576 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



Mix the ascitic fluid and agar at as 

 low a temperature as possible. This 

 aids the production of a transparent 

 medium. 

 Sterilization: Sterilize (10) in the auto- 

 clave. 

 Use: Cultivation of meningococci and 



other pathogenic cocci. 

 Variants : 



(a) Fildes substituted serum for ascitic 

 fluid. The serum was prepared as 

 follows: 



(1) Collect horse or ox blood in tall 

 sterile jars from the slaughter 

 house. 



(2) Allow to stand until all the cor- 

 puscles have deposited. It may 

 require 14 days for this to take 

 place. After 5 days examine the 

 jars and pipette or siphon off the 

 clear serum. Do not disturb the 

 sediment. 



(3) Distribute the clear serum in 200.0 

 cc. lots in perfectly fitting glass 

 stoppered flasks. 



(4) Add chloroform to each bottle 

 until 0.5% chloroform has been 

 added. 



(5) Add a drop of sterile oil to the 

 stopper and fasten a dust cover 

 tightly over the stopper. 



(6) Place the bottles ip the water 

 bath for one hour at 45°C. Shake 

 occasionally. 



(7) When the bottles have cooled, 

 remove a sample under aseptic 

 conditions and mix with agar. 

 Incubate the mixture at 37°C. for 

 five days to test sterility. 



(b) Fildes also prepared the medium as 

 follows: 



(1) Collect ox or horse blood from a 

 slaughter house. 



(2) Defibrinate the blood by strirring 

 with a sterilized large wooden 

 stick wrapped in gauze. 



(3) Lake the blood by the addition of 

 an equal volume of distilled water. 



(4) Distribute in 200.0 cc. lots into 

 perfectly fitting glass stoppered 

 bottles. (The author mentions 

 that the medium should contain 

 no suspended material, or it will 



not sterilize with the method 

 employed.) 



(5) Add chloroform to each bottle 

 until 0.5% chloroform has been 

 added. 



(6) Add a drop of sterile oil to the 

 stopper and fasten a dust cover 

 tightly over the stopper. 



(7) Place in the air incubator at 37 °C. 

 for 24 hours, shaking constantly. 



(8) After this time, remove a sample 

 under aseptic conditions and test 

 its sterility by mixing it with agar 

 and incubating. (Time not speci- 

 fied.) If sterile do not heat longer 

 for heating tends to darken the 

 medium. 



(9) Prepare ordinary lemco (or meat 

 infusion) agar containing 2.5 to 

 3.0% agar. 



(10) Measure (9) into 200.0 cc. flasks in 

 150.0 cc. lots. 



(11) Autoclave (time and pressure not 

 given). 



(12) Add 25.0% of sterile laked blood 

 (8) to (11). (Method of distribu- 

 tion not specified.) 



Reference: Fildes (1917 p. 492). 



1844. Haner and Frost's Milk Body Fluid 

 Agar 



Constituents : 



1. Nutrient agar 1.0% 

 (dehydratedDif co) . 1000.0 cc. 



2. Milk, sterile 1000.0 cc. 



3. Serum (horse or 



rabbit 6,0 to 12.0%). 120.0 to 240.0 cc. 

 Preparation : 



(1) Prepare 1.0% agar from Difco de- 

 hydrated product. 



(2) Add equal part of sterile milk to 

 sterile (1). 



(3) Add 6.0% to 12.0% of sterile horse or 

 rabbit serum to (2). 



Sterilization: Method of sterilization not 



given. 

 Use: Cultivation of pneumococci and 



streptococci. 

 Variants: The authors used whole blood 



instead of serun. 

 Reference: Haner and Frost (1921 



p. 270). 



