BCG vaccine bacilli that seemed less virulent, and changed the course of their research to see if repeated subculturing would produce a strain that was attenuated enough to be considered for use as a vaccine. The BCG strain was isolated after subculturing 239 times during 13 years from virulent strain on glycerine potato medium. The research continued throughout World War I until 1919, when the now avirulent bacilli were unable to cause tuberculosis disease in research animals. Calmette and Guerin transferred to the Paris Pasteur Institute in 1919. The BCG vaccine was first used in humans in 1921. Public acceptance was slow,
Inoculation needle is uncapped. The open end of the uncapped slant culture is then flamed. The slant will be positioned to move up the needle until the inoculation needle tip lightly come into contact with the base surface of the sterile media. The inoculation needle inoculates the sterile agar by the manipulation of the media so that the needle tip grazes the agar surface in a zigzag pattern. Aseptic technique is then applied to the withdrawn inoculation needle. Inoculation of a plate culture is done through the streaking technique to make a streak plate. After lifting the lid so that it hovers