Further Studies on the Paired Fibrous Structure of Mycobacterial Cell Wall

A "paired fibrous structure" was found in "ghost cells" of myeobacteria by Takeya et el. (l) and was supposed to be related to the cell envelope. The present paper will describe further observations on this structure. Cells of Mycobacterium Jucho strain I grown on a thin collodion membrane deposited on 4 per cent glycerol agar were replaced onto the same medium containing isoniazid or streptomycin 1 rag. per ml., or mitomycin C 0.1 to 100 #g per ml. After 1 to 3 days of incubation at 37°C., the collodion film was removed from the medium on copper mesh screen and "ghost cells" thus obtained were examined by the electron microscope. For further treatment of the "ghost cells," the collodion film with the "ghost cells" was again floated off on the surface of the solution containing various chemicals and incubated for I to 24 hours. A droplet of each solution was put on the cells on the collodion film before incubation. After treatment, the collodion film was taken on distilled water for "washing" and removed on copper mesh screen to be examined by the electron microscope. Repeated freezing and thawing, ultrasonic vibration of cell suspensions, and grinding of cells with glass beads were also used to prepare "ghost cells" of Mycobacterium Jucho strain and Mycobacterium tuberculosis hominis (H37Rv). Leproma of rats which had been infected with Mycobacterium lepraemurium was cut into small pieces and ground with sea sand. After removing sea sand and host cell debris by low speed centrifugation, the bacterial cells were washed and collected by high speed centrifugation. About 5 to I0 per cent of the cells collected were found to be "ghost cells" by electron microscopy.