CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



639 



1995. Savini and Savini-Castano's Bacteria 

 Blood Agar 



Same as medium 977 but use nutrient 

 agar instead of bouillon. 



1996. Bieling's Optochin Hydrochloride 

 Blood Agar 



Constituents: 



1. Distilled water 100.0 cc. 



2. Optochin hydrochloride 0.1 g. 



3. Nutrient agar (3.0%) 



4. Blood, horse 60.0 cc. 



Preparation : 



(1) Dissolve 0.1 g. optochin hydrochloride 

 in 10.0 cc. distilled water (should 

 not be kept more than 3 days). 



(2) Prepare 3.0% nutrient agar. 



(3) Mix 1.0 cc. of (1) with 150.0 cc. of (2). 



(4) Mix 60.0 cc. horse blood with 90.0 cc. 

 distilled water. 



(5) Heat (4) for one hour at 60°C. 



(6) Melt (3) and cool to 60°C. 



(7) Mix equal parts (6) and (5), both 

 being at 60 ''C. 



Sterilization: Not specified. 



Use: Differentiation of streptococci and 

 pneumococci. Author reported that 

 pneumococci showed no growth. Hemo- 

 lytic streptococci colonies flat, and did 

 not change the color of the medium. 

 May be easily removed from the medium. 

 Streptococcus mitior colonies were dry 

 and brown and hard to remove from the 

 medium. 



References: Bieling (1921 p. 264), Klim- 

 mer (1923 p. 226). 



1997. Heim's Hemoglobin Agar 



Constituents: 



1 . Distilled water 90.0 cc. 



2. Hemoglobin 10.0 g. 



3. KOH (10%,) 10.0 cc. 



4. Nutrient agar 

 Preparation : 



(1) Mix 1, 2 and 3. 



(2) Mix 1.0 cc. sterile (1) with 7.0 cc. 

 nutrient agar. 



(3) Pour into plates or slant. 

 Sterilization: Sterilize (1) in streaming 



steam. 

 Use: Cultivation of pneumococci. Also 

 used by other investigators for the culti- 

 vation of influenza bacilli. 



Variants : 



(a) Savini and Savini-Castano reported 

 influenza bacilli developed only upon 

 the following medium when other 

 organisms were growing side by side 

 with it. Staphylococcus aureus gave 

 best results for this purpose. 



(1) Prepare a solution of 5.0% hemo- 

 globin in N/10 NaOH or soda solu- 

 tion. Boil until the solution is 

 complete. 



(2) Filter and sterilize (1). (Method 

 of sterilization not given.) 



(3) Prepare nutrient agar. 



(4) Melt sterile (3) and cool to about 

 50°C. 



(5) To each 10.0 cc. of (4) add 1.0 cc. 

 of (2) and mix well without shaking 

 violently. 



(6) Allow the agar to solidify over 

 night in a cool place. 



(b) Thalhimer cultivated Bac. influenzae 

 on the following medium. He re- 

 ported that a slightly better medium 

 was obtained if a few cc. of hydrogen 

 peroxide were added before filtering. 

 Growth was even more luxuriant than 

 on ordinary blood agar. 



(1) Dissolve amorphous powdered 

 hemoglobin (Merck & Co., and 

 Eimer and Amend) in 100.0 cc. of 

 water until a deep mahogany brown 

 color is obtained. (About 10.0 g. 

 hemoglobin.) 



(2) Filter thru a Reichel porcelain 

 filter. 



(3) Add enough of (2) to sterile fluid 

 agar to obtain the same intensity 

 of color as ordinary blood agar. 



(4) Tube and slant. 



References: Heim (1907 p. 1587), Savini 

 and Savini-Castano (1911 p. 493), Thal- 

 himer (1914 p. 189). 



1998. Esch's Alkaline Hemoglobin Agar 



Constituents : 



1. Distilled water. 



2. Hemoglobin (Merck's). 



3. Neutral agar. 

 Preparation: 



(1) Triturate hemoglobin. 



(2) Add 5.0 g. hemoglobin to 15.0 cc. N/1 

 NaOH + 15.0 cc. distilled water. 



