492 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(9) Add (1) to (8). 



(10) Boil until albumin is coagulated 

 and floats in the clear liquid. 

 Restore to weight. 



(11) Readjust the reaction if neces- 

 sary. 



(12) Add 0.5% of normal HCl. 



(13) Filter thru cotton supported on 

 a coil of wire using a suction 

 pump to hasten filtration. 



(14) Tube. 



(15) Sterilize in steam for 15 minutes 

 on 3 successive days or in the 

 autoclave for 20 minutes at 

 120°C. Slant some of the tubes 

 after final sterilization. 



(n) Frost (1923). 



(1) Add 15.0 g. agar to 500.0 cc. of 

 water and dissolve by heating to 

 120°C. in the autoclave, closing 

 off the gas, and allowing to cool 

 or boil until the agar is dissolved 

 (about 30 minutes) and make up 

 the loss of water due to evap- 

 oration. 



(2) Free 500.0 g. of lean beef from 

 fat and connective tissue and 

 mince. 



(3) Add 500.0 cc. of distilled water. 



(4) Place in a vessel for cooking and 

 then cook for 30 minutes at 70 °C. 



(5) Filter thru paper and make up the 

 volume to 500.0 cc. 



(6) Add 10.0 g. of peptone and 5.0 g. 

 of NaCl. 



(7) Heat until solution is complete. 



(8) Neutralize to phenolphthalein. 



(9) Cool to 60°C. 



(10) Add (1) to (9). 



(11) Boil until albumin is coagulated 

 and floats in the clear fluid. 

 Restore the weight. 



(12) Readjust the reaction if neces- 

 sary. 



(13) Add 0.5% of normal HCl. 



(14) Filter thru cotton supported on a 

 coil of wire using a suction pump 

 to hasten filtration. 



(15) Tube. 



(16) Sterilize in steam for 15 minutes 

 on 3 successive days or in the 

 autoclave for 20 minutes at 120°C. 

 Slant some of the tubes after 

 sterilization. 



(o) Smith (1905). 



(1) Add 1000.0 cc. distilled water to 

 500.0 g. of finely minced lean beef. 



(2) Infuse for 24 hours in the ice 

 chest or heat for one hour in the 

 water bath at 55°C. 



(3) Boil for 60 minutes in the steamer 

 or in a covered dish. 



(4) Filter thru a clean cloth in a 

 meat press. 



(5) Cool and remove the fat by filter- 

 ing thru S and S filter paper. 



(6) Make up the volume to 1000.0 cc. 

 by the addition of water. 



(7) Add 1.0% Witte's peptone and 

 0.5% NaCl. 



(8) Steam for one-half hour. 



(9) Filter. 



(10) Cool. 



(11) Adjust the reaction. 



(12) Steam again for 30 minutes. 



(13) Filter. 



(14) Add 10.0 g. powdered agar to the 

 filtrate. 



(15) Steam for 30 minutes and cool 

 to 58°C. 



(16) Add the whites of 2 eggs (beaten 

 thoroly and neutralized to litmus 

 by dilute HCl). 



(17) Steam one hour and filter while 

 hot thru S & S filter paper which 

 has been thoroly warmed with 

 boiling distilled water. The agar 

 may be filtered thru absorbent 

 cotton. 



(18) Tube. 



(19) Sterilize by heating in the auto- 

 clave at 110°C. for 10 minutes or 

 by short steamings in the steamer 

 on each of 3 successive days. 



(p) Guillemard (1906) cultivated anae- 

 robes on the following medium: 



(1) Mix 500.0 g. of chopped meat,, 

 10.0 g. peptone and 5.0 g. NaCl 

 in 1000.0 cc. water. 



(2) Bring to a boil and boil 10 min- 

 utes. 



(3) Filter while hot. 



(4) Allow to cool for 12 hours. 



(5) Filter while cold. 



(6) The solution is clear and of an 

 acid reaction. 



(7) Make up to a liter if necessary. 



(8) Add a piece of red litmus paper. 



