Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended Use
Recommended for the isolation and cultivation of denitrifying bacteria from soil samples.
Composition**
| Ingredients | Gms / Litre |
|---|---|
| Potassium nitrate | 1.000 |
| L-Asparagine | 1.000 |
| Sodium citrate | 8.500 |
| Potassium dihydrogen phosphate | 1.000 |
| Magnesium sulphate | 1.000 |
| Calcium chloride | 0.200 |
| Ferric chloride | 0.0001 |
| Agar | 15.000 |
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 27.7 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
Asparagine Nitrate Medium is formulated as per Subba Rao (3). Nitrogen transformation in soil results in the loss of molecular nitrogen. The conversion of nitrate and nitrite into molecular nitrogen or nitrous oxide through microbial processes is known as denitrification. Denitrification of bound nitrogen to gaseous nitrogen is mediated by numerous species of bacteria, which normally use oxygen as hydrogen acceptor (aerobic). These bacteria also posses the ability to use nitrate and nitrite in the place of oxygen as the hydrogen acceptor (anaerobically).
Asparagine is source of organic nitrogen and is readily available for microbial energy and growth while the salts in the medium help for growth of microorganisms.
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
Light amber coloured clear to slightly opalescent gel forms in Petri plates
Cultural Response
Cultural characteristics observed after an incubation at 25-30°C for upto 7 days.
| Organism | Growth |
|---|---|
| Achromobacter denitrificans ATCC 14648 | luxuriant |
| Bacillus subtilis subsp. spizizenii ATCC 6633 (00003*) | luxuriant |
| Micrococcus luteus ATCC 10240 | luxuriant |
| Pseudomonas aeruginosa ATCC 27853 (00025*) | luxuriant |
| Thiobacillus denitrificans ATCC 29685 | good |
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 (1,2).
| Product Name | Asparagine Nitrate Medium |
|---|---|
| SKU | M724 |
| Product Type | Regular |
| Physical Form | Powder |
| Origin | Chemically defined |
| Packaging type | HDPE |
| References | 1. Subba Rao N. S., 1977, Soil Microorganisms and Plant Growth, Oxford and IBH Publishing Co., New Delhi. |
| Customized Product Available | No |
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