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CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(g) Roddy prepared the medium as 

 follows: 



(1) Mix 300.0 cc. distilled water with 

 100.0 cc. of beef blood serum. 



(2) Add 4.0 g. of glucose or any other 

 desired carbohydrate, alcohol, etc., 

 to (1). 



(3) Add a saturated aqueous solution 

 of litmus to give a blue solution. 



(4) Sterilize in the steam sterilizer for 

 20 minutes on each of 3 successive 

 days. 



(h) Baeslack and Keane used a medium 

 prepared as follows for the cultiva- 

 tion of Spirochaeta pallida from tis- 

 sue. The tissue was removed from 

 the patient and pushed into the 

 medium from one-half to two-thirds 

 the length of the tube. Incubate 

 at 37°C. for 3 to 5 days and remove a 

 portion with a sterile pipette on a 

 slide for dark field e.xamination. 



(1) Dilute normal horse serum free 

 from preservatives with sterile 

 distilled water in the proportion 

 of3:l. 



(2) Fill sterile tubes within an inch 

 from the top with (1) and close the 

 tube with a sterile rubber stopper. 



(3) Heat for an hour in a water bath at 

 60 °C. 



(4) On the following day heat for one 

 hour at 70°C. and on the third day 

 at 70°C. until the medium takes on 

 the consistency of syrup. 



(i) Giltner prepared the medium as 

 follows: 



(1) Dilute beef or sheep serum with 

 three times its volume of distilled 

 water. 



(2) Heat in the Arnold for 15 minutes. 



(3) Distribute into desired quantities. 



(4) Add 1.0% of any desired carbo- 

 hydrate, alcohol, etc. and sufficient 

 litmus to give a deep purple color. 



(5) Sterilize by the fractional method 

 in the steamer. 



(j) Abbott gave the following method of 

 preparation: 



(1) Mix one part blood serum with 3 

 parts distilled water. 



(2) Neutralize (indicator not specified). 



(3) Heat in an Arnold steamer until the 

 mixture becomes opalescent. 



(4) Add 1.0% of a 5.0% aqueous solu- 

 tion of litmus to (3). 



(5) Add 1.0% of any desired carbo- 

 hydrate, alcohol, etc., to (4). 



(6) Tube. 



(7) Sterilize in the Arnold and allow the 

 steamer to remain uncovered during 

 the process to avoid over heating. 

 Length of time of sterilization not 

 specified. 



(k) Harvey prepared the medium as 

 follows: 



(1) Mix one part clear serum (ox) with 

 3 parts water. 



(2) Steam for 15 minutes (or heat for 20 

 minutes at 118°C.) 



(3) Add sufficient litmus solution to 

 give a deep blue tint and 1.0% of 

 any desired carbohydrate, alcohol, 

 etc. The litmus solution may be 

 omitted. Heat at 118°C. in step 

 (2) when omitting the litmus. 



(4) Sterilize at 100°C. for 20 minutes on 

 each of 3 successive days, or by 

 filtration. 



(1) Gildemeister prepared a medium as 

 follows and used it as a substitute for 

 nutrose in Barsiekow's medium: 



(1) Mix 5.0 to 10.0 cc. of beef serum 

 with 90 to 95.0 cc. of distilled water. 



(2) Sterilize for one hour in a steamer. 



(3) Dissolve 1.0 g. of glucose, lactose or 

 mannitol in 5.0 cc. of Kubel-Tie- 

 mann litmus solution. 



(4) Add (3) to (2). 



(5) Distribute in tubes. 



(6) Sterilize on 3 successive days for 15 

 to 20 minutes each day. 



(m) Klimmer used the same basic solution 

 as Gildemeister in variant (1), but 

 used 2.0% mannitol instead of 1.0%, 

 and used 2.0% sucrose and 2.0% 

 maltose also as added nutrients, 

 (n) Park, Williams and Krumwiede gave 

 the following method of preparation: 



(1) Dilute serum with two or three 

 times its volume of distilled water. 



(2) Sterilize in the Arnold. 



(3) Prepare 10.0 or 20.0% solutions of 

 any desired carbohydrate, alcohol, 

 etc. 



(4) Heat (3) in small containers in the 

 Arnold sterilizer for 30 minutes on 3 



