BAM Media M10a: ALOA (Agar Listeria Ottavani & Agosti) Medium

January 2003

Meat peptone18 g
Tryptone6 g
Yeast extract10 g
Sodium pyruvate2 g
Glucose2 g
Magnesium glycerophosphate1 g
Magnesium sulphate0.5 g
Sodium chloride5 g
Lithium chloride10 g
Disodium hydrogen phosphate anhydrous2.5 g
5-bromo-4-chloro-3-indolyl-β-D-glucopyranoside0.05 g
Agar, according to gelation-strength12 g to 18 g
Water, according to volume of fungistat supplement925-930 ml

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.

Advisory for BAM Users on Reported Supply Problems for Cycloheximide

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