00030*) | 50-100 | good-luxuriant | >=50% | light pink |
| #Klebsiella aerogenes ATCC 13048 (00175*) | 50-100 | good-luxuriant | >=50% | pink-red |
| Staphylococcus aureus subsp.aureus ATCC 25923 (00034*) | >=104 | inhibited | 0% | |
| Staphylococcus aureus subsp.aureus ATCC 6538 (00032*) | >=104 | inhibited | 0% |
Key: (*) Corresponding WDCM numbers.
(#) - Formerly known as Enterobacter aerogenes
Storage and Shelf Life
Store between 10-30°C in a tightly closed container and the prepared medium at 20-30°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 (13,14).
| Product Name | Violet Red Bile Glucose HiCynth™ Agar w/o lactose |
|---|---|
| SKU | MCD581 |
| Product Type | HiCynth™ |
| Physical Form | Powder |
| Origin | Chemically defined (HiCynth™) |
| Packaging type | HDPE |
| References | 1.American Public Health Association, Standard Methods for the Examination of Dairy Products, 1978, 14th Ed., WashingtonD.C. 2.Baird R.B., Eaton A.D., and Rice E.W., (Eds.), 2015, Standard Methods for the Examination of Water and Wastewater, 23rded., APHA, Washington, D.C. 3.Corry J. E. L., Curtis G. D. W. and Baird R. M., (Ed.), 1995, Culture Media for Food Microbiology, Vol. 34, Progress inIndustrial Microbiology, Elsevier, Amsterdam. 4.International Organization for Standardization (ISO), 1993, Draft ISO/DIS 7402. 5.Isenberg, H.D. Clinical Microbiology Procedures Handbook. 2nd Edition. 6.Jorgensen,J.H., Pfaller , M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W. (2015) Manual ofClinical Microbiology, 11th Edition. Vol. 1. 7.MacConkey A., 1905, J. Hyg., 5, 333-379. 8.MacFaddin J. F., 1985, Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria, Vol. 1, Williamsand Wilkins, Baltimore9.Mossel D. A. A., Eclderink I., Koopmans M. and Van Rossem F., 1978, Lab. practice, 27 No. 12: 1049.10.Mossel D. A. A. and Vega C. L., 1973, Hlth. Lab. Sci., 11:303 13.Salfinger Y., and Tortorello M.L. Fifth (Ed.), 2015, Compendium of Methods for the Microbiological Examination of Foods,5th Ed., American Public Health Association, Washington, D.C. 14.Wehr H. M. and Frank J. H., 2004, Standard Methods for the Microbiological Examination of Dairy Products, 17th Ed.,APHA Inc., Washington, D.C. |
| Customized Product Available | No |
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