294 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



Variants: Kraus, Zia and Zubrzciky incu- 

 bated the flasks for 3 hours at 50°C. for 

 24 hours at 37°C. before use. 



References : Dieudonnc (1909 p. 108), Kraus, 

 Zia and Zubrzciky (1911 pp. 1084-1085). 



960. Orcutt and Howe's Fat Blood Bouillon 



Constituents : 



1. Bouillon. 



2. Blood, whole defibrinated. 



3. Cream. 

 Preparation : 



(1) Distribute sterile bouillon in 1.0 cc. 

 lots in sterile test tubes. 



(2) Add 1.0 cc. of defibrinated blood to 

 each tube. 



(3) Allow milk to stand in sterile test 

 tubes. 



(4) Remove the cream for (3). 



(5) Dilute sterile (3) 1:10 with sterile 

 bouillon. 



(6) Add 0.02 cc. of (5) to each tube of (3). 

 Sterilization: Sterilization of bouillon not 



specified. Sterilize (4) by steaming at 

 100°C. for 20 minutes on each of 3 suc- 

 cessive days. 



Use: To study the influence of fat on 

 hemolysis by staphylococci. Authors 

 reported that with the organism studied 

 hemolysis took place in the presence of 

 whole milk, cream, butter, olive oil and 

 triolein, but not in the presence of fat 

 free milk, nut butter, triacetin, tri- 

 butyrin or pork fat. 



Variants: The authors used one of the 

 following instead of cream: 



(a) Remove the cream from milk by 

 centrifugation. Add 0.01 cc. of the 

 milk, after it has been sterilized by 

 the fractional method in steam, to 

 each tube of (3) above. 



(b) Make a suspension of one of the fats 

 (olive oil, butter, nut butter, tri- 

 acetin, triolein, tributyrin and pork 

 fat were used) by shaking 1.5 cc. of 

 the fat in 10.0 cc. bouillon for one to 

 2 hours using a mechanical shaker. 

 Add sufficient of one of these emul- 

 sions to bouillon so that each cubic 

 cm. of bouillon shall contain 0.002 

 cc. of fat. 



Reference: Orcutt and Howe (1922 p. 412). 



961. Fildes' Pepsinized Blood Bouillon 



Constituents : 



1. Bouillon 



2. Saline solution 150.0 cc. 



3. Blood, defibrinated, sheep... 50.0 cc. 

 Preparation : 



(1) Place 150.0 cc. of saline solution, not 

 distilled water, 6.0 cc. pure HCl, 

 50.0 cc. defibrinated sheep blood and 

 1.0 g. pepsin (B.P. granulated) in a 

 250.0 cc. bottle fitted with a well 

 ground glass stopper. Add ingre- 

 dients in order specified. 



(2) Shake thoroughly. 



(3) Place in a water bath at 55 °C. for 2 

 to 24 hours, shaking occasionally at 

 first. 



(4) Add 12.0 cc. of 20.0% NaOH. 



(5) Adjust the reaction to pH 7.6 using 

 cresol red as an indicator (per- 

 manganate color). 



(6) Add pure HCl, drop by drop until 

 pH 7.0 to 7.2 is reached, using phenol 

 red as an indicator (red). 



(7) Add 0.25% chloroform and dissolve 

 by shaking. 



(8) When ready for use, shake the flask 

 and remove the pepsinized blood with 

 a sterile pipette. Add from 2.0 to 

 5.0% to bouillon. 



Sterilization: Not specified. 

 Use: Cultivation of B. influenza. 

 Reference: Fildes (1920 p. 129). 



962. Dunham's Glucose Serum Bouillon 



Constituents : 



1. Water 300.0 cc. 



2. Bouillon. 



3. Glucose 10.0 g. 



4. Serum (sheep) 100.0 cc. 



5. Litmus (1.0% solution of Kahlbaum) 

 Preparation : 



(1) Prepare bouillon. 



(2) Dissolve 10.0 g. of glucose in (1). 



(3) Dilute 1 part sheep serum with 3 

 times its volume of water. 



(4) Mix 1 part of sterile (3) with 3 parts 

 sterile (2). 



(5) Add a sufficient quantity of sterile 

 1.0% Kahlbaum's purified litmus 

 solution to impart a distinct color to 

 the whole. 



