Bio-205L Microbiology Lab – Topic 2: Methods of Culture Transfer, Pure Culture Isolation, and Antimicrobial Testing (Online – Bacterial Staining)

Manipulation of Microorganisms - Methods of Culture Transfer, Pure Culture Isolation, and Antimicrobial Testing: After participating in this lab, students will be able to use aseptic technique to transfer bacteria from one medium to another, use the streak-plate method to isolate bacterial colonies, and use the Kirby-Bauer disk diffusion assay to test for bacterial sensitivity to antibiotics, antiseptics, and disinfectants. The results of antimicrobial testing can help determine if a bacterial isolate is susceptible or resistant to various antibiotics, and may help direct a clinician to an effective treatment. (Online) Bacterial Staining: After participating in this topic, students will be able to perform simple stains and several differential staining techniques that are used to observe unique molecules and structures found in various bacterial genera. The Gram stain, which aids in differentiating bacterial cell wall structure, the acid-fast stain, which detects mycobacteria, and the endospore stain, which detects endospore-forming bacteria, are useful for identification of these unique microorganisms.

Objectives:

  1. Demonstrate the aseptic transfer of bacteria from broth culture to agar slant media, or from an agar slant culture to broth media.
  2. Demonstrate the isolation and purification of bacteria using the streak-plate method.
  3. Describe the cultural characteristic of bacterial species growth based on macroscopic appearance in slant, plate, and broth media.
  4. Determine the zone of inhibition caused by various antibiotics.

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