588 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(18) Tube and slant. 



(19) Leave the tubes in a slanted position 

 for 72 hours at room temperature 

 until the contents have well set. 

 Then seal the tubes with cork and 

 paraffin. 



Sterilization: See under preparation. 

 Use: Cultivation of B. tuberculosis. 

 Reference: Roddy (1917 p. 46). 



1870. Chapin's Egg Yolk Urine Agar 



Constituents : 



1. Meat infusion 1000.0 cc. 



2. Urine 200.0 cc. 



3. Peptone 50g 



4. N/1 H2SO4 (1.2%) 12.0 cc. 



5. Agar to solidify 



6. Pea flour (0.5%) 5.0 g. 



7. Glucose (0.5%) 5.O g. 



8. Egg yolk (5.0%) 50.0 cc. 



Preparation : 



(1) Prepare meat infusion. 



(2) Heat to remove coagulated proteid. 



(3) Add 2, 3, and 4 (a normal dilution of 

 H2SO4, 1.2%). 



(4) Divide into 5 flasks and add varying 

 amounts of N/1 NaOH, 0.0, 1. 2/2.8, 

 3.7 and 4.7% respectively having the 

 following reactions: 3.4, 2.6, 1.8, 

 0.5 and alkaline. 



(5) Add a sufficient quantity of agar, 

 0.5% of 6 and 0.5% of 7. 



(6) Add 5.0% egg yolk to the cooled 

 medium just before pouring the 

 plates. 



Sterilization: Method not specified. 



Use: To study influence of CO2 and reac- 

 tion of medium on cultivation of gono- 

 coccus. The author incubated the 

 cultures in a 10,0% CO2 atmosphere. He 

 reported good growth in the medium con- 

 taining 4.7% NaOH the reaction being 

 alkaline, and also in media whose reac- 

 tions were -|-2.6 and 1.8. A lighted candle 

 left in the jar beside the plates at the time 

 of sealing the chamber gave a favorable 

 atmosphere. 

 Reference: Chapin (1918 p. 342). 



1871. Harvey's Sucrose Egg Agar 



Constituents : 



1. Sucrose (1.0%,) Agar 150.0 cc. 



2. Water 10.0 cc. 



3. NasCOs (6.5% Solution) 10.0 cc. 



4. Neutral red (0.5% soln.) 



(1-0%) l.Scc. 



5- Egg 10.0 cc. 



Preparation : 



(1) Mix the contents of one egg with an 

 equal quantity of water. 



(2) Add to this mixture an equal amount 

 of 6.5% anhydrous NasCOg. 



(3) Add 1 volume of the sterile mixture to 

 5 volumes neutral 1.0% sucrose agar. 



(4) Add to the mixture while melted 1.0% 

 freshly prepared 0.5% neutral red. 



(5) Slope or pour on plates. 

 Sterilization: Sterilize (2) for one hour at 



100°C. Method of sterilization of 1.0% 



sucrose agar not given. 

 Use: Isolation of cholera vibrio. Harvey 



reported that the medium had orange tint 



and V. cholerae colonies had deep red 



centers. 

 Reference: Harvey (1921-22 p. 86). 



1872. Leboeuf's Egg White Liver Infusion 



Agar 

 Constituents : 



1- Water looO.O cc. 



2. Liver, horse or beef 500.0 g. 



3. Peptone (Chapoteaut) 20.0 g. 



4. Egg white 



5. Starch, potato 



6. Agar 

 Preparation : 



(1) Boil 500.0 g. of finely chopped horse 

 or beef liver in 1000.0 cc. of water 

 slowly for 90 minutes. 



(2) Filter thru paper and make up the 

 filtrate to the original volume. 



(3) Add 20.0 g. of Chapoteaut's peptone 

 and heat to boiling. 



(4) Neutralize to litmus. 



(5) Filter thru paper. 



(6) Mix one part egg white with ten 

 parts distilled water. 



(7) Boil (6) and filter. 



(8) Mix 100.0 cc. of sterile (7) with 

 1000.0 cc. of (5), and add 5.0 g. 

 potato starch. 



(9) Readjust the reaction to neutral if 

 necessary. 



(10) Add 20.0 g. of agar per 1000.0 cc. of 

 (9) and autoclave at 115°C. for 35 

 minutes. 



(11) Tube. 



