THE TOXINE. 
PKEPARATION OF THE TOXINE. 
The culture . — For the preparation of the toxine we use a culture 
known as “Park’s bacillus No, 8.” This culture has now become 
famous in laboratories both in this country and abroad on account of 
its remarkable power of retaining its virulence and producing strong 
poisons. The culture was isolated in 189d b}^ Dr. Anna W. Williams, 
of the New York health department, and is sometimes known as the 
Park- Williams bacillus. 
The best results are obtained by growing the organism as a surface 
growth in the special bouillon presently to be described. We cany it 
over from test tube to test tube eveiy day, taking only the surface 
film with a flattened platinum needle. 
The strongest poisons are obtained when the surface growth is 
heavy and the bouillon remains clear. A precipitate of the old and 
dead organisms always collects at the bottom of the tube. 
The houillon . — The bouillon is a special alkaline medium prepared 
from fresh lean beef freed of muscle sugar and all other sugars, and to 
which a small quantity of dextrose (glucose) is added, which seems to 
favor the production of a strong poison. 
The bouillon is prepared largely in accordance with the instructions 
laM down by Theobald Smith in a paper entitled “The relations of 
dextrose to the production of toxin in bouillon cultures of the diph- 
theria bacillus.” “ This culture medium, which we call “Smith’s 
bouillon,” is prepared by us as follows: 
Cut out all the fat and tendon of the beef. Pass through the meat 
grinder, catching all the expressed juice. Weigh and add twice the 
weight of water. Place in the cool room at 15° C. foi- twenty-four 
hours, then strain through cloth, pressing firmly. Weigh the amount 
of meat infusion thus obtained. Take the reaction and neutralize 
with sodium h}"drate to 1.5 per cent acidit}^ to phenol-phthalin in 
order that the colon bacillus, which is now planted in it, may grow 
well. Inoculate with a reliable culture of T. coll communis^ using 10 
c. c. of a twenW-four-hour-old bouillon culture for each liter of meat 
infusion. Grow at 37° C. for twent 3 "-four hours. Add the white of 
«Journ. Exper. Med., vol. 4, nos. 3-4, 1899. 
(35) 
