CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



293 



Preparation : 



(1) Mix the yolks of 2 eggs, the whites of 

 4 eggs and 12.0 cc. normal NaOH 

 in 1000.0 cc. of water. 



(2) Add 1 part (1) to 5 parts bouillon. 

 Sterilization: Method not specified. 

 Use: Cultivation of cholera organism. 

 Reference: Park, Williams and Krumwiede 



(1924 p. 126). 



957. Piorkowski's Alkaline Egg Albumin 

 Bouillon 



Constituents : 



1. Bouillon 1000.0 cc. 



2. Glucose (1.0%) 10.0 g. 



3. Peptone (2.0%) 20.0 g. 



4. Egg albumin (2.0% dry) 



5. NaOH 

 Preparation : 



(1) Prepare a 1.0% glucose bouillon con- 

 taining 2.0% peptone. 



(2) Distribute in 5.0 cc. lots. 



(3) Prepare a 2.0% solution of dry egg 

 albumin in tap water. 



(4) Add 1.0 cc. (3) and 20.0% of a N/10 

 NaOH solution. 



Sterilization: Method not given. 



Use: Enrichment of streptococci from the 



blood. 

 Reference: Piorkowski (1922 p. 69). 



958. Hibler's Blood Bouillon 



Constituents : 



1. Bouillon. 



2. Blood, dog. 

 Preparation : 



(1) Collect dog blood in sterile test tubes 

 in about 5 cc. lots. (Blood may be 

 collected in beaker and then the finely 

 divided clot with a corresponding 

 amount of serum, be distributed in 

 tubes. 



(2) Add a few drops of bouillon (or 

 water). 



Sterilization: Sterilize by heating at 58 or 

 97°C. Before use heat the tubes for 15 

 minutes in flowing steam. 



Use: Cultivation of anaerobes and various 

 parasitic forms. Author reported that 

 the anaerobes grew better in a medium 

 sterilized at 58°C. Fresh blood gave 

 best results. 



Variants : 



(a) Davis mixed one-third volume of 



fresh rabbit blood with two thirds 

 volume of bouillon to cultivate 

 Ducrey's bacillus (chancroid ba- 

 cillus), 

 (b) Wollstein cultivated Pfeiffer bacillus 

 on a medium prepared as follows: 



(1) Boil fresh rabbit blood for 2 minutes 

 in a water bath. 



(2) Centrifuge. 



(3) Add 0.5 cc. of the pale pink or yel- 

 low fluid resulting from (2) to 

 20.0 cc. of bouillon adjusted to 

 pH = 7.8. 



(c) Tanner and Besson added 0.5, 0.3 or 

 0.25 the volume of blood (or serum) 

 to bouillon. 



(d) Besson prepared a medium for the 

 cultivation of pneumococci, meningo- 

 cocci and gonococci as follows: 



(1) Collect 400.0 cc. of beef blood in a 

 sterile flask containing 30.0 cc. of a 

 1.0% ammonium citrate. 



(2) Shake thoroly. 



(3) Store for several hours at room 

 temperature. 



(4) Dilute (3) with 3 volumes of sterile 

 physiological salt solution. 



(5) Add 1 part (4) to 15 parts sterile 

 bouillon. 



(e) Thjotta and Avery boiled whole blood 

 for 10 minutes, centrifuged and 

 diluted the supernatant fluid 1 to 10, 

 to 1:10,000 with bouillon adjusted to 

 pH = 7.8. He reported no growth 

 with dilutions of 1:1000 or greater. 



References: Hibler (1899 p. 604), Davis 

 (1903 p. 405), Wollstein (1919 p. 556), 

 Tanner (1919 p. 47), Besson (1920 p. 31), 

 Thjotta and Avery (1921 p. 109). 



959. Dieudonne's Alkaline Blood Bouillon 



Constituents: 



1. Bouillon 500.0 cc. 



2. Blood (defibrinated beef).... 250.0 cc. 



3. KOH (normal) 250.0 cc. 



Preparation : 



(1) Add 250.0 cc. of normal KOH to 

 250.0 cc. defibrinated beef blood. 



(2) Adjust nutrient bouillon (or peptone 

 solution) to neutral point to litmus. 



Sterilization: Sterilization not specified. 



Author mentioned that (1) might be 



sterilized in the autoclave. 

 Use: Enrichment of cholera vibrio. 



