The Diagnosis of Kala-Azar. 



301 



spleen and blood have not yet been attempted at higher and lower 

 concentrations than those indicated. Flagellates grow on medium 

 at least as alkaline as P H 9.0. In all of these cultures post-flagellate 

 forms may be found including definite Leishman-Donovan bodies. 

 Other forms suggesting Cornwall's second type of "thick tails" 1 

 are occasionally met with. Cultures on buffered medium remain 

 viable for a considerable time. A successful subculture has been 

 made after 56 days although no flagellates were found in a drop of 

 the fluid used for inoculation. Growth took place on the surface 

 of the medium above the water of condensation, to a height of 

 four centimeters in one instance. The surface of the medium was 

 slightly dulled. Such surface growth seemed especially rich in 

 rosettes and active flagellates. 



The results from peripheral blood thus far cultured are as 

 follows: Ten samples of blood have been taken from five different 

 patients and distributed into forty tubes of buffered N.N.N, 

 medium of various hydrogen-ion concentrations and incubated at 

 22 0 C. Twenty-nine of these cultures were positive (72.5 per 

 cent.). One or more tubes from nine of the ten samples showed 

 flagellates (90 per cent.). At least one culture was positive from 

 each patient. One case gave positive results after having received 

 20 c.c. of 0.2 per cent, colloidal antimony sulphide eight hours 

 before the blood was taken. Before the culture she had received 

 in all 121 c.c. of the suspension, equivalent to 0.173 gram of 

 metallic antimony injected intravenously over a period of fourteen 

 days. Another patient had received intravenously 377 c.c. of a 

 similar suspension, equivalent to 0.539 gram of metallic antimony, 

 over a period of thirty-three days. The last dose of 30 c.c. was 

 given three days before the culture was taken. 



Summary. 



1. Human red cells and serum are unfavorable to the growth 

 of Leishmania donovani. 



2. A method is given for removing the red cells and serum from 

 blood before planting. 



3. By this method blood cultures have been obtained from 

 nine out of ten samples of blood from five patients, some of them 

 after considerable antimony treatment. 



1 Cornwall and LaFrenais, loc. cit., p. 299. 



