PRODUCTION OF TETANUS TOXIN 413 



1915. The Pasteur strain was the one formerly in use at the 

 Pasteur Institute, Paris, for the production of tetanus toxin. 

 That this strain did not produce so potent a toxin (1:25,000) 

 as the Research and Goadby strains (1:35,000) was somewhat 

 surprising as it had been kept on a medium made with Martin 

 pepton for years, whereas the Goadby and Research strains had 

 been accustomed to this medium for several months only. 



That Martin pepton broth as made by us is capable occasion- 

 ally of giving potent toxin, is evidenced by the results obtained 

 in experiment 1 (table 4) and in experiment 4 (table 5.) The 

 other preparations of toxin broth were made with the same pre- 

 cautions and care but, as is shown, with far different results. 

 When it was apparent that equal parts of Martin pepton and 

 veal infusion were not giving satisfactory toxin, tests were made 

 with broth prepared by usmg more concentrated Martin pepton 

 solution with ordinary veal infusion (experiment 4, table 5); 

 also by using the usual amount of Martin pepton solution with 

 a more highly concentrated veal infusion. In neither case was 

 the result so satisfactory as with the usual preparation. Sub- 

 sequent broths were prepared according to the original method, 

 that is to say using equal parts Of infusion and pepton, but the 

 toxin was far too low in potency to be used. 



Word was received in January, 1918, with regard to the modi- 

 fication of Martin pepton broth used with favorable results for 

 tetanus toxin at the Pasteur Institute. The variation between 

 this procedure and the one we were using lay in the concentration 

 of the pepton solution, in the shortened incubation period and also 

 in the absence of glucose. A preparation of toxin broth (see 

 experiment 6, table 6), was made according to this modification, 

 one-half of the broth having 1 per cent glucose added to it while 

 the other half was prepared without any sugar. After seven 

 days incubation instead of the usual period of fifteen days, the 

 cultures were filtered and tested. The results of the animal tests 

 were most disappointing as pigs inoculated with 1 cc. of a dilution 

 of 1:5,000 from each preparation showed an absence of tetanic 

 symptoms. In his article on the preparation of this pepton, 

 Martin advises that not less than five stomachs should be used 



