606 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(5) Mix one part of (4) at 45°C. with five 

 parts melted infusion agar at 45°C. 

 (See variant (v) medium 1661 for 

 preparation.) 

 Sterilization: Not specified. 

 Use: Isolation of B' influenzae. 

 Variants: Author reported that Brilliant 

 green which inhibits the gram + bacteria 

 may be added. 

 Reference: Harvey (1921-22 p. 76). 



1911. Kristensen's Hemoglobin Infusion 

 Agar 



Constituents : 



1. Water 25,000.0 cc. 



2. Meat 25,000.0 g. 



3. Peptone 1.0% 



4. NaCl 0.5% 



5. Hemoglobin 1.0% 



Preparation : 



(1) Mince 25,000.0 g. beef . 



(2) Add 25 liters of water and allow to 

 stand in the cold until the next day. 



(3) Boil for a short time. 



(4) Press the fluid from the meat. 



(5) Add about 16 liters of water to the 

 meat, heat to boiling once more and 

 press the juice again from the meat. 



(6) Mix the fluid from (4) and (5). 



(7) Add 1.0% peptone and 0.5% NaCl. 



(8) Adjust to pH of about 7.8 to 7.9. 



(9) Dissolve 2.0% agar in (8). (Agar 

 may be omitted giving a liquid 

 medium.) 



(10) Filter thru cotton wool and distri- 

 bute in 600.0 cc. flasks. 



(11) The pH of the sterile agar should 

 be about 7.2. 



(12) Allow a flask containing defibrinated 

 blood to stand several hours to per- 

 mit the corpuscles to sink. 



(13) Pour off the supernatant fluid and 

 replace with distilled water. 



(14) Distribute the hemoglobin solution 

 thus obtained in 20.0 cc. quantities. 



(15) Store in the ice box until ready for 

 use. 



(16) Add 20.0 cc. of (15) to 600.0 cc. of 

 sterile melted (11) cooled to 40 to 

 50°C. 



(17) Pour into plates. 

 Sterilization: Sterilize (10) in the steamer 



on each of 3 successive days. 



Use: Cultivation of hemoglobinophilic 



bacteria. 

 Reference: Kristensen (1922 p. 231). 



1912. Brown and Orcutt's Hemoglobin 

 Infusion Agar 



Constituents : 



1. Veal infusion agar. 



2. Hemoglobin. 

 Preparation : 



(1) Prepare veal infusion agar. 



(2) Defibrinate blood and wash re- 

 peatedly with sterile physiological 

 salt solution. 



(3) Lake the washed corpuscles with dis- 

 tilled water and remove the cor- 

 puscles stroma by centrifugation. 

 (Centrifuge the laked blood cor- 

 puscles until the supernatant hemo- 

 globin solution no longer gives a 

 clouding reaction with salt.) 



(4) Add the hemoglobin solution thus 

 obtained to melted agar (amounts not 

 specified) and pour in plates. 



Sterilization: Not specified. 



Use: To determine food requirements for 



Bacillus pyogenes. 

 Reference: Brown and Orcutt (1920 p. 223). 



1913. Esch's Alkaline Hemoglobin Infusion 



Agar (Tanner) 



Constituents : 



1. Infusion agar 85.0 cc. 



2. Hemoglobin (horse) (Merck).. 5.0 g. 



3. KOH (N/2) 30.0 cc. 



Preparation : 



(1) Dissolve 5.0 g. of Merck's horse hemo- 

 globin in 30.0 cc. of half normal KOH. 



(2) Steam for 30 minutes. 



(3) Mix 15.0 cc. of (2) with 85.0 cc. of 

 neutral (litmus) infusion agar. 



(4) Pour in plates. 



(5) Dry at room temperatures with covers 

 removed. 



Sterilization: Not specified. 



Use: Cultivation of Microspira cholerae. 



Reference: Tanner (1919 p. 70). 



1914. Sherwood and Downs' Basal Serum 



Agar 



Constituents: 



1. Infusion agar 



2. Serum 



3. Andrade's Indicator 



1000.0 cc. 

 20.0 cc. 



