CULTURE MEDIA FOR CULTIVATION' OF MICROORGANISMS 



359 



(2) Cover brain material with Ringer's 

 solution (see medium 180). 



(3) Cover with sterile oil. 

 Sterilization: Method not given. 

 Use: Cultivation of Negri bodies. 

 Reference: Moon (1913 p. 233). 



1232. Weiss' Brain Medium 



Constituents: 



1. Water 1000.0 cc. 



2. Sheep brain 2000.0 g. 



Preparation : 



(1) Mix two parts finely ground sheep 

 brain and one part water. 



(2) Do not adjust the reaction. 

 Sterilization: Sterilize in the autoclave. 

 Use: To develop B. botulinus spores and to 



study thermal death rates of Clostridia. 



Variants: Tanner and McCrea mixed two 

 parts water and one part sheep brain. 

 The pH normally was from 6.8 to 7.0. 

 The medium was sterilized in the auto- 

 clave at 15 pounds pressure for 30 minutes 

 in 125 to 150 cc. lots in 250 cc. Erlenmeyer 

 flasks. 



References: Weiss (1921 p. 71), Tanner and 

 McCrea (1923 p. 270). 



1233. Zotta's Brain Infusion Solution 



Constituents: 



1. Saline solution (0.75% 



NaCl) 200.0 cc. 



2. Brain (beef) 100.0 g. 



Preparation : 



(1) Shake 100.0 g. of fresh beef brain 

 freed from its meninges with broken 

 glass until a homogenous mixture is 

 formed. 



(2) Add 200.0 cc. of saline solution 

 (0.75% NaCl) to (1) and mix 

 thoroly. 



(3) Filter thru gauze. 



(4) Heat for 20 minutes. 



(5) Tube the supernatant liquid. 

 Sterilization: Sterilize at 115°C. for 20 min- 

 utes. Final pH 6.9. 



Use: Cultivation of Leptomonns pyr- 



rhocoris. 

 Reference: Zotta (1923 p. 281). 



1234. Harvey's Heart Medium 



Constituents: 



1. Water 500.0 cc. 



2. Heart, beef 500.0 g. 



Preparation : 



(1) Cut up 500.0 g. of fat-free ox or horse 

 heart. 



(2) Add water to just cover the meat. 



(3) Cook thoroly and slowly in a closed 

 vessel over a small flame. 



(4) Pour off the liquid portion into a 

 beaker. 



(5) Mince the solid residue very finely. 



(6) Add the minced residue to the liquid 

 in the beaker. 



(7) Bring up the weight of the contents 

 of the beaker to 1000.0 g. 



(8) Break down any small lumps of meat 

 between the fingers. 



(9) Make faintly alkaline to litmus. 



(10) Distribute into test tubes. 



(11) Cover with a layer of paraffin. 

 Sterilization: Sterilize 45 minutes at 100°C. 

 Use: Cultivation of anaerobes. 

 Variants : 



(a) Stitt gave the following method of 

 preparation: 



(1) Grind 250.0 g. of beef heart muscle. 



(2) Suspend (1) in 250.0 cc. of water. 



(3) Heat slowly to cook the meat 

 thoroly. 



(4) Add NaOH to make the reaction 

 neutral to litmus. 



(5) Distribute in test tubes. 



(6) Sterilize in the autoclave. 



(b) Park, Williams and Krumwiede pre- 

 pared the medium as follows: 



(1) Mince 8.0 oz. bullock's heart and 

 grind in a mortar with 8.0 oz. of 

 tap water. 



(2) Cook (time not given). 



(3) Make alkaline. 



(4) Sterilize (method not given). 



(5) Tube. 



References: Harvey (1921-22 p. 94), Stitt 

 (1923 p. 42), Park, Williams and Krum- 

 wiede (1924 p. 125). 



1235. Harde and Hauser's Fish Medium 



Constituents: 



1. Water. 



2. Fish (whiting). 

 Preparation : 



(1) Cut the flesh of a whiting fish in about 

 1 centimeter cubes and place one cube 

 in a test tube. 



(2) Add 8.0 cc. of water to each tube 

 of (1). 



