Porphyromonas circumdentaria Love et al.
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2. Under anaerobic conditions, withdraw 0.5 ml of the 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. Additional tubes may be inoculated with 0.5 ml each from the suspension. Streak several blood plates to check for colonial morphology and purity.
4. Incubate tubes under an anaerobic atmosphere at 37oC. Incubate one agar plate anaerobically for colony formation, and one aerobically for aerobic contamination check.
5. Within 24 to 48 hours, growth is evident by turbidity in the broth and circular, smooth colonies on the anaerobic agar surface. No growth should occur on the blood agar plate incubated aerobically.
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.
Always use freshly prepared pre-reduced media or pre-reduced media that has been previously prepared but stored under anaerobic conditions. Resazurin in the media is a color indicator for anaerobic conditions. Observance of pink color in medium before use or during incubation shows anaerobic conditions have not been met and oxidation has occurred. Medium should be discarded.
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Love DN, et al. Description of Porphyromonas circumdentaria sp. nov. and reassignment of Bacteroides salivosus (Love, Johnson, Jones, and Calverley 1987) as Porphyromonas (Shah and Collins 1988) salivosa comb. nov [published errata appear in Int J Syst Bacteriol 1992 Oct;42(4):660 and 1993 Jul;43(3):630]. Int. J. Syst. Bacteriol. 42: 434-448, 1992. PubMed: 1503973