HiMedia Laboratories GmbH, Marie-Curie-Str. 3, 64683, Einhausen, Germany.
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Intended Use
Recommended for selection and subculture of Staphylococcus aureus in accordance with the harmonized method of USP/EP/BP/JP/IP.
Composition
| Ingredients | g/L |
|---|---|
| Peptone # | 5.000 |
| Tryptone ## | 5.000 |
| HM Peptone B ### | 1.000 |
| Sodium chloride | 75.000 |
| D-Mannitol | 10.000 |
| Phenol red | 0.025 |
| Agar | 15.000 |
| pH after sterilization (at 25°C) | 7.4±0.2 |
- # Peptic digest of animal tissue
- ## Pancreatic digest of casein
- ### Equivalent to Beef extract
Formula adjusted, standardized to suit performance parameters
Directions
Mannitol Salt Agar is a ready to use solid media in glass bottle. The medium is pre-sterilized, hence it does not need sterilization. Medium in the bottle can be melted either by using a pre-heated water bath or any other method. Slightly loosen the cap before melting. When complete melting of medium is observed dispense the medium in tubes as butts /slants or in plates as desired and allow to solidify. If on plate, either streak, inoculate or surface spread the test inoculum (50-100 CFU) aseptically.
Principle And Interpretation
Staphylococci are widespread in nature, although they are mainly found on the skin, skin glands and mucous membranes of mammals and birds. The coagulase-positive species i.e. Staphylococcus aureus is well documented as a human opportunistic pathogen. The ability to clot plasma continues to be the most widely used and accepted criterion for the identification of pathogenic staphylococci associated with acute infections (1). Staphylococci have the unique ability of growing on a high salt containing media (2). Isolation of coagulase-positive staphylococci on Phenol Red Mannitol Agar supplemented with 7.5%NaCl was studied by Chapman (3). The resulting Mannitol Salt Agar Base is recommended for the isolation of coagulase-positive staphylococci from cosmetics, milk, food and other specimens (1,4,5,6,7). It is also used in the performance of microbial limit tests for the selective isolation of Staphylococcus. The formulation is in accordance with the harmonization of USP/EP/BP/JP/IP (8-12).
The medium contains HM peptone B, tryptone and peptone which makes it very nutritious as they provide carbon, nitrogen compounds, long chain amino acids, vitamins and other essential growth factors and trace nutrients. Many other bacteria except Staphylococci are inhibited by 7.5% sodium chloride. Mannitol is the fermentable carbohydrate fermentation of which leads to acid production, detected by phenol red indicator.
S.aureus ferment mannitol and produce yellow coloured colonies surrounded by yellow zones. Presumptive coagulase-positive yellow colonies of S.aureus should be confirmed by performing the coagulase test [tube or slide (1)]. A possible S.aureus must be confirmed by the coagulase test. Also the organism should be subcultured to a less inhibitory medium not containing excess salt to avoid the possible interference of salt with coagulase testing or other diagnostic tests (e.g. Nutrient Broth)(M002) (13). Few strains of S.aureus may exhibit delayed mannitol fermentation. Negative results should therefore be re-incubated for an additional 24 hours before being discarded (14).
Type of specimen
Pharmaceutical samples
Specimen Collection and Handling
For pharmaceutical samples, follow appropriate techniques for sample collection, processing as per guidelines (8-12).
Enrichment: Sample is initially enriched in Tryptone Soya Broth (MH011) and incubated at 30-35°C for 18-24 hours.
Selection and subculture : The enriched culture is then subcultured on Mannitol Salt Agar (MPH118) and incubated at 30-35°C for 18-72 hours.
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
- This medium is a selective medium, some strains of Staphylococcus aureus may exhibit a delayed fermentation of mannitol.
- Certain other bacteria are also mannitol fermenting other than Staphylococcus, therefore further biochemical testing is required for identification.
- 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.
- It is recommended to store the plates at 24-30°C to avoid minimum condensation.
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 Mannitol Salt Agar in bottles
Colour of medium
Red coloured medium
Quantity of medium
100ml of medium in bottles
pH
7.20-7.60
Sterility Check
Passes release criteria
Growth Promotion Test
Growth Promotion was carried out in accordance with the harmonized method of USP/EP/BP/JP/IP, 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.
Growth promoting properties
Growth of microorganism comparable to that previously obtained with previously tested and approved lot of medium occurs at the specified temperature for not more than the shortest period of time specified inoculating <=100 cfu(at 30-35° C for <=18 hours).
Indicative properties
Colonies are comparable in appearance and indication reaction to those previously obtained with previously tested and approved lot of medium occurs for the specified temperature for a period of time within the range specified inoculating <=100cfu (at 30-35°C for 18-72 hours).
Inhibitory properties
No growth of the test microorganism occurs for the specified temp for not less than longest period of time specified inoculating >=100cfu (at 30-35°C for >= 72 hours).
Cultural Response
| Organism | Inoculum (CFU) | Growth | Recovery | Colour of colony |
|---|---|---|---|---|
| Growth Promoting + Indicative | ||||
| Staphylococcus aureus subsp. aureus ATCC 6538 (00032*) | 50-100 | luxuriant | >=50% | yellow/white colonies surrounded by yellow zone |
| Inhibitory | ||||
| Escherichia coli ATCC 8739 (00012*) | >=103 | inhibited | 0% | |
| Additional Microbiological testing | ||||
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) | 50-100 | luxuriant | >=50% | yellow/white colonies surrounded by yellow zone |
| Staphylococcus epidermidis ATCC 12228 (00036*) | 50 -100 | fair - good | 30-40% | red |
| Staphylococcus epidermidis ATCC 14990 (00132*) | 50-100 | fair - good | 30-40% | red |
| Proteus mirabilis ATCC 12453 | 50-100 | none-poor | 0-10% | yellow |
| Escherichia coli ATCC 25922 (00013*) | >=103 | inhibited | 0% | |
| #Klebsiella aerogenes ATCC 13048 (00175*) | >=103 | inhibited | 0% | |
Key: (*) Corresponding WDCM numbers
(#) Formerly known as Enterobacter aerogenes
Storage and Shelf Life
On receipt store between 15-30°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 sample must be decontaminated and disposed of in accordance with current laboratory techniques (13,14).
| Product Name | Mannitol Salt Agar |
|---|---|
| SKU | SMH118 |
| Customized Product Available | No |
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