Cultivation 565 



All of the bodies are intracellular, as can easily be determined 

 by examining sections of tissue, but in smears of splenic pulp the 

 cells are broken and many free bodies may appear. The cells in 

 which they occur are lymphocytes, endothelial cells, and peculiar 

 large cells whose histogenesis is obscure. They are rarely to be 

 found in polymorphonuclear leukocytes, and though there has 

 been much discussion upon this point, probably never appear in the 

 red blood-corpuscles. 



The bodies divide by binary and multiple fission, without rec- 



^^m 







^^ 



Fig. 221. — Leishman-Donovan bodies from the spleen of a case of Kala-azar. 

 X about 1000 (From Beattie and Dickson's "A Text-book of. General Path- 

 ology," by kind permission of Rebman, Limited, publishers). 



ognizable mitotic changes. When multiple fission occurs, the 

 nucleus divides several times before the protoplasm breaks up. 

 The organism is not motile and at this stage has no flagella. 



Cultivation. — The organism was first cultivated artificially 

 by Rogers* in citrated splenic juice at 17° to 24°C. It can also be 

 cultivated in the blood-serum agar medium used by Novy, McNeal, 

 and Hall for trypanosomes, and in the N. N. N. medium of NicoUe, 

 which has the following composition : 



Water 900 cc. 



Salt (NaCl) 6 gm. 



Agar-agar 16 gm. 



Dissolve, distribute in tubes, sterilize, and add to the medium in each tube after 

 uquefying and cooling to 40°-so°C., one-third of its volume of rabbit's blood 

 obtained by cardiac puncture. Slope the tubes for twelve hours, incubate at 

 37 C. for five days to test the sterility of the medium, then keep at the ordinary 

 temperature of the laboratory, sealed to prevent evaporation. 



It is imperative that the material planted be sterile so far as bacteria 



* "Brit. Med. Jour.," 1904, 11, 645. 



