THE TRANSJnSSION' OP LEISHMANIOSIS BY CULTURES. 375 
the peritoneum a very active phagocytosis of the flagellate 
forms takes place, so that after a short time they are broken 
up and completely destroyed. 
Continuing’ previous researches left incomplete, I have 
again undertaken inoculations of guinea-pigs and white rats 
with cultures, having at my disposition besides the strain 
obtained by myself from a young patient, Rocca V — , of 
Bovalino Calabro, other strains also which I owe to the kind- 
ness of Mesnil and Jemma. While the cultures of the 
Pasteur Institute of Paris, originating from Tunis, had been 
kept alive for some years by successive transplantations, mine 
and tliose of Jemma were vei’y recent and still in the first 
subcultures. The cultures used were at the height of their 
development, some in the first days of the subculture (3, 5 
or 7 days), others older (10, 15 or 20 days) and others very 
old (1, 2 or 3 months). The object was to inject all the 
various forms which have been described in the cultures of 
Lei sh mania. The quantity injected was almost always 
2 C.C., sometimes 4 c.c., for animals of medium size, guinea- 
pigs weighing about 300 gr. and young rats. In some cases 
1 c.c. of the culture was injected, to try the effect of injecting 
the animals once, twice, thrice or four times. Twenty guinea- 
pigs and 10 rats were inoculated. 
The method of injection was for the most part intra-peri- 
toneal, and only in a few cases was the material of the 
culture injected under the skin or directly into the current of 
the cii’cnlation, by the vein of the tail of the rats or by 
means of heart-puncture — a method which seems to me pre- 
ferable since it does not present excessive difficulties of 
technique. 
Some of the animals died spontaneously, two guinea-pigs 
and two rats, in the course of the investigations, as happens 
often with these animals in the laboratory from causes 
extraneous to the experiments, and of these I made a rigorous 
examination in search of the Leishmanias ; only one rat and 
one guinea-pig were lost without examination, and of these, 
of course, I have taken no account. The others were killed 
