117 



more than one, and at least three anti- 

 gens in the reaction, all of them titrated 

 and studied as to their fixing power: 

 the WASSERMANN test should never 

 be practised with one antigen, because 

 this might bring about grave errors. 

 Cholesterinised antigens should be used 

 with care and cholesterinising should 

 be made by a previous titiation with 

 known sera and comparatively with an- 

 tigens already tried: thus the optimum 

 quantity of cholesterin to add to a given 

 voliune of antigen is reached. This titra- 

 tion is lengthy and the antigen must 

 only be made use of after it has been 

 well verified with numerous knovsri sera 

 and compared with antigens already 

 tried. 



We tested the serum of 60 patients 

 of Leisiimaniosis americana in the 19th 

 Ward of the Santa Casa de Misericordia 

 of Rio de Janeiro, thanks to the cour- 

 tesy of the chief, Prof. Dr. FERNANDO 

 TERRA to whom v.'e are very much 

 indebted for his kindness. 



In all patients we searched for sy- 

 philis most painstakingly, so as to be 

 able to come to e.\act conclusions. 



The clinical diagnostic of leishma- 

 niosis. especially in the mucous form of 

 the disease, lends itself to confusion 

 with syphilis in its similar lesions: it 

 is a capital fact that syphilis attacks 

 the bony tissue wliereas leishmaniosis 

 may destroy the tissues and the carti- 

 lage but ahvays leaves the bone intact. 



The clinical precedents of each of 

 the cases of great importance in re- 

 vealing or excluding concomitant syphi- 

 lis. 



The serologic examination renders, 

 we think, invaluable services, for out of 

 the 60 cases we observed, not a single 

 one of pure leishmaniosis, free from 

 syphilis deviated the complement when 

 jnixed with the antigens used in the 

 ¡ft'ASSERMANN test. 



Technic employed in carrying out these 

 reactions. 



Antigens: — The antigens used were 

 six in number: luetic liver (alcoholic 

 extract of liver of foetus with heredita- 

 ry syphilis); human heart (cholesteri- 

 nised alcoholic extract); ox-heart (cho- 

 lesterinised alcoholic extract); antigens 

 of acetone-insoluble lipoids, according to 

 NOGUCHTs technic; antigen of BORDET 

 and RUELENS (used in the Pasteur 

 Institute of Brussels, a modification of 

 NOGUGHI's process of extracting ace- 

 tone-insoluble lipoids); antigen of LES- 

 SER rethereal extract of ox-heart). Ace- 

 tone-insoluble lipoids, both for NOGU- 

 GHI's and for BORDET and RUELENS' 

 process were extracted from ox-heart. 

 All these antigens were perfectlj' 

 known having been tried on numerous 

 known syphilitic and known normal se- 

 ra. Nevertheless, to make sure, they 

 were all titrated again on the day of the 

 reaction as to their inhibitory power 

 and controlled with known positive and 

 known negative sera. For verifying the 

 inhibitory power we used two or three 

 times the dose employed in reactions 

 together with a fixed dose of comple- 

 ment and sheep-corpuscles sensitised 

 with two or three hemolysing units. 

 Complement:— We employed fresh 

 guinea-pig seriun; to obtain a more ho- 

 mogeneous complement we employed 

 from eight to ten guinea-pigs which we- 

 re bled on the same day the 

 reaction was carried out. Complement 

 titration was done by mixing with a 

 fixed dose of hemolysins and the dose 

 of antigen to be used in the reaction. 

 Hemolytic sijstem: — Rabbit anti-red 

 corpuscles of sheen, the notencv of the 

 serum for the corpuscles being always 

 controlled on the day it is made use of. 

 We also verified all the other ele- 

 ments, complement, hemolj^sins. antigens 

 and saline solution, taken one by one 

 and mixed with sheep corpuscles to see 



