596 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



and the tube covered with a rubber 

 cap, after which it is placed at 25°C. 

 for about one week, 

 (c) Behrens gave the following method 

 of preparation : 



(1) Digest 125.0 g. choiaped beef in 

 250.0 cc. water over night in the 

 cold, or for one hour at 55 °C. 



(2) Strain (1), boil the extract and 

 filter. 



(3) Dialyze the filtrate in a large col- 

 lodium sac against running distilled 

 water for 24 to 48 hours. 



(4) Dilute (3) to 1000.0 cc. 



(5) Dissolve 20.0 g. peptone, 5.0 g. 

 NaCl, 0.1 g. CaCh, 10.0 cc. N/1 

 NajCOa solution and 20.0 g. agar 

 in (4). 



(6) Distribute in test tubes in 10.0 cc. 

 lots. 



(7) Sterilize in autoclave by heating to 

 105-108° for 15 minutes. 



(8) Shortly before use, the desired 

 number of agar tubes are melted in 

 the water bath, cooled to 60°C. 

 and two volumes of defibrinated 

 rabbit's blood are added. 



(9) Mix well and solidify in slanting 

 position. 



References: MacNeal (1904 p. 535), Tobey 

 (1906 p. 125), Novy and MacNeal (1905 

 p. 265), Thompson (1908 p. 77), Behrens 

 (1914 p. 27). 



1893. Smedley's Blood Infusion Agar 



Constituents : 



1. Bullock heart infusion 1000.0 cc. 



2. Peptone 10.0 g. 



3. NaCl 10.0 g. 



4. Agar 20.0 g. 



5. Blood, defibrinated rabbit.. 1000.0 cc. 

 Preparation : 



(1) Prepare an infusion of bullock's heart. 



(2) Dissolve 1.0% peptone and 1.0% 

 NaCl and 2.0% agar in (1). 



(3) Clear in the usual way by the addition 

 of the white of an egg. 



(4) Tube. 



(5) Collect blood from the heart of a 

 rabbit by means of sterilized Pasteur 

 bulbs and then defibrinated in steri- 

 lized bottles containing a little broken 

 glass. 



(6) Cool sterile (3) to 45°C. and add an 

 equal volume of (5). 



(7) Mix and slant. 



Sterilization: Method of sterilization of 

 (4) not given. 



Use : Cultivation of Trypanosomata Lewisi 

 and other trypanosomata. Other in- 

 vestigators cultivated a number of differ- 

 ent organisms on similar media. 



Variants : 



(a) Duval (Gurd) prepared a medium as 

 follows: 



(1) Prepare a beef infusion. 



(2) Dissolve 10.0 g. peptone, 20.0 g. 

 agar and 5.0 g. NaCl in (1). 



(3) Adjust reaction to 0.6% acid to 

 phenolphthalein (hot titration). 



(4) Tube. 



(5) Sterilize in the autoclave. 



(6) Cool to 52°C. and add a small 

 quantity of defibrinated sterile 

 human blood. From 4 to 7 drops 

 of blood are added to each 6 to 

 10.0 cc. of agar. 



(7) Shake thoroly. 



(8) Slant or pour in sterile Petri dishes. 

 Gurd used the medium to isolate 

 gonococci. He reported that gono- 

 cocci colonies were raised worty- 

 looking, bluish gray or almost color- 

 less semi-transparent and round. 

 Gave a bright crimson almost trans- 

 parent medium. If agar was above 

 60°C. when mixed with blood, the 

 hemoglobin was destroyed and me- 

 dium assumed a dirty brown color. 



(b) Duval cultivated B. leprae on a 

 medium prepared as indicated. He 

 reported that cultures without the 

 blood and using B. typhosus and 

 Entameba coli gave growth of B. 

 leprae. 



(1) Prepare nutrient infusion agar. 



(2) Sterilize (Method not given). 



(3) Add 1.0% human defibrinated 

 blood to sterile (2). 



(4) Slope. 



(5) Inoculate the surface with pure 

 growth cultures of B. influenza 

 and meningococcus. 



(6) Incubate at room temperature for 

 48 hours. 



(7) Inoculate the slant with an emul- 

 sion prepared from leprous tissue. 



