"THE PREPARATION OF TETANUS ANTITOXIN | 
By E. H. RUEDIGER 
(From the Section of Sera and Prophylactics, Biological Laboratory, 
Bureau of Science, Manila, P. I.) 
EIGHTY-FIVE TEXT FIGURES 
Notwithstanding the fact that tetanus antitoxin is extensively 
used in practically all countries of the world, very little has been 
written about its preparation in recent years. 
Eisler and Pribram? advise the injection of tetanus toxin 
and iodine trichloride for the first three months of the time 
that a horse is being immunized against tetanus toxin. This 
is followed by another three months’ treatment with tetanus 
toxin alone. Although weakening the tetanus toxin by mixing 
it with iodine trichloride saves the lives of many serum horxses 
and is almost indispensable when tetanus antitoxin is not avail- 
able, it is now rarely used; the horses can be, and now usually 
are, fortified with the antitoxin. 
In order to obtain good antitoxin, good toxin is indispensable. 
Good toxin usually can be obtained by growing suitable tetanus 
bacilli in glucose broth under anaérobic conditions. In my ex- 
perience the following procedure has given fairly good results: 
To 500 grams of chopped lean beef add 1,000 cubic centimeters of dis- 
tilled water and-boil for one hour. Enough water should be added to 
allow for evaporation. Allow the infusion to cool; strain and add the 
following: Witte’s pepton, 20 grams; sodium chloride, 5 grams; glucose, 
10 grams. 
Prepare the broth in the usual way and with sodium hydrate solution 
reduce the acidity to about 0.5 per cent normal acid. Pass the broth 
through a paper filter, sterilize it in the autoclave, cool it rapidly in run- 
ning water, inoculate with tetanus bacilli, and incubate it under hydrogen 
at a temperature of from 36° C. to 37° C. for from seven to ten days 
(zarely fourteen days). After having been incubated sufficiently long, the 
broth is rendered germ-free by passing it through a sterile germ-proof 
filter. Phenol, in the proportion of 0.5 cubic centimeter per 100 cubic centi- 
meters of filtrate, may be added. The reaction of the filtrate will be 
about 2 per cent normal acid. This should be reduced to practically 
neutral by adding sodium hydrate solution. An acid filtrate produces 
severe local reaction on subcutaneous injection, while after the injection 
* Read at the annual meeting of the Philippine Islands Medical Associa- 
tion, November 4-7, 1914. 
* Kraus und Levaditi, Handbuch der Technik und Methodik der Immuni- 
tatsforschunge. Gustav Fischer, Jena (1909), 2, 139. 
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