CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



533 



lactose and a sufficient quantity of a 

 saturated alcoholic solution of brom 

 cresol purple (generally about 0.75 

 CO.) to give the desired depth of 

 color. 

 (8) Flask in 200.0 cc. quantities. 



Sterilization: Autoclave at 15 pounds pres- 

 sure for 10 minutes. 



Use: Analysis of fecal flora. 



Reference: Torrey (1926 p. 353). 



1736. Abbott's Glycerol Infusion Agar 



Constituents : 



1. Water 1000.0 cc. 



2. Beef 500.0 g. 



3. Peptone (1.0%) 10.0 g. 



4. NaCl (0.5%) 5.0 g. 



5. Agar (1.0 to 1.5%) . . . 10.0 to 15.0 g. 



6. Glycerol (5.0 to 7.0%). 50.0 to 70.0 g. 

 Preparation: 



(1) Add 500.0 g. of chopped lean beef to 

 1 liter of water and soak for 24 hours, 

 kept at ice box temperature. 



(2) Strain thru a coarse towel and press 

 until a liter of fluid is obtained. 



(3) Dissolve 10.0 g. (1.0%) peptone, and 

 5.0 g. (0.5%) NaCl. 



(4) Dissolve 1.0 to 1.5% agar in (3) by 

 boiling in a porcelain lined iron 

 vessel. Add 250 to 300.0 cc. water 

 and boil until this volume of water 

 has evaporated, leaving one liter 

 volume. 



(5) Place the vessel in a large dish of 

 cold water. 



(6) Stir the agar constantly until cooled 

 to 68 to 70°C. 



(7) Add the white of one egg which has 

 been beaten up in about 50.0 cc. of 

 water, (a 10.0% dry albumin solution 

 may be used). 



(8) Mix (7) thoroly with (6). 



(9) Allow to boil slowly for 30 minutes. 

 Do not allow the volume of the 

 liquid to fall below the liter mark. 



(10) Filter thru a heavy folded filter 

 paper. 



(11) Add 5.0 to 7.0% glycerol to (10). 

 Sterilization: Sterilize by steam (method 



not given). 

 Use: General culture medium. 

 Variants : 



(a) Harvey added 10.0 cc. glycerol, 3.0 

 cc. normal NaOH to 87.0 cc. of 



infusion agar (see variant (v) medium 



1661). The medium was used to 



cultivate V. cholerae. 

 (b) Harvey added 50.0 cc. of glycerol to 



1000.0 cc. of infusion agar (see 



variant (v) medium 1661). 

 Reference: Abbott (1921 p. 132), Harvey 

 (1921-22 p. 87). 



1737. Kowalski's Glycerol Lung Infusion 

 Agar 



Constituents : 



1. Water 2500.0 cc. 



2. Lung, calf 1000.0 g. 



3. NaCl 18.0 g. 



4. Potassium phos- 

 phate 9.0 g. 



5. (NH4)2S04 9.0g. 



6. Na2S04 25.0 g. 



7. Sug.ar 90.0 g. 



8. Peptone 25.0 g. 



9. Agar 40.0 g. 



10. Glycerol 20.0 to 25.0 g. 



Preparation : 



(1) Remove the lung of a calf imme- 

 diately after death. Grind it in a 

 meat grinding machine, keeping as 

 clean as possible during the 

 procedure. 



(2) Add two liters of water to 1000.0 g. 

 of finely ground (1 ) . Place in a glass 

 container and boil. 



(3) After a half hour, strain thru a sieve. 

 Press out the residue, and refilter 

 the liquid obtained. It is then added 

 to the original filtrate. 



(4) Add to the quite clear fluid, 18.0 g. 

 NaCl, 9.0 g. potassium phosphate, 

 9.0 g. (NH4)2S04, 25.0 g. Na2S04, 

 90.0 g. sugar, 25.0 g. peptone. 



(5) After complete solution of the mate- 

 rials, add 2.0% agar which has been 

 previously soaked for 24 hours in 

 water. 



(6) Boil to dissolve the agar, stirring 

 continually. 



(7) Neutralize very carefully with equal 

 portions of KOH and NaOH. 



(8) Make up to 2500.0 cc. and cool to 

 58°C. 



(9) Add the whites of four hen eggs, 

 beaten to a foam. 



(10) Boil a minute, and filter thru a hot 

 water funnel. 



