Actinobacillus pleuropneumoniae (Shope) Pohl et al.

1. Open vial according to enclosed instructions or visit www.atcc.org for instructions.
2. Rehydrate the entire pellet with approximately 0.5 mL of #814 broth. Aseptically transfer the entire contents to a 5-6 mL tube of #814 broth. Additional test tubes can be inoculated by transferring 0.5 mL of the primary broth tube to these secondary broth tubes.
3. Use several drops of the primary broth tube to inoculate a #814 plate and/or #814 agar slant.
4. Incubate at 37°C for 24-48 hours in an atmosphere of 5% CO₂.

If a broth culture is desired, use Medium #814 formulation without the agar, or HTM (Haemophilus Test Broth). To make HTM, first a fresh hematin stock solution is prepared by dissolving 50 mg of hematin powder in 100 mL of 0.01N (0.01 mol/L) NaOH with heat and stirring until the powder is dissolved thoroughly. Thirty milliliters of the hematin stock solution is added to 1 L of Mueller-Hinton Broth with 5 g of yeast extract. After autoclaving and cooling, aseptically add 3 mL of a NAD (nicotinamide adenine dinucleotide) stock solution (50 mg of NAD dissolved in 10 mL of distilled water; filter-sterilized).

Additional information on this culture is available on the ATCC^® web site at www.atcc.org.

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