638 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



dark brown or brownish violet, dry and 

 hard to remove from the agar. Pneumo- 

 cocci colonies were larger than strepto- 

 cocci, about 5 mm. in diameter, colonies 

 were dark brown or brownish, forming a 

 smooth, coherent mucilaginous mem- 

 brane. A light yellow decolorization of 

 the medium took place when pneumococci 

 growth was luxuriant. If agar and blood 

 were mixed at 50°C. a clear ruby red 

 transparent medium was obtained. 

 Heating above this temperature gave a 

 darker medium. When mixed at 60°C. 

 the medium was not transparent in thick 

 [layers. 

 References: Bieling (1921 p. 262), Klimmer 

 (1923 p. 226). 



1993. Mandelbaum and Heinemann's Glyc- 

 erol Blood Agar (KoUe and Wasser- 

 mann 



Constituents : 



1. Glycerol agar. 



2. Blood human. 



Preparation: (1) Streak glycerol agar with 

 human blood. 



Use: Cultivation of diphtheria bacilli. 

 Author reported that diphtheria colonies 

 were light with yellowish brown ring. 

 Pseudo diphtheria colonies red. Similar 

 media were used to cultivate a large 

 number of organisms by a number of 

 investigators. 



Variants : 



(a) Sick prepared a medium as follows 

 for clinical purposes: 



(1) Draw blood under aseptic condi- 

 tions directly from a vein into a 

 sterile Erlenmeyer flask containing 

 glass beads, using a sterile needle 

 and sterile tube. 



(2) Shake the flask to defibrinate the 

 blood. 



(3) Prepare 3.0% agar containing 2.0 

 to 3.0% glycerol. 



(4) Mix one part (2) with 4 or 5 parts 

 sterile (3), cooled to 50°C. 



(5) Distribute into sterile test tubes. 



(b) Besson (Tanner) gave the following 

 method of preparation: 



(1) Melt tubes of glycerol agar and cool 

 to 40°C. 



(2) Add 1.0 CO. of blood from a rabbit's 

 artery. 



(3) Mix the tubes without shaking. 



(4) Cool in a slanting position. 



(c) Bezangon and Griffon (Besson) gave 

 the following method of preparation: 



(1) Add 5.0% glycerol to nutrient agar. 



(2) Liquify sterile (1) and cool to 45°C. 



(3) Add about 1.0 cc. of fresh rabbit 

 blood. (A 1.0% solution of com- 

 mercial hemoglobin may replace 

 the blood.) 



(4) Mix well without shaking. 



(5) Slant and cool. 



(d) Cantani (Besson) prepared a medium 

 as follows: 



(1) Mix equal parts of sterile blood and 

 sterile glycerol. 



(2) Allow to stand several hours. 



(3) Add 0.5 to 0.75 cc. of (2) to tubes of 

 liquified sterile agar. 



(e) Stitt cultivated pneumococci, strep- 

 tococci, gonococci and meningococci 

 on a medium prepared by smearing 

 the surface of a glycerol agar slant 

 with a platinum loop of blood ob- 

 tained from the lobe of a well cleaned 

 ear. 



References: KoUe and Wasserman (1912 

 p. 414), Sick (1912 p. Ill), Tanner (1919 

 p. 69), Besson (1920 pp. 53, 54), Stitt 

 (1923 p. 47). 



1994. Wasserman's Nutrose Blood Agar 

 (Abel) 



Constituents: 



1. Water 30.0 to 40.0 cc, 



2. Blood (hog) 15.0 cc. 



3. Glycerol 2.0 or 3.0 cc. 



4. Nutrose 0.8 g. 



5. Agar (2.0% peptone) 

 Preparation : 



(1) Mix 15.0 cc. of hog blood, 30.0 to 

 40.0 cc. of water, 2.0 to 3.0 cc. of 

 glycerol and 0.8 g. nutrose. 



(2) Shake the mixture constantly and 

 boil for 15 minutes. 



(3) Repeat the boiling and shake on the 

 following day for 15 minutes. 



(4) When desired for use heat to 50 to 

 60°C. and mix with an equal quantity 

 of sterile 2.0% peptone agar. 



(5) Pour into plates. 

 Sterilization: Not specified. 

 Use: Cultivation of gonococci. 

 Reference: Abel (1912 p. 162). 



