Cultivation 527 



In addition to these bodies the protoplasm may contain one or two 

 vacuoles. 



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- 

 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. 



Fig. 217. 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.) 



Cultivation. The organism was first cultivated artificially 

 by Rogers in citrated splenic juice at 17 to 24C. 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 Nicolle, 

 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 liquefying and cooling to 4o-5oC., one- third of its volume of rabbit's 

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

 at 37C. 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 



