The effect of purification on the ultrastructure and infectivity of egg-attenuated Chlamydia psittaci (6BC)

Abstract

A procedure is described for the purification of mixed populations of the three different morphological forms of Chlamydia psittaci (6BC) from infected yolk sac membranes. Elementary bodies and small intermediate bodies are not perceptibly damaged during purification which involves homogenization of the host cells, differential centrifugation, sedimentation through 20% sucrose, and treatment with trypsin. The observation that elementary bodies undergo plasmolysis in 20% sucrose is interpreted as indicating that the cytoplasmic membrane of these particles is intact at that stage in the purification. Reticulate bodies and large intermediate bodies are damaged, to a degree, by the homogenization step. This damage takes the form of discontinuities of the outer envelope membrane, and results in the loss of the regular coccobacillary shape of these particles and in an increase in their size. Treatment with a combination of RNase and DNase was found to cause profound damage to all three morphological forms of the chlamydiae.