CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



699 



(5) Filter the whey thru filter paper 

 and then thru an asbestos filter. 



(6) Distribute in sterile flasks. 



(7) Sterilize by the addition of 1.5% 

 chloroform. 



(8) Pour in plates. 



(e) Klimmer solidified whey, prepared 



from milk by coagulating the casein 



at 40^C. in the presence of rennet, 



with agar. 



References: Stutzer and Hartleb (1897 p. 



403), Huss (1907 p. 58), Meier (1918 p. 



435), Fulmer and Grimes (1923 p. 585), 



Klimmer (1923 pp. 172,203). 



2163. Bronfenbrenner et al. Whey Agar 



Constituents : 



1. Water 200.0 cc. 



2. Milk 100.0 cc. 



3. CR indicator 1.5 cc. 



4. Agar (3.0%) 6.0 g. 



Preparation : 



(1) Use fresh milk. Syphon the milk 

 from under the cream. 



(2) Bring it to boiling and add 2.5 cubic 

 centimeters of 10.0% MnCU solution 

 to each 100 cubic centimeters of milk. 



(3) Cool the mixture as soon as clot is 

 formed and filter thru a single layer 

 of cloth. 



(4) Titrate an aliquot portion hot and 

 adjust the bulk of medium to neutral 

 reaction (1 xlO~^). 



(5) Bring quickly to boiling, cool and 

 filter thru paper. 



(6) At this time prepare an agar jell of 

 3.0% concentration in plain water, 

 sterilize and dilute with an equal 

 volume of neutralized milk-whey. 



(7) While the mixture is hot add CR 

 indicator at the rate of 0.5 cubic 

 centimeters for each 100 cubic centi- 

 meters of medium distributed into 

 sterile tubes. 



Sterilization: Method of sterilization of 

 agar solution (see step (6) above) not 

 given. Sterilize the final medium for 10 

 minutes at 15 pounds pressure. 



Use: Cultivation of colon-typhoid group. 

 Authors reported that the medium gave 

 as good results as lactose peptone agar 

 as a medium for colon-typhoid bacteria 

 and was cheaper and more easily 

 prepared. 



Reference: Bronfenbrenner, Davis and 

 Morishima (1918-19 p. 347). 



2164. Ayres' Casein Agar (Tanner) 



Constituents: 



1. Water 1000.0 cc. 



2. Casein 10.0 g. 



3. NaOH (N/1) 7.0 cc. 



4. Agar 10-0 S- 



Preparation : 



(1) Add 10.0 g. casein (Eimer and Amend 

 C.P. casein prepared according to 

 Hammersten) and 7.0 cc. of normal 

 NaOH to 300.0 cc. of water. 



(2) Boil'until solution is complete. It is 

 desirable to allow the mixture to 

 stand several hours. 



(3) Make up to 500.0 cc. volume. 



(4) Bring the reaction between -|-0.1 and 

 +0.2 Fuller's scale. Do not allow the 

 solution to become alkaline to phenol- 

 phthalein or over +0.2. 



(5) Filter. 



(6) Dissolve 10.0 g. agar in 500.0 cc. of 

 water. 



(7) Filter. 



(8) Mix (5) and (7). 



(9) Tube. 



Sterilization: Sterilize in the autoclave 

 under pressure for 20 minutes and cool the 

 tubes in cold water or ice water. 



Use: General culture medium. Harvey 

 cultivated soil bacteria on a medium 

 prepared in a similar manner. 



Variants : 



(a) Harvey prepared the medium as 

 follows: 



(1) Mix 10.0 g. casein with 100.0 cc. 

 distilled water. 



(2) Add N/1 sodium hydroxide 7.0 cc. 



(3) Steam until the casein is dissolved. 



(4) Dissolve 10.0 g. agar in 900.0 cc. 

 distilled water. 



(5) Mix the agar solution with the 

 casein solution. 



(6) Filter thru thick filter paper. 



(7) Adjust the reaction to 1.5% acid to 

 phenolphthalein. 



(8) Distribute into test tubes. 



(9) Sterilize in autoclave. 

 Reference: Tanner (1919 p. 70), Harvey 



(1921-22 p. 96). 



