Legionella israelensis Bercovier 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 48-72 hours.
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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Bercovier H, et al. Isolation of legionellae from oxidation ponds and fishponds in Israel and description of Legionella israelensis sp. nov.. Int. J. Syst. Bacteriol. 36: 368-371, 1986.