CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



609 



(7) Cool and solidify in slanting 

 position. 



(8) Inoculate tubes and place in a 

 10.0% atmosphere of CO2. 



(i) Kinsella, Brown and Garcia used the 

 following medium for the isolation of 

 gonococci: 



(1) Prepare 1.6% beef or veal infusion 

 agar. 



(2) Adjust to pH = 7.6. 



(3) Distribute into 100.0 cc. quanti- 

 ties in Erlenmeyer flasks and 

 sterilize (method not given). 



(4) When ready for use melt (3) and 

 add to each flask containing 

 100.0 cc. of hot agar (90-100°C.) 

 30.0 cc. of (sterile) beef seriun. 



(5) Mix thoroly, distributing the small 

 particles of coagulated serum thru 

 the medium. 



(6) Pour into sterile Petri dishes, 

 cover with porous terracotta lids 

 and place in incubator over night 

 (18 hours). 



(7) Replace terra-cotta lids by glass 

 lids and the medium is read for 

 inoculation. 



Authors reported that gonococci 



reached maximum growth in 48 hours. 



The type of serum used had little or 



no influence on the value of the 



medium. Plates should never dry 



under the terra-cotta lids longer than 



24 hours. 



References: Veillon (1898 p. 22), Joos 



(1899 p. 303), Behrens (1914 p. 29), Ferry 



and Davis (1918 p. 295), Ball (1919 p. 82), 



Harvey (1921-22 pp. 79, 80, 82), Jones 



(1922 p. 363), Fitch (1922 p. 234), Kinsella, 



Brown and Garcia (1923 p. 1). 



1917. Kutscher's Serum Placenta Agar 



Constituents: 



1. Water 1000.0 cc. 



2. Placenta 500.0 g. 



3. Agar 25.0 g. 



4. NaCl 5.0 g. 



5 Glucose 10.0 g. 



6. Xutrose 20.0 g. 



7. Peptone (Chapoteaut) 20.0 g. 



8. Serum (Beef) 335.0 cc. 



Preparation : 



(1) Cut fresh placenta into small pieces. 



(2) Weigh the pieces and juice and add a 

 double weight of water. 



(3) Prepare 2.5% agar in the usual way, 

 adding 0.5% NaCl, 1.0% glucose, 

 2.0% nutrose and 2.0% Chapoteaut's 

 peptone. The reaction is slightly 

 alkaline. 



(4) Distribute in 100.0 cc. lots. 



(5) To 3 parts sterile (4) add 1 part sterile 

 beef serum. 



(6) Mix well and pour into plates. The 

 reaction is slightly alkaline. 



Sterilization: Method of sterilization of (4) 

 not given. Sterilize beef serum in 

 50.0 cc. lots by placing it at 60''C. for one 

 hour on 4 successive days. 



Use: Cultivation of meningococci. Author 

 reported that after 18-24 hours at 37°C. 

 meningococci colonies were circular, light 

 transparent, grayish-blue and about 

 2-3 mm. in diameter. 



Variants : 



(a) Scott gave the following method of 

 preparation: 



(1) Mince the placenta and prepare a 

 boiled extract as with meat (exact 

 method not given). 



(2) Add 2.0% nutrose, 1.0% glucose, 

 1.5% peptone (Chapoteaut) and 

 2.5% agar. Dissolve by steaming. 



(3) Adjust the reaction to a -1-8 (Eyre). 



(4) Flask in 75.0 cc. lots. 



(5) Sterilize (method not given). 



(6) When ready for use melt and cool 

 to 50 °C. 



(7) Add 25.0 cc. of sterile (filtered) ox 

 serum to each flask. 



(8) Mix and pour at once into plates. 



(b) Klimmer specified the use of 2.0% 

 agar instead of 2.5%. 



References: Kutscher (1908 p. 287), Scott 

 (1915-17 p. 466), Harvey (1921-22 p. 79), 

 Klimmer (1923 pp. 202, 225). 



1918. Bezanfon's Serum Placenta Agar 



Constituents : 



1. Water 1000.0 cc. 



2. Placenta 500.0 g. 



3. Agar 25.0 g. 



4. Glucose 10.0 g. 



5. Peptone 20.0 g. 



6. NaCl 5.0 g. 



7. Serum 300.0 cc. 



Preparation : 



(1) Soak 500.0 g. of finely chopped pla- 

 centa in 1000.0 cc. of water in the 

 ice box over night. 



