Legionella pneumophila Brenner et al.
ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.
ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
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 entire pellet.
3. Aseptically transfer this aliquot back into the broth tube. Mix well.
4. Use several drops of the suspension to inoculate a #1099 agar slant and/or plate.
5. Incubate the tubes and plate at 37°C in an atmosphere of 5% CO2 for 24-48 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.
Additional information on this culture is available on the ATCC® web site at www.atcc.org.
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