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Scientific Proceedings (128) 



tive cultures from one patient. At first Rogers merely citrated 

 the patient's blood but Nicolle was much more successful with 

 his "N.N.N." medium, a rabbit blood agar. The experiments of 

 Rogers 15 and of Cornwall and LaFrenais 14 showed that culture 

 media made with human blood were not favorable to the growth 

 of Leishmania donovani. 



Two series of experiments were made using salt agar as in 

 N.N.N, medium but with washed red cells and serum, both un- 

 heated and heated at 56 C° for one-half hour, from man, horse, 

 sheep and rabbit. Human red cells were somewhat inhibitory; 

 human serum much more so. Rabbit cells and serum were less 

 unfavorable to growth but were inferior to whole defibinated rab- 

 bit blood. In action the cells and serum of the horse and sheep 

 lay between those of man and the rabbit. As a result of these 

 experiments, a technic for blood culture was devised as follows : 

 10 c.c. of blood was drawn from a vein into 2 c.c. of 1 per cent, 

 citrated Locke's solution in a syringe and immediately expelled 

 into 50-70 c.c. of the same fluid in a flask. On return to the 

 laboratory, the diluted blood was divided between two 50 c.c. 

 centrifuge tubes and spun slowly (about 750 r.p.m.) for five 

 minutes. The blood was thus divided into a supernatant cloudy 

 fluid and a sediment of red cells. The cloudiness of the first is 

 caused by blood platelets and some red cells. Extracellular Leish- 

 man-Donovan bodies were occasionally found in this fraction. 

 After rapid centrifugation (about 1400 r.p.m. for five minutes) 

 the sediment from this upper portion was planted into N.N.N, 

 medium of about P„ 7.6. Using this technic, 115 culture tubes 

 were made from 34 blood specimens from untreated cases of kala- 



0 Wylie, J. H., China Med. Jour., 1920, xxxiv, 593. 



7 Knowles, R., Indian Jour. Med. Res., 1920, viii, 177. 



8 Rogers, Sir Leonard, Fevers in the Tropics, Third Ed., p. 48. 



o Bramachari, A. N., Kala-azar and Its Treatment, Butterworth, 1920, 

 p. 99. 



io Mayer, M., and Werner, H., Deutsch. Med. Wchnschr., 1914, xl, 67. 

 iiWenyon, C. M., Jour. Trop. Med. and Hyg., 1914, xvii, 49. 

 12 Row, R., Indian Jour. Med. Res., 1914, ii, July editorial. 

 i-'tKorke, V., noted in Cornwall and LaFrenais, Indian Jour. Med. Res., 

 1915-16, iii, 398. 



i-t Cornwall, S., and LaFrenais, H. M., Indian Jour. Med. Res., 1915-16, 

 iii, 398. 



i- r > Rogers, Sir Leonard, Lancet, 1919, i, 505. 



