Prolonged survival of virulent Treponema pallidum (Nichols strain) in cell-free and tissue culture systems

Abstract

Survival of Treponema pallidum was found to be prolonged in the presence of tissue culture. Of the 12 cultures studied, cottontail rabbit epithelium (Sf1Ep) supported T. pallidum for the longest time. In horizontal Leighton tubes with reduced medium and an atmosphere of 5% CO2 in N2, the 50% survival time (ST50) was 5 to 6 days for treponemes associated with monolayers of Sf1Ep cells. Comparable cell-free tubes had ST50 values of less than 4 days. In vertical Leighton tubes containing 6 ml of prereduced medium incubated aerobically, gradients of O2 tension and redox potential were established. Attachment and survival of T. pallidum were greatest at a depth of about 10 to 20 mm. Motility was between 70 and 95% in this area throughout the first 14 days of incubation. Occasionally, greater than 50% motility was observed for as long as 21 days. The redox potential and O2 tension in the optimal area of gradient cultures were reproduced by adjusting the medium depth in a shell vial culture system containing cells on a horizontal cover slip. Treponemes associated with the cell monolayer in both gradient and shell vial cultures were still virulent after 21 days in vitro. The dilution of testis extract and the concentration of T. pallidum were found to be important factors in survival of T. pallidum.