Dehydrated culture media
LIM Medium (Lysine, Indole, Motility Medium)
For identification of Enterobacteriaceae.
- Cat. No.
- LC-1389
- Category
- Dehydrated culture media
- Format
- 500 g
- Storage
- 2 to 25 °C
- Minimum guaranteed shelf life
- 15 months
LIM Medium (Lysine - Indole - Motility) is a semisolid medium used for the differentiation of the Enterobacteriaceae group by motility, lysine decarboxylase activity and indole production.
Key features
Applications
Differentiation of enterobacteria.
Industry
General cultivation.
Preparation
30.02 g in one liter of distilled water; boil for one minute, sterilize in autoclave at 121 °C for 15 minutes.
Principles and uses
Peptone provides nitrogen, vitamins, minerals and amino acids essential for growth. Yeast extract is a source of vitamins, particularly of the B-group. Dextrose is the fermentable carbohydrate. Bromocresol purple is the pH indicator. Lysine is added to detect the production of the specific enzyme. The addition of tryptophan to the medium allows the performance of the indole test. The low concentration of bacteriological agar is for motility detection.
Technical specifications
| Solubility | Without residues |
|---|---|
| Appearance | Fine powder |
| Color of the dehydrated medium | Light beige |
| Color of the prepared medium | Purple |
| Final pH (25 °C) | 6.7 ± 0.2 |
| Amount per litre | 30.02 g |
| Storage | 2 to 25 °C |
Formula
| Ingredient | g/L |
|---|---|
| Bacteriological agar | 2.7 |
| Bromocresol purple | 0.02 |
| Dextrose | 1 |
| L-Lysine | 10 |
| Peptone | 12.8 |
| Yeast extract | 3 |
| L-Tryptophan | 0.5 |
Preparation and use
Preparation
Suspend 30.02 grams of the medium in one liter of distilled water. Mix well and dissolve by heating with frequent agitation. Boil for one minute until complete dissolution. Dispense into tubes and sterilize in autoclave at 121 °C for 15 minutes.
Instructions for use
- Inoculate by stabbing the LIM medium and incubate in an aerobic atmosphere for 24 hours at 37 ± 2 °C.
- Read the motility and lysine decarboxylase reactions before adding the Kovac's Reagent (LC-5205) for the indole test.
- The motility is indicated by cloudiness in the medium or growth extending away from the line of inoculation.
- Lysine decarboxylation is indicated by a purple color in the medium. A negative lysine reaction produces a yellow color at the bottom of the tube.
- The test for indole should be conducted by adding a few drops of Kovac's Reagent (LC-5205). A positive test is indicated by the formation of a pink-to-red color in the reagent layer after a few minutes.
- If the indole reaction is negative, incubate for an additional 24 hours.
- Compare the results with a non-inoculated test tube.
Quality control
Microbiological test
Incubation conditions: 37 ± 2 °C / 24 h.
| Microorganism | Specification | Characteristic reaction |
|---|---|---|
| Enterobacter cloacaeATCC 13047 | Good growth | Lysine (-), Indole (-), Motility (+) |
| Proteus mirabilisATCC 25933 | Good growth | Lysine (-), Indole (-), Motility (+) |
Bibliography
- Igarashi H. et al. (1969) A new differential medium for enteric pathogens, lysine-indole-motility (LIM) medium.
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