CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



295 



(6) Incubate 2 or 3 days to detect any- 

 accidental contamination. 

 Sterilization: Sterilize (2), (3) and litmus 



solution separately in the Arnold on each 



of 3 successive days. 

 Use: To study fermentation by diplococci. 

 Reference: Dunham (1906 p. 19). 



963. Shmamine's Liver Serum Bouillon 



Constituents : 



1. Bouillon 100.0 cc. 



2. Sodium nucleate 1.0 to 2.0 g. 



3. Physiological salt soln. 20.0 cc. 



4. Serum, horse 100.0 cc. 



5. Liver, rabbit 

 Preparation : 



(1) Dissolve 1 to 2.0 g. of the sodium salt 

 of nucleic acid in 20.0 cc. of physio- 

 logical salt solution. 



(2) Boil in the water bath for 15 minutes. 



(3) Mix (2) with 100.0 cc. of sterile clear 

 transparent horse serum. 



(4) Add a small piece of rabbit liver that 

 has been burned off in the flame. 



(5) Mix equal parts sterile bouillon 

 and (4). 



(6) Following final sterilization pass C0> 

 thru the tubes for 2 minutes and then 

 seal with rubber stoppers and para- 

 ffin. (The fact that the medium was 

 to be tubed and a small piece of rab- 

 bit liver added to each tube was not 

 specified.) 



Sterilization: Method of sterilization of 

 serum or bouillon not specified. Sterilize 

 (5) for one hour on each of 3 successive 

 days at 60 °C. 



Use: Cultivation of spirochetes. 



Reference: Shmamine (1912 p. 323). 



964. Besson's Serum Bouillon 



Constituents : 



1. Bouillon. 



2. Serum. 



Preparation: (1) Mix one-half, one-third or 

 one-fourth the amount of sterile serum 

 with ordinary sterile bouillon under 

 aseptic conditions. 



Sterilization: Not specified. 



Use : General culture medium. 



Variants : 



(a) Kligler and Robertson cultivated 

 Spirochaeta obermeieri on a medium 

 prepared as follows: 



(1) Mix one part rabbit serum with 2 

 parts physiological salt solution. 



(2) Prepare a bouillon containing 10.0% 

 peptone. 



(3) Add 1.0% of (2) to (1). 



(4) Adjust (3) to pH = 7.2. 



(5) Distribute into 3 to 4.0 cc. lots into 

 test tubes approximately 1.0 cm. in 

 diameter. 



(6) Inoculate with one drop of infected 

 blood, or if subcultures are being 

 made, add a drop of fresh rabbit 

 blood and rotate the tube gently. 

 (This furnishes the necessary fibrin. 

 Instead of fibrin, 0.05 to 0.1% agar 

 may be used.) 



(7) Cover with a layer of oil about 

 1.5 cm. high. 



(8) Incubate at 28-30°C. 



(b) Klimmer added one part serum to 3 

 to 20 parts bouillon, under aseptic 

 conditions. 



(c) Stitt cultivated Borrelia recurrentis 

 on a medium prepared as follows: 



(1) Mix three parts of a 1.0% peptone 

 bouillon with one part rabbit or 

 horse serum (ascitic fluid may be 

 used). 



(2) Tube in tall tubes. 



(3) Cover with a layer of oil not to e.x- 

 ceed 1.5 cm. in height. 



References: Besson (1920 p. 31), Kligler 

 and Robertson (1922 p. 315), Klimmer 

 (1923 p. 200), Stitt (1923 p. 54). 



965. Veillon's Ascitic Fluid Bouillon 



Constituents: 



1. Bouillon 1000.0 cc. 



2. Ascitic fluid 1000.0 cc. 



Preparation : 



(1) Adjust the reaction of bouillon to be 

 slightly alkaline. 



(2) Add an equal amount of sterile ascitic 

 fluid to sterile (1). 



Sterilization: Method of sterilization of 

 bouillon or ascitic fluid not given. 



Use: Cultivation of gonococci and other 

 highly pathogenic organisms. 



Variants : 



(a) The author mixed bouillon with § its 

 volume of ascitic fluid. 



(b) Elser and Huntoon isolated meningo- 

 cocci from blood or spinal fluid in a 

 mixture of 1000.0 cc. of ascitic fluid 



