Legionella waltersii Benson et al.
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2. Aseptically transfer the entire contents of the vial into a single broth tube (5 6 ml) of #1099 broth. Mix well. Additional tubes can be inoculated by transferring 0.1 ml to slants and 0.5 ml to broth tubes.
3. Use several drops of the suspension to inoculate a #1099 plate.
4. Incubate the tubes and plate at 37°C in an atmosphere of 5% CO2 for 72 to 96 hours.
Legionella waltersii is nutritionally fastidious and is 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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