BAM Media M10a: ALOA (Agar Listeria Ottavani & Agosti) Medium
|Meat peptone||18 g|
|Yeast extract||10 g|
|Sodium pyruvate||2 g|
|Magnesium glycerophosphate||1 g|
|Magnesium sulphate||0.5 g|
|Sodium chloride||5 g|
|Lithium chloride||10 g|
|Disodium hydrogen phosphate anhydrous||2.5 g|
|Agar, according to gelation-strength||12 g to 18 g|
|Water, according to volume of fungistat supplement|
Dissolve the dehydrated components or dehydrated complete base in the water by boiling.
Adjust the pH, if necessary, so that after sterilization it is 7.2 ± 0.2.
Sterilize by autoclaving for 15 minutes at 121°C.
Supplements: Dissolve 2 g of L-α-Phosphatidylinositol (Sigma P 6636) in 50 ml of cold distilled water.
Stir for about 30 minutes until a homogeneous suspension is obtained.
Autoclave at 121°C for 15 minutes. Cool to 48-50°C.
Dissolve the following compounds individually in the specified solvents and filter-sterilize the solutions: 0.02 g nalidixic acid (sodium salt) in 5 ml of water; 0.02 g of ceftazidime in 5 ml of water; 76700 U of polymyxin B in 5 ml of water; 0.05 g of cycloheximide in 2.5 ml of ethanol and add 2.5 ml of water; 0.01 g of amphotericin B in a mixture of 2.5 ml HCl (1 M) and 7.5 ml dimethylformamide.
Complete medium: Agar base 925 ml (or 930 ml); Nalidixic Acid solution; 5 ml; Ceftazidime solution, 5 ml; Cycloheximide solution, 5 ml (or Amphotericin B solution 10 ml); Polymyxin B solution, 5 ml;
L-α-Phosphatidylinositol solution, 50 ml.
This medium is available commercially as the ready-to-use agar medium.