Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended Use
Recommended for promoting sporulation in Bacillus species.
Composition**
| Ingredients | Gms / Litre |
|---|---|
| HM peptone B # | 3.000 |
| Gelatin peptone | 5.000 |
| Manganese sulphate | 0.030 |
| Agar | 15.000 |
| Final pH (at 25°C) | 6.8±0.2 |
**Formula adjusted, standardized to suit performance parameters
# - Equivalent to Beef extract
Directions
Suspend 23.03 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
Nutrient Agar w/Manganese, conventionally abbreviated as NAMn favours culture and sporulation of aerobic Bacillus species especially from canned foods. HM peptone B and gelatin peptone provide necessary nutrients required for growth of Bacillus species. Manganese is known to influence and enhance sporulation in Bacillus species (1,2,5,6). It has been reported that organisms recovered from spoilage of foods such as fruit drinks, tomatoes, acidified onions and other canned foods sporulate well aerobically on Nutrient Agar with added manganese (7). Thermophilic bacteria such as B. stearothermophillus are capable of growth at 55-65°C while an incubation temperature of 30 to 35°C is favorable for culture and sporulation of mesophilic spore formers (7). This property is exploited to grow and therefore differentiate mesophilic and thermophilic spoilage bacteria. As recommended by APHA, in routine diagnosis for spoilage in canned foods, microbiological cultural procedures involve the use of primary recovery media and subculture media to identify spoilage bacteria and study its growth characteristics. Recovery media for aerobes generally include DTA (Dextrose Tryptone Agar) (M092) or DTB (Dextrose Tryptone Bromocresol Broth) (M122). Use of Cooked M Medium (M149) is recommended for recovery of anaerobic organisms. NAMn is widely used as subculture media for aerobes. When rod shaped aerobes in pure culture are isolated on DTA (or DTB) media (M092/ /M122) and sporulation is not evident, the isolates should be subcultured on Nutrient Agar with Manganese, at the temperature of initial isolation. After incubation for upto 10 days, if spore production has taken place, the spores are heat shocked to destroy all vegetative cells and cultured on Nutrient Agar w/ Manganese at both 30-35°C and 55°C. The temperature at which outgrowth occurs from the spore state indicates whether the isolate is an obligate mesophile (growth at 30 to 35°C), an obligate thermophile (growth at 55°C) or a facultative thermophile (growth at 30 to 35°C and at 55°C). After use, contaminated materials must be sterilized by autoclaving before discarding. Nutrient Agar with Manganese supports growth and enhances sporulation by aerobic spore-formers and can be used primarily to differentiate mesophilic from thermophilic Bacillus species.
Type of specimen
Food samples
Specimen Collection and Handling
For food samples, follow appropriate techniques for sample collection and processing as per guidelines (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 should be followed while handling specimens. Safety guidelines may be referred in individual safety data sheets.
Limitations
- Further biochemical and serological tests must be carried out for complete identification.
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 yellow homogeneous free flowing powder
Gelling
Firm, comparable with 1.5% Agar gel
Colour and Clarity of prepared medium
Light amber coloured clear to slightly opalescent gel forms in Petri plates
Reaction
Reaction of 2.3% w/v aqueous solution at 25°C. pH: 6.8±0.2
pH
6.60-7.00
Cultural response
Cultural characteristics observed after an incubation at 35-37°C for upto 5 days.
| Organism | Inoculum (CFU) | Growth | Recovery |
|---|---|---|---|
| Bacillus stearothermophilus ATCC 7953 | 50-100 | luxuriant (incubated at 55°C for upto 5 days) | >=50% |
| Bacillus coagulans ATCC 8038 | 50-100 | luxuriant (with sporulation) | >=50% |
| Bacillus licheniformis ATCC9945a | 50-100 | luxuriant (with sporulation) | >=50% |
| Bacillus megaterium ATCC 9855 | 50-100 | luxuriant (with sporulation) | >=50% |
| Bacillus polymyxa ATCC 8526 | 50-100 | luxuriant(with sporulation) | >=50% |
| Bacillus subtilis subsp. spizizenii ATCC 6633 (00003*) | 50-100 | luxuriant (with sporulation) | >=50% |
Key: (*) Corresponding WDCM numbers.
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. 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 (3,4).
Reference
- Charney, J., Fisher, W. P. and Hegarty C. P., 1951, J. Bacteriol., 62:145.
- Curran, H. R. and Evans F. R., 1954, J. Bacteriol., 67: 489.
- Isenberg, H.D. Clinical Microbiology Procedures Handbook 2nd Edition.
- 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.
- Maunder D. T., 1970, "Examination of canned foods for microbial spoilage." Microbiology, Metal Div. R. and D,
- Continental Can Co., Inc., Oak Brook, Ill.
- Penna T. C., Machoshvili I. A., Taqueda, M. E and Ferraz, C. A. 1998, PDA J. Pharm. Sci. Technol., 52 (5):198.
- Salfinger Y., and Tortorello M.L., 2015, Compendium of Methods for the Microbiological Examination of Foods, 5th Ed., American Public Health Association, Washington, D.C.
| Product Name | Nutrient Agar w/ Manganese |
|---|---|
| SKU | M931 |
| Product Type | Regular |
| Physical Form | Powder |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1. Charney, J., Fisher, W. P. and Hegarty C. P., 1951, J. Bacteriol., 62:145. |
| Customized Product Available | No |
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