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the animal developing symptoms of the disease on the fourth day after 

 subcutaneous injection of culture material. From this animal the 

 contents of a fresh vesicle was p aced in an agar tube, and in two days 

 the fluid was swarming with the same kinds of organisms that had 

 been introduced from the earher culture tube. 



From the standpoint of protozoology these results are entirely 

 consistent with the established facts of the life histories of other cell- 

 invading flagellated parasites. It is regrettable that the technique was 

 such that no definite conclusions can be drawn in regard to the finer 

 structures of the parasite. All of the tissues and smears were fixed 

 in 70 per cent alcohol which is far from satisfactory for the interpretation 

 of delicate structural details. Observations on the living organisms 

 leave no doubt of the flagellum, but we do doubt a statement to the 

 effect that this flagellum "is not a flagellum in the usual sense of the 

 word, but rather a flagellum-hke appendage to the cell." A better 

 fixation and more careful staining would probably reveal the nucleus 

 and blepharoplast and the insertion of the flagellum base in or near the 

 blepharoplast. The polymorphic structure of the organisms is not an 

 uncommon feature of allied forms of protozoa; short and stumpy 

 types mingled with long thread-like forms are characteristic of cul- 

 tural stages of Crithidia, Leptomonas, Trypanosoma and Leishmania, 

 and similar differences in form are met with in the normal hosts. 



The intra-cellular bodies with their varied forms, which, however, 

 are reducible to one general type, are strikingly suggestive of Leish- 

 mania donovani of kala azar. But here again, the finer structures 

 must remain unknown until a better method of fixation is employed. 



On the whole, we are incUned to accept Stauffacher's interpretation 

 of the organism which he names Aphthomonas infestans, as belonging 

 to the group of simple flagellated protozoa (Monadida) and closely 

 related to the genus Leishmania. If this work is confirmed on material 

 fixed with better methods, we should naturally expect the next step 

 to be the discovery of the transmitting agent in some form of tick or 

 biting fly. 



Gary N. Calkins. 



Columbia University, 

 March, 1916. 



