7.8: Procedures

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ORGANISMS

Trypticase Soy broth cultures of Pseudomonas aeruginosa and Staphylococcus aureus. Caution: handle these organisms as pathogens.

MEDIUM

Motility Test medium (2 tubes)

PROCEDURE (to be done individually and in pairs)

1. Observe the phase-contrast microscopy demonstration of motile Pseudomonas aeruginosa.

Movie of motile Pseudomonas from YouTube.

2. Observe the dark-field microscopy demonstration of motile Pseudomonas aeruginosa.

3. Take 2 tubes of Motility Test medium per pair. Stab one with Pseudomonas aeruginosa and the other with Staphylococcus aureus. Stab the bacterium about 1/2 - 3/4 of an inch into the agar, taking care not to tilt or twist the loop so that the loop comes up through the same cut as it went down. Incubate the tubes in your test tube rack at 37°C until the next lab period.

4. Observe the flagella stain demonstrations of a Vibrio species (monotrichous), Proteus vulgaris (peritrichous) and Spirillum undula (amphitrichous) as well as the dark-field photomicrograph of the spirochete Leptospira. When observing flagella stain slides, keep in mind that flagella often break off during the staining procedure so you must look carefully to observe the true flagellar arrangement.