Inoculation of Specimen to Primary Media
- Choose media
- Inoculate specimen to media
- Streak to obtain isolated colonies
Incubate Immediately- Use one of the several anaerobic methods described (see next page) to meet and maintain proper atmospheric conditions.
- Always use care to ensure that media does not sit outside of the anaerobic conditions for extended periods.
- Incubate at 35° to 37°C for 5–7 days to ensure recovery of slow-growers (up to 14 days, especially in cases of joint/prosthetic device infection, jaw/mandible abscesses).
Examination
- Examine plates at 24 hours to determine if growth has occurred; reincubate using anaerobic methods. Reexamine plates every day—always refer to the laboratory's standard operating procedure (this step may be dependent upon the type of anaerobic environment utilized).
- If growth has occurred, aerotolerance testing (subcultures) is performed on each isolate recovered from the primary medium. This is done to recover the anaerobes, as some aerobic bacteria grow anaerobically. (More on this aspect in forthcoming pages.)
- Document colony morphology (appearance) and perform Gram stains as necessary.
- Always Gram stain any growth observed in broth. The findings must be compatible with what is growing on the plates; subculturing this same broth should be performed whenever the findings do not match what is expected.
