CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



597 



(c) Hagemeister cultivated pathogenic 

 trypanosomes in the following 

 medium: 



(1) Infuse one pound of finely chopped 

 beef or horse meat with one liter 

 of water for 24 hours. 



(2) Boil for one hour. 



(3) Filter thru a towel. 



(4) Make up to 1 liter by the addition 

 of water. 



(5) Add 10.0 g. peptone (Sice), 20.0 g. 

 agar and 5.0 g. NaCl to (4). 



(6) Heat to boiling, stirring con- 

 stantly. 



(7) Neutralize by the addition of N/1 

 NaOH using litmus as an in- 

 dicator. 



(8) Add 5.0 cc. of N/1 NaOH per liter. 



(9) Heat in the autoclave at 100°C. 

 for one hour. 



(10) Filter thru a double filter or cotton 

 and distribute in sterile tubes. 



(11) Sterilize in the steamer for one 

 hour on each of three successive 

 days. 



(12) Add 2 or 3 times the volume of 

 sterile defibrinated blood to each 

 tube of agar, melted and cooled 

 to 60°C. Mix well. 



(13) Solidify in a slanted position. 



(14) Seal the tubes with paraffin after 

 inoculation. 



(d) Warden isolated and cultivated gono- 

 cocci on the following medium: 



(1) Dissolve 22.5 g. agar in 1000.0 cc. 

 of salt free broth. 



(2) Neutralize to phenolphthalein. 



(3) Add 25.0 cc. of sterile defibrinated 

 rabbit blood at 60°C. to 1000.0 cc. 

 of sterile (2) cooled to 60°C. 

 (The blood may be omitted.) 



Warden reported that growth was 

 better if agar was completely dry 

 (in desiccators over H2SO4) before 

 inoculation. 



(e) Holman studied hemolysis by strep- 

 tococci on a medium prepared as 

 follows : 



(1) Plain infusion agar (+0.6) is 

 sterilized in 100.0 cc. quantities 

 in flasks. 



(2) Heat a flask of (1) in the autoclave 

 and place in the paraffin oven at 

 5S°C. for some time. 



(3) Cool the flask to 50°C. and thoroly 

 mix 5.0 cc. of defibrinated human 

 blood with the fluid agar. 



(4) Pour this mixture into petri dishes 

 for blood agar plates on the surface 

 of previously prepared agar. 



(5) The agar for (4) is prepared by 

 dissolving 1.5% agar in normal 

 saline (0.85% NaCl), filtered, 

 tubed and sterilized. 



(6) Pour the blood agar mixture (3) 

 on the plain agar (5) to a depth of 

 about 2 millimeters. 



He reported that it was advisable to 

 add the serum with the red blood cells 

 as the serum is an important aid in 

 hemolysin production. 



(f) Kohman studied the oxygen tension 

 of meningococci on the following 

 medium : 



(1) Prepare a 2.5% meat infusion agar. 



(2) Dissolve 10.0 g. of Bacto peptone 

 in 1000.0 cc. of (1). 



(3) Add 40.0 cc. of human defibrinated 

 blood to (2). 



(g) Bernstein and Lowe used the follow- 

 ing medium for the isolation of 

 influenza bacilli: 



(1) Prepare infusion agar with a 

 reaction of pH = 7.1. 



(2) Add gentian-violet so that the 

 content be 1:5,000, taking a con- 

 centrated alcoholic solution of 

 dye as unity. 



(3) Method of sterilization not given. 



(4) Add sterile defibrinated human 

 blood to sterile melted (3). 



(5) Pour in sterile plates and store in 

 refrigerator for use. 



(6) Streak surface with washed 

 sputum or material from naso- 

 pharynx. 



The author reported that pneumo- 

 cocci, streptococci and staphylococci 

 were nearly all inhibited. If ascitic 

 fluid be added, the gram + organisms 

 were not inhibited by the dye. 

 (h) Wahl, White and Lyall cultivated 

 influenza bacilli on a medium pre- 

 pared as follows: 



(1) Prepare neutral beef infusion agar. 



(2) To every 100.0 cc. of sterile (1) 

 add 5.0 cc. of sterile defibrinated 

 sheep blood at 45 to 50°C. 



