CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



489 



(10) Place in a steamer for 12-15 hours. 



(11) Filter thru a hot funnel. 



(12) The steaming for 12-15 hours is 

 sufficient for sterilization. 



(b) Schultz (1891). 



(1) Place 3 to 4 liters of distilled 

 water in an enamel container and 

 add 20.0 g. finely cut up agar- 

 agar to it. 



(2) Boil two hours in a closed vessel. 



(3) Place 500.0 g. of the best quality 

 meat, without fat or tendons in a 

 glass container fitted with a lid. 



(4) Pour 1300.0 cc. of distilled water 

 over the meat. 



(5) Store in a cool place until the 

 ne.xt day. 



(6) Filter thru 4 thicknesses of 

 cloth and press the remaining 

 meat to obtain as much fluid as 

 possible. 



(7) Pour the filtrate into a kettle, 

 add 10.0 g. peptone (siccum), 

 5.0 g. NaCl and the whites of two 

 eggs, beaten up in 2 or 3 volumes 

 of water. 



(8) Boil under a gas flame for 15 

 minutes. 



(9) Adjust to faint alkalinity, using 

 phenolphthalein as an indicator, 

 and the end point being a faint 

 red color. 



(10) Pour into an iron kettle, add 

 100.0 cc. distilled water, boil 

 strongly for 5 minutes and filter. 



(11) Add 1 liter of (10) to (2). 



(12) Boil until there remains only 

 1 liter of medium. 



(13) Filter thru a hot funnel. 



(14) Distribute into test tubes and 

 steam on 3 successive days for 

 one hour. 



<c) Frothingham (1895). 



(1) Add one pound of finely chopped 

 lean meat to 1000.0 cc. water, and 

 let stand for about one hour. 



(2) Heat in the water bath to about 

 60°C. for 3 hours. 



(3) Boil 30 minutes. 



(4) Filter and when cool make 

 slightly alkaline (3.0 to 5.0 g. of 

 Liebig's meat extract may be sub- 

 stituted for the meat infusion). 



(5) Dissolve 5.0 g. NaCl and 10.0 g. 

 dried peptone in (4). 



(6) Boil 15 minutes, preferably over 

 a free flame. 



(7) Add 10.0 to 20.0 g. agar to (6) and 

 boil until dissolved. 



(8) Neutralize if necessary and add 

 glycerol if desired. 



(9) Filter. 



(10) Sterilization not specified, 

 (d) Frothingham (1895). 



(1) Add one pound of finely chopped 

 lean meat to 1000.0 cc. of water 

 and allow to stand from 12 to 

 24 hours in a cool place. 



(2) Strain thru a cheese cloth or a 

 coarse towel and squeeze in a 

 meat press or by twisting the 

 ends of the cloth until 1000.0 cc. 

 of the meat juice is obtained. 

 Make up to 1000.0 cc. by the addi- 

 tion of water if necessary. 



(Three to 5.0 g. of Liebig's meat 

 extract may be substituted for 

 the meat infusion.) 



(3) Dissolve 5 g. NaCl and 10.0 g. 

 dried peptone in (2). 



(4) Boil 15 minutes either in steam 

 or over a flame. 



(5) Dissolve 10.0 to 20.0 g. of agar in 

 (5) by boiling. 



(6) Neutralize, if necessary and add 

 glycerol if desired. 



(7) Boil from 45 minutes to an hour. 



(8) Filter. The filtrate should be 

 clear. If not add the yolks of 

 2 eggs, mix rapidly and boil for a 

 quarter to a half hour. Filter. 



(9) Sterilization not specified, 

 (e) Ravenel (1898-1900). 



(1) Chop 500.0 g. meat and mix with 

 500.0 cc. water. 



(2) Place in cool place over night 

 and then strain. 



(3) Chop agar-agar and add to 

 500.0 cc. of water. 



(4) Put (3) in autoclave and run up 

 to 2 atmospheres of pressure. 



(5) Cool below 100 °C. before opening. 



(6) Cool (5) to 75°C. 



(7) Mix (2) and (6). 



(8) Add 1.0% dried peptone and 

 0.5% salt. 



(9) Boil 3 to 5 minutes. 



