Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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MacConkey HiVeg™ Agar w/ CV, vNaCl, 0.005% NR and 1.5% Agar
To identify Enterobacteriaceae in the presence of coliforms and lactose non-fermenters from water, sewage, food products etc.
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Intended Use
Recommended to identify Enterobacteriaceae in the presence of coliforms and lactose non-fermenters from water, sewage, food products and other samples.
Composition
| Ingredients | g/L |
|---|---|
| HiVeg® peptone | 20.000 |
| Lactose | 10.000 |
| Sodium chloride | 5.000 |
| Synthetic detergent | 1.500 |
| Neutral red | 0.050 |
| Crystal violet | 0.001 |
| Agar | 15.000 |
| Final pH (at 25°C) | 7.2±0.2 |
Formula adjusted, standardized to suit performance parameters
Directions
Suspend 51.55 grams of medium in 1000 ml purified / distilled water. Heat to boiling with gentle swirling to dissolve the agar completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Avoid overheating. Cool to 45 - 50° C. Mix well and pour into sterile Petri plates. The surface of the medium should be dry when inoculated.
Principle And Interpretation
MacConkey Agar is the earliest selective and differential medium for cultivation of enteric microorganisms from a variety of clinical specimens (1,2). Subsequently MacConkey Agar and Broth have been recommended for use in microbiological examination of foodstuffs (3) and for direct plating / inoculation of water samples for coliform counts (4). These media are also accepted by the Standard Methods for the Examination of Milk and Dairy Products (5). Original medium contains protein, bile salts, sodium chloride and two dyes.
The selective action of this medium is attributed to crystal violet and bile salts, which are inhibitory to most species of gram-positive bacteria. Gram-negative bacteria usually grow well on the medium and are differentiated by their ability to ferment lactose. Lactose fermenting strains grow as red or pink and may be surrounded by a zone of acid precipitated bile. The red colour is due to production of acid from lactose, absorption of neutral red and a subsequent colour change of the dye when the pH of medium falls below 6.8. Lactose non-fermenting strains, such as Shigella and Salmonella are colourless and transparent and typically do not alter appearance of the medium. Yersinia enterocolitica may appear as small, non-lactose fermenting colonies after incubation at room temperature.
MacConkey HiVeg® Agar w/ CV, NaCl, 0.005% NR and 1.5% Agar is prepared by using vegetable peptones in place of animal based peptones which make the media free of BSE/TSE risks.
HiVeg® peptone serves as a source of carbon, nitrogen, long chain amino acids, vitamins and other essential growth nutrients. Lactose serves as the carbon source by being the fermentable carbohydrate. Synthetic detergent and crystal violet serves to make the medium selective by inhibiting accompanying gram-positive bacteria. Neutral red is the pH indicator dye while sodium chloride maintains the osmotic equilibrium of the medium.
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 (3,5,6).
For water samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards (4). 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
- Though the medium is recommended for selective isolation, further biochemical and serological testing must be carried out for complete identification.
- The surface of the medium should be dry when inoculated.
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 pink homogeneous free flowing powder
Gelling
Firm comparable with 1.5% Agar gel.
Colour and Clarity of prepared medium
Red with purplish tinge coloured clear to slightly opalescent gel forms in Petri plates.
Reaction
pH of 5.15% w/v aqueous solution at 25°C (after sterilization).
pH
7.00-7.40
Cultural Response
Cultural response was observed after an incubation at 35-37°C for 18-24 hours.
| Organism | Inoculum (CFU) | Growth | Recovery | Colour of colony |
|---|---|---|---|---|
| Escherichia coli ATCC 25922 (00013*) | 50-100 | luxuriant | >=50% | pink to red with bile precipitate |
| # Klebsiella aerogenes ATCC 13048 (00175*) | 50-100 | luxuriant | >=50% | pink to red |
| Enterococcus faecalis ATCC 29212 (00087*) | 50-100 | none-poor | 0-10% | colourless to pale pink |
| Salmonella Enteritidis ATCC 13076 (00030*) | 50-100 | luxuriant | >=50% | colourless |
| Salmonella Paratyphi A ATCC 9150 | 50-100 | luxuriant | >=50% | colourless |
| Salmonella Paratyphi B ATCC 8759 | 50-100 | luxuriant | >=50% | colourless |
| Salmonella Typhi ATCC 6539 | 50-100 | luxuriant | >=50% | colourless |
| ##Proteus hauseri ATCC 13315 | 50-100 | luxuriant | >=50% | colourless |
| Shigella flexneri ATCC 12022 (00126*) | 50-100 | fair to good | 30-40% | colourless |
| Staphylococcus aureus subsp. aureus ATCC 6538 (00032*) | >=104 | inhibited | 0% |
Key: (*) Corresponding WDCM numbers
(##)Formerly known as Proteus vulgaris (#) 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 (7,8).
| Product Name | MacConkey HiVeg™ Agar w/ CV, vNaCl, 0.005% NR and 1.5% Agar |
|---|---|
| SKU | MV081A |
| Product Type | HiVeg™ |
| Physical Form | Powder |
| Origin | Animal Free (Veg), Lactose |
| Packaging type | HDPE |
| References | 1.Murray P. R, Baron E, J., Jorgensen J. H., Pfaller M. A., Yolken R. H., (Eds.), 8th Ed., 2003, Manual of Clinical Microbiology,ASM, Washington, D.C. 2.Wehr H. M. and Frank J. H., 2004, Standard Methods for the Microbiological Examination of Dairy Products, 17th Ed.,APHA Inc., Washington, D.C. 3.Salfinger Y., and Tortorello M.L. Fifth (Ed.), 2015, Compendium of Methods for the Microbiological Examination ofFoods, 5th Ed., American Public Health Association, Washington, D.C. 4.FDA Bacteriological Analytical Manual, 2005, 18th Ed., AOAC, Washington, D.C. 5.Baird R.B., Eaton A.D., and Rice E.W., (Eds.), 2015, Standard Methods for the Examination of Water andWastewater, 23rd ed., APHA, Washington, D.C. 6.The United States Pharmacopoeia, 2018, The United States Pharmacopeial Convention, Rockville, M.D. 7.Williams, (Ed.), 2005, Official Methods of Analysis of the Association of Official Analytical Chemists, 19th Ed., AOAC,Washington, D.C8.Medrek T. F and Barnes Ella M., 1962, J. Appl. Bacteriol., 25(2),159-1689.Barnes Ella M. and Shrimpton D. H., 1957, J. Appl. Bacteriol., 20(2),273-285.10.Thornley Margaret J., 1957, J. Appl. Bacteriol., 20(2), 273-285. 11.Eddy B. P., 1960, J. Appl. Bacteriol., 23(2).216-249. 12.MacConkey A., 1905, J. Hyg., 5:333. 13.MacConkey A., 1900, The Lancet, ii:20. 14.British Pharmacopoeia, 2016, The Stationery office British Pharmacopoeia.1 5.Isenberg, H.D. Clinical Microbiology Procedures Handb0ook. 2nd Edition |
| Customized Product Available | No |
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