Dehydrated culture media
Asparagine Broth
For the presumptive identification and enumeration (MPN) of Pseudomonas aeruginosa.
- Cat. No.
- LC-1207
- Category
- Dehydrated culture media
- Format
- 500 g
- Storage
- 2 to 25 °C
- Minimum guaranteed shelf life
- 15 months
Asparagine Broth is an excellent enrichment broth for Pseudomonas aeruginosa. This medium is recommended for its enumeration and identification based on the hydrolysis of asparagine to aspartic acid performed by the bacteria.
Key features
Applications
Detection and selective enumeration of Pseudomonas.
Industry
Environmental monitoring.
Preparation
13.5 g in one liter of distilled water, add 8 mL of glycerol; boil for one minute, sterilize in autoclave at 121 °C for 15 minutes.
Principles and uses
Pseudomonas aeruginosa is an opportunist pathogen for humans, capable of growing in water with a low concentration of nutrients. This is why natural mineral water and spring water are free of Pseudomonas aeruginosa at the time of their commercialization. This microorganism can also be found in swimming pool water.
The formula contains a strictly mineral base with asparagine as the sole source of nitrogen, and glycerol as the carbon source. Potassium salts act as a buffer system and magnesium sulfate provides magnesium ions required in a large variety of enzymatic reactions, including DNA replication, and also acts as a buffer.
Since P. aeruginosa hydrolyzes asparagine to aspartic acid, the appearance of growth with or without fluorescent pigmentation is considered a presumptive test for its presence. Counts are determined using the MPN tubes. Confirmation is made by subculturing a loopful from each turbid tube into Acetamide Broth.
Technical specifications
| Solubility | Without residues |
|---|---|
| Appearance | Fine powder |
| Color of the dehydrated medium | White |
| Color of the prepared medium | Colorless |
| Final pH (25 °C) | 7.0 ± 0.2 |
| Amount per litre | 13.5 g |
| Storage | 2 to 25 °C |
Formula
| Ingredient | g/L |
|---|---|
| Dipotassium phosphate | 1 |
| DL-Asparagine | 2 |
| Magnesium sulfate | 0.5 |
| Monopotassium phosphate | 10 |
Preparation and use
Preparation
Suspend 13.5 grams of the medium in one liter of distilled water. Add 8 mL of glycerol. Mix well and dissolve by heating with frequent agitation. Boil for one minute until complete dissolution. Dispense into appropriate containers and sterilize in autoclave at 121 °C for 15 minutes. To obtain a double-strength broth, dissolve 27 grams of the medium and add 16 mL of glycerol.
Instructions for use
Enumeration is carried out by the MPN method.
Incubate at 35 ± 2 °C for 24-48 hours.
Quality control
Microbiological test
Incubation conditions: 35 ± 2 °C / 24-48 h.
| Microorganism | Specification |
|---|---|
| Pseudomonas aeruginosaATCC 10145 | Good growth |
| Pseudomonas aeruginosaATCC 27853 | Good growth |
Bibliography
- APHA. Standard Methods for Examination of Water and Wastewater, 14th ed. 1975.
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