CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



607 



Preparation : 



(1) Prepare meat infusion agar and adjust 

 to a reaction of +0.2% to phenol- 

 phthalein. 



(2) Dilute serum with distilled water 1:3. 



(3) Add 6.0 cc. of (2) to each 100.0 cc. of 



(4) Add under aseptic conditions a suffi- 

 cient amount of a sterile solution of 

 one of the added nutrients so that it 

 be present in 2.0% strength in 

 sterile (3). 



(5) Add Andrade's indicator. 



(6) Distribute directly into sterile test 

 tubes in 8.0 cc. lots and slant and 

 inoculate as in Russell's medium. 



(7) Incubate to determine contamination. 

 Sterilization: Sterilize (3) in the autoclave. 



Method of sterilization of solutions of 

 added nutrients not given. 



Use: To determine fermentation of carbo- 

 hydrates by streptococci. Authors re- 

 ported that this medium supported the 

 growth of pneumococci and quite a num- 

 ber of the delicate growing streptococci. 



Added nutrients: The authors added 2.0% 

 of any desired carbohydrate. 



Reference: Sherwood and Downs (1919 

 p. 135). 



1915. Harvey's Placenta Blood Serum Agar 



Constituents: 



1. Infusion agar. 



2. Serum, Placental blood. 

 Preparation: (1) Mix equal parts of sterile 



or sterilized placental blood at 45°C. 

 with infusion agar at 45°C., 1.0% acid to 

 phenolphthalein. (See variant (v) me- 

 dium 1661 for preparation.) 



Sterilization: Not specified. 



Use: General culture medium. 



Reference: Harvey (1921-22 p. 78). 



1916. Veillon's Serum Agar 



Constituents : 



1. Infusion agar 100.0 cc. 



2. Serum (human) 100.0 cc. 



Preparation : 



(1) Prepare infusion agar and cool to 

 40 °C. 



(2) Heat serum at 38 to 40°C. 



(3) Mix equal parts of (1) and (2). 

 Sterilization: Not specified. 



Use: Cultivation of gonococci. Other in- 

 vestigators used similar media for the 

 cultivation of a large variety of organisms. 

 Variants: The following authors prepared 

 media as indicated below: 

 (a) Joos used the following medium for 

 the diagnosis of diphtheria. He re- 

 ported that diphtheria colonies may 

 appear after 4-5 hours as small grey 

 white colonies, of moist appearance 

 which can easily be recognized. 

 Streptococci and staphylococci were 

 inhibited. Serum need not be col- 

 lected under aseptic conditions. 



(1) Extract 500.0 g. beef (several days 

 old) with 1000.0 cc. water. 



(2) Filter and dissolve 20.0 g. peptone 

 and 5.0 g. NaCl in (1). 



(3) Make alkaline with N/1 NaOH 

 by adding (about 7.0 to 8.0 cc. of 

 normal solution per liter) to neu- 

 tral medium. 



(4) To 300.0 cc. of blood serum add 

 50.0 cc. of N/1 NaOH solution and 

 150.0 cc. of (3). (Distilled water 

 may be used instead of (3). 



(5) Place the mixture in a flat bot- 

 tomed flask and place the flask in 

 a water bath at about 60° to 70 ""C. 

 for 2 to 3 hours. 



(6) Raise the temperature to 100°C., or 

 better, place in a steamer for 30 

 to 45 minutes. 



(7) Then add an equal amount 

 (500.0 cc.) of (3) and 20.0 g. of 

 agar agar. 



(8) Dissolve the agar as quickly as 

 possible. 



(9) When solution is complete, filter 

 in hot condition and sterilize for 

 15 minutes at 100 to 110°C. in an 

 autoclave. 



(10) Pour into sterile Petri dishes. 



(b) Behrens cultivated Trypanosoma 



Brucei on the following medium: 



(1) Digest chopped beef in 250.0 cc. 

 water over night in the cold, or for 

 one hour at 55°C. 



(2) Strain (1), boil the extract and 

 filter. 



(3) Dialyze the filtrate in a large col- 

 lodium sac against running dis- 

 tilled water for 24 to 48 hours. 



