616 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



3. Maltose 10. o g. 



4. Ascitic fluid 250.0 cc. 



Preparation : 



(1) Prepare infusion agar, containing 

 1.0% Witte's peptone. 



(2) Mix 75 parts (2) with 25 parts ascitic 

 fluid. 



(3) Add 1% maltose to (2). 

 Sterilization: Not specified. 

 Use: Cultivation of meningococci. 

 Reference: Esch (1909 p. 152). 



1932. Steinschneider's Hydrocele 

 Fluid Agar 



Constituents : 



1. Infusion agar 300.0 cc. 



2. Hydrocele fluid 100.0 cc. 



Preparation : 



(1) Mix three parts sterile infusion agar 

 with one part sterile liquid hydrocele 

 fluid. 



(2) Pour into sterile Petri dishes. 

 Sterilization: Method not given. 

 Use: Cultivation of gonococci. 

 Reference: Steinschneider (1890 p. 533). 



1933. Heiman's Pleuritic Serum Agar 

 Constituents : 



1. Infusion agar (2.0%) 200.0 cc. 



2. Pleuritic serum 100.0 cc 



3. Peptone (1.0%) 2.0 g ' 



4. NaCl (0.5%) 1.0 g. 



Preparation: (1) Add one part sterilized 



liquid pleuritic (chest) serum to 2.0% 



agar containing 1.0% peptone and 5% 



NaCl. 

 Sterilization: Sterilize on 9 successive days 



at 65°C. 

 Use: Cultivation of gonococci. 

 Reference: Heiman (1896 p. 888). 



1934. Lentz and Tietz's Malachite Green 



Bile Agar (Klimmer) 

 Constituents : 



1. Meat infusion 2000.0 cc. 



2- Agar 60.0 g. " 



3. Peptone 20.0 g. 



4. NaCl 10.0 g' 



5- Bile 30.0 cc. 



6. Malachite Green I (Hochst) 



Preparation : 



(1) Soak 60.0 g. of agar in 2000.0 cc. of 

 meat water. 



(2) Boil for 3 hours. 



(3) Dissolve 3, 4 and 5 in 250.0 cc. of 

 water by heating slightly. 



(4) Mix (3) and (2). 



(5) Neutralize to litmus or adjust to 

 1.0 to 1.5 acid to phenolphthalein. 



(6) Boil for an hour. 



(7) Readjust the reaction if necessary. 



(8) Distribute in flasks. 



(9) The reaction of the sterile agar when 

 ready for use should be 1.8%. 



(10) Melt sterile (9) and add 30.0 cc. of 

 sterile beef bile and 1.0 cc. of a solu- 

 tion of 1.0 g. malachite green I 

 (Hochst) in 60.0 cc. distilled water. 

 It is preferable to mix 20.0 cc. of agar 

 with 0.05, 0.1, 0.15, 0.2, etc., cc. of a 

 2.0% malachite green solution and 

 pour into plates. Inoculate with 

 colon and typhoid bacilli and use 

 that dilution of malachite green that 

 inhibits the colon organisms but not 

 the typhoid organisms. Add a 

 corresponding amount of malachite 

 green and bile to the 100.0 cc. of agar. 



Sterilization: Method of sterilization of (8) 

 not given. 



Use: Cultivation of colon-typhoid group. 



Reference: Klimmer (1923 p. 212). 



1935. Lentz and Tietz's Malachite Green 

 Nutrose Bile Agar (Klimmer) 



Constituents : 



1. Meat infusion 2000.0 cc. 



2- Agar 60^0 g. " 



3. Peptone 20 g 



4- NaCl io:og: 



5. Nutrose 20.O g. 



6. Bile, beef 30.0 cc. 



7. Malachite green 

 Preparation : 



(1) Soak 60.0 g. of agar in 2000.0 cc. of 

 meat water. 



(2) Boil for three hours. 



(3) Dissolve 3, 4 and 5 in 250.0 cc. of 

 water by heating slightly. 



(4) Mix (3) and (2). 



(5) Neutralize to litmus or adjust to 

 1.0 to 1.5 acid to phenolphthalein. 



(6) Boil for an hour. 



(7) Readjust the reaction if necessary. 



(8) Distribute in flasks. 



(9) The reaction of the sterile agar when 

 ready for use should be a -3.5%. 



(10) Melt sterile (9) and add 3.0 cc. of 



