CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



513 



(6) Beat the white of an egg in 

 100.0 cc. of water and add to (5). 



(7) Mix well. 



(8) Autoclave at 120°C. for one hour. 



(9) Filter thru moistened Chardin 

 filter using a hot water funnel. 



(10) Tube. 



(11) Sterilize at 115° for 20 minutes, 

 (m) Giltner (1921). 



(1) Prepare an ordinary agar (method 

 not given) from 5.0 g. NaCl, 5.0 g. 

 Liebig's extract, 10.0 g. peptone 

 and 30.0 g. agar in sufficient water 

 to make 1000.0 cc. 



(2) Adjust the reaction to or +0.2%. 



(3) Store in 100 cc. quantities in 

 Erlenmeyer flasks. 



(n) Wolf and Shunk (1921). 



(1) Dissolve 10.0 or 20.0 g. agar, 3.0 g. 

 Liebig's beef extract, 10.0 g. Pep- 

 tone (Armour's) and 5.0 g. NaCI 

 in 1000.0 cc. water by heating in 

 the autoclave. 



(2) Flask. 



(3) Sterilize at 15 pounds for 15 

 minutes. 



(4) Coolto50°C. 



(5) Pipette with sterile pipette, 

 10.0 cc. quantities into sterile test 

 tubes and add appropriate quanti- 

 ties of strong acid or alkali to give 

 desired reaction. (Authors used 

 HCl sp. gr. 1.20 or 39.11% and 

 NaOH sp. gr. 1.226 or approxi- 

 mately 20.0%.) 



(6) Mix the tubes thoroly and cool. 

 Do not sterilize. 



(o) Giltner (1921). 



(1) Place 500.0 g. of water in an agate 

 water pail and add 15.0 g. of agar. 



(2) Wash the agar well, separating the 

 shreds and squeezing thru the 

 hands. Wash until clean. 



(3) Make up to the original volume 

 the addition of tap water. 



(4) Heat over a free flame until the 

 agar is dissolved, stirring con- 

 stantly. 



(5) Add 3.0 g. of standard meat ex- 

 tract (or use 500.0 cc. meat infu- 

 sion) to 500.0 cc. of tap water. 



(6) Add 1,0%, peptone and 0.5% NaCl. 



(7) Cool (4) to 60°C. and mix with (6). 



(8) Add 10.0 g. of albumin mixed with 

 100.0 cc. water. 



(9) Autoclave at 15 pounds pressure 

 for 2 hours. 



(10) Adjust the medium to the desired 

 reaction, by addition of normal 

 NaOH or HCl. 



(11) Filter while boiling hot thru 

 plaited filter paper. 



(12) Distribute as desired. 



(13) Sterilize (method not given), 

 (p) Stitt (1923). 



(1) Place 3.0 g. Liebig's meat extract, 

 10.0 g. peptone, Agar 2.0 or 3.0%, 

 and 5.0 g. NaCl in 1000.0 cc. water 

 in a rice cooker, or any other large 

 straight sided granite-iron re- 

 ceptacle. The whites of one or 

 two eggs may or may not be dis- 

 solved in the water. 



(2) Place in the chamber of the 

 dressing sterilizer and heat to be- 

 tween 5 and 10 pounds (110 to 

 115°) for at least 45 minutes. 



(3) Adjust the pH as desired. 



(4) Put back in the sterilizer for 

 30 minutes at the same tempera- 

 ture as in (2). The greater the 

 pressure applied the darker the 

 medium will be, so that a pressure 

 not to exceed 3 pounds is used when 

 a light medium is desired. 



(5) Leave the medium in the sterilizer 

 over night with the steam turned 

 off. 



(6) In the morning dump out the 

 jelly mass and cut off and discard 

 the bottom portion containing the 

 sediment. 



(7) Melt the remainder of the agar. 



(8) Distribute in flasks (or nursing 

 bottles). 



(9) Sterilize (method not specified), 

 (10) Store until ready for use. 



(q) Stitt (1923). 



(1) Dissolve 15.0 g. of agar in 500.0 cc. 

 of water in the inner compartment 

 of a rice cooker. 



(2) Cool to 55°C. 



(3) Dissolve the whites of one or two 

 eggs in 500.0 cc. of water. 



(4) Prepare a paste from 3.0 g. Liebig's 

 meat extract, 10.0 g. peptone and 

 5.0 g. NaCl by adding (3) little 

 by little. 



(5) Add the remainder of (3) to (4). 



(6) Heat (5) to 50 to 55°C. 



