MacConkey Agar w/o CV, NaCl w/ 0.5% Bile Salts

Intended use

For the 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.

Composition

**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.

Note:For the cultivation of Vibrio species, 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 Agar w/o CV, NaCl W/ 0.5% Bile Salts is a modification of the original formulation with the exception of crystal violet and sodium chloride. 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 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.

Type of specimen

Clinical - faeces, urine and other pathological material, foodstuffs and dairy samples, water samples, pharmaceutical samples.

Specimen Collection and Handling

For clinical samples follow appropriate techniques for handling specimens as per established guidelines (7,8).

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

In Vitro diagnostic use. For professional use only. 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. 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.

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. Use before expiry date on the label. 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 clinical sample must be decontaminated and disposed of in accordance with current laboratory techniques (7,8).

Please refer disclaimer Overleaf.

Reference

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 of Foods, 5th Ed., American Public Health Association, Washington, D.C.
4. Lipps WC, Braun-Howland EB, Baxter TE, eds. Standard methods for the Examination of Water and Wastewater, 24th ed. Washington DC:APHA Press; 2023.
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-National Formulatory (USP-NF), 2022.
7. Isenberg, H.D. Clinical Microbiology Procedures Handbook. 2nd Edition
8. 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.