LEISHMANIA DONOVANI 



409 



intact, and where the cytoplasm is dotted over with pairs of nuclei and 

 kinetoplasts, just as they are in the supposed multiple segmentation forms. 

 Still more doubtful are the forms which Archibald (1913, 1914), Smallman 

 (1913), and Statham and Butler (1913) have described. These are more 

 or less spherical portions of cytoplasm containing granules, which have 

 no such definite arrangement as the nuclei and kinetoplasts in the bodies 

 just discussed. They bear some faint resemblance to schizogony stages 

 of malarial parasites or other organisms as seen in dried smears. Here, 

 again, the origin of these structures is in the cytoplasm of large cells with 

 granular cytoplasm, portions of which have been broken off. They are 

 merely fragmentation bodies, and have no relation to the leishmania. 



CULTURE. — The greatest interest attaches to the culture of leish- 

 mania. Rogers (1904) demonstrated that flagellates of the leptomonas 



Fig. 190. — Culture Forms of Leishmania donovani fixed with Schaudinn's 

 Fluid and stained with Iron H.^matoxylin (x 2,000). (Original.) 



type appeared in citrate solution, to which material from spleen puncture 

 of cases of kala azar had been added, an observation which proved con- 

 clusively the flagellate nature of the puzzling Leishman-Donovan body 

 (Fig. 190). Though flagellates developed and multiplied in the medium 

 employed at the ordinary laboratory temperature, this was not always 

 the case, and subculture Was not satisfactorily obtained. Rogers obtained 

 better results with citrated human blood acidified with citric acid, but it 

 was Nicolle (1908c) who demonstrated the possibility of culture at a tem- 

 perature of 22° C. in the water oi condensation in tubes of Novy and 



