500 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(e) Meier prepared a similar medium as 

 follows: 



(1) Boil 500.0 g. of fat and tendon free 

 beef in 1 liter of water. 



(2) Filter. 



(3) Dissolve 15.0 g. agar, 10.0 g. pep- 

 tone and 5.0, 20.0 or 40.0 g. of lac- 

 tose or glucose in the filtrate. 



(4) Neutralize by the addition of 

 KOH. Add KOH until turmeric 

 paper is turned quite weakly 

 brownish red. 



(5) Sterilization not specified. 

 References: Smith (1897 p. 480), Com- 

 mittee A. P. H. A. (1901 p. 384), (1905 

 p. 108), (1905 Sup. #1), (1909 p. 285), 

 Meier (1918 p. 436). 



1665. Francis' Basal Infusion Agar 



Constituents: 



1. Infusion agar (1.5%) 1000.0 cc. 



2. Peptone (1.0%) 10.0 g. 



Preparation : 



(1) Prepare a stock beef infusion agar 

 using fresh beef, 1.0% agar and 1.0% 

 peptone. 



(2) Adjust (1) to pH-7.6. 



(3) Add one of the added nutrients in the 

 manner indicated below. 



(4) Incubate 24 to 48 hours to test 

 sterility. 



Sterilization: Method not given. 



Use: Cultivation of Bacterium tularense. 



Variants : 



(a) The author used 1.0% agar in prepa- 

 ration of media as indicated under 

 added nutrients. 



(b) Francis adjusted the reaction of the 

 stock agar to pH-7.3 when preparing 

 serum glucose cystine agar, serum 

 glucose cysteine hydrochloride agar, 

 and cysteine hydrochloride agar. 



Added nutrients: Francis prepared the 

 media listed below as indicated. 



(a) Serum glucose agar. Add 1.0% glu- 

 cose from a sterile 50.0% glucose solu- 

 tion and 5.0% serum to the melted 

 agar cooled to 45°C. 



(b) Glucose blood agar. Substitute 5.0% 

 defibrinated blood for serum in (a). 



(c) Blood agar. Same as (b) but add no 

 glucose. 



(d) Mediums (a), (b) and (c) plus spleen 

 tissue. 



(1) Remove spleen from a healthy rab- 

 bit and cut into pieces 3 mm. in 

 diameter under aseptic conditions. 



(2) Rub one piece of (1) on each agar 

 slant of (a), (b) or (c) and allow 

 the spleen to remain on the surface 

 of each slant just above the water 

 of condensation. 



(e) Cystine agar. Add 0.02% cystine to 

 solid sterile 1.0% stock agar and 

 steam to melt agar and sterilize the 

 cystine. 



(f) Spleen agar. Rub pieces of spleen 

 over 1.0% stock agar slants in the 

 manner as indicated under (d) (1) 

 and (2) above. 



(g) Serum glucose cystine agar. 



(1) Add 0.1% cystine and 1.0% glucose 

 to solid sterile 1.0% stock agar. 



(2) Steam to melt the agar and to steri- 

 lize the cystine and glucose. 



(3) Cool to 5b°C. 



(4) Add 5.0% horse serum. 



(h) Serum glucose cysteine hydrochloride 

 agar. Substitute cysteine hydro- 

 chloride for cystine in medium (g) 

 above. 



(i) Cystine hydrochloride agar. Add 

 0.1% cystine hydrochloride and 1.0% 

 glucose to sterile solid 1.0% stock 

 agar and heat to melt the agar and 

 sterilize the cysteine hydrochloride 

 and glucose. 

 Reference: Francis (1922 p. 102, 987, 988) 



(1923 p. 1398). 



1666. Deycke's Alkali Albuminate Agar 



Constituents: 



1. Water 1200.0 cc. 



2. KOH (3.0%) 36.0 g. 



3. Veal 1000.0 g. 



4. NaCl 



5. Peptone 

 " 6. Agar 

 Preparation : 



(1) Digest 1000.0 g. of finely ground fat 

 free veal in 1200.0 cc. of 3.0% KOH 

 solution for 2 days at 37°C. 



(2) Heat (1) for one hour on a water bath 

 at 60 to 70°C. until all the protein 

 has dissolved. 



(3) Add HCl, carefully to precipitate 

 the albuminates. 



(4) Collect on a filter. 



