288 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



pass thru a Berkefeld filter but were 

 absorbed by charcoal. Broth containing 

 extract alone, however, did not give con- 

 tinued growth of the influenza bacilli. 

 Primary inoculations were naade from 

 blood broth. 

 Reference: Thjotta and Avery (1921 

 p. 100). 



941. Thjotta Pea or Bean Bouillon 



Constituents : 



1. Distilled water 100.0 cc. 



2. Green peas or beans 100.0 g. 



3. Plain broth 

 Preparation : 



(1) Preparation of bouillon not given. 



(2) Adjust the reaction to pH = 7.8. 



(3) Flame the surface of green pea or 

 string bean pods. 



(4) Open the pods with sterile forceps 

 and crush out the separate seeds into 

 different sterile dishes. 



(5) Add an equal weight of distilled 

 water to the emulsion and adjust the 

 reaction of pH = 4.6. 



(6) Boil 10 minutes. 



(7) Strain thru glass wool and filter thru 

 a Berkefeld filter (N). 



(8) Test sterility, and store on ice until 

 ready for use. Readjust the reaction 

 from pH = 7.3 to 7.5. 



(9) Dilute each of the extracts in (2) 

 in the proportions 1:10 and 1:100. 



Sterilization: Sterilization of pea or bean 

 extract effected in steps (6) and (7) in 

 preparation. Sterilization of bouillon 

 not given. 



Use: To study bacterial nutrition of 

 Bacillus influenzae. Author reported 

 that the medium supported growth of 

 B. influenzae. Growth accessory ma- 

 terials resisted boiling for 10 minutes, 

 were destroyed by autoclaving, would 

 pass thru a Berkefeld filter, but were 

 absorbed by charcoal. Broth containing 

 extracts alone, however, did not give 

 continued growth of the influenza bacilli. 

 Primary inoculations were made from 

 blood broth. 



Reference: Thjotta and Avery (1921 

 p. 100). 



942. Thjotta and Avery's Potato Bouillon 



Constituents: 

 1. Distilled water 400.0 cc. 



2. Brewers yeast 100.0 g. 



3. Plain broth 



4. Potato 

 Preparation : 



(1) Preparation of bouillon not given. 



(2) Adjust the reaction of (1) to pH 



= 7.8. 



(3) Emulsify 100.0 g. of brewers yeast 

 in 400.0 cc. of distilled water with 

 the reaction of pH = 4.6. 



(4) Boil over a free flame for 10 minutes. 



(5) Allow to sediment at room tem- 

 perature. 



(6) Pipette off the clear supernatant 

 extract and test sterility. 



(7) Store in ice chest until ready for use. 

 Readjust the reaction of the extract 

 from pH = 7.3 to 7.5 just before use. 



(8) Procure sterile pieces of potato from 

 the center of an old potato without 

 abrasions or imperfections. Take 

 all possible precautions to remove 

 the pieces in as asterile a condition 

 as possible. 



(9) Distribute the sterile pieces of 

 potato into (2). 



(10) Add yeast extract (7) amount not 

 given. 



Sterilization: Sterilization of yeast extract 

 effected in step (4) in the preparation. 

 Sterilization of bouillon not given. 



Use: To study bacterial nutrition of Bacil- 

 lus influenzae. Author reported that 

 growth in unheated potato occurred 

 equally well without the addition of yeast 

 extract. When the potato was heated at 

 120° for 45 minutes no growth occurred 

 unless the yeast extract was added. 

 Medium did not give continued growth. 

 Initial inoculation made from blood 

 broth. 



Variants: The authors omitted the yeast 

 extract. 



Reference: Thjotta and Avery (1921 

 p. 109). 



943. Thjotta's Bacterial Emulsion Bouillon 



Constituents : 



1. Bouillon. 



2. Bacterial emulsion. 

 Preparation : 



(1) Prepare bouillon with a reaction of 

 pH = 7.8. 



(2) Grow bacteria (mucoid bacillus 

 (Friedlander's), Bacillus ozoenoe and 



