Optimal conditions for most mycobacterial cultures are:
a. 25°C to 30°C in the dark in an atmosphere of 2% carbon dioxide (CO2) and high hu-midity.
b. 35°C in the dark in an atmosphere of 5% to 10% CO2 and high humidity.
c. 35°C in the dark in a microaerophilic at-mosphere of 10% CO2 and high humidity.
d. 32°C in the dark in an atmosphere of de-creased CO2 and increased humidity.
B
For optimal cultivation of Mycobacterium spp. from most specimens, cultures are incubated at 35°C in the dark in an atmosphere of 5% to 10% CO2 and high humidity. If specimens are ob-tained from skin or superficial lesions suspected to contain Mycobacterium marinum or Myco-bacterium ulcerans, then an additional set of solid media should be inoculated and incubated at 25°C to 30°C. In addition, a chocolate agar plate or the placement of an X-factor (hemin) disk on conventional media and incubation at 25°C to 33°C are needed for the recovery of Mycobacte-rium haemophilum.
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