42 LOCAL TISSUE REACTIVITY 



taiK'oiis injcdion ol I he filtrate rei^iilarly pioduccd a severe kjcal 

 iiillaniniation, \vlii(li in this respect, sharply differed from the 

 insi^nilicant primary effect produced by active principles of other 

 bacterial species. 



Jotikow-Werejnikow aird Lipatova (1933-34) were the first to 

 reproduce the phenomenon of local skin reactivity to B. pestis. 

 The cultures were grown on plain agar, pH 6.6 at 37° c. for 

 thirty-six hours. LInder these conditions of cultivation, as shown 

 by these authors, the organisms developed abimdant capsular ma- 

 terial. The growth was washed off in distilled water and frozen 

 and thawed ten times. The rabbits were prepared by intradermal 

 inoculation of 0.25 c.c. of the material and twenty-four hours 

 later, received an intravenous injection of the same. According 

 to the description of these authors, the reactions were of extreme 

 severity following the intravenous injection. If the preparatory 

 injections ^vere made next to a large vein of the skin of the 

 abdominal Avail, there also occtnred a swelling of the vein, in 

 addition to the typical reaction at the prepared site. Subsequently 

 a large portion of the vein became necrosed. 



ACTIVE PRINCIPLES OF VARIOUS MEMBERS OF HEMOGEOBINOPHILIC 



GROUP 



Frisch (1930) elicited the phenomenon to B. infiiieiizae with 

 filtrates of cultures of the organism on chocolate agar. The po- 

 tency of the filtrates was not determined. 



L. Gross (1930) observed the phenomenon of local skin reac- 

 tivity to B. pertussis. The filtrates were made from washings of 

 growth on chocolate agar Kolle flasks. The concentration of the ac- 

 tive principles wms never of high potency and seemed to vary for 

 unknown reasons. In the best preparations not more than 50 re- 

 acting units per 1 c.c. were obtained. Mishulow, MoAvry and Scott 

 (1930) modified the method of preparation of these active prin- 

 ciples and seemed to have obtained positive reactions in a large 

 percentage of rabbits. They used the follo\ving method: 



Cultures were grown on horse blood chocolate agar and trans- 

 planted at twenty-four hour intervals for two or three generations. 

 Blake bottles of chocolate agar were inoculated with growth from 

 the slants (one slant for each bottle) and 4 c.c. of 1 per cent 

 horse serum beef heart broth, j)H 7.2 to 7.4, were added to each 

 bottle. Bottles were incubated at 37^^ c. for tAventy to t\venty-two 

 hours. The growth was washed off with a buffer solution (47 gms. 



