CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



629 



growth occurred. Crystals of salts of 

 fatty acids could not be demonstrated. 

 Variants: The author gave the following 

 variants : 



(a) Used beef fat instead of lanolin. 



(b) Vierling reported that on the follow- 

 ing medium the fat was saponified 

 causing the agar to become turbid 

 and the crystalline salts of fatty acids 

 to form. The layer of fat was not 

 destroyed in any place. 



(1) Pour a thin layer of beef fat into 

 Petri dishes. 



(2) When the fat has solidified pour a 

 layer of glucose agar that has been 

 cooled to 45°C. over the layer 

 of fat. 



Reference: Vierling (1920 p. 205). 



1969. Abe's Meat Water Infusion Agar 

 Constituents: 



1. Water 1000.0 cc. 



2. Nutrient agar (2.0%) 



3. Beef 500.0 g. 



Preparation : 



(1) Chop 500.0 g. of fat free beef in a 

 meat chopping machine. 



(2) Mix (1) and 1000.0 cc. of water, and 

 allow to stand for 18 to 24 hours in 

 an ice box. 



(3) Filter the liquid thru filter paper 

 and then thru a Chamberland filter. 



(4) Distribute this sterile reddish liquid 

 into sterile test tubes. 



(5) Melt tubes of 2.0% sterile nutrient 

 agar. 



(6) Heat (4) to 45-50°C. 



(7) To each 5.0 cc. of (5) add 2.0 cc. of (6). 

 (S) Mix thoroly. 



(9) Pour into sterile Petri dishes. 



Sterilization: Method of sterilization of 

 nutrient agar not given. The infusion is 

 sterilized by filtration, see step (3) above. 



Use: Cultivation of gonococci. Author re- 

 ported that gonococci colonies appeared 

 after 18 hours at 37°C. as light gray small 

 typical colonies. Kligler used a similar 

 medium to study effect of extracts on 

 growth of pathogenic organisms. 



Variants : 



(a) Kligler prepared media as given 

 below to study effect of extracts on 

 growth of pathogenic organisms: 

 (1) Prepare nutrient agar. 



(2) Mascerate equal weights of fresh 

 lean beef heart. 



(3) Take up each portion of (2) with 

 9 volumes of saline solution. 



(4) Treat each of these suspensions in 

 one of the following ways: 



(a) Keep in the ice box over night 

 and steam in an Arnold sterilizer 

 for one hour. 



(b) Keep in the ice box over night and 

 filter thru a Berkefeld candle. 



(c) Keep at 55°C. over night and heat 

 in an Arnold sterilizer for one 

 hour. 



(d) Keep at 55°C. over night and 

 filter thru a Berkefeld candle. 



(e) Extract by boiling one hour and 

 then sterilize in an Arnold steri- 

 lizer for one hour. 



(f) Extract by boiling for one hour. 



(5) Take 1.0 cc. of one of the extracts 

 obtained and add to 5.0 cc. of sterile 

 nutrient agar. 



Kligler reported that heat destroyed 

 the growth stimulating material in 

 the extract. Best growth in (4) (b), 

 no stimulated effects in (4) (e) 

 and (f). 

 (b) Kligler also used the following media 

 to study the effects of growth acces- 

 sory substances on pathogenic bac- 

 teria: 



(1) Prepare nutrient agar. 



(2) Obtain beef heart, rabbit tissues, 

 rabbit organs, cat organs, cat tis- 

 sues or mucosa of various organs 

 as free from blood as possible under 

 aseptic conditions. 



(3) Wash (2) with saline solution to 

 remove all visible traces of blood. 



(4) Weigh and chop into small bits and 

 suspend in 9 times the weight of 

 saline solution. 



(5) Shake thoroly and place in the ice 

 box over night. 



(6) Centrifuge and filter thru a Berke- 

 feld candle. 



(7) Test sterility before using. 



(8) Tube (1) in 5.0 cc. lots. 



(9) Add amounts of (7) to each tube of 

 (8) varying from 0.01 cc. to 1.0 cc. 



References: Abe (1907 p. 707), Kligler 

 (1919 pp. 32, 43). 



