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Lactobacillus MRS Agar (MRS Agar)
Recommended for the isolation and enumeration of lactic acid bacteria from meat and meat products. The composition and performance criteria of this medium are as per the specifications laid down in ISO 1995, Draft ISO/DIS 13720:2010.
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Lactobacillus MRS Agar is recommended for the isolation and enumeration of lactic acid bacteria from meat and meat products. The composition and performance criteria of this medium are as per the specifications laid down in ISO 13721:1995.
Composition
As per ISO 13721:1995
Ingredients (As per ISO 13721:1995)
| Ingredients | g/L |
|---|---|
| Meat extract | 8.000 |
| Peptone | 10.000 |
| Yeast extract | 5.000 |
| Tri-ammonium citrate | 2.000 |
| Sodium acetate | 5.000 |
| Magnesium sulphate, heptahydrate | 0.200 |
| Manganese sulphate, tetrahydrate | 0.050 |
| Dipotassium Hydrogen phosphate | 2.000 |
| Glucose | 20.000 |
| Sorbitan monooleate (Tween 80) | 1.000 |
| Agar | 12.00-18.00 |
| Final pH (at 25°C) | 5.7±0.2 |
Lactobacillus MRS Agar (MRS Agar) M6411 Ingredients
| Ingredients | g/L |
|---|---|
| HM extract# | 8.000 |
| Peptone | 10.000 |
| Yeast extract | 5.000 |
| Ammonium citrate | 2.000 |
| Sodium acetate | 5.000 |
| Magnesium sulphate, heptahydrate | 0.200 |
| Manganese sulphate, tetrahydrate | 0.050 |
| Dipotassium phosphate | 2.000 |
| Glucose (Dextrose) | 20.000 |
| Polysorbate 80 | 1.000 |
| Agar | 12.000 |
| Final pH (at 25°C) | 5.7±0.2 |
**Formula adjusted, standardized to suit performance parameters
# - Equivalent to Meat extract
Directions
Suspend 65.13 grams (the equivalent weight of dehydrated medium per litre) 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 medium is based on the formulation of deMan, Rogosa and Sharpe (1) with slight modification. It supports luxuriant growth of all Lactobacilli from oral cavity (1), dairy products (2), foods (3), faeces (4) and other sources (5). Lactobacillus MRS Agar is recommended by ISO Committee (6).
Peptone and meat extract B supplies nitrogenous and carbonaceous compounds, long chain amino acids and other essential growth nutrients. Yeast extract provides vitamin B complex and glucose 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. Phosphates provide good buffering action in the media.
Lactobacilli are microaerophillic 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 (5). Lactobacilli isolated on MRS Agar should be further confirmed biochemically.
Type of specimen
Food and dairy samples
Specimen Collection and Handling
For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (6,7). 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 shall be followed while handling specimens. Safety guidelines may be referred in individual safety data sheets.
Limitations
- Individual organisms differ in their growth requirement and may show variable growth patterns on the medium.
- 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.
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: Cream to light yellow homogeneous having tendency to form soft lumps which can be easily broken down to powder form free flowing powder
Gelling: Firm, comparable with 1.2% Agar gel.
Colour and Clarity of prepared medium: Medium to dark amber coloured slightly opalescent gel forms in Petri plates
Reaction: Reaction of 6.51% w/v aqueous solution at 25°C. pH : 5.7±0.2
pH: 5.50-5.90
Cultural Response: Cultural characteristics observed after an incubation at 25°C for 72 h± 3 h
| Organism | Inoculum (CFU) | Growth | Recovery |
|---|---|---|---|
| Lactobacillus acidophilus ATCC 4356 | 50-100 | luxuriant | >=50% |
| Lactobacillus casei ATCC 9595 | 50-100 | luxuriant | >=50% |
| Lactobacillus fermentum ATCC 9338 | 50-100 | luxuriant | >=50% |
| # Lactiplantibacillus plantarum ATCC 8014 | 50-100 | luxuriant | >=50% |
| Lactococcus lactis subsp. lactis ATCC 19435 | 50-100 | None-poor | <=10% |
| Lactococcus sakei ATCC 15521 | 50-100 | luxuriant | >=50% |
| Pediococcus damnosus ATCC 29358 | 50-100 | luxuriant | >=50% |
| Pediococcus pentosaceus ATCC 33316 | 50-100 | luxuriant | <=10% |
| Bifidobacterium bifidum** ATCC 11863 | 50-100 | luxuriant | <=10% |
| Escherichia coli ATCC 25922 (00013*) | 50-100 | none-poor | <=10% |
Key ** -Growth under anaerobic conditions for 72 hours
(*) Corresponding WDCM numbers.
#Formerly known as Lactobacillus plantarum
Storage and Shelf Life
Store below 8°C in tightly closed container and the prepared medium at 2 - 8°C. Use before expiry date on the label.
| Product Name | Lactobacillus MRS Agar (MRS Agar) |
|---|---|
| SKU | M641I |
| Product Type | Regular |
| Physical Form | Powder |
| 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. 6.Marshall R.T. (Ed.), 1992, Standard Methods for the Examination of Dairy Products, 16th ed., APHA, Washington,D.C. 7.Sabine and Vaselekos, 1965, Nature, 206:960 8.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 9.Wehr H. M. and Frank J. H., 2004, Standard Methods for the Microbiological Examination of Dairy Products, 17th Ed.,APHA Inc., Washington, D.C |
| Customized Product Available | No |
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