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MacConkey HiVeg™ Agar w/o CV and NaCl, w/ 0.0075% NR and 1.2% Agar

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MV082A
For cultivation and differentiation of enteric bacteria, restricting swarming of Proteus species from specimens such as urine which may contain large number of Proteus species as well as potentially pathogenic Gram-positive organisms.

Intended use

Recommended for cultivation and differentiation of enteric bacteria, restricting swarming of Proteus species from specimens which may contain large number of Proteus species as well as potentially pathogenic gram-positive organisms.

Composition

Ingredients g/L
HiVeg® peptone 23.000
Lactose 10.000
Synthetic detergent 2.000
Neutral red 0.075
Agar 12.000
Final pH (at 25°C) 7.4±0.2

Formula adjusted, standardized to suit performance parameters

Directions

Suspend 47.07 grams in 1000 ml purified/distilled water. Heat to boiling to dissolve the medium 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.

For the cultivation of Vibriospecies, add 5 gram per litre of Sodium chloride before sterilization.

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 was 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) and pharmaceutical preparations (6). The original medium contains peptone, bile salts, sodium chloride and two dyes. MacConkey HiVeg® Agar w/o CV and NaCl, w/ 0.0075% NR and 1.2% Agar is a modification of the original formulation with the exception of crystal violet and sodium chloride. MacConkey HiVeg® Agar w/o CV and NaCl, w/0.0075% NR and 1.2% Agar is same as MacConkey Agar w/o CV, NaCl w/0.5% Bile Salts except that the animal based peptones are completely replaced with vegetable peptones to avoid the BSE/TSE risks associated with animal peptones.

This medium prevents the swarming of Proteus species that are generally encountered in pathological specimens. Also potentially pathogenic gram-positive organisms can be isolated using this medium.

The selective action of this medium is attributed to synthetic detergent, 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 synthetic detergent. 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

Foodstuffs and dairy samples, Water samples, Pharmaceutical samples.

Specimen Collection and Handling

For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (3,5). 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

  1. Though the medium is recommended for selective isolation, further biochemical and serological testing must be carried out for further confirmation.
  2. 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 1.2% Agar gel.

Colour and Clarity of prepared medium: Orange red coloured clear to slightly opalescent gel forms in Petri plates

Reaction: Reaction of 4.7% w/v aqueous solution at 25°C. pH: 7.4±0.2

pH: 7.20-7.60

Cultural Response: Cultural characteristics 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 fair to good 30-40% 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*) 50-100 fair-good 30-40% pale pink-red

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 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).

More Information
Product Name MacConkey HiVeg™ Agar w/o CV and NaCl, w/ 0.0075% NR and 1.2% Agar
SKU MV082A
Product Type HiVeg™
Physical Form Powder
Origin Animal Free (Veg)
Packaging type HDPE
References 1. MacConkey, 1900, The Lancet, ii:20.
2.MacConkey, 1905, J. Hyg., 5:333.
3.Downes F. P and Ito K. (Ed.), 2001, Compendium of Methods for the Microbiological Examination of Foods, 4th ed., APHA,Washington, D.C.
4.Greenberg A. E., Clesceri L. S. and Eaton A. D., (Eds.), 2005, Standard Methods for the Examination of Water andWastewater, 21st 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.The United States Pharmacopoeia XXI and the National Formulary, 16th ed., 1985, United States PharmacopoeialConvention, Inc., Washington, D.C.
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