734 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



finely chopped fat and tendon 

 free beef. 



(2) Allow to stand in the ice box for 

 12 hours, or heat at 50 to 55° for 

 30 minutes. 



(3) Heat slowly to boiling". 



(4) Boil slowly for 10 minutes stirring 

 constantly. 



(5) Strain thru a cloth. 



(6) Heat slowly (not above 80°C.) 

 and add 20.0 g. peptone, 5.0 g. 

 NaCl and 120.0 g. of gelatin per 

 liter of filtrate. 



(7) Shake until solution is complete. 



(8) Add soda solution to neutralize 

 to litmus. 



(9) Heat in the autoclave at 112° for 

 10 minutes. 



(10) Filter thru paper. 



(11) Distribute. 



(12) Sterilize at 110° for 10 minutes. 

 (Do not heat over 110°C.) 



(13) Store in a cool place. 

 ^t) Harvey (1921-22). 



(1) Proceed as in the preparation of 

 infusion broth, see variant (bb) 

 of medium 779, step (1) thru (9). 



(2) Add 10.0 g. peptone, 5.0 g. sodium 

 chloride and 120.0 g. gelatin. 



(3) Heat gently with constant stirring. 



(4) Steam or boil 45 minutes to obtain 

 complete solution. 



(5) Bring the volume up to 1000.0 cc. 

 by the addition of hot water. 



(6) Adjust reaction to a definite pH 

 value, or faintly alkaline to litmus 

 or 1.0% acid to phenolphthalein. 



(7) Steam 30 minutes. 



(8) Filter, while hot, thru well-wetted, 

 thick filter paper by placing filter 

 funnel, stand and receptacle for 

 filtrate in the steam sterilizer, and 

 steaming till filtration is complete. 



(9) Distribute into flasks or test tubes. 



(10) Sterilize. 



(11) Cool the flasks or test tubes on 

 final removal from the sterilizer. 



(12) Store in a cool place, 

 (u) Pitfield (1922). 



(1) Cover 500.0 g. of finely cut fat 

 free beef with 1000.0 cc. water. 



(2) Shake well and place on ice over 

 night. 



(3) Squeeze out the fluid by means of a 



cloth and make up the volume to 

 1 liter. 



(4) Inoculate with a culture of the 

 colon bacillus. 



(5) Allow to stand at room tempera- 

 ture over night. 



(6) Boil and add 10.0 g. Witte's pep- 

 tone and 5.0 g. NaCl. 



(7) Weigh the saucepan and contents 

 and heat to 60°C. 



(8) Make up the loss in weight by the 

 addition of water. 



(9) Neutralize to litmus, 

 (v) Klimmer (1923). 



(1) Prepare meat infusion. 



(2) Dissolve 10.0 g. peptone, 5.0 g. 

 NaCl and 100.0 g. gelatin in (1) 

 by heating between 50 and 60°C. 

 Do not heat above 60°C. 



(3) Neutralize to litmus. 



(4) Filter. 



(5) Add 5.0 cc. of normal soda solu- 

 tion. 



(6) Sterilize by heating for 40 minutes 

 on each of 3 successive days for 

 15 minutes in streaming steam. 



(w) Park, Williams and Krumwiede 

 (1924). 



(1) Dissolve 100.0 g. gelatin, 10.0 g. 

 peptone and 5.0 g. NaCl in 

 1000.0 cc. meat juice. 



(2) Coagulate the albumin present in 

 the meat juice to clarify. 



(3) The coagulation of the albumin 

 present in the meat juice clears 

 the medium. 



(4) Filter thru cotton. 



(5) Tube and sterilize in the Arnold 

 sterilizer 20 to 30 minutes on 3 

 successive days. 



(x) Park, Williams and Krumwiede 

 (1924). 



(1) Add "10.0 g. (1.0%) peptone and 

 sodium chloride 0.5 g.," to 

 1000.0 cc. of meat infusion. 



(2) Heat nearly to boiling and add 

 100.0 g. gelatin. Dissolve with 

 as little heat as possible. 



(3) Adjust the reaction neutral to 

 litmus (pH = 6.8 to 7.0). 



(4) Cool to 50°C. and clarify by heat- 

 ing for 45 minutes in an Arnold 

 sterilizer. 



(5) Filter. 



