Improved detection of oral spirochetes with an anaerobic culture method

A method for routine culturing of oral spirochetes was studied in periodontitis patients and subjects with healthy gingivae. Subgingival plaque bacteria, including oral spirochetes were cultured anaerobically using the steel wool jar method, the glove-box procedure and the plate-in-bottle method. They were also observed directly with a phase-contrast microscope. The number of colonies of recovered spirochetes was significantly higher with the plate-in-bottle method when Medium 10 (M10) or M10 supplemented with 10% rumen fluid was used. In almost all cases, spirochetes were detected with this culture method. The ratios of spirochetes from patient samples, cultured by the plate-in-bottle method, and of other microorganisms, cultured in Eggerth-Gagnon (EG) medium by the steel wool jar method, were almost identical of those observed by phase-contrast microscopy. These results indicate that it can be just as possible to culture certain oral spirochetes as other anaerobic bacteria.