MacConkey Agar RS Plate

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SKU:
MP1702


Intended Use

Recommended for isolating and differentiating Gram negative enteric bacilli from specimens containing swarming strains of Proteus species.

Composition**

Ingredients g/L
Peptone 17.000
Proteose peptone 3.000
Lactose 10.000
Bile salts 5.000
Sodium chloride 5.000
Neutral red 0.030
Crystal violet 0.001
Agar 13.500
Final pH (at 25°C) 7.1±0.2

**Formula adjusted, standardized to suit performance parameters

Directions

Either streak, inoculate or surface spread the test inoculum (50-100 CFU) aseptically on the plate.

Principle And Interpretation

MacConkey agars are slightly selective and differential plating media mainly used for the detection and isolation of gram-negative organisms from clinical (1,2), dairy(3), food(4,5), water(6), and industrial sources (7). It is also recommended for the selection and recovery of the Enterobacteriaceae and related enteric gram-negative bacilli. These agar media is less selective since the concentration of bile salts, which inhibit gram-positive microorganisms, is low in comparison with other enteric plating media. Other than that this medium is also used for count of coli-aerogenes bacteria in cattle and sheep faeces (1), the count of coli-aerogenes and non-lactose fermenters in poultry carcasses (8), bacterial counts on irradiated canned minced chicken (9).

MacConkey Agar is the earliest selective and differential medium for cultivation of enteric microorganisms from a variety of clinical specimens (10,11). The 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 colonies 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, transparent and typically do not alter appearance of the medium.

Peptones are sources of nitrogen and other nutrients. Lactose is a fermentable carbohydrate, bile salts and crystal violet are selective agents that inhibit growth of gram-positive organisms. Neutral red is the pH indicator dye.

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 (1,2).

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(6).

For industrial samples, follow appropriate techniques for sample collection, processing as per guidelines (7).

After use, contaminated materials must be sterilized by autoclaving before discarding.

Warning and Precautions

In Vitro diagnostic 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

Limitations

  1. Individual organisms differ in their growth requirement and may show variable growth patterns on the medium
  2. Each lot of the medium has been tested for the organisms specified on the COA. It is recommended to users to validate the medium for any specific microorganism other than mentioned in the COA based on the user's unique requirement.
  3. It is recommended to store the plates at 24-30°C to avoid minimum condensation.
  4. Though the medium is recommended for selective isolation, further biochemical and serological testing must be carried out for further confirmation.
  5. 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
Sterile MacConkey Agar RS Plate in 90 mm disposable plates with smooth surface and absence of black particles/cracks/bubbles

Colour of medium
Orange red coloured medium

Quantity of medium
25ml of medium in 90mm disposable plate

pH
6.90-7.30

Cultural Response
Cultural response was observed after an incubation at 30-35°C for 18-72 hours. Recovery rate is considered as 100% for bacteria growth on Soybean Casein Digest Agar.

Organism Inoculum (CFU) Growth Recovery Colour of colony
Corynebacterium diphtheriae type gravis >=103 inhibited 0%
Shigella flexneri ATCC 12022 (00126*) 50-100 fair to good 30-40% colourless
Salmonella Paratyphi A ATCC 9150 50-100 luxuriant >=50% colourless
Salmonella Abony NCTC 6017 (00029*) 50-100 luxuriant >=50% colourless
## Proteus hauseri ATCC 13315 50-100 luxuriant >=50% colourless
Salmonella Typhi ATCC 6539 50-100 luxuriant >=50% colourless
Staphylococcus epidermidis ATCC 12228 (00036*) >=103 inhibited 0%
Escherichia coli ATCC 8739 (00012*) 50-100 luxuriant >=50% pink-red with bile precipitate
Staphylococcus aureus subsp.aureus ATCC 6538 (00032*) >=103 inhibited 0%
Salmonella Paratyphi B ATCC 8759 50-100 luxuriant >=50% colourless
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
Salmonella Typhimurium ATCC 14028 (00031*) 50-100 luxuriant >=50% colourless
Enterococcus faecalis ATCC 29212 (00087*) 50-100 none - poor <=10% colourless to pale pink
Salmonella Enteritidis ATCC 13076 (00030*) 50-100 luxuriant >=50% colourless
Staphylococcus aureus subsp.aureus ATCC 25923 (00034*) >=103 inhibited 0%

Key :-
* Corresponding WDCM numbers
# Formerly known as Enterobacter aerogenes
## Formerly known as Proteus vulgaris

Storage and Shelf Life

On receipt store between 20-30°C 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 (1,2).

More Information
Product Name MacConkey Agar RS Plate
SKU MP1702
Customized Product Available No
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