CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



321 



(4) Digest at 40 to 42 °C. for 24 hours. 



(5) Filter thru cloth and paper, and 

 adjust the reaction to the correct 

 alkalinity. 



(6) Steam for one hour, filter and add 

 0.5% glucose and 2.0% glycerol. 

 (Agar and gelatin may be prepared 

 from this medium.) 



(7) Add 3 drops of blood to supply vita- 

 mines just before use. 



Sterilization: Method not given. 



Use: Used in human microparasitology. 

 The addition of blood may be unnecessary 

 with subcultures. Author claimed good 

 results with anaerobes as well as aerobes. 



Reference: Celozzi (1918 p. 291). Taken 

 from (1919 p. 147). 



1123. Douglas Trypsin Broth (Hartley) 



Constituents: 



1. Water 250.0 cc. 



2. Meat, horse 150.0 g. 



3. XazCOa (0.8% soln.) 250.0 cc. 



Preparation : 



(1) Mi.x 150.0 g. of lean minced horse 

 meat with 250.0 cc. tap water and 

 heat at 80°C. in a steamer. 



(2) Add 250.0 cc. of an 0.8% NajCOa 

 (anhydrous) and cool to 45^0. 



(3) Add 5.0 cc. of chloroform and 5.0 cc. 

 of Cole and Onslow's pancreatic 

 extract. 



(4) Incubate at 37°C. for 6 hours shaking 

 frequently. 



(5) Add 40.0 cc. normal HCl and heat in 

 the steamer for 30 minutes. 



(6) Cool and filter. 



(7) Adjust to pH 8.0. 



(8) Distribute as desired. 

 Sterilization: Pass steam thru the auto- 

 clave for one hour then raise the pressure 

 slowly to 10 pounds and turn off the 

 steam. For sterilization of larger quan- 

 tities (one liter in a flask) maintain the 

 pressure at 10 pounds for 30 minutes. 



Use: Preparation of diphtheria to.xin. 

 Reference: Hartley (1922 p. 482). 



1124. Harvey's Basal Trypsinized Casein 

 Solution 



Constituents: 



1. Water 1000.0 cc. 



2. Casein 100.0 g. 



3. NaCl 2.5 g. 



4. CaCl2 0.125 g. 



Preparation : 



(1) Add 100.0 g. commercial casein to 

 500.0 cc. boiling water containing 

 0.8% anhydrous NaaCOj. 



(2) Raise the temperature to the boiling 

 point. 



(3) Add 500.0 cc. of cold water. 



(4) Make faintly alkaline to litmus. 



(5) Coolto45°C. 



(6) Add 400.0 cc. of pancreatic extract. 



(7) Incubate 4 hours at 37°C. 



(8) Raise the temperature to the boil- 

 ing point. 



(9) Filter thru thick wet clean cloth. 



(10) Filter thru thick wet filter paper. 



(11) Bring up the volume to 1000.0 cc. by 

 the addition of water. 



(12) Add 2.5 g. NaCl and 0.125 g. CaCl,. 



(13) Adjust the reaction. 



(14) Steam 30 minutes. 



(15) Filter while hot thru paper. 



(16) Disolve 1.0% of one of the added 

 nutrients in sterile (15). 



(17) Distribute to a depth of one inch in 

 small test tubes containing fer- 

 mentation tubes. 



(18) Cover with 0.25 inch of liquid 

 paraffin. 



(19) Add 3 drops of egg albumin mixed 

 with bouillon (see variant 848 for 

 preparation) to each tube. (This 

 serves as an indicator.) 



(20) Place the tubes in boiling water for 

 20 minutes. 



Sterilization: Sterilize (15) in the auto- 

 clave. 



Use: Determine fermentation of sugars by 

 anaerobic organisms. 



Added nutrients: Author added 1.0% glu- 

 cose or any other desired carbohydrate, 

 alcohol, etc. 



Reference: Harvey (1921-22 p. 111). 



1125. Teruuchi and Hida's Trypsinized 

 Casein Solution 



Constituents: 



1. Water 1000.0 cc. 



2. Casein 100.0 g. 



3. NaCl (0.5%) 5.0 g. 



Preparation : 



(1) Dissolve 100.0 g. of pure casein in 

 1 liter of 0.8% NazCOj (anhydrous). 



(2) Add 5.0 to 10.0 g. of pancreatin 

 (Gehe and Co.) and shake with 

 chloroform. 



