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MacConkey HiVeg™ Agar w/o CV, w/ 0.003% NR and 1.5% Agar

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SKU:
MV008
For selective isolation and differentiation of lactose fermenting and lactose non-fermenting enteric bacteria.

Intended use

Recommended for selective isolation and differentiation of lactose fermenting and lactose non fermenting enteric bacteria.

Composition**

Ingredients g/L
HiVeg® peptone 17.000
HiVeg® peptone No. 3 3.000
Lactose 10.000
Synthetic detergent 1.500
Sodium chloride 5.000
Neutral red 0.030
Agar 15.000
Final pH (at 25°C) 7.1±0.2

**Formula adjusted, standardized to suit performance parameters

Directions

Suspend 51.53 gms 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 (4). MacConkey HiVeg® Agar w/o CV w/ 0.003% NR and 1.5% Agar is prepared by completely replacing animal based peptone with vegetable peptones to avoid BSE/TSE risks associate with animal peptones.

HiVeg® peptone and HiVeg® peptone No. 3 serves as a source of carbon, nitrogen, long chain amino acids and other essential growth nutrients. The selective action of this medium is attributed to synthetic detergents, which is 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 colonies. 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.

Type of specimen

Food and dairy sample; Water samples

Specimen Collection and Handling:

For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (4,5).

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 specimens. Safety guidelines may be referred in individual safety data sheets

Limitations

  1. Although this medium is selective for gram negative organisms, biochemical identification and serological testing using pure cultures is recommended for complete identification.
  2. It is advised to incubate for recommended period and temperature to avoid misintepretation of results.
  3. It is advised to read the results immediately after icubation, as overgrowth of Proteus species may mask other colonies.

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 temmperature.

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
Orange red coloured clear to slightly opalescent gel forms in Petri plates

Reaction
Reaction of 5.15% w/v aqueous solution at 25°C. pH: 7.1±0.2

pH
6.90-7.30

Cultural Response

Cultural characteristics observed after an incubation at 35 - 37°C for 18 - 24 hous.

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 <=10% pale pink to red
## Proteus hauseri ATCC 13315 50-100 luxuriant >=50% colourless
Salmonella Paratyphi A ATCC 9150 50-100 luxuriant >=50% colourless
Shigella flexneri ATCC 12022 (00126*) 50-100 fair to good 30-40% colourless
Salmonella Paratyphi B ATCC 8759 50-100 luxuriant >=50% colourless
Salmonella Enteritidis ATCC 13076 (00030*) 50-100 luxuriant >=50% colourless
Salmonella Typhi ATCC 6539 50-100 luxuriant >=50% colourless
Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) >=104 inhibited 0%
Corynebacterium diphtheriae type gravis >=104 inhibited 0%
Staphylococcus epidermidis ATCC 12228 (00036*) >=104 inhibited 0%

Key: * Corresponding WDCM numbers.

# Formerly known as Enterobacter aerogenes

## 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 inorder 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 (3,7).

More Information
Product Name MacConkey HiVeg™ Agar w/o CV, w/ 0.003% NR and 1.5% Agar
SKU MV008
Product Type HiVeg™
Physical Form Powder
Origin Animal Free (Veg), Lactose
Packaging type HDPE
References 1. MacConkey, 1900, The Lancet, ii:20.
2.MacConkey, 1905, J. Hyg., 5:333.
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.Baird R.B., Eaton A.D., and Rice E.W., (Eds.), 2015, Standard Methods for the Examination of Water and Wastewater,23rd ed., APHA, Washington, D.C.
5.Wehr H. M. and Frank J. H., 2004, Standard Methods for the Microbiological Examination of Dairy Products, 17th Ed.,APHA Inc., Washington, D.C.
6.Isenberg, H.D. Clinical Microbiology Procedures Handbook. 2nd Edition.
7.Jorgensen,J.H., Pfaller , M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W. (2015)Manual of Clinical Microbiology, 11th Edition. Vol. 1.
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