Actinomyces israelii (Kruse) Lachner-Sandoval
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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. Under anaerobic conditions, withdraw 0.5 mL of recommended broth from a single test tube (5 to 6 mL) and rehydrate the entire vial contents.
3. Aseptically transfer this aliquot back into the broth tube. A slant and additional broth tubes may be inoculated with 0.2 mL each of the cell suspension. Blood plates may be streaked to check for colonial morphology and purity.
4. Incubate tubes and one plate under anaerobic conditions at 37°C. Incubate one blood plate in 5% CO2 or in air at 37°C.
5. Within 48-72 hours, growth should be evident by sediment in the broth and growth on agar surfaces. On #260 agar incubated anaerobically, colonies are tan, punctate, pulvinate and dry. Colony size is variable, ranging from pinpoints to about 1 mm in diameter.
Anaerobic conditions for transfer may be obtained by either of the following:
· Use of an anaerobic gas chamber, or
· Placement of test tubes under a gassing cannula system hooked to anaerobic gas.
Anaerobic conditions for incubation may be obtained by any of the following:
· Loose screw caps on test tubes in anaerobic chamber,
· Loose screw caps on test tubes in an activated anaerobic gas pack jar, or
· Use of sterile butyl rubber stoppers on test tubes so that an anaerobic gas headspace is retained.
Additional information on this culture is available on the ATCC web site at www.atcc.org.
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