Lactobacillus MRS Agar, Granulated (MRS Agar, Granulated)

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SKU:
GM641
For isolation and cultivation of Lactobacilli.


Intended use

Recommended for isolation and cultivation of all Lactobacilli from food, dairy and clinical samples.

Composition

Ingredients g / L
Proteose peptone 10.000
HM Peptone B # 10.000
Yeast extract 5.000
Dextrose (Glucose) 20.000
Tween 80 (Polysorbate 80) 1.000
Ammonium citrate 2.000
Sodium acetate 5.000
Magnesium sulphate 0.100
Manganese sulphate 0.050
Dipotassium hydrogen phosphate 2.000
Agar 12.000
Final pH (at 25°C) 6.5±0.2

**Formula adjusted, standardized to suit performance parameters
# Equivalent to Beef extract

Directions

Suspend 67.15 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. Cool to 45-50°C. Mix well and pour into sterile Petri plates.

Principle And Interpretation

Lactobacilli MRS media are based on the formulation of deMan, Rogosa and Sharpe (1) with slight modification. It supports luxuriant growth of all Lactobacilli from oral cavity (2), dairy products (3), foods (2), faeces (4,5) and other sources (6). Proteose peptone and HM peptone B supply nitrogenous and carbonaceous compounds. Yeast extract provides vitamin B complex and dextrose is the fermentable carbohydrate and energy source. Polysorbate 80 supplies fatty acids required for the metabolism of Lactobacilli. Sodium acetate and ammonium citrate inhibit Streptococci, moulds and many other microorganisms. Magnesium sulphate and manganese sulphate provide essential ions for multiplication of lactobacilli. Phosphates provide good buffering action in the media. Lactobacilli are microaerophilic and generally require layer plates for aerobic cultivation on solid media. When the medium is set, another layer of un-inoculated MRS Agar is poured over the surface to produce a layer plate. Lactobacilli isolated on MRS Agar should be further confirmed biochemically.

Type of specimen

Clinical samples - urine, faeces, etc.; Food and dairy samples

Specimen Collection and Handling

For clinical samples follow appropriate techniques for handling specimens as per established guidelines (4,5). For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (2,7,8). 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. Individual organisms differ in their growth requirement and may show variable growth patterns on the medium.
  2. Further Biochemical and serological testing is required for complete identification.

Performance and Evaluation

Performance of the medium is expected when used as per the direction on the label within theexpiry period when stored at recommended temperature.

Quality Control

Appearance
Cream to light yellow coloured granular medium

Gelling
Firm, comparable with 1.2% Agar gel.

Colour and Clarity of prepared medium
Medium to dark amber coloured, clear to slightly opalescent gel forms in Petri plates

Reaction
Reaction of 6.71% w/v aqueous solution at 25°C. pH: 6.5±0.2

pH
6.30-6.70

Cultural Response

Cultural characteristics observed after an incubation at 35-37°C for 18-24 hours or longer.(with 5% CO2)

Organism Inoculum (CFU) Growth Recovery
Lactobacillus rhamnosus ATCC 9595 50-100 luxuriant >=50%
Lactobacillus fermentum ATCC 9338 50-100 luxuriant >=50%
^Lactobacillus delbrueckii subsp. lactis ATCC 7830 50-100 luxuriant >=50%
#Lactiplantibacillus plantarum ATCC 8014 50-100 luxuriant >=50%
Lactobacillus sakei ATCC 15521(00015*) 50-100 luxuriant >=50%
Lactobacillus lactis ATCC 19435(00016*) 50-100 luxuriant >=50%
Pediococcus pentosaceus ATCC 33316 (00158*) 50-100 luxuriant >=50%
Escherichia coli ATCC 25922 (00013*) >=104 inhibited 0%
Bacillus cereus ATCC 11778 (00001*) >=104 inhibited 0%

Key: (*) Corresponding WDCM numbers.
#Formerly known as Lactobacillus plantarum ^Formerly known as Lactobacillus leichmannii

Storage and Shelf Life

Store dehydrated powder 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 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 clinical sample must be decontaminated and disposed of in accordance with current laboratory techniques (4,5).

More Information
Product Name Lactobacillus MRS Agar, Granulated (MRS Agar, Granulated)
SKU GM641
Product Type Granulated
Physical Form Granular
Origin Animal
Packaging type HDPE
References 1.deMan J., Rogosa M. and Sharpe M., 1960, J. Appl. Bacteriol., 23:130.
2.Marshall R.T. (Ed.), 1992, Standard Methods for the Examination of Dairy Products, 16th ed., APHA, Washington,D.C.
3.Downes F. P. and Ito K., (Eds.), 2001, Compendium of MethodsFor the Microbiological Examination of Foods, 4th Ed., APHA, Washington, D.C.
4.Sabine and Vaselekos, 1965, Nature, 206:960.
5.MacFaddin J.,1985, Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria, Vol.1, Williams andWilkins, Baltimore.
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