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CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(8) Collect the fluid which drains 

 thru the cloth together with that 

 obtained by squeezing the meat 

 in the cloth. 



(9) Filter the fluid collected thru 

 well-wetted, thick filter paper. 



(10) Add to the filtrate: peptone 

 10.0 g., sodium chloride 5.0 g. 



Note: The peptone should be 

 worked into a paste or suspension 

 by gradual addition of a little of 

 the meat extract before addition 

 to the filtrate. 



(11) Steam or boil 45 minutes. 



(12) Bring the volume up to 1000.0 cc. 

 by the addition of water. 



(13) Estimate and adjust reaction to a 

 pH value or slightly alkaline to 

 litmus or 1.0% acid to phenol- 

 phthalein. 



(14) Steam 30 minutes. 



(15) Clarify if necessary, and filter 

 while hot thru well wetted, thick 

 filter paper, or thru two layers of 

 absorbent cotton wool. 



Note: If simple filtration thru 

 thick paper alone is not sufficient 

 to give a clear medium, clearing 

 should be effected by means of 

 white of egg or other clearing 

 agent. 



(a) Beat up the white of one or two 

 eggs along with the crushed 

 shells in about 20.0 cc . water. 



Note: Raw meat juice, 15.0 cc. 

 per liter of medium may be sub- 

 stituted for white of egg. 



(b) Add to the medium little by 

 little before filtration and at a 

 temperature not exceeding 60°C. 



(c) Stir to mix. 



(d) Steam for 30 minutes. 



(e) Remove from steamer and shake 

 up well to mix. 



(f) Steam again 15 minutes. 



(g) Filter in the steamer thru thick 

 filter paper, or thru 2 layers of 

 absorbent cotton wool. 



(h) Refilter, if necessary, the first 

 portion of the filtrate. 



(16) Distribute the filtrate into 

 or test tubes. 



(17) Sterilize. 



(cc) Harvey (1921-22). 



Prepare the medium like variant 

 (aa) (1) thru (15). 



(16) Inoculate bouillon with B. coli. 



(17) Incubate 24 hours. 



(18) Boil 20 minutes. 



(19) Make a paste or suspension of 

 about 15 g. purified talc with a 

 little of the dead bouillon culture. 



(20) Add the suspension to the culture. 



(21) Filter and refilter thru filter 

 paper till clear. 



(22) Distribute into test tubes. 



(23) Sterilize. 



(dd) Harvey (1921-22). 



(1) Mince finely fat-free beef. 



(2) Add 500.0 g. to 1000.0 cc. distilled 

 water or tap water. 



(3) Keep in a cool place over night. 



(4) Skim off fat floating on the 

 surface. 



(5) Pour the mixture on to a wet, 

 thick, clean cloth. 



(6) Collect the fluid which drains 

 thru the cloth together with that 

 obtained by squeezing the meat 

 in the cloth. 



(7) Bring the volume up to 1000.0 cc. 

 by the addition of water. 



(8) Add peptone 10.0 g., sodium 

 chloride 5.0 g. 



(9) Estimate and adjust the reaction 

 to a definite pH or faintly alka- 

 line to litmus or 1.0% acid to 

 phenolphthalein. 



(10) Steam or boil 45 minutes. 



(11) Filter, while hot, thru well- 

 wetted, thick filter paper, or thru 

 2 layers of absorbent cotton 

 wool. 



(12) Distribute the filtrate into flasks 

 or test tubes. 



(13) Sterilize (S9.5, 9.6 according to 

 Harvey). 



(ee)Heinemann (1922). 



(1) Clean one pound of beef or veal 

 of adhering fat, etc., and grind in 

 a meat chopper. 



(2) Cover with a liter of water. 



(3) Digest over night at room tem- 

 perature. 



(4) Heat to 60°C. and digest at this 

 temperature for two hours. 



