264 proceedings of the indiana academy of science. 

 The Cultivation of Trypanosoma Brucei.* 



Charles Behrens. 



From the Bacteriological Laboratory, Purdue University, Lafayette, 

 Indiana. 



The first successful cultivation of Trypanosoma Brucei was announced 

 by Novy and MacNeal in 1903.' The medium which gave the best results 

 was prepared by adding to a 1 to 8 meat extract 2 per cent peptone; 0.5 per 

 cent sodium chloride; 1 per cent normal sodium carbonate; and 2 per cent 

 agar. Two parts of defibrinated rabbit blood was added and the mixture 

 was allowed to solidify in an inclined position. The medium was then in- 

 oculated with two drops of white mouse, rat, or guinea-pig blood, very rich 

 in trypanosomes. An invisible growth occurred in the water of condensation 

 at 25°C. Novy and MacNeal after testing fifty animals (mice, rats, and 

 guinea-pigs) found that only four gave positive results, or 8 per cent. In 

 1905 Smedley^ reported that three out of ten attempts or 30 per cent were 

 positive. 



Because of the inconsistent results a number of experiments were made 

 using media of quite a different nature. The first attempt along these lines 

 was to use a medium in Avhich the m(>at constituent was rejilaced by an ex- 

 tract made of beans and peas. This was obtained by macerating and boiling 

 1 per cent of each with distilled water. The usual anu^unts of sodium 

 chloride, normal sodium carl)onate and agar were added to this extract. 

 One part of bean and pea agar was mixed with two parts of defibrinated rab- 

 bit })lood, well agitated and solidified in an inclined position. On inoculating 

 the medium with two drops of defibrinated trypanosomal rat blood and sub- 

 sequent inculcation at 25°C more constatil results Averc obtained than with 

 the original Novy-MacNeal medium. 



A second medium was made similar to that employed by Nicolle'' in the 

 cultivation of Leishman-Donovan bodies. In this medium no meat extract 

 or alkali was added. It was prepared by dissolving 2 per cent of agar, 2 

 per cent of peptone, and 0.5 per cent of sodium chloride in distilled water. 

 To this modified Nicolle medium two parts of defibrinated rabbit blood was 

 added, the same as in the case of the other media. 



A third medium Avas employed and was made as follows: 125 gms. 

 finely chopped lean beef, and 250 cc. of water were allowed to digest over 

 night in the cold, or for one hour at 55°C, boiled and filtered. Th(^ filtrate 

 was then dialyzed in a large lliin collodiuin sac against running distilled 



*The strain of Trypanosoma Brucei usotl in these experiments was supplied thru 

 the kindness of Dr. F. O. Now. Ann Arbor. Miehifian. 



1. .Jour. Am. Mefl. .Assn. 1903, p. 1266. "Contributions to Medical Kesearcli" dedi- 

 cated to V. C. Vaugiian, .\nn Arbor, 190.3, p. .549. .Jour. Infect. Dis., 1904, 1 p. 1. 



2. Jour. HyR., 190.5. .5 p. 38. 



3. Arch, de L'Inst. Pasteur de Tunis. 1908. p. 55. .\nn. deJL'Inst. Pasteur, 1909, 

 23, p. 361. 



