368 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



Added nutrients: The author used 1.0% 

 glucose, 1.0% lactose, 2.0% sucrose, 2.0% 

 mannitol, etc. 



Variants: The author suggested the use of 

 Czaplewski's alkaline serum (10.0 cc. 

 normal NaOH per 100.0 cc. serum) instead 

 of the alkaline serum given above . Czap- 

 lewski's mixture may be sterilized in the 

 autoclave. 



Reference: Leuchs (1920 p. 1415), Klein 

 (1920, p. 297). 



1256a. Martin, Pettit and Vaudremer's 

 Serum Solution. 



Constituents: 



1. Physiological salt solution. 1000.0 cc. 



2. Serum, bovine 100.0 cc. 



Preparation : 



(1) Dilute bovine serum 1 to 10 with 

 physiological salt solution. 

 Sterilization: Not given in the abstract. 

 Use: Cultivation of Spirochaeta ictero- 



haemorrhagiae. 

 Variants : 



(a) Rabbit serum diluted 1:16 proved 

 to be a better medium. 



(b) Griffith cultivated Spirochaeta 

 icterohaemorrhagiae and other or- 

 ganisms on a medium prepared by 

 diluting one part beef serum with two 

 parts physiological salt solution and 

 heating at 70°C. until the mixture 

 became slightly viscous. He covered 

 the medium with a thin layer of 

 paraffin oil after inoculation. 



(c) Harvey mixed 5 parts sterile 0.85% 

 NaCl solution with one part rabbit 

 serum heated at 56°C. for 30 minutes. 



(d) Harvey mixed 1 part ox serum heated 

 to 56°C. for 30 minutes with 9 parts 

 sterile 0.85% NaCl solution, or a 

 sterile solution obtained by dissolv- 

 ing 9.2 g. NaCl, 0.05 g. NajCOa, 

 0.1 g. KCl, 0.1 g. CaCh and 10.0 g. 

 sodium citrate in a liter of water. 

 This solution is Locke's solution. 



(e) Hogue cultivated Spirochaeta eury- 

 grata on a medium prepared as 

 follows: 



(1) Dilute serum (best results obtained 

 with pig serum) 1:4. 



(2) Tube 0.85%, sterile salt solution in 

 15.0 cc. lots in sterile tubes. 



(3) AddO.3 cc. of (1) to each tube of (2). 



(4) A pH = 7.0 gave the best results. 



(5) Cover with a layer of paraffin oil. 

 (f) Stitt cultivated Balantidium coli 



intestinal protozoa and amoeba in a 

 medium prepared as follows: 



(1) Mix one part inactivated human 

 blood serum with 16 parts 0.5% 

 salt solution. 



(2) The reaction is faintly alkaline to 

 litmus. 



(3) Place 8.0 cc. in tubes having a 

 diameter of 10 mm. and a length of 

 150 mm. giving the medium a depth 

 of about 100 mm. 



(4) Sterilization not specified. 



(5) Inoculate with 0.1 cc. of undiluted 

 feces containing mucus, by means 

 of the tube capillary pipette into 

 the bottom of the tube. 



References : Martin, Pettit and Vaudremer 

 (1917 p. 197), Griffith (1919-20 p. 60), 

 Harvey (1921-22 p. 79), Hogue (1922 p. 

 619), Stitt (1923 p. 52). 



1257. Davis' Serum Medium. 



Constituents: 

 1. Serum, rabbit. 



Preparation : 



(1) Tube sterile rabbit's blood serum into 

 sterile test tubes. 



Sterilization: Method not given. 



Use: Used in uncoagulated form by Davis 

 to cultivate the Ducrey bacillus (chan- 

 croid bacillus). Other investigators cul- 

 tivated spirochetes, pneumococci, 

 streptococci, etc., in similar media. 



Variants : 



(a) Longcope cultivated pneumococci 

 and streptococci in a medium pre- 

 pared as follows: 



(1) Collect 20.0 cc. of blood from the 

 arm vein and allow to clot in a cool 

 place. 



(2) Draw off the serum after from 24 to 

 48 hours. 



(3) Use from 2 to 5.0 cc. serum as 

 culture medium. 



(b) Schereschewsky heated horse serum 

 at 60°C. until it was brought to a 

 gelatinous consistency, then incu- 

 bated at 37°C. until partial autolysis 

 was effected. The medium was used 

 for the cultivation of the syphillus 

 spirochaete. 



