Legionella quateirensis Dennis et al.
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2. Using a single tube of #1099 broth (5 to 6 ml), withdraw approximately 0.5 to 1.0 ml with a Pasteur or 1.0 ml pipette. Rehydrate the pellet.
3. Aseptically transfer this aliquot back into the broth tube. Mix well.
4. Use several drops of the suspension to inoculate an agar slant and/or plate.
5. Incubate at 37oC in an atmosphere of 5% CO2 for 5-7 days. Loosen the screw caps during the incubation time.
Strain shows growth by uniform turbidity throughout the broth.
Legionella species are nutritionally fastidious and are inhibited by media of poor caliber, so strict attention should be paid to the quality of the ingredients. The pH of the medium should be checked when cool, and is most critical. Unbuffered medium should not be used. Exposure of medium to light, particularly when hot, may also result in the accumulation of peroxides which can be inhibitory to the bacteria.
Before rehydration, vials may be stored at +2 to +8oC for extended periods with minimal loss of viability. Subculturing to maintain culture is discouraged because of the possibility of mutation, selection of variants, or contamination. To minimize changes, it is recommended that cells be harvested and stored at ‑70oC or below.
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Dennis PJ, et al. Five new Legionella species isolated from water. Int. J. Syst. Bacteriol. 43: 329-337, 1993. PubMed: 8494743