538 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



solution containing 6.0% peptone. The 

 lactose, litmus and caffeine were added to 

 the hot sterile nutrient agar. 

 References: Lubenau (1907 p. 249), 

 Harvey (1921-22 p. 92). 



1750. Harvey's Caffeine Endo Agar 



Add 33.0 cc. of 10.0% caffeine to 1000.0 cc. 

 of Harvey's modification of Endo agar (see 

 variant (e) medium 1729). 



1751. Gathgen's Caffeine Fuchsin Sulphite 

 Agar (Bezanson) 



Constituents : 



1. Infusion agar (3.0%) 1000.0 cc. 



2. Lactose (10.0% solution). . . 120.0 cc. 



3. Fuchsin 



4. Na.SOa (10.0% soln.) 25.0 cc. 



5. Caffeine 3.3 g. 



Preparation: 



(1) Add 120.0 cc. of a 10.0% soda solution 

 to a liter of neutral 3.0% infusion 

 agar. 



(2) Add 120.0 cc. of a 10.0% lactose 

 solution to (1). 



(3) Add 10.0 g. of crystalline fuchsin to 

 100.0 cc. of 96.0% alcohol and allow to 

 stand for 2 hours. 



(4) Decant (3) and add 2.5 cc. of the 

 supernatant fluid to sterile (2). 



(5) Prepare a 10.0% solution of NazSOa- 



(6) Add 25.0 cc. of sterile (5) to (4). 



(7) Add 0.33 g. of pure caffeine to each 

 100.0 cc. quantity. 



(8) Pour in plates. 



Sterilization: Sterilize (2) at 100°C. for one 

 hour. Sterilize (5) by heating at 80°C. 

 for one hour. 



Use: Differentiation of colon typhoid 

 group. 



Variants : 



(a) Harvey added 33.0 cc. of a 10.0% 

 caffeine solution to 1000.0 cc. of 

 Harvey's modification of Endo agar 

 (see variant (e) 1729). 



(b) Klimmer prepared a similar medium 

 as follows: 



(1) Soak 30.0 to 40.0 g. agar in meat 

 water or a 1.0% meat extract solu- 

 tion for several hours. 



(2) Boil for 3 hours. 



(3) Dissolve 10.0 g. peptone and 5.0 g. 

 NaCl in (2). 



(4) Neutralize to litmus. 



(5) Add 7.0 cc. of normal soda solution 

 or 10.0 cc. of a 10.0% soda solution. 



(6) Boil. 



(7) Filter. 



(8) Add 10.0 g. c.p. lactose, 5.0 cc. of a 

 concentrated filtered alcoholic 

 fuchsin solution and 25.0 cc. of a 

 10.0% freshly prepared sodium 

 sulphite solution. 



(9) Add 0.33% crystalline caffeine to 

 (8). 



Klimmer reported that typhoid, 



paratyphoid and cholera colonies 



were blue; colon colonies were red. 



References: Bezangon (1920 p. 345), 



Harvey (1921-22 pp. 92, 93), Klimmer 



(1923 p. 210). 



1752. Viehoever's Basal Glucose Extract 

 Agar 



Constituents : 



1. Water 500.0 cc. 



2. Peptone (Witte) 6.0 g. 



3. Meat extract (Liebig's) 4.0 g. 



4. NaCl 1.0 g. 



5. Glucose 50 g. 



6. Agar 10.0 g. 



Preparation : 



(1) Dissolve 2, 3, 4, 5 and 6 in 1. 



(2) Dissolve one of the added nutrients 

 in (1) and adjust the reaction as 

 indicated. 



(3) Tube in 5.0 cc. lots. 

 Sterilization: Method not given. 



Use: To study spore production by urea 

 splitting organisms, Bac. probatus. 



Added nutrients: The author prepared one 

 of the mediums as indicated: 



(a) Neutral basal medium + 0.25% 

 NasCOs. 



(b) Neutral basal medium + 1.0% 

 CaCOa. 



(c) Neutral basal medium + 2.0% urea 

 + 0.3% (NH4)2C03. 



Variants: The author gave the following 

 variants: 



(a) Basal medium with 0.3, 0.2 or 0.1 

 concentration of nutrients + 0.1% 

 NasCOa. 



(b) Basal medium with 0.3 concentration 

 of nutrients + 0.05, 0.25 or 1.0% 

 Na.COs. 



(c) Basal medium with 0.3 concentration 



