Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Ashby’s Mannitol Agar
Recommended for isolation of Azotobacter species from soil that can use mannitol and atmospheric nitrogen as source of carbon and nitrogen respectively.
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Azotobacter#CC293D
Intended Use
Recommended for isolation of Azotobacter species from soil that can use mannitol and atmospheric nitrogen as source of carbon and nitrogen respectively.
Composition**
| Ingredients | Gms / Litre |
|---|---|
| Mannitol | 20.000 |
| Dipotassium hydrogen phosphate | 0.200 |
| Magnesium sulphate | 0.200 |
| Sodium chloride | 0.200 |
| Potassium sulphate | 0.100 |
| Calcium carbonate | 5.000 |
| Agar | 15.000 |
| Final pH (at 25°C) | 7.4±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 40.7 grams in 1000 ml purified / distilled water. Heat just to boiling. 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.
Note: Due to presence of calcium carbonate, the prepared medium forms opalescent solution with white precipitate.
Principle And Interpretation
Azotobacter is a genus of free-living diazotrophic bacteria which have the highest metabolic rate compared to any other microorganisms. Azotobacters are chemoorganotrophic, using sugars, alcohols and salts of organic acids for growth. Azotobacters can non-symbiotically fix atmospheric nitrogen aerobically due to their unique mode of metabolism. Besides the ability to fix atmospheric nitrogen, Azotobacter also synthesize biologically active substances, which attributes to improving seed germination, plant growth etc.
Ashby's Mannitol Agar are formulated as described by Subba Rao (3). It is used for isolation of Azotobacter, a non-symbiotic nitrogen fixing bacteria which uses mannitol as a carbon source and atmospheric nitrogen as nitrogen source. Dipotassium phosphate provides buffering to the medium. Various essential ions required for promoting growth of Azotobacter are also available in this medium
Type of specimen
Soil samples
Specimen Collection and Handling
For soil samples, follow appropriate techniques for sample collection and processing as per guidelines (3). 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
- Due to variable nutritional requirements, some strains show poor growth on this medium.
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
White to cream homogeneous free flowing powder
Gelling
Firm, comparable with 1.5% Agar gel
Colour and Clarity of prepared medium
Whitish opalescent gel forms in Petri plates
Reaction
Reaction of 4.07% w/v aqueous solution at 25°C. pH : 7.4±0.2
pH
7.20-7.60
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for upto 5 days.
| Organism | Growth |
|---|---|
| Azotobacter nigricans ATCC 35009 | good-luxuriant |
| Azotobacter vinelandii ATCC 478 | good-luxuriant |
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 (1,2).
| Product Name | Ashby’s Mannitol Agar |
|---|---|
| SKU | M706 |
| Product Type | Regular |
| Physical Form | Powder |
| Origin | Chemically defined |
| Packaging type | HDPE |
| References | 1. Subba Rao, 1977, Soil Microorganisms and Plant Growth, Oxford and IBH Publishing Co., India. |
| Customized Product Available | No |
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