A MODIFICATION OF THE HYGIENIC LABORA- 

 TORY METHOD FOR THE PRODUCTION 

 OF TETANUS TOXIN 



HARRIET LESLIE WILCOX 



Research Laboratories, New York City Department of Health 



In the November, 1915, issue of the Journal of Medical Re- 

 search, ^ Anderson and Leake briefly describe the method used 

 at the Hygienic Laboratory, Washington, D. C, for the produc- 

 tion of a uniformly potent toxin. The method in use at the 

 Research Laboratory is essentially that given by Anderson and 

 Leake with a few shght variations. As there have been many 

 inquiries as to how we obtain our potent toxin it was thought 

 that the details of the exact procedure might be of sufficient 

 value to pubhsh. 



STOCK CULTURES 



The stock cultures are grown on a semi-sohd medium made 

 in the following way. 



Veal broth 1000 cc. 



Agar 5 grams 



Witte's Peptone 10 grams 



NaCl 5 grams 



Reaction Neutral to phenolphthalein 



About 8 CC. to 10 cc. of this medium are put into tubes which 

 are autoclaved at 15 pounds pressure for one-half hour, the tubes 

 being then ready for use. 



To transfer cultures, one of the semi-solid agar cultures is 

 melted and 1 cc. is added to a freshly melted semi-solid agar tube, 

 at least ten sub-cultures being thus made from one stock culture. 

 After inoculation, the tubes are cooled, the plugs inmiersed in 



' Anderson and Leake, 1915. A Method of producing Tetanus Toxin, Jour- 

 nal of Medical Research, 33, 239. 



333 



