PROTOCOL
I. Oxidative-fermentative test using OF media with glucose
A. Inoculation of media
Two tubes of oxidative-fermentative medium are inoculated by stabbing “half way to the bottom” (8) or ¼-inch from the bottom (6) with the test organism. Overlay one of the two tubes with 1 cm of mineral oil (8) (Fig. 1, 2, 3). This overlay prevents the diffusion of oxygen into the medium and creates an anaerobic condition in the tube.
B. Incubation conditions
Incubation at 35°C for 48 hours is recommended for most gram-negative rods (5, 6). Slow growing bacteria may take 3 to 4 days before results can be observed (6).
C. Interpretation of results