Microbiology · General Microbiology (Bacterial Genetics, Culture Media, Stains, Sterilization)

During heat sterilization in an autoclave, the D-value (decimal reduction time) concept is used to validate sterilization efficacy. The standard autoclaving cycle is 121°C for 15 minutes at 15 psi pressure. Which organism is used as the biological indicator (BI) to validate autoclave performance?

  • A Bacillus cereus spores (ATCC 11778)
  • B Clostridium sporogenes spores (ATCC 7955)
  • C Geobacillus stearothermophilus spores (formerly B. stearothermophilus)
  • D Bacillus subtilis var. niger spores
Correct answer: C. Geobacillus stearothermophilus spores (formerly B. stearothermophilus)

Explanation

Geobacillus stearothermophilus (formerly Bacillus stearothermophilus) spores are the biological indicator for moist heat (autoclave) sterilization because they are highly resistant to moist heat with a D121 value of ~1.5 minutes; commercial strips contain 10^6 spores. After autoclaving, the strips are incubated at 56°C — no growth (no color change in medium) confirms successful sterilization. Bacillus subtilis var. niger (globigii) spores are used for dry heat (160°C hot air oven) and ethylene oxide gas sterilization validation. C. sporogenes ATCC 7955 is used for steam-in-place validation at lower temperatures.

Reference: Ananthanarayan & Paniker's Textbook of Microbiology, 11th ed.

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